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A B S T R A C T B O O K – C O N T E N T 

Foreword............................................................................... 2 

Organization ......................................................................... 3 

Program of the Workshop .................................................... 4 

Abstracts of Talks ................................................................. .9 

Abstracts of Posters ............................................................ 53 

List of Participants ............................................................... 91 

Sponsors ............................................................................. 96 

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X X I V S P P S C O N G R E S S 2 0 1 1 

Content

Foreword 

eword 

ord 

A B S T R A C T B O O K – F O R E W O R D 

It is a great pleasure to welcome you to the XXIV Scandinavian Plant Physiology 

Society Congress in Stavanger, Norway which will take place from the 21 st to the 25 th 

of August 2011. The congress will be held at Stavanger Forum, a conference and 

exhibition centre close to downtown Stavanger. 

The XXIV Scandinavian Plant Physiology Society Congress represents a prime 

opportunity for young as well as established scientists to present and discuss their 

research in an intimate setting. The aim of the congress is to highlight cutting edge, 

fundamental research within plant biology but also to draw attention to the growing 

plant biotechnology industry sector and the relationship between basic and applied 

research efforts. In addition, the congress has education sessions where strategies 

for the education and training of the next generation of plant scientists will be 

discussed. 

The XXIV Scandinavian Plant Physiology Society Congress has an extensive and 

exciting list of invited speakers together with multiple sessions covering numerous 

topics within plant biology and plant biotechnology. The sessions are organized 

consecutively to give all participants the opportunity to attend all presentations. 

The congress also benefits from several social networking events ranging from a 

welcome reception to a museum visit and hikes in the mountains surrounding 

Stavanger. We hope these events will allow you to reacquaint yourselves with old 

friends and colleagues, to meet new fellow scientists and to build new networks 

within your research area. 

We are grateful for financial support from The Scandinavian Plant Physiology Society 

and from the exhibitors present at the congress. 

On behalf of the organizing committee of the XXIV Scandinavian Plant Physiology 

Society Congress we welcome you to Stavanger and hope that you will enjoy and 

benefit from the congress. 

Simon Geir Møller 

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X X I V S P P S C O N G R E S S 2 0 1 1

SIMON GEIR MØLLER 

University of Stavanger, Norway 

SIGRUN REUMANN 


JODI MAPLE 


EVEN HEIEN 


SVEIN GRIMSTAD 

Bioforsk Vest, Norway 

KARSTEN FISCHER 

University of Tromsø, Norway 

MIKE BUSHELL 

Syngenta Limited, UK 

JEAN-DAVID ROCHAIX 

University of Geneva, Switzerland 

JIM AJIOKA 

University of Cambridge, UK 

POUL ERIK JENSEN 

University of Copenhagen, Denmark 

BEN SCHERES 

Univerisity of Utrecht, Netherlands 

A B S T R A C T B O O K – O R G A N I Z A T I O N 

JUERGEN SOLL 

Ludwig-Maximillian University of Munich, Germany 

Organizing Committee 

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ATLE BONES 

Norwegian University of Science and 

Technology, Norway 

POUL ERIK JENSEN 

University of Copenhagen, Denmark 

EVA-MARI ARO 

University of Turku, Finland 

JURGEN SOLL 

Ludwig-Maximilinas University, 

Germany 

KATHARINA PAWLOWSKI 

University of Stockholm, Sweden 

Invited Speakers 

KEARA A. FRANKLIN 

University of Bristol, UK 

SAM ZEEMAN 

ETH, Switzerland 

GUNTIS ABOLTINS-ABOLINS 

Statoil, Norway 

KEN KEEGSTRA 

Michigan State University, USA 

ROLAND VON BOTHMER 

Svalbard Global Seed Vault, Norway 

JANE PARKER COUPLAND 

Max Planck Institute for Plant 

Breeding Research, Germany 

Organization

Program of the Congress 

gram of the Congress 

A B S T R A C T B O O K – P R O G R A M O F T H E C O N G R ESS 

m of the Congress 

Saturday20 th August 

JSPS SPONSORED COLLOQUIUM 

(Find more at http://www.spps2011.no/index.cfm?id=328096) 

Sunday 21 st August 

9.00 – 18.00 EXCURSIONS FOR DELEGATES 

� Pulpit Rock (4 hour hike) – 6 hours in total 

� Kjerag (6 hours hike) – 9 hours in total 

17.00 REGISTRATION AND MOUNTING OF POSTERS 

20.00 WELCOME RECEPTION AT STAVANGER FORUM 

Monday 22 nd August 

8.00 REGISTRATION 

8.45 – 9.00 OPENING OF THE CONGRESS 

PLENARY SESSIONS 

Chair:SIMON GEIR MØLLER (University of Stavanger, Norway) 

9.00 – 9.45 PLENARY SESSION 1 

Biotechnology and industry 

MIKE BUSHELL (Syngenta Limited, UK) 


The biology of algae 

JEAN-DAVID ROCHAIX (University of Geneva, Switzerland) 

10.30 – 11.00 COFFEE BREAK 

of the Congress 

INVITED TALKS 


11.00 – 11.30 INVITED TALK 1 

Synthetic biology 

JIM AJIOKA (University of Cambridge, UK) 

11.30 – 12.00 INVITED TALK 2 

Photosynthesis – photosystem I in synthetic biology 

POUL ERIK JENSEN (University of Copenhagen, Denmark) 

the Congress 

12.00 – 14.00 LUNCH AND POSTER SESSION 

SESSION 1 – BIOTECHNOLOGY AND SYNTHETIC BIOLOGY 

Chair:PETER RUOFF (University of Stavanger, Norway) 

14.00 – 14.20 Heterogonous expression and characterization of human mini-insulin 

protein in tobacco plant 

KY YOUNG PARK (Sunchon National University, South Korea) 

14.20 – 14.40 Studies on targeted silencing of two paralogous ACC oxidase genes in 

banana 

PUNG-LING HUANG (National Taiwan University, Taiwan) 

14.40 – 15.00 Phytoremediation of nitroesters 

RADKA PODLIPNÁ (Czech Academy, Czech Republic) 

4 


Congress 

ngress 

ess


SESSION 2 – SYSTEMS BIOLOGY AND -OMICS 

Chair:JUSTIN PACHEBAT (IBERS/University of Aberystwyth, UK) 

15.00 – 15.20 Indentification of novel plant peroxisomal targeting signals by a 

combination of machine learning methods and in vivo subcellular targeting 

analyses 

SIGRUN REUMANN (University of Stavanger, Norway) 

15.20 – 15.40 Arabidopsis global stress regulation 

PANKAJ BARAH (Norwegian University of Science and Technology, Norway) 

15.40 – 16.00 Chromatin organization in Arabidopsis thaliana 

LARS HENNING (Swedish University of Agricultural Sciences, Sweden) 

16.00 – 16.20 RAD sequencing of plants 

JUSTIN PACHEBAT (IBERS/University of Aberystwyth, UK) 

16.20 – 17.00 COFFEE BREAK 

SESSION 3/4 – PHOTOBIOLOGY, PHOTOSYNTHESIS, 

PHOTORESPIRATION, ALGAEAND MARINE BIOLOGY 

Chair:POUL ERIK JENSEN (University of Copenhagen, Denmark) 

17.00 – 17.20 Engineering glycolate oxidation in Arabidopsis chloroplasts to improve 

photosynthesis 

ALEXANDRA MAIER (Heinrich Heine University, Germany) 

17.20 – 17.40 High efficiency of photosynthesis can be achieved in chlorophyll b-less barley 

mutant Chlorina 3613 

ELENA TYUTEREVA (Komarov Botanical Institute RAS, Russian Federation) 

17.40 – 18.00 Thylakoid membrane tethering of ferredoxin-NADP + oxidoreductase: Redoxregulated 

interaction with Tic62 

MINNA LINTALA (University of Turku, Finland) 

18.00 – 18.20 Thioredoxin interactions in the chloroplast lumen of Arabidopsis thaliana 

THOMAS KIESELBACH (University of Umeå, Sweden) 

18.20 FINISH 

Tuesday 23 rd August 

PLENARY SESSIONS 

Chair:SIGRUN REUMANN (University of Stavanger, Norway) 


Integrated molecular circuits for stem cell activity in Arabidopsis 

roots 

BEN SCHERES (University of Utrecht, Netherlands) 


Chloroplast protein import 

JUERGEN SOLL (Ludwig-Maximillian University of Munich, Germany) 

10.30 – 11.00 COFFEE BREAK 

INVITED TALKS 


11.00 – 11.30 INVITED TALK 3 

Light and temperature signal crosstalk in plant development 

KEARA A. FRANKLIN (University of Bristol, UK) 

5 



11.30 – 12.00 INVITED TALK 4 

The role of reversible glucan phosphorylation in the control of 

starch degradation 

SAM ZEEMAN (ETH, Switzerland) 


SESSION 5 – CELL BIOLOGY 


14.00 – 14.20 Chloroplast differentiation by secretory protein 

YASUO NIWA (University of Shizuoka, Japan) 

14.20 – 14.40 Network motifs leading to plant nitrate homeostasis and oscillatory nitrate 

assimilation 

PETER RUOFF (University of Stavanger, Norway) 

14.40 – 15.00 Gravitropism of Arabidopsis thaliana roots requires the polarization of PIN2 

toward the root tip in meristematic cortical cells 

ABIDUR RAHMAN (Iwate University, Japan) 

SESSION 6 – ORGANELLE BIOLOGY 

Chair:KARSTEN FISCHER (University of Tromsø, Norway) 

15.00 – 15.20 Peroxisome degradation pathways in plants 

OLGA VOITSEKHOVSKAJA (Komarov Botanical Institute RAS, Russian Federation) 

15.20 – 15.40 Regulation of chloroplast biogenesis by chloroplast NADPH-dependent 

thioredoxin system 

EEVI RINTAMÄKI (University of Turku, Finland) 

15.40 – 16.00 Molecular identification of the channel protein mediating the diffusion of 

photorespiratory metabolites across the membrane of plant peroxisomes 

PRADEEP SONI (University of Stavanger, Norway) 


SESSION 7 – DEVELOPMENT 

Chair:JAAKKO KANGASJARVI (University of Helsinki, Finland) 

16.30 – 16.50 Xylan biosynthesis – characterization of glycosyltransferase 43 in Populus 

CHRISTINE RATKE(Swedish University of Agricultural Sciences, Sweden) 

16.50 – 17.10 Oxidative processes are involved in the early events leading to shoot 

gravitropic bending by mediating auxin redistribution 

SONIA PHILOSOPH-HADAS(The Volcani Center, Israel) 

SESSION 8 – SIGNALLING 

Chair:KATHARINA PAWLOWSKI (University of Stockholm, Sweden) 

17.10 – 17.30 BBX21 integrates light and ABA signals during germina 

MAGNUS HOLM (University of Gothenburg, Sweden) 

17.30 – 17.50 Members of the gibberellin receptor gene family (GIBBERELLIN INTENSIVE 

DRAFT1) play distinct roles during Lepidium sativum and Arabidopsis 

thaliana seed germination 

ANTJE VOEGELE (Albert Ludwig University, Germany) 

17.50 – 18.10 Structurally similar but functionally distinct B subunits of protein 

phosphatase 2A 

GRZEGORZ KONERT (University of Turku, Finland) 

18.10 FINISH 

19.30 – 21.00 VISIT OF THE NORWEGIAN PETROLEUM MUSEUM 

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Wednesday 24 th August 


Renewable energy and industry 

GUNTIS ABOLTINS-ABOLINS (Statoil, Norway) 


Cell wall biosynthesis and biofuels 

KEN KEEGSTRA (Michigan State University, USA) 

PLENARY SESSIONS AND INVITED TALK 

Chair:LUTZ EICHACKER (University of Stavanger, Norway) 

10.30 – 11.00 INVITED TALK5 

Plant diversity 

ROLAND VON BOTHMER (Svalbard Global Seed Vault, Norway) 

11.00 – 12.00 SPPS GENERAL ASSEMBLY 


12.00 – 14.00 Teaching tools workshop 

MARY WILLIAMS (Features Editor, The Plant Cell, USA) 

Topic: How to be a great teacher 

Limited to 24 participants (priority to Postdocs and graduate students) 

SESSION 11 – CLIMATE CHANGE AND ADAPTATION TO ABIOTIC STRESS 


14.00 – 14.20 Functional analysis of a novel membrane protein family, COR413, controlled 

by an Arabidopsis transcription factor DREB1A 

MOTOKI KANAI (University of Tokyo, Japan) 

14.20 – 14.40 Analysis on the diversity of drought tolerance of Panicum grasses 

TANJA ZIMMERMANN (Goethe University Frankfurt, Germany) 

14.40 – 15.00 Influence of salinity on antioxidant activity in canola (Brassica napus) 

cultivars 

ESLAM SHAHBAZI (Isfahan University of Technology, Iran) 


PARALLEL SESSION 9 – EDUCATION 

Chair:LISBETH JONSSON (University of Stockholm, Sweden) 

15.30 – 15.45 SHORT PRESENTATION OF THE PROVISIONAL SPPS EDUCATION COMMITTEE 

15.45 – 16.30 Lessons to be learned from the ASPB education committee 

JANE ELLIS (Former chair of the Education Committee of the American Society of Plant 

Biology) 

16.30 – 17.00 SHORT PRESENTATIONS OF EDUCATION AND OUTREACHED PROJECTS 

17.00 – 17.30 PLANS AND IDEAS FOR THE SPPS EDUCATION COMMITTEE. PRESENTATION AND 

DISCUSSION 

SESSION 10/14 – BIOENERGY AND PRIMARY AND SECONDARY MATABOLISM 

Chair:KEN KEEGSTRA (Michigan State University, USA) 

15.30 – 15.50 Apoplastic H2O2 generation mechanisms during extracellular lignin 

formation in Norway spruce cell culture; Effect of H2O2 removal on phenolic 

metabolism 

ANNA KÄRKÖNEN (University of Helsinki, Finland) 

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15.50 – 16.10 The bioimprove programme aims at improved biomass and bioprocesssing 

properties of wood 

HANNELE TUOMINEN (Umeå University, Sweden) 

16.10 – 16.30 What is the transport mechanism of monolignols in lighifying xylem of 

Norway spruce? 

KURT V. FAGERSTEDT (University of Helsinki, Finland) 

16.30 – 16.50 Carotenoid biosynthesis, stabilization and degradation, which determine 

color intensity in petals of yellow-pigmented cut roses, are regulated by 

application of methyl jasmonate 

SHIMON MEIR (The Volcani Center, Israel) 

17.30 FINISH 

19.00 CONGRESS DINNER AT FLOR OG FJÆRE 

Thursday 25 th August 

PLENARY SESSION 

Chair:PETER RUOFF (University of Stavanger, Norway) 


Fine-tuning of plant immune responses to host-adapted 

pathogens 

JANE PARKER COUPLAND (Max Planck Institute for Plant Breeding Research, Germany) 


SESSION 13 – PATHOGEN DEFENSE AND PLANT DISEASE 

Chair:ATLE BONES (Norwegian University of Science and Technology, Norway) 

10.20 – 10.40 Protein phosphates 2A as a regulator of stress responses in Arabidopsis 

SAIJALIISA KANGASJÄRVI (University of Turku, Finland) 

10.40 – 11.00 Impact of P. syringae pv. phaseolicola vs P. syringae pv. Tomato DC3000 on 

photosynthesis and associated processes of the host plant Phaseoulus 

vulgaris 

MARIA LUISA PEREZ BUENO (Estación Experimental del Zaidín, CSIC, Spain) 

11.00 – 11.20 Cell death and resistance 2 gene is a negative regulator of cell death and 

encodes an AAA-type ATPase 

HANN LING WONG (University Tunku Abdul Rahman, Malaysia) 

11.30 – 12.00 POSTER AWARDS AND CLOSING OF THE CONGRESS 

12.00 – 13.00 LUNCH 

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A B S T R A C T B O O K – A B S T R A C T S O F T A L K S 

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Abstracts of Talks

Plenary session 1 


BIOTECHNOLOGY AND INDUSTRY 

Mike Bushell 1 , Jeremy Godwin 2 

1 Syngenta International Research Centre, Jealott’s Hill, Bracknell, UK 

2 Syngenta R&D, Stein, Switzerland 

E-mail: mike.bushell@syngenta.com 

There are many global challenges for which sustainable solutions are urgently required. 

Taken together, tackling issues of poverty reduction, food security, water and energy 

security, in the face of climate change and a world population rising beyond 10 billion 

people before 2100, presents a daunting prospect - well described as “a perfect storm”. 

All of these challenges relate to agriculture. We need to grow more food, yet availability 

of new land for agriculture is limited. Crop and livestock production accounts directly for 

about 14% of all Greenhouse Gas emissions. An additional 17% GHG emissions come from 

land use changes such as deforestation, which also impacts negatively on biodiversity. 

Agriculture consumes about 70% of all fresh water usage. Water is already a limiting 

resource in many parts of the world, and is often being used unsustainably. Diffuse 

pollution from farming activities can also affect water quality. Energy is used to create 

farm inputs, such as synthetic fertilisers, and land is used to grow crops for biofuel, or 

wood for burning. Raising the economic wellbeing of people in poor rural communities is 

critical to addressing all the millennium development goals. 

In short we need to “grow more from less”, meeting the demands for food using land 

and natural resources much more efficiently. We need local solutions that strike the right 

balance between environmental, economic and social issues. There is a growing body of 

opinion that the Sustainable Intensification of Agriculture is a critical approach, taking an 

holistic view of farming – a systems approach at field, farm and landscape levels. 

Biotechnology is a vital tool for agriculture, and will increase in importance in the future. 

Plant biotechnology is often seen as synonymous to Genetic modification of crop plants, 

which is a controversial area for some people. Use of GM organisms in producing many 

other products, such as enzymes or pharmaceuticals, does not raise the same level of 

passionate debate. With the exception of Europe and most of Africa, GM crops are a 

valuable and widely accepted technology, grown on around 10% of the global arable crop 

area. 15 years experience and a huge body of scientific evidence show that GM crops are 

as safe to grow and eat as non-GM crops. Yet Plant Biotechnology is not all about GM 

crops; it is much more than that. 

Unlocking the secrets of the plant genome has given us sophisticated tools to accelerate 

the breeding of superior new plant varieties through the use of genetic markers that 

correlate with desirable properties. “Marker assisted breeding” is a key technology for 

crop improvement. Diagnostics are widely used in human health care, and similar tools 

can help with plant disease prediction and resistance monitoring. Engineered microorganisms 

are used for precision screens and mode of action diagnosis. A systems 

approach to farm productivity will integrate agronomy knowledge with the best available 

agricultural technologies, including Plant Biotechnology . This presentation will look at 

integrated approaches to Sustainable Intensification using examples of approaches for 

abiotic stress tolerance and water use efficiency, insect and fungal disease control on a 

range of globally important crops. 

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SYNTHETIC BIOLOGY: CELL-CELL COMMUNICATION TOOLS FOR 

ENGINEERING MORPHOGENESIS IN PLANTS AND MICROBES 

James Ajioka, Jim Haseloff, James Brown, PJ Steiner, Tim Rudge, Fernan Federici 

University of Cambridge, Cambridge, United Kingdom 

E-mail: ja131@hermes.cam.ac.uk 

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Invited talk 1 

Synthetic Biology consciously employs engineering principles such as modularity, 

abstraction and standardization where biological/genetic parts (e.g. protein coding 

sequences), devices (e.g. bacterial operons) and systems (e.g. sporulation) are 

characterized. Using computational modeling and a standardized method of DNA 

assembly, a myriad of genetic designs can be constructed from a basic set of modular 

parts. The notion of modularity extends to higher order phenomena in biology such as 

multicellular development (e.g. plant organs) and multi-organism interactions (e.g. 

microbial biofilms). Rational engineering of neomorphic structures requires the ability to 

control self-organized pattern formation systems. Central to this process is defining 

proliferating cell cohorts via intercellular signaling, so cell-cell communication becomes an 

essential property for the control and rational design of multicellular assemblages. 

Bacterial quorum sensing systems are well-characterized cell-cell communication systems 

that can be de-constructed in a modular fashion. Gram-negative bacteria use homoserine 

lactones as intercellular signals to initiate cellular changes in response upon achieving a 

specific cell density. In contrast, Gram-positive bacteria employ small peptides as 

signaling molecules in quorum sensing. Examples of each type of signaling system have 

been de-constructed into signal sender and receiver devices.

Plenary session 2 

THE BIOLOGY OF ALGAE 

Jean-David Rochaix 


Departments of Molecular Biology and Plant Biology, University of Geneva, Geneva, Switzerland 

E-mail: Jean-David.Rochaix@unige.ch 

The alga Chlamydomonas reinhardtii has emerged as a powerful model system because of 

its small size, fast growth and short sexual cycle. Its nuclear, chloroplast and 

mitochondrial genomes have been entirely sequenced. This alga is amenable to both 

forward and reverse genetic analysis and efficient transformation methods for the 

nuclear, chloroplast and mitochondrial compartments have been established. It is thus 

well suited for fundamental biological research. Here I will first focus on the biogenesis of 

the photosynthetic apparatus, a process which depends on the concerted interplay 

between the chloroplast and nuclear genetic systems and show how a newly developed 

repressible chloroplast gene expression system has been used to elucidate the role of 

essential chloroplast genes. Because the functioning of the photosynthetic apparatus is 

strongly influenced by environmental conditions, we also study how algae adjust to 

changes in the quality and quantity of light. One of these adjustments, called state 

transitions, involves an adaptive reconfiguration of the light-harvesting complex within 

the thylakoid membrane and a reorganization of the photosynthetic electron transport 

chain. It leads to the balancing of the absorbed light excitation energy between 

photosystem I and photosystem II and allows algal cells to respond to their metabolic 

needs for ATP. 

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PHOTOSYNTHESIS -PHOTOSYSTEM I IN SYNTHETIC BIOLOGY 

P. E. Jensen, K. Jensen, L. M. M. Lassen, A. Zygadlo Nielsen, B. L. Møller 

Department of Plant Biology and Biotechnology, KU-LIFE, Frederiksberg, Copenhagen, Denmark 

E-mail: peje@life.ku.dk 

13 


Invited talk2 

Photosystem I (PSI) from plants, algae and cyanobacteria utilizes energy harvested from 

sunlight to mediate light-driven electron transport across the thylakoid membrane and 

produce reducing equivalents for the metabolic reactions of the organism. 

Cytochrome P450s constitute a very large and highly versatile superfamily of membranebound 

enzymes that catalyse a wide variety of different reactions often difficult to 

achieve using the approaches of chemical synthesis. In this project we aim at coupling PSI 

directly to a cytochrome P450 (P450) to develop a system in which the enzymatic 

reaction of P450s is driven directly by the energy of solar light. 

In nature, electrons for photosynthetic electron transport are derived from water 

oxidation at PSII. At PSI, the electrons are donated to ferredoxin (Fd) on the stromal side 

of the thylakoid membrane. The electrons supplied by PSI may via the soluble electron 

carrier Fd be donated to ferredoxin NADP + oxidoreductase (FNR), which reduces 

NADP + to NADPH. In vivo, the electrons required for the reactions of the P450s are taken 

from NADPH through the NADPH-cytochrome P450 oxidoreductase enzyme. Our recent 

results demonstrate that the delivery of electrons to the P450 can circumvent the 

production and oxidation of NADPH, and instead directly transfer electrons from PSI 

viaFd to an adjacent P450 [1]. 

A supra-metabolon containing PSI and a cytochrome P450 are being constructed to 

facilitate highly efficient electron transfer. We aim at linking the P450 enzyme directly to 

subunits of PSI. Ultimately, the aim is to generate a system in which the fusion protein is 

stably expressed in the thylakoid membrane of cyanobacteria or plants and utilized to 

carry out the desired enzymatic reactions in vivo. 

References 

[1]Jensen et al., ACS Chem. Biol. 2011

Session 1. Biotechnology and synthetic biology 


HETEROGONOUS EXPRESSION AND CHARACTERIZATION OF HUMAN MINI- 

INSULIN PROTEIN IN TOBACCO PLANTS 

Ji Su Kim, Yu Jin Choi, Ky Young Park 

Sunchon National University, Jeollanam-do, South Korea 

E-mail: plpm@sunchon.ac.kr 

This study is focus on the development for plant-derived biopharmaceuticals using 

molecular farming by the attempt to propose prototype and search expression system 

that plant-derived pharmaceutical protein can be produced. The mini-proinsulin is 

designed by minimizing C-peptide between A and B chain of insulin. Ala-Ala-Lys (AAK) 

processing site, trypsin cleavage site, is added in the end of Lys residue of B chain and in 

the front of Gly residue A chain. Also, His6-tag is re-combined to purify insulin in front of a 

C-terminal ER retention signal (KDEL) sequence which induces protein synthesis with high 

efficiency by accumulating in the form of protein. Especially, it is made possible to purify 

insulin protein by one-time trypsin treatment after extraction with His6-tag. Arg(R), 

Arg(R) dibasic processing site was added between A chain and His6-tag linked with Cpeptide. 

N. tabacum plants were transformed with this binary expression vector miniinsulinpTRAkt-rfp. 

After regeneration of transgenic plants in selection medium, the selected 

transgenic plants (T0) were confirmed the miniinsulin gene by PCR. After purification of 

miniinsulin protein from transgenic plants (T0), it will be determined whether it has an 

accurate molecular weight and processing, and then it has a physiological activity for 

insulin in vitro. 

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15 



STUDIES ON TARGETED SILENCING OF TWO PARALOGOUS ACC OXIDASE 

GENES IN BANANA 

Yi-Yin Do, Yu-Chen Liao, Mei-Lun Yeh, Pung-Ling Huang 

Department of Horticulture, National Taiwan University, Taipei, Taiwan 

E-mail: pungling@ntu.edu.tw 

Banana (Musa spp.) is one of the typical climacteric fruits with great economic 

importance. Ethylene production is essential for the ripening of banana and 1aminocyclopropane-1-carboxylic 

acid oxidase (ACO), the last key enzyme in the ethylene 

biosynthesis pathway, is encoded by two or more paralogous genes in the banana 

genome. Two ACO genes, Mh-ACO1 and Mh-ACO2, have been cloned from banana and 

their expression patterns during fruit ripening have been characterized. To elucidate the 

respective roles of these two genes a series of RNA interference (RNAi) lines that showed 

reduced expression of either of these two genes were obtained. A significant delay in 

change of peel color, and a considerably reduction in ethylene production and respiration 

rate were observed in the Mh-ACO2 RNAi lines. The characteristics of these phenotypes 

suggest that Mh-ACO2 plays pivotal role in fruit ripening of banana. The silencing of Mh- 

ACO1 seemed affect growth and development at the whole plant level and resulted in a 

dwarf phenotype characterized by slow growth in the early growth stage. The 

phenotypes related to the role of Mh-ACO1 involved in fruit ripening will be the subject of 

a future, more detailed investigation of these plants.



PHYTOREMEDIATION OF NITROESTERS 

Radka Podlipná 1 , Eliška Nachlingerová 2 , Tomáš Vaněk 1 

1 Laboratory of Plant Biotechnologies, Experimental Botany CAS, Prague, Czech Republic 

2 Faculty of Environmental Technology, Institute of Chemical Technology, Prague, Czech Republic 

E-mail: podlipna@ueb.cas.cz 

Explosives are toxic, recalcitrant to degradation and contaminate large areas of land and 

ground water. Remediation of these xenobiotics is difficult and an enormous logistical 

task. Phytoremediation is a technique that offers an environment friendly, low-cost 

alternative to current remediation techniques. However this approach is hindered by the 

low metabolic abilities of plants towards these xenobiotic compounds and the 

phytotoxicity of these compounds. Pentaerythritol tetranitrate (PETN) is one of the most 

powerful detonated explosives. The ecotoxicity of PETN was characterized by the test 

based on the inhibition of elongation of primary roots 3days-old mustard and hemp 

seedlings. The fate of PETN on exposure to plant was investigated in hydroponic and in 

vitro systems. Medium and extracts of plants were analyzed by HPLC (High-performance 

liquid chromatography). This study showed that PETN was taken up by plant species in 

both systems. The accumulation of this nitroester was observed more in the roots than in 

the aerial parts of the plants. 

Acknowledgments 

The authors thank for financial support of projects 1MPO6030 and grant MYES of CR n. OC1002. 

16 



17 


Session 2. Systems biology and -omics 

IDENTIFICATION OF NOVEL PLANT PEROXISOMAL TARGETING SIGNALS BY 

A COMBINATION OF MACHINE LEARNING METHODS AND IN VIVO 

SUBCELLULAR TARGETING ANALYSES 

Thomas Lingner 1,2 , Amr R. Kataya 2 , Gerardo E. Antonicelli 2,3 , Aline Benichou 2 , Kjersti 

Nilssen 2 , Xiong-Yan Chen 2 , Tanja Siemsen 3 , Burkhard Morgenstern 1 , Peter Meinicke 1 , 

Sigrun Reumann 2,3 

1 Department of Bioinformatics, Institute for Microbiology, Goettingen, Germany 

2 Centre for Organelle Research, University of Stavanger, Stavanger, Norway 

3 Department of Plant Biochemistry, Georg-August-University of Goettingen, Goettingen, Germany 

E-mail: sigrun.reumann@uis.no 

In the post-genomic era, accurate prediction tools are essential for identification of the 

proteomes of cell organelles. Prediction methods have been developed for peroxisometargeted 

proteins in animals and fungi, but are missing specifically for plants. For 

development of a predictor for plant proteins carrying peroxisome targeting signals type 

1 (PTS1), we assembled more than 2,500 homologous plant sequences, mainly from EST 

databases. We applied a discriminative machine learning approach to derive two different 

prediction methods, both of which showed high prediction accuracy and recognized 

specific targeting-enhancing patterns in the regions upstream of the PTS1 tripeptides. 

Upon application of these methods to the Arabidopsis genome, a total of 392 gene 

models were predicted to be peroxisome-targeted. These predictions were extensively 

tested in vivo, resulting in a high experimental verification rate of Arabidopsis proteins 

previously not known to be peroxisomal. The prediction methods were able to correctly 

infer novel PTS1 tripeptides, which even included novel residues. Twenty-three newly 

predicted PTS1 tripeptides were experimentally confirmed, and a high variability of the 

plant PTS1 motif was discovered. These prediction methods will be instrumental in 

identifying low-abundance and stress-inducible peroxisomal proteins and defining the 

entire peroxisomal proteome of Arabidopsis and agronomically important crop plants.



ARABIDOPSIS GLOBAL STRESS REGULON 

Pankaj Barah 1 , Simon Rasmussen 2 , Maria Cristina Suarez-Rodriguez 3 , Laurent Gautier 2 , 

John Mundy 3 , Henrik Bjørn Nielsen 2 , Atle M. Bones 1 

1 

Department of Biology, Norwegian University of Science and Technology, Trondheim, Norway 

2 

Center for Biological Sequence Analysis, Department of Systems Biology, Technical University of Denmark, Kongens 

Lyngby, Denmark 

3 

Department of Biology, University of Copenhagen, Copenhagen, Denmark 

E-mail: pankaj.barah@bio.ntnu.no 

Co-expressed or co-regulated genes can indicate their involvement in similar biological 

processes, meaning that individual modules can be attributed to specific biological 

processes. Using this basic concept together with information about signal transduction 

and metabolic pathways, genes that share a similar expression profile across multiple 

spatial, temporal, environmental and genetic conditions could be considered as 

eukaryotic regulon. 

Plants have developed elaborate networks of defense mechanisms against different 

types of stresses, but the level of network crosstalk makes it challenging to correlate 

various types of responses to a particular stress. Meta-analysis of the Arabidopsis 

transcriptome offers the potential to identify regulons. As there are many standards on 

how to grow plants and to conduct transcription experiments, it is difficult to extract 

compatible information across data sets. To overcome this problem of incompatibility of 

independent microarray experiments, 23 different genotypes (10 ecotypes and 13 

mutants) were subjected to a set of 5 individual stress treatments and 8 combinations of 

stress treatments under same experimental conditions. This was a part of ERA-NET Plant 

Genomics, MultiStress project (http://www.erapg.org/). Using this ERA-PG MultiStress 

global dataset, we have explored the organization of stress regulons in the model plant 

Arabidopsis. 

18 



CHROMATIN ORGANISATION IN ARABIDOPSIS THALIANA 

Huan Shu 1 , Wilhelm Gruissem 2 , Lars Hennig 1 

1 SLU Uppsala, Uppsala, Sweden 

2 ETH Zurich, Zirich, Switzerland 

E-mail: lars.hennig@slu.se 

19 



In a eukaryotic nucleus, DNA is packaged into chromatin, which is commonly classified as 

hetero- and euchromatin. Heterochromatin, as opposed to euchromatin, has been first 

identified at the cytological level as chromosomal regions that remained strongly 

condensed through interphase, and the condensed heterochromatin often proved to be 

resistant to nuclease treatments. Heterochromatin of the model plant Arabidopsis is 

thought to be of low complexity and to contain mainly centromeric and pericentromeric 

repeat regions and nucleolar organizers. However, these conclusions were often based 

on fluorescence in situ hybridization (FISH) experiments, which show only a local 

snapshot but cannot easily reveal genome-wide information. Here, we present a genomewide 

map of candidate heterochromatin sequences in Arabidopsis using relative 

resistance to DNaseI as an operational criterion for the heterochromatic state. 

Heterochromatic regions in Arabidopsis were identified at genomic level and with high 

resolution by DNase-array, a technique to quantify chromatin sensitivity to DNase I as a 

function of genome position We will present a detailed characterization of DNaseIresistant 

heterochromatic regions of the Arabidopsis genome in relation to chromosomal 

localization, transcriptional activity and chromatin modifications; and we will discuss how 

these results help to illuminate heterochromatin - a poorly understood dark matter of the 

genome.



RAD SEQUENCING OF PLANTS 

Justin Pachebat 

IBERS, Aberystwyth University, Aberystwyth, United Kingdom 

E-mail: jip@aber.ac.uk 

RAD sequencing is a form of reduced representation sequencing that utilises the highthroughput 

illumina sequencing platform to generate SNPs for genotyping large numbers 

of individuals in a population. The procedure involves digestion of genomic DNA with a 

rare cutting restriction enzyme, ligation of a barcoded illumina compatible sequencing 

adapter to the digested DNA, followed by pooling of samples, sonication to frgament the 

DNA, and subsequent ligation of a second sequencing adapter and PCR amplification of 

the multiplex library. These multiplexed samples are sequenced on an Illumina platform 

and reads assigned to individual plants via the barcodes, and SNPs identified in the reads. 

Here we describe how RAD sequencing can be applied to plants, such as Lolium perenne 

for SNP genotyping, despite the lack of a reference genome. 

20 



Session 3/4. Photobiology, photosynthesis, photorespiration, algae and marine biology 

ENGINEERING GLYCOLATE OXIDATION IN ARABIDOPSIS CHLOROPLAST 

TO IMPROVE PHOTOSYNTHESIS 

Alexandra Maier, HolgerFahnenstich, Sussane von Caemmerer, Ulf-Ingo Flügge, 

Veronica G. Maurino 

Institute for Developmental- and Molecular Biology of Plants Heinrich-Heine-University, Düsseldorf, Germany 

E-mail: alexandra.maier@uni-duesseldorf.de 

In C3-plants, the oxygenation reaction of RubisCO directs the flow of carbon through the 

photorespiratory pathway, which can result in a loss of up to 50% of the carbon previously 

fixed. In this work, a complete glycolate catabolic cycle was established into chloroplasts 

of Arabidopsis by which one molecule of glycolate is completely oxidized within the 

chloroplast to two molecules of CO2. For a functionally, operational pathway three 

transgenic and two endogenous enzyme activities were requiered. Plants expressing the 

novel pathway produced rossettes with more leaves and higher fresh and dry weight but 

individual leaves were thinner than the wild type. The photosynthetic rates were higher 

on a dry weight and chlorophyll basis and there were no differences in the apparent 

CO2compensation point. In addition, transgenic plants showed lower glycine/serine ratios 

than the wild type, indicating a reduction of the flux through the photorespiratory 

pathway. In this way, a photorespiratory bypass was created which resulted in increased 

efficiency of CO2 assimilation and enhanced biomass production. Efforts to develop of an 

optimized version of the basic approach using only two transgene enzymes will also be 

presented. 

21 




HIGH EFFICIENCY OF PHOTOSYNTHESIS CAN BE ACHIEVED IN 

CHLOROPHYLL B-LESS BARLEY MUTANT CHLORINA 3613 

Elena V.Tyutereva, A.N.Ivanova, Yu. ZhelninskayaI, Olga V. Voitsekhovskaja 

Komarov Botanical Institute RAS, St. Petersburg, Russian Federation 

E-mail: tuterlena@mail.ru 

Field-grown chlorophyllb-less barley chlorina 3613 plants were shaded to 800 µmolm -2 s -1 

(i.e. to 40% of 2000-2500 µmol m -2 s -1 PAR measured at noon) for a week, followed by 

restoration of natural radiation regime. Under the shading gauze, the yellow-pale chlorina 

plants turned bright-green while their chloroplast ultrastructure, leaf and stem 

dimensions, vegetative and seed production changed significantly. After removal of the 

gauze, the chlorophyll a content increased twice which was accompanied by a threefold 

increase in CO2 absorption rate as compared with control plants, while no restoration of 

chlorophyll b synthesis occurred. The high efficiency of photosynthesis persisted in these 

plants until the end of vegetation. The experiment was carried out in 2006 and 

successfully reproduced in 2007 but failed to reproduce in cold and cloudy summer in 

2008 showing that the results could be achieved only in a narrow range of light and 

temperature conditions. The dynamics of LHC proteins and of photosystems reaction 

centres in control and shaded chlorina 3613 plants was studied by western blot analyses. 

The effects of the shading on restoration of chloroplast structure and functions in fieldgrown 

chlorophyll b-less chlorina 3613 plants are discussed. 

22 




THYLAKOID MEMBRANE TETHERING OF FERREDOXIN-NADP + 

OXIDOREDUCTASE (FNR): REDOX-REGULATED INTERACTION WITH TIC62 

Minna Lintala 1 , Philipp Benz 2,3,4 , Anna Stengel 2,3 , Jürgen Soll 2,3 , Bettina Bölter 2,3 , Paula 

Mulo 1 

1 Molecular Plant Biology, Department of Biochemistry and Food Chemistry, University of Turku, Turku, Finland 

2 Munich Center for Integrated Protein Science CiPSM, Ludwig-Maximilians-UniversitätMünchen, Munich, Germany 

3 Department of Biology I, Botany, Ludwig-Maximilians-UniversitätMünchen, Planegg-Martinsried, Germany 

4 Present address: Energy Biosciences Institute, University of California at Berkeley, Berkeley, USA 

E-mail: mikali@utu.fi 

Ferredoxin-NADP + oxidoreductase (FNR) has a well defined role in catalyzing the final 

step of linear photosynthetic electron transfer chain. By performing electron transfer 

from ferredoxin to NADP + , FNR is an important mediator of reducing power from 

membrane-bound light reactions to the stromal metabolic pathways. In Arabidopsis 

thaliana, two chloroplast-targeted FNR isoforms FNR1 and FNR2 can be found attached to 

the thylakoid and inner envelope membranes, as well as in the stroma. We have shown 

that Tic62 protein binds FNR to thylakoid membranes. Formation of the large Tic62-FNR 

complexes is mediated by light and stromal redox-state. As photosynthesis is not 

affected in Arabidopsis tic62 mutant plants, which lack most of the thylakoid associated 

FNR, and because most of the FNR is released from the thylakoids in light, we presume 

that the soluble pool of FNR is highly capable of performing photosynthetic activity. We 

conclude that Tic62 represents a major FNR interaction partner not only at the inner 

envelope membrane but also at the thylakoid membrane of Arabidopsis thaliana. We 

furthermore propose that correct allocation of FNR is used to efficiently regulate Fddependent 

electron partitioning in the chloroplast. 

23 




THIOREDOXIN INTERACTIONS IN THE CHLOROPLAST LUMEN OF 

ARABIDOPSIS THALIANA 

Michael Hall, Wolfgang Schröder,Thomas Kieselbach 

Department of Chemistry, Umeå University and Umeå Plant Science Center, Umeå, Sweden 

E-mail: michael.hall@chem.umu.se 

Thioredoxins, originally discovered as a link between photosynthesis and chloroplast 

metabolism, are a family of small disulfide-reductaseproteins found in all organisms. 

Arabidopsis thaliana has at least 44 known genes coding for thioredoxins or thioredoxinlike 

proteins, out of which 21 are predicted or experimentally shown to be located in 

plastids. Within the thylakoid membrane, the focal point of oxygenic photosynthesis, lays 

the thylakoid lumen, a small sub-organelle compartment containing a distinct protein 

population. The proteome of the thylakoid lumen of Arabidopsis thaliana is comprised of 

approximately 80 proteins. While many of the proteins have unknown functions, a 

number of them are important enzymes regulating processes such as oxygen-evolution 

and the xanthophyll cycle. Although no thioredoxin has so far been identified in the 

thylakoid lumen, several observations strongly indicate the presence of a thiotransduction 

pathway in the lumen: an x-ray structure study of the luminal immunophilin 

FKBP13 by Gopalan et al. showed that this protein contained two disulfide bonds which 

could be reduced by E.coli thioredoxin and Marchand et al. found that the extrinsic 

Photosystem II proteins PsbO1 and PsbO2, and also the luminal pentapeptide protein TL17 

could be reduced by thioredoxin h3. 

We have identified putative thioredoxin targets in the chloroplast lumen of Arabidopsis 

thaliana using three complementary proteomic methods, fluorescence two-dimensional 

gel-electrophoresis, two-dimensional gel-electrophoresis coupled with differential 

alkylation and Trx affinity chromatography. In total we have identified 19 Trx target 

proteins, thus covering more than 40% of the currently known lumenal chloroplast 

proteome. We also show that the redox state of thiols is decisive for degradation of the 

extrinsic PsbO1 and PsbO2 subunits of photosystem II. Our current work is focused on 

functional and structural characterization of target proteins of unknown function, 

including the PsbP-domain proteins and pentapeptide repeat proteins. 

24 



INTEGRATED MOLECULAR CIRCUITS FOR STEM CELL ACTIVITY IN 

ARABIDOPSIS ROOTS 

Ben Scheres 

University of Utrecht, Utrecht, The Netherlands 

E-mail: b.scheres@uu.nl 

25 


Pleanary session 3 

Plant stem cells reside in niches and are maintained by short-range signals emanating 

from organizing centres. The Arabidopsis PLETHORA genes encode transcription factors 

required for root stem cell specification [1, 2]. PLT expression is induced by the indolic 

hormone auxin, depends on auxin response factors and follows auxin accumulation 

patterns. The PLT gene clade extensively regulates expression of the PIN facilitators of 

polar auxin transport in the root and this contributes to a specific auxin transport route 

that maintains stem cells at the appropriate position[3]. We are addressing the properties 

of the PLT-PIN feedback loop by gene and protein network analysis and computational 

modelling. The emerging picture is one in which flexible feedback circuits translate auxin 

accumulation into region- and cell type specification patterns. 

Stem cells and their daughters in the root display specific asymmetric divisions at fixed 

locations. We investigate how such divisions are spatially regulated. The SHORTROOT- 

SCARECROW transcription factor pathway plays a role in patterning the quiescent center 

and cortex/endodermis stem cells and provides mitotic potential to the stem cell 

daughters that form the proximal meristem. This activity involves the conserved 

RETINOBLASTOMA-RELATED (RBR) pocket protein[4], and we have established 

molecular links between the RBR pathway and SCARECROW action that form a feedback 

control system. In addition, RBR activity is modulated by auxin abundance, itself 

regulated through an intercellular distribution system, and by cell cycle progression. 

Formal analysis of this feedback circuit indicates that it acts as a bistable switch that 

ensures the occurrence of an asymmetric division at fixed positions. Our work illustrates 

how formative divisions that shape plant tissues can be robustly positioned by dynamic 

regulatory circuits that combine intracellular and extracellular loops. 

References 

[1]Aida et al.,Cell 2004 

[2] Galinhaet al., Nature 2007 

[3] Grieneisen et al., Nature2007 

[4] Wildwater et al., Cell2005



CHLOROPLAST PROTEIN IMPORT 

Jürgen Soll 

Ludwig-Maximilinas University, München, Germany 

E-mail: soll@lmu.de 

Chloroplasts must import several thousand different proteins to obtain and maintain their 

biochemical identity and function. Protein turnover, environmental adaptation and 

cellular division of labour require a constant adjustment of the protein complement in 

chloroplasts. Protein import into chloroplasts is regulated at all stages of the process, 

ranging from co- or early posttranslational events in the cytosol to late stages in the 

translocation process which reflect the metabolic as well as the redox status of the 

organelle. 

26 



27 



LIGHT AND TEMPERATURE SIGNAL CROSSTALK IN PLANT DEVELOPMENT 

Kerry Franklin 

University of Bristol, Bristol, United Kingdom 

E-mail: kerry.franklin@bristol.ac.uk 

Light and temperature are two of the most important environmental cues regulating 

plant development. Our work investigates crosstalk between these signalling pathways. 

Plants monitor their ambient light environment using specialised information-transducing 

photoreceptors. A major role of the phytochrome photoreceptors in natural 

environments is the detection of neighbouring vegetation and initiation of architectural 

responses to avoid shading. Modest changes in ambient growth temperature can, 

however, dramatically affect shade avoidance strategy. We are currently investigating the 

regulatory mechanisms controlling shade avoidance at different temperatures. This work 

has identified crosstalk between light quality and freezing tolerance signalling pathways 

in cooler conditions. Growth at elevated temperatures also has dramatic effects on plant 

flowering time, stature and biomass. The mechanisms through which plants sense 

temperature are less understood. We previously showed that high temperature and light 

quality signalling pathways converge on a shared transcription factor, PHYTOCHROME 

INTERACTING FACTOR 4 (PIF4) to control elongation growth. Our recent work has now 

provided mechanistic insight in to PIF4 function at elevated temperatures. Upstream and 

downstream targets in this signalling pathway will be discussed and parallels with shade 

avoidance explored.



THE ROLE OF REVERSIBLE GLUCAN PHOSPHORYLATION IN THE CONTROL 

OF STARCH DEGRADATION 

Samuel C. Zeeman 

Department of Biology, ETH Zürich, Zürich, Switzerland 

E-mail: szeeman@ethz.ch 

Starch, a major product of photosynthesis in chloroplasts, is comprised of glucans that 

form semi-crystalline granules. Starch made during the day is degraded at night to 

support metabolism. Glucan phosphorylation, mediated by glucan water dikinases (GWD 

and PWD), is required for starch degradation. Phosphorylation disrupts the starch granule 

surface, rendering the glucans accessible for degrading enzymes. The chloroplastic 

phosphatase SEX4 (Starch EXcess4) is also required for starch degradation. 

Phosphorylated intermediates of starch breakdown accumulate in sex4 mutants as the 

phosphate groups, while necessary to disrupt the granule surface, obstruct locally 

enzymes of starch degradation, such as beta-amylases. Starch granule degradation in 

vitro is increased by simultaneous phosphorylation and dephosphorylation, corroborating 

the hypothesis that reversible glucan phosphorylation and glucan hydrolysis are 

synergistic processes. Plants have two homologues of SEX4 - LSF1 and LSF2 (Like Sex 

Four). With collaborating labs, we recently demonstrated that the loss of LSF1 also causes 

a starch-excess phenotype. However, the roles of LSF1 and SEX4 differ; phosphooligosaccharides 

do not accumulate in lsf1 and recombinant LSF1 protein has no 

phosphatase activity. These findings indicate additional complexity in the process of 

transient phosphorylation of the granule during starch degradation, which is the subject 

of ongoing research in our lab. 

28 



CHLOROPLAST DIFFERENTIATION BY SECRETORY PROTEIN 

Yasuo Niwa, Akiko Ogino, Hironori Kageshima, Shingo Goto, Hirokazu Kobayashi 

University of Shizuoka, Shizuoka, Japan 

E-mail: niwa@u-shizuoka-ken.ac.jp 

29 


Session 5.Cell biology 

The plant hormones such as auxin and cytokinin can control plant cell de-differentiation 

and re-differentiation. It is possible to induce green callus from white callus by high 

concentration of cytokinin treatment. In this study, white callus induced from roots of 

Arabidopsis were used for mutant screening by activation tagging. Mutant candidates 

were selected as a greening phenotype from the white callus without high levels of 

cytokinin treatment. As a result, three candidates can be obtained and named ces101, 102, 

and 103. To identify the gene for ces102 phenotype, TAIL-PCR (thermal asymmetric 

interlaced-PCR) was performed. T-DNAs were found to be inserted into two locations in 

Arabidopsis genome. The expression level of two genes located near the insertion points 

was increased. From the results of phenotypic analysis by over-expression of candidate 

genes, the gene encoding 119 amino acids corresponds to ces102. According to the 

characteristics of the amino acid sequence, CES102 was expected to have a signal 

peptide. Localization analysis of GFP fusion protein, CES102-GFP could be detected at ER. 

From these results, plastid differentiation might be controlled through the secretory 

pathway.



NETWORK MOTIFS LEADING TO PLANT NITRATE HOMEOSTASIS AND 

OSCILLATORY NITRATE ASSIMILATION 

Yongshun Huang, Ingunn W. Jolma, Tormod Drengstig, Peter Ruoff 

Centre for Organelle Research, University of Stavanger, Stavanger, Norway 

E-mail: peter.ruoff@uis.no 

Homeostasis is an essential property of all living systems. While themolecular details of 

homeostatic component processes such as uptake, storage,efflux and assimilation are 

becoming well understood our knowledge how theprocesses integrate and lead to 

robust homeostatic behavior in the presence ofenvironmental perturbations is still poor. 

Here we describe a complete set oftwo-component negative feedback motifs, which, 

based on thecontrol-engineering concept of integral feedback can maintain 

robusthomeostasis both at steady state and at oscillatory or pulsatile workingconditions. 

By using cytosolic nitrate homeostasis in plants as an example we demonstrate how 

these controller motifs integrate uptake, storage,efflux, and assimilation, and how 

switching between different controller typescan occur during remobilization of nitrate 

from the vacuole. 

30 



GRAVITROPISM OF ARABIDOPSIS THALIANA ROOTS REQUIRES THE 

POLARIZATION OF PIN2 TOWARD THE ROOT TIP IN MERISTEMATIC 

CORTICAL CELLS 

31 



Abidur Rahman 1 , Maho Takahashi 1 ,Kyohei Shibasaki 1 , Shuang Wu 2 , Takehito Inaba 3 , Seiji 

Tsurumi 4 , Tobias I. Baskin 2 

1 Cryobiofrontier Research Center, Faculty of Agriculture, Iwate University, Morioka, Iwate, Japan 

2 Departmentof Biology, University of Massachusetts, Amherst, Massachusetts, USA 

3 Interdisciplinary Research Organization, Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan 

4 Center for Supports to Research and Education Activities Isotope Division, Kobe University, Nada, Kobe, Japan 

E-mail: abidur@iwate-u.ac.jp 

In the root, the transport of auxin from the tip to the elongation zone, referred to here as 

shootward, governs gravitropic bending. Shootward polar auxin transport, and hence 

gravitropism, depends on the polar deployment of the PIN-FORMED auxin efflux carrier 

PIN2. In Arabidopsis thaliana, PIN2 has the expected shootward localization in epidermis 

and lateral root cap; however, this carrier is localized toward the root tip (rootward) in 

cortical cells of the meristem, a deployment whose function is enigmatic. We use 

pharmacological and genetic tools to cause a shootward relocation of PIN2 in 

meristematic cortical cells without detectably altering PIN2 polarization in other cell types 

or PIN1 polarization. This relocation of cortical PIN2 was negatively regulated by the 

membrane trafficking factor GNOM and by the regulatory A1 subunit of type 2-A protein 

phosphatase (PP2AA1) but did not require the PINOID protein kinase. When GNOM was 

inhibited, PINOID abundance increased and PP2AA1 was partially immobilized, indicating 

both proteins are subject to GNOM-dependent regulation. 

Shootward PIN2 specifically in the cortex was accompanied by enhanced shootward polar 

auxin transport and by diminishedgravitropism. These results demonstrate that auxin 

flow in the root cortex is important for optimal gravitropic response.

Session 6.Organelle biology 


PEROXISOME DEGRADATION PATHWAYS IN PLANTS 

Olga V. Voitsekhovskaja 1,2 , Andreas Schiermeyer 3 , Sigrun Reumann 1,4 

1 Department of Plant Biochemistry, Georg-August-University of Goettingen, Goettingen, Germany 

2 Komarov Botanical Institute, Russian Academy of Sciences, St. Petersburg, Russian Federation 

3 Fraunhofer-Institut fürMolekularbiologie und AngewandteOekologie, Aachen, Germany 


E-mail: ovoitse@yandex.ru 

In fungi and mammals, peroxisomes are reported to be degraded by micro- and/or 

macroautophagy but such knowledge is yet lacking for plants. To study peroxisome 

degradation pathways in plants, stable transgenic suspension-cultured cell lines of 

tobacco cv Bright Yellow 2 were generated that expressed a peroxisome-targeted version 

of enhanced yellow fluorescent protein (EYFP). Autophagy was induced by carbohydrate 

withdrawal from the culture medium. Biochemical and cytological analyses demonstrated 

that application of the inhibitor of macroautophagy, 3-methyladenine (3-MA), caused a 

significant accumulation of EYFP-SKL and native peroxisomal proteins and an increase in 

cellular peroxisome numbers, indicating that peroxisomes are degraded by 

macroautophagy. The subcellular localization of peroxisomes was shown to coincide with 

autolysosomes and autophagic bodies, consistent with the transport of peroxisomes to 

the vacuole for proteolytic degradation by macroautophagic compartments. 

Unexpectedly, 3-MA caused a significant accumulation of peroxisomes also under 

nutrient-rich conditions, demonstrating that plant peroxisomes are turned over by 

constitutive macroautophagy under standard growth conditions. The rate of peroxisome 

turnover exceeded that of plastids and mitochondria under both conditions. The results 

suggest that peroxisomal matrix proteins are prone to oxidative damage even under nonstressed 

conditions and that specific yet unknown signaling pathways exist for selective 

degradation of dysfunctional peroxisomes. 

32 



33 



REGULATION OF CHLOROPLAST BIOGENESIS BY CHLOROPLAST NADPH- 

DEPENDENT THIOREDOXIN SYSTEM 

Eevi Rintamäki 1 , Anna Lepistö 1 , Jouni Toivola 1 , Florence Vignols 2 

1 Department of Biochemistry and Food Chemistry, University of Turku, Turku, Finland 

2 IRD, UMR5096 GénomeetDéveloppement des Plantes, Montpellier, France 

E-mail: evirin@utu.fi 

Thioredoxins are small regulatory proteins containing redox-active cysteine pair. In the 

reduced state, thioredoxins control the function of cellular target proteins by reducing 

the disulfide bridges in the redox active sites of proteins. Subsequently, the oxidized 

thioredoxins are reduced by thioredoxinreductases (NTR), forming together a 

thioredoxin system. Chloroplast NADPH-thioredoxin reductase (NTRC) is a unique 

enzyme with dual domains comprising both the NTR and thioredoxin sequences within 

the protein. We have dissected the impact of NTRC in chloroplast development. 

Knockout of NTRC severely reduced the growth of Arabidopsis. The number of 

chloroplasts per cell and the level of chlorophyll in leaves were significantly reduced in 

the T-DNA insertion lines of NTRC (ntrc). Genes encoding critical regulatory enzymes in 

chlorophyll biosynthesis were differentially expressed in ntrc plants. Transmission 

electron microscopy illustrated a distorted biogenesis of chloroplasts in ntrc plants. A 

series from the regular to seriously aberrant chloroplasts with abnormally distributed 

thylakoid membranes existed in a single ntrc mesophyll cell. A preliminary Y2H assay 

indicated that the thioredoxin domain of NTRC can interact with FtsZ protein that forms a 

core component of the inner division machinery of plastids. We propose that NTRC 

contributes the redox regulation of chlorophyll biosynthesis and/or to plastid division.



MOLECULAR IDENTIFICATION OF THE CHANNEL PROTEIN MEDIATING THE 

DIFFUSION OF PHOTORESPIRATORY METABOLITES ACROSS THE 

MEMBRANE OF PLANT PEROXISOMES 

Pradeep Soni 1 , Elke Maier 2 , Roland Benz 2 , Sigrun Reumann 1 


2 Rudolf-Virchow-Zentrum, Wuerzburg, Germany 

E-mail: pradeep.soni@uis.no 

Plant peroxisomes are essential cell organelles that primarily carry out oxidative 

metabolic reactions. Major metabolic pathways of plant peroxisomes such as 

photorespiration have been described fairly well in the past years regarding the soluble 

matrix proteins involved. However, transport proteins for metabolic intermediates have 

not been identified in any plant species to date. Their identification and biochemical 

characterization are highly important, for instance, to allow for kinetic modelling of 

photorespiration under abiotic stress conditions. By electrophysiological means, a porinlike 

channel has been characterized in the membrane of spinach leaf peroxisomes and 

castor bean glyoxysomes in the 90s. To take advantage of the molecular and genomic 

tools available for Arabidopsis, we isolated peroxisomes from mature Arabidopsis leaves. 

Indeed, an anion-selective channel of similar single channel conductance could be 

detected in a standard electrolyte such as 1 M KCl. The selectivity properties were 

examined using different types of ions, and binding assays have been carried out using 

different photorespiratory substrates. Taken together, the channel protein appears wellsuited 

to mediate the diffusion of small carboxylates such as intermediates of 

photorespiration, fatty acid beta-oxidation, and the glyoxylate cycle (C2-C6, e.g. glycolate, 

malate, citrate). Candidate proteins including the Arabidopsis ortholog of a mammalian 

metabolite channel of peroxisomes have been subcloned and are presently being overexpressed 

in Pichia to allow for further purification by affinity chromatography. The 

purified proteins will be reconstituted in planar lipid bilayers and their permeability 

properties be investigated in different inorganic and organic ion solutions of different 

structural properties. 

34 



35 


Session 7.Development 

XYLAN BIOSYNTHESIS - CHARACTERIZATION OF GLYCOSYLTRANSFERASE 

43 IN POPULUS 

Christine Ratke, Marcel Naumann, Barbara Terebieniec, Ewa Mellerowicz 

Department of Forest Genetics and Plant Physiology, SLU, UPSC, Umeå, Sweden 

E-mail: christine.ratke@slu.se 

Woody tissues are formed by thickening of the secondary cell wall, consisting of cellulose, 

xylan and lignin. Consequently xylan represents a big fraction of Earths biomass, 

prompting an interest in its biosynthesis and function. 

Xylan deficient mutants in Arabidopsis show a reduced growth and the irregular xylem 

(irx) phenotype. Two of these mutants are defective in two different glycosyltransferases 

(GT) from family 43. The xylan in these mutants has a reduced chain length suggesting 

that these glycosyltransferases are involved in xylan backbone elongation. 

We are characterizing the role of GT43 family in wood development in Populus. The family 

has seven genes forming three distinct clades. RT-PCR and promoter-GUS-fusions were 

used to study the tissue-specific expression patterns of the genes. Hybrid aspen was used 

to downregulate all three GT43 clades individually and in combinations by RNAi, using 

both constitutive and wood-specific promoters. We are analyzing the effects on plant 

growth, cell morphology and fiber lengths, cell wall composition, saccharification / 

fermentation ability and mechanical properties. 

This work on xylan biosynthesis is increasing our basic understanding regarding wood 

development and properties, potentially leading to industrial applications.

Session 7.Development 


OXIDATIVE PROCESSES ARE INVOLVED IN THE EARLY EVENTS LEADING TO 

SHOOT GRAVITROPIC BENDING BY MEDIATING AUXIN REDISTRIBUTION 

Neta Bashan 1 , Shimon Meir 1 , Haya Friedman 1 , Elisha Tel-Or 2 ,Sonia Philosoph-Hadas 1 

1 Department of Postharvest Science of Fresh Produce, ARO, TheVolcani Center, Bet-Dagan, Israel 

2 The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, The Robert H. Smith Faculty of Agriculture, 

Food & Environment, The Hebrew University of Jerusalem, Rehovot, Israel 

E-mail: vtsoniap@volcani.agri.gov.il 

The stimulus transduction events occurring during shoot gravitropism are mediated 

through differential changes in the level and action of auxin, associated with differential 

growth leading to shoot curvature. Garvistimulated tomato (Solanumlycopersicum) 

shoots showed a visual upward bending after a lag of 5 h, which was preceded by an 

auxin gradient in favor of the lower shoot side. Both the auxin redistribution across the 

shoot and the subsequent bending were prevented by the antioxidant N-acetyl-cysteine 

(NAC), suggesting the involvement of reactive oxygen species (ROS) in the process. Our 

microarray analysis of tomato shoots, using the Affymetrix Tomato GeneChip, revealed 

differential changes in expression of 266 genes, occurring during the initial 0.5-5 h of 

gravistimulation prior to bending. The differential changes in expression of auxin-related 

genes in favor of the lower shoot side occurred already following 0.5 h of 

gravistimulation, while those of cell wall-related genes, associated with shoot bending, 

occurred only 3 h following gravistimulation. Among the identified genes, 11 were related 

to auxin, and 36 were associated with oxidative processes. It seems therefore, that ROS 

mediate the early gravity-induced lateral auxin movement across the shoot, which is 

necessary for theauxin asymmetric distribution leading to shoot bending. 

36 



BBX21 INTEGRATES LIGHT AND ABA SIGNALS DURING GERMINATION 

Magnus Holm, DongqingXu, ChamariHettiarachchi 

Göteborg University, Göteborg, Sweden 

E-mail: magnus.holm@gu.se 

37 


Session 8.Signalling 

Light stimulates germination of Arabidopsis seeds whereas the phytohormoneAbscisic 

acid (ABA) inhibits germination. We have previously identified the B-box encoding 

transcriptional regulator BBX21 as a positive regulator of light signaling during seedling 

development and shown that it interacts both genetically and physically with COP1 and 

HY5 in photomorhogenesis. Here we show that the bbx21 null mutant is hyper-sensitive 

to ABA during seed germination and seedling growth and that this sensitivity depends on 

the HY5 protein since a bbx21hy5 double mutant lacks discernible ABA phenotypes. We 

will present results from a genetic analysis of double mutants between bbx21 and abi1 to 

abi5 and from a functional analysis of how the BBX21 protein repress ABA responses.



MEMBERS OF THE GIBBERELLIN RECEPTOR GENE FAMILY (GIBBERELLIN 

INSENSITIVE DWARF1) PLAY DISTINCT ROLES DURING LEPIDIUMSATIVUM 

AND ARABIDOPSIS THALIANA SEED GERMINATION 

Antje Voegele, Ada Linkies, Kerstin Müller, Gerhard Leubner 

Institute for Biology II, Faculty of Biology, Albert-Ludwigs-University, Biology II, Freiburg, Germany 

E-mail: antje.voegele@biologie.uni-freiburg.de 

A plant species survival depends on appropriate timing of seed germination. The seeds of 

Arabidopsis thaliana and its close relative Lepidium sativum are highly similar in physiology 

and structure, with Lepidium seeds being much bigger, facilitating tissue-specific 

analyses. Both species possess a thin endosperm layer surrounding the embryo, which 

regulates germination by acting as constraint to radicle protrusion. Gibberellin (GA) 

signaling controls germination at least partly by regulating endosperm weakening by cellwall 

modifying proteins. Arabidopsis has three known GA receptors: GID1a, GID1b and 

GID1c. Using phylogenetic analysis we have shown that the angiosperm GID1s cluster in 

two eudicot (GID1ac, GID1b) groups, indicating their distinct roles during plant 

development. We used a comparative Brassicaceae approach, combining gid1 mutants 

and whole-seed analyses in Arabidopsis with seed-tissue-specific analyses in Lepidium to 

show that GA signaling via GID1ac is required for Arabidopsis seed germination and that 

GID1 transcript expression patterns differed, with GID1b being distinct from GID1ac. We 

identified putative cell-wall modifying expansins and xyloglucanendotransglycosylases/hydrolases 

(XTHs) in a SSH cDNA library from Lepidium endosperm 

tissue. Their transcript expression patterns strongly suggest their regulation by distinct 

GID1-mediated GA signaling pathways: the GID1b and the GID1ac pathway, fulfilling 

distinct regulatory roles in Brassicaceae seed germination. 

38 



39 



STRUCTURALLY SIMILAR BUT FUNCTIONALLY DISTINCT BSUBUNITS OF 

PROTEIN PHOSPHATASE 2A 

GrzegorzKonert, Andrea Trotta,MoonaRahikainen, Michael Wrzaczek, Eva-Mari Aro, 

SaijaliisaKangasjärvi 

University of Turku, Turku, Finland 

E-mail: grzkon@utu.fi 

Serine/threonine protein phosphatase 2A (PP2A) family members carry out crucial 

functions in the regulation of signaling through phosphorelay cascades in animals and 

plants. The predominant form of PP2A is heterotrimer, consisting of a catalytic subunit C, 

a scaffold subunit A, and a highly variable regulatory subunit B, which is thought to 

determine the target specificity of subunit C in the PP2A holoenzyme. We have taken a 

reverse genetic approach to identify PP2A subunits that specifically modulate the ability 

of plants to tolerate environmental stresses. A specific Bsubunit of PP2A was identified as 

a component in the cross-talk between light acclimation, disease resistance and ageing in 

the model plant Arabidopsis thaliana. Knock-out mutants for PP2A-Bshow constitutive 

disease resistance, alterations in antioxidant metabolism and age-dependent cell death 

when grown under moderate light regimes. Preliminarily, another highly similar PP2A 

subunit seems to have a distinct function in the regulation of cellular integrity. 

Intriguingly, however, genetic analysis indicates that these two PP2A regulatory Bsubunits 

are functionally interconnected with each other. Currently, we are working on 

the subunit composition and signalling interactions of the PP2A holoenzymes to reveal 

the functional specificities of the regulatory Bsubunits in Arabidopsis thaliana.



CELL WALL BIOSYNTHESIS AND BIOFUELS 

David Cavalier, Linda Danhof, Jonathan Davis, Benjamin Fode, Jacob Jensen, Barbara 

Reca, Yan Wang, Curtis Wilkerson, Kenneth Keegstra. 

Great Lakes Bioenergy Research Center, Michigan State University, East Lansing, USA 

E-mail: Keegstra@msu.edu 

Hemicellulosic polysaccharides constitute a significant portion of plant biomass and need 

to be fully utilized to make biofuel production sustainable. While the hemicellulosic 

fraction of most common forms of biomass consists mainly of xylan polymers containing 

pentose monomers, some biomass contains hemicellulosic polysaccharides that are rich 

in hexoses. An important prelude to manipulation of hemicellulose quality and quantity is 

to understand both the biochemical details of hemicelluose biosynthesis as well as the 

regulatory events that control hemicellulose synthesis and deposition. Considerable 

progress has been made in recent years in identifying the genes and proteins responsible 

for hemicellulose biosynthesis. The Cellulose Synthase-Like (CSL) proteins are involved in 

making the backbone of many hemicellulosic polysaccharides, while several classes of 

glycosyltransferases are required for addition of the side chain residues. This progress will 

be reviewed by providing an overview of the strategies that have been employed in 

studying this problem and the major conclusions that have been reached. However, 

many important issues, especially related to the regulation of hemicellulose biosynthesis 

still need to be elucidated. Some of these issues and our approaches to resolving them 

will be presented. 

40 


PLANT DIVERSITY 

Roland von Bothmer 


The Global Seed Vault, NordGen, Svalbard, Norway 

E-mail: Roland.von.Bothmer@slu.se 

41 



The wild biodiversity has attained the largest attention but the cultivated diversity has 

caused most international obstacles for its exploitation. The domestication process 

starting some 10 000 years ago initiated major genetic changes in the wild gene pool. To 

go from a wild to a cultivated state triggered a number of large shifts in morphology, 

anatomy and physiology. The new crops gradually adapted to new habitats and climates, 

which further increased the diversity. Modern plant breeding built on selections in the 

older landraces, which became obsolete and gradually vanished. Genetic erosion has 

escalated and much diversity has disappeared and the urgent need for further gene bank 

activities increases. In a global perspective of major human population growth and 

demand for drastically increased food production the pressure on genetic resources 

becomes a major issue. Efficient conservation and utilization of germplasm are thus 

important. We are nevertheless loosing diversity in the genebanks since there has not 

been a good back up system. The aim for The Svalbard Global Seed Vault is to be a global 

back up system for long term storage of all unique genebank accessions in the world. Is 

this feasible?


Session 11.Climate change and adaptation to abiotic stress 

FUNCTIONAL ANALYSIS OF NOVEL MEMBRANE PROTEIN FAMILY, COR413 

CONTROLLED BY AN ARABIDOPSIS TRANSCRIPTION FACTOR DREB1A 

Motoki Kanai, Kyonoshin Maruyama, Koji Yamada, Satoshi Kidokoro, Kazuo Shinozaki, 

Kazuko Yamaguchi-Shinozaki 

Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo, Japan 

E-mail: aa097014@mail.ecc.u-tokyo.ac.jp 

When plants are exposed to various abiotic stresses, they respond and adapt to these 

stresses. DREB1A/CBF3 is one of cold-inducible genes encoding transcription factors in 

Arabidopsis. Overexpression of DREB1A caused increased tolerance to cold stress in 

Arabidopsis, indicating that the DREB1A protein functions in cold stress-responsive gene 

expression. Many protein products regulated by DREB1A are probably responsible for the 

stress tolerance of plants. 

In this study, we analyzed genes for novel membrane protein family, COR413s whose 

expression is regulated by DREB1A. We confirmed that the COR413 genes, COR413-IM1, 

COR413-IM2.1, and COR413-PM1 were cold inducible by using RNA gel blot analysis. We 

observed the subcellular localization patterns of the proteins by using GFP fusion genes. 

COR413-IM1 and COR413-IM2.1 were localized at the chloroplast membrane, while 

COR413-PM1 was thought to be localized at ER. We analyzed phenotypes of T-DNA 

insertion mutants of these genes. There is little difference in cold tolerance between WT 

plants and T-DNA insertion mutants. However we found that cor413-im1 and cor413-im2.1 

mutants accumulated more anthocyanin as compared to the wild-type plants under cold 

stress condition. We performed microarray analyses using these mutants. 

42 



43 



ANALYSIS ON THE DIVERSITY OF DROUGHT TOLERANCE OF PANICUM 

GRASSES 

Tanja Zimmermann, Thomas Berberich, Wolfgang Brüggemann, Hideo Matsumura, 

Ryohei Terauchi 

Working Group of Prof. Dr. Brüggemann, Goethe University, Frankfurt, Germany 

E-mail: t.zimmermann@sciencemail.org 

Due to the predicted climate change, increasingly severe droughts and temperatures will 

have a great impact on biodiversity. Plants using C4 photosynthesis have a competitive 

advantage over those using the C3 photosynthetic pathway because of lower 

photorespiration and a three fold higher water use efficiency. Additionally, the optimal 

growth temperature for C4 plants ranges from 30 - 45 °C while that for C3 plants is about 

15 - 25 °C. These parameters favour C4 grasses in the West African Savannah. Within the 

genus Panicum the C3 type and the C4 NAD-ME type of photosynthesis are realized. 

Members of this genus not only dominate the landscape in the West African Savannah 

but also serve as crop and pasture grasses. 

To ascertain the cause for differences in the ability to adapt to drought stress, three 

species of the genus Panicum will be analysed by proteomic and transcriptomic 

approaches under control and drought conditions. P. turgidum (C4) is known to be 

extremely drought tolerant, P. laetum (C4) is less drought tolerant and P. bisulcatum (C3) 

serves as a control being sensitive to drought. By comparing these three species, 

differences in gene expression and protein profiles can help understanding the 

mechanisms of drought adaptation and resistance.



INFLUENCE OF SALINITY ON ANTIOXIDANT ACTIVITY IN CANOLA 

(BRASSICA NAPUS L.) CULTIVARS 

Eslam Shahbazi Tangkloureh, A. Arzani, G. Saeidi 

Department of Agronomy and Plant Breeding, College of Agriculture, Isfahan University of Technology, Isfahan, Iran 

E-mail: es_shahbazi@yahoo.com 

The effects of salt stress on germination, seedling growth parameters (root and shoot 

lengths, root and shoot fresh and dry weights) and activity of antioxidant enzymes in 

leaves of six cultivars of canola (Brassica napus L.) were investigated. Two F1 hybrids 

(Hyola401, Hyola330) and four open pollinated cultivars (Zarfam, Okapi, RGs003 and 

Sarigol) were used in this study. Seeds were germinated under various level of salinity 0, 

50, 100, 150 and 200 mM NaCl concentrations. Results showed that an increase in NaCl 

concentrations progressively inhibited seed germination. Hyola401 showed the greatest 

germination percentage at all salinity levels. Seedling growth parameters were affected 

by salt stress particularly at 150 mM and 200mM. Leaf antioxidant activities of SOD, APX 

and GR were increased by salinity increment up to 150mM while they were decreased in 

200 mMNaCl concentration. Although constitutive levels of activity of antioxidative 

enzymes were almost the same among canola cultivars, Hyola401 induced antioxidant 

enzyme activities more efficiently when subjected to NaCl treatment. Among the tested 

cultivars, F1 hybrid 'Hyola401' could be considered as salt tolerance as possessing superior 

germination percentage, seedling growth parameters, antioxidant activities under salinity 

stress. On the other hand, F1 hybrid 'Hyola330'performed inferior in those aspects and 

was the most susceptible cultivars to salinity stress. 

44 



45 


Session 10/14. Bioenergy and primary and secondary metabolism 

APOPLASTIC H2O2 GENERATION MECHANISMS DURING EXTRACELLULAR 

LIGNIN FORMATION IN NORWAY SPRUCE CELL CULTURE; EFFECT OF H2O2 

REMOVAL ON PHENOLIC METABOLISM 

Anna Kärkönen 1,2 , T. Pehkonen 1 , T. Warinowski 1 , S. Holmström 1 , G. Brader 3 , C.N. 

Meisrimler 4 , S. Lüthje 4 , T.H. Teeri 1 

1 Department of Agricultural Sciences, University of Helsinki, Finland 

2 MTT Agrifood Research Finland, Department of Agricultural Sciences, University of Helsinki, Finland 

3 Department of Biosciences, University of Helsinki, Finland 

4 University of Hamburg, Biozentrum Klein Flottbek, Hamburg, Germany 

E-mail: anna.karkonen@helsinki.fi 

Apoplastic H2O2 is required for extracellular lignin production in Norway spruce tissue 

culture as removal of H2O2 with potassium iodide (KI) repressed extracellular lignin 

synthesis. This suggests that peroxidases activate monolignols for lignin polymerisation. 

At least two mechanisms for H2O2 formation were present in spruce apoplast: the one 

having characteristics of a haem-containing enzyme, and the other of that of a flavincontaining 

enzyme [1]. 

Purified spruce plasma membranes contained several enzymes able to generate 

superoxide that can dismutate to H2O2.Naphthoquinones, juglone and menadione 

strongly stimulated superoxide production. Full-length gene for spruce respiratory burst 

oxidase homologue (Parboh1, NADPH oxidase) was cloned. It had a stable expression 

during lignin formation and was two-fold induced by elicitation [1]. 

Phenolic dimers accumulated in both cells and in the culture medium when lignin 

biosynthesis was inhibited. Cells also started to divide which is in contrast to ligninforming 

conditions where cells died soon after lignin formation. Interestingly, removal of 

KI after a 3-week-treatment restored extracellular lignin formation. The inducible cell 

culture system enables us to study regulation behind lignin and dilignol formation. These 

data will be discussed. 

References 

[1] Kärkönen et al., Planta 2009



THE BIOIMPROVEPROGRAMMEAIMS AT IMPROVED BIOMASS AND 

BIOPROCESSING PROPERTIES OF WOOD 

Rishi Bhalerao, Leif Jönsson, EwaMellerowicz,BjörnSundberg, HanneleTuominen 

Umeå Plant Science Centre, Umeå University, Umeå, Sweden 

E-mail: hannele.tuominen@plantphys.umu.se 

"Bioimprove-Improved biomass and bioprocessing properties of wood" is a research 

programme financed by the Swedish Research Council Formasfor the years 2011-2014. The 

programme aims at identification of some of the molecular mechanisms that control 

biomass production and chemical composition of the wood in forest trees, and how this 

knowledge can be utilised to improve production of materials and green chemicals from 

the lignocellulosic raw material in biorefinery type of applications. We have produced a 

large number of genetically modified hybrid aspen trees in order to improve biomass 

production and to test the performance of selected lines also in field conditions. In 

addition to the biomass production capacity of forest trees, an important factor for 

bioprocessing of lignocellulosic material in biorefineries is its susceptibility to the acid and 

enzymatic treatments required to break down the cellulose to glucose molecules. The 

susceptibility is largely determined by the chemical properties of the lignocellulosic raw 

material. We have therefore focused on modification of the composition and content of 

cellulose, hemicellulose and lignin in transgenic hybrid aspen trees, and started analysing 

the effects on the bioprocessing properties on a laboratory-scale. The expected outcome 

of the programme is the creation of a new research environment where wood biologists, 

wood chemists and ecologists interact to create a sustainable production of trees with 

new and enhanced properties to meet the requirements for new products, sustainable 

energy use and decreased CO2 emissions. 

46 



47 



WHAT IS THE TRANSPORT MECHANISM OF MONOLIGNOLS IN LIGNIFYING 

XYLEM OF NORWAY SPRUCE? 

Kurt V. Fagerstedt 1 , Junko Takahashi 2 , Enni Väisänen 1,3 , Anna Kärkönen 3,4 

1 Department of Biosciences, Division of Plant Biology, University of Helsinki, Finland 

2 Umeå Plant Science Center, Umeå University, Umeå, Sweden 

3 Department of Agricultural Sciences, University of Helsinki, Helsinki, Finland 

4 MTT Agrifood Research, University of Helsinki,Helsinki, Finland 

E-mail: kurt.fagerstedt@helsinki.fi 

While the biosynthesis of monolignols is relatively well characterized, the transport of 

monolignols into the apoplastic space in developing xylem is known to a far lesser extent. 

Our aim is to understand the cell and molecular biology of the transport mechanisms. We 

can envisage three possibilities for the transport: Golgi vesicle-mediated transport, 

plasma membrane-located ABC-type transporter proteins or membrane channel proteins, 

and diffusion based on the hydrophobic properties of the monolignols. While the Golgi 

vesicle-based transport has been shown plausibly not to be the transport mechanism [1], 

we are testing the other possibilities by using 14 C-labelled phenylalanine and monolignols 

containing a radioactive or fluorescent label together with transporter inhibitors in a 

Norway spruce (Picea abies) tissue culture system that produces extracellular lignin [2]. 

Also plasma membranes isolated from lignifying xylem of mature trees are used in the 

experiments. As a background for the present study, we have done an extensive 

database search on genes related to transport phenomena and their expression levels in 

different tissues of conifers. The latest results of the ongoing work will be discussed. 

References 

[1] Kaneda et al., Plant Physiol. 2008 

[2] Kärkönen et al., Physiol. Plant. 2002



CAROTENOID BIOSYNTHESIS, STABILIZATION AND DEGRADATION, WHICH 

DETERMINE COLOR INTENSITY IN PETALS OFYELLOW-PIGMENTED CUT 

ROSES, ARE REGULATED BY APPLICATION OF METHYL JASMONATE 

Alon Glick 1 , Sonia Philosoph-Hadas 1 , Alexander Vainstein 2 ,Shimon Meir 1 

1 

Department of Postharvest Science of Fresh Produce, ARO, TheVolcani Center, Bet-Dagan, Israel 

2 

The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 

Jerusalem, Israel 

3 

Department of Genetics, The Hebrew University of Jerusalem, Jerusalem, Israel 

4 

Department of Plant Pathology and Weed Research, TheVolcani Center, Bet-Dagan, Israel 

E-mail: shimonm@volcani.agri.gov.il 

Various yellow-pigmented cut rose (Rosa hybrida) cultivars such as 'Frisco' show color 

fading. The color intensity and the petal carotenoid content increased during the first two 

days of vase life, and gradually decreased thereafter. Application of methyl jasmonate 

(MJ) to the cut roses retained their petal color intensity and carotenoid content 

throughout vase life. Our analysis show that the decrease in carotenoid content resulted 

from both decreased carotenoid biosynthesis and increased carotenoid degradation, 

mainly of the yellow pigments violaxanthin, antheraxanthin and neoxanthin. The 

reduction in carotenoid biosynthesis resulted from decreased expression of two genes 

encoding two key enzymes in the pathway,phytoene synthase (Psy) and Ζcarotenedesaturase 

(Zds). The increase in carotenoid degradation resulted from 

increased expression of the plastidic carotenoid cleavage dioxygenase (CCD4) gene, 

which coincided with decreased content of the plastoglobulin chromoplast protein C 

(CHRC) and plastoglobule decomposition. MJ treatment inhibited petal color fading by 

increasing carotenoid biosynthesis and inhibiting carotenoid degradation. Taken 

together, our results suggest that MJ treatment regulates the three different processes 

that determine color intensity in yellow-pigmented rose petals, namely: carotenoid 

biosynthesis by increasingPsy and Zds expression; stabilizing the plastoglobules by 

increasing CHRC content, and carotenoid degradation by inhibiting CCD4 expression. 

48 



FINE-TUNING OF PLANT IMMUNE RESPONSES TO HOST-ADAPTED 

PATHOGENS 

Jane Parker, Ana Garcia, Servane Blanvillain-Baufumé, Katharina Heidrich, Lennart 

Wirthmüller, Steffen Rietz 

Department of Plant-Microbe Interactions, Max-Planck Institute for Plant Breeding Research, Cologne, Germany 

Email: parker@mpipz.mpg.de 

49 



Plants have evolved a multi-layered innate immune system to recognize and respond to 

potentially destructive microbes in the environment. Resistance to host-adapted 

biotrophic pathogens often involves receptor-mediated transcriptional mobilization of 

defense pathways and programmed death of host cells at infection sites. However, these 

induced defenses are energetically costly and disturb normal metabolism and growth and 

therefore have to be tightly controlled. Fine-control is achieved via a stress signaling 

network which allows flexibility in responsiveness to pathogens and other environmental 

stimuli, depending on the prevailing conditions. While components of individual stress 

signaling pathways have been identified and to some extent characterized our 

understanding of how these pathways intersect at the molecular, cellular and tissue 

levels is relatively poor. We’re attempting to unravel resistance processes triggered by 

intracellular immune receptor-mediated recognition of a bacterial type III secreted 

effector and position them within the larger stress signaling network. I’ll describe our 

analysis of resistance signaling dynamics within host cells and how we think intracellular 

reconfigurations of protein complexes governing transcriptional reprogramming 

contribute to a balanced and flexible immune response.

Session 13.Pathogen defense and plant desease 


PROTEIN PHOSPHATASE 2A AS A REGULATOR OF STRESS RESPONSES IN 

ARABIDOPSIS 

Andrea Trotta 1 , Grzegorz Konert 1 , Moona Rahikainen 1 , Michael Wrzaczek 2 , Mikko 

Tikkanen 1 , Eva-Mari Aro 1 , Saijaliisa Kangasjärvi 1 

1 Department of Biochemistry and Food Chemistry, University of Turku, Turku, Finland 

2 Department of Biosciences, University of Helsinki, Helsinki, Finland 

E-mail: saijaliisa.kangasjarvi@utu.fi 

Chloroplasts perform essential signaling functions in cellular networks that modulate 

various stress responses in plants. We have taken a reverse genetic approach to identify 

components in the cross-talk of chloroplast signaling. Mutants deficient in subunits of 

protein phosphatase 2A (PP2A) showed peculiar light-intensity-dependent phenotypes, 

and were chosen for further analysis. A PP2A regulatory subunit B was identified as a 

cytoplasmic component in the cross-talk between light acclimation and disease resistance 

in Arabidopsis thaliana. Under moderate light intensity, knock-down pp2a-b mutants show 

disintegration of chloroplasts, premature yellowing and constitutive activation of salicylic 

acid and jasmonic acid dependent defense responses, but no increases in hormone levels. 

Promoter::GUS analysis indicates activity of PP2A-B in patches that highly resemble the 

yellowings on pp2a-b leaves. Genetic analysis suggests that PP2A-B is functionally 

connected with CONSTITUTIVE EXPRESSION OF PR GENES 5 (CPR5), which acts as a 

negative regulator disease resistance and senescence in Arabidopsis leaves. Yeast twohybrid 

screening for candidate PP2A-B interacting proteins also identified components 

that operate on defense-associated pathways. Moreover, the constitutive defense 

response associates with hypomethylation of DNA and increased levels of methioninesalvage 

pathway components in pp2a-b'γ leaves. Studies are underway to specify the 

mechanisms of PP2A dependent signaling functions in plants. 

50 



51 



PSEUDOMONAS SYRINGAE PV.PHASEOLICOLA AND DICKEYA DADANTII 3937: 

TWO PATHOGENIC LIFE-STYLES ANALYSED BY IMAGING TECHNIQUES 

M. Luisa Pérez-Bueno 1 , Emilia López-Solanilla 2 , Mónica Pineda 1 , Elena Díaz Casado, Cayo 

Ramos 3 , Pablo Rodríguez-Palenzuela 2 , Matilde Barón 1 

1 Estación Experimental del Zaidín, CSIC, Granada, Spain 

2 Centro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid - Instituto Nacional de Investigación 

y Tecnología Agraria y Alimentaria, Madrid, Spain 

3 Area of Genetics, University of Malaga, Malaga, Spain 

E-mail: marisa.perez@eez.csic.es 

P. syringaepv. phaseolicola (Pph) and D. dadantii (D.d.) 3937 are model organisms for two 

very different “pathogenic life-styles”. The first one is the causal agent of halo blight 

disease of beans, characterized by small watersoaked spots which gradually turn 

necrotic, and become surrounded by a wide chlorotic halo (caused by the effect of a nonhost 

specific toxin produced by the bacteria). The second one is the causal agent of 

bacterial soft rot in vegetables. This disease occurs most commonly on fleshy storage 

tissues. Both pathogens cause great losses in crop production worldwide. 

Chlorophyll fluorescence (Chl-F) and blue-green fluorescence (BGF) techniques are 

sensitive and specific tools which provide information about the primary and secondary 

metabolism in plants, respectively. Chl-F, BGF and temperature leaf patterns obtained by 

imaging techniques were shown to be in correspondence with pathogen distribution and 

symptom development during viral pathogenesis. In the present work, the systemic 

infection of D.d. 3937 in N. benthamiana and Pph in Phaseolus vulgaris have been studied 

by these imaging techniques to understand the impact of these pathogens in plant 

physiology. 

Acknowledgements 

Financial support: MCINN-FEDER/AGL2008-00214, CVI 03475 (Junta de Andalucía). MLPB was recipient of a 

CSIC-JAE Doc contract.



CELL DEATH AND RESISTANCE 2 GENE IS A NEGATIVE REGULATOR OF CELL 

DEATH AND ENCODES AN AAA-TYPE ATPASE 

Hann Ling Wong 1 , Masayuki Isshiki 2 , Tsutomu Kawasaki 3 , Ko Shimamoto 4 

1 Department of Biological Science, Universiti Tunku Abdul Rahman, Kampar, Malaysia 

2 Kihara Institute for Biological Research, Yokohama City University, Yokohama, Japan 

3 Department of Bioscience, Kinki University, Nara, Japan 

4 Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology, Ikoma, Japan 

E-mail: hlwong2007@gmail.com 

The cell death and resistance (cdr2) mutant rice plants display a spontaneous cell death 

phenotype that resembles disease symptoms even without pathogen infection. This 

mutant belongs to the initiation class of lesion mimic mutant. Previously, we found that 

cdr2 mutant plants exhibit enhanced resistance to rice blast fungus infection. Suspensioncultured 

cdr2 cells also showed elevated defense-related genes expression and higher 

calyculinA-induced H2O2 production. In this study, we have localized the cdr2 mutation by 

map-based cloning. We further sequenced the candidate ORFs around the localized 

region and revealed that the cdr2 mutation is probably caused by a single-base (G to A) 

substitution, resulting in a Gly to Arg change in an ORF encoding an AAA-type ATPase. We 

analyzed the expression pattern of the Cdr2 gene by RT-PCR and found that its 

expression is ubiquitous in all rice tissues examined. Overexpression of Cdr2 gene in the 

cdr2 mutant complemented the lesion mimic phenotype and suppressed the cdr2 

mutation-induced elevation defense-related genes. Furthermore, co-expression of Cdr2 

gene suppressed Bax-induced cell death in Nicotiana benthamiana. Taken together, Cdr2 

may function as a negative regulator in regulating cell death and defense response. 

52 


A B S T R A C T B O O K – A B S T R A C T S O F P O S T E R S 

53 


Abstracts of Posters


THE INDUCTION AND GROWTH OF POTATO (SOLANUM TUBEROSUM. L) MICROTUBERS, 

SANTE CULTIVAR, RESPONSE TO THE DIFFERENT CONCENTRATIONS OFBAP AND 

SUCROSE 

Alireza Iranbakhsh 1 , Mostafa Ebadi 2 

1 Islamic Azad University, Aliabad Katool, Golestan Province, Iran 

2 Islamic Azad University, Damghan Branch, Semnan Province, Iran 

E-mail:iranbakhshar@yahoo.com 

The effects of different concentrations of Banzil Amino Purin (BAP) and sucrose as induction combinations 

on microtuber formation and the time of this process, numbers and fresh and dry weight of microtubers 

were investigated. A two-stage culture was used to study the effect of hormonal and sucrose treatments. 

In the first stage, a liquid MS medium containing 0.5 mgl -1 BAP + 0.4 mgl -1 GA3 + 20 gl -1 sucrose was used for 

the increase of branches. The cultures of single node were grown against white light (4000 - 5000 LUX) and 

on sucrose for one month. In the second stage, microtuber formation induced on fluid MS medium 

containing different concentrations of sucrose (30, 40, 60, 80 mgl -1 ) and BAP (1, 2, 5, 10 mgl -1 ) was used in 

continuous darkness. Microtuber formation was investigated within 10 weeks after induction. 

CHARACTERISATION OF SECRETED COMPOUNDS IN NORWAY SPRUCE TISSUE CULTURE 

MEDIUM BY CHROMATOGRAPHY AND MASS SPECTROMETRY 

Gwe G.Chenyi 1 , S.Chong 2 , S.Koutaniemi 2 , M.Tenkanen 2 , S.C.Fry 3 , A.Kärkönen 1,4 

1 Department of Agricultural Sciences, University of Helsinki, Finland 

2 Department of Food and Environmental Sciences, University of Helsinki, Finland 

3 Institute of Molecular Plant Sciences, University of Edinburgh, United Kingdom 

4 MTT Agrifood Research Finland, Jokioinen, Finland 

E-mail: gilbert.gwe@helsinki.fi 

Oligosaccharides secreted into the culture medium of suspension cultures have been shown to be similar to 

those in cell walls [1, 2]. Some secreted oligosaccharides may play a crucial role in plant growth and 

development. For example, fungal cell wall-derived oligosaccharides elicit the generation of a defenceassociated 

H2O2 burst in extracellular lignin-producing Norway spruce cell cultures [3, 4]. We hypothesise 

that soluble oligosaccharides present in the culture medium of spruce cells regulate the constant low levels 

of H2O2 present during extracellular lignin formation. Cultured spruce cells were fed with [U-14C]glucose 

and treated subsequently with H2O, elicitor, KI or KCl. The culture medium compounds were separated by 

paper chromatography and high voltage paper electrophoresis. The results show varying amounts of 

certain compounds in different treatments. The positively charged compounds have been further purified 

by cation exchange chromatography. The identification (mass spectrometry) of compounds is underway 

and their involvement in plant growth and development will be investigated. 

References 

[1] McNeil et al., Annu. Rev. Biochem. 1984 

[2] York et al., Methods Enzymol. 1985 

[3] Kärkönen and Fry, J. Exp. Bot. 2006 

[4] Kärkönen et al., Planta 2009 

POLYPHOSPHATES IN PLANTS 

Hélène Pélissier Combescure, Elizabeth Nees Ahm, Tom Hamborg Nielsen 

Department of Plant Biology and Biotechnology, Faculty of Life Sciences, Copenhagen University, Copenhagen, Denmark 

E-mail: hepel@life.ku.dk 

Organisms can store phosphate as polyphosphate, a linear polymer of phosphate residues linked by highenergy 

bonds. Polyphosphate serves as an energy reservoir, and chelates heavy metal ions. In bacteria, it 

plays a role in stress responses, gene regulation, motility and virulence. The polyphosphate kinase (PPK1), 

54 



mostly studied in bacteria, catalyzes the synthesis of polyphosphate from ATP and the degradation of 

polyphosphate. Putative PPK1 have been identified in the genome of Physcomitrella. Our aim is to 

understand the role of polyphosphate in plants, using Physcomitrella as a model and engineer higher plants 

expressing PPK1 to create a new form of P storage. These plants may serve in the phytoremediation of 

excess phosphate. Two putative PPK1 genes (PpPPK1A and PpPPK1B) were isolated from Physcomitrella. 

PPK1A and PPKB encode proteins of 90 and 98 kDa, respectively. In order to biochemically characterize the 

proteins and confirm their function purification of recombinant proteins in E.coli is under way. To further 

understand polyphosphate metabolism, gene expression was analyzed in Physcomitrella under various 

growth conditions. Arabidopsis overexpressing PPK1s are currently being analyzed for polyphosphate 

content, and phosphate metabolism. In order to analyze polyphosphate levels in plants we have 

successfully purified the recombinant E.coli PPK1 using a polyHis-tag. 

DEHYDRIN IN BUCKWHEAT SEEDS 

Michiko Momma 

National Food Research Institute, Tsukuba, Ibaraki, Japan 

E-mail: michiko@affrc.go.jp 

Dehydrin is a class of LEA proteins that are expressed in the late stage of seed maturation in response to 

water stress and formation of abscisic acid. While involvement of dehydrin in the desiccation tolerance of 

plants has been extensively studied, little is clarified about the property and function of dehydrin proteins 

in seeds. We have shown protective activity of dehydrin proteins in soybean and rice seeds to the 

freezes/thaw denaturation of enzyme proteins. Studies also suggested that the accumulation of dehydrin in 

peanut relate to human allergenic response. In this study, the presence of dehydrin proteins in buckwheat 

was indicated for the first time, using immunoblotting for antibody against its highly conserved lysine-rich 

motif. A major (20 kDa) and a minor (16 kDa) band were found in buckwheat flours, noodles and boiled 

water used for noodle cooking. The recognition of the major dehydrin protein (20 kDa) was difficult in the 

SDS-PAGE analysis, since its molecular weight was very close to that of legumin, most abundant storage 

protein of buckwheat. The minor dehydrin-like protein at 16 kDa showed moderate resistance to pepsindigestion. 

Results suggested that buckwheat and its product contain dehydrin-like proteins, which 

associated with pepsin resistance. 

METABOLIC PROFILING COMBINED WITH MULTIVARIATE STATISTICAL ANALYSIS CAN 

BE APPLICABLE FOR THE DISCRIMINATION OF SALT TOLERANCE OF RICE (ORYZA SATIVA 

L.) CULTIVARS. 

Yu Ran Kim 1 , In Sun Yoon 2 , Taek Ryun Kwon 2 , Hyun Ju Kim 1 , Eunjung Bang 1 , Myung Hee Nam 1 

1 Korea Basic Science Insitute, Yuseong-gu, Daejeon, South Korea 

2 Department of Agricultural Biotechnology, National Academy of Agricultural Science, Gwonseon gu, Suwon, South Korea 

E-mail: nammh@kbsi.re.kr 

Plant responses to salinity involve changes in the activity of genes and proteins, which consequently lead to 

the changes of metabolism. Metabolome represents final phenotype of cells caused by environment or 

gene expression. We conducted metabolic profiling approach to screen and characterize genetic diversity 

of salt tolerance on the metabolite level. For this purpose, we selected 40 kinds of salt resistant and salt 

sensitive rice cultivars according to their physiological response to NaCl. Low concentration of NaCl was 

treated for 13 days. Metabolic profiling was conducted with NMR spectroscopy. The partial least squares 

discriminant analysis (PLS-DA) of the 1H-NMR spectra showed a clear discrimination between salt exposed 

and control groups. The discrimination between salt sensitive and salt resistant cultivars under salt 

untreated condition could also obtained by PLS-DA analysis. Major peaks in 1H-NMR spectra contributing to 

the discrimination were assigned as choline, allantoin, sugars and amino acids like alanine, gutamine, 

glutamateand aspartate. These results represent that metabolome analysis can be applicable to the 

discovery of specific phenotype between genetic variations concerning salt tolerance. 

55 



ENHANCING METHIONINE ACCUMULATION IN TOBACCO SEEDS EXPOSES A 

PHYSIOLOGICAL LINK BETWEEN METHIONINE AND GLUTATHIONE AND ITS IMPACT ON 

SEED GERMINATION 

Itamar Godo, Ifat Matityahu, Rachel Amir 

1 Laboratory of Plant Science, Migal Galilee Technology Center, Kiryat Shmona, Israel 

2 Tel Hai Collage, Upper Galilee, Israel 

E-mail: rachel@migal.org.il 

Methionine is an essential amino acid whose level limits the nutritional value of crop plants; however, to 

date, efforts to increase its content had limited success. We used tobacco plants to express a feedback 

insensitive Arabidopsis cystathionine γ-synthase (AtCGS), the first committed enzyme in the methionine 

biosynthesis pathway, under a seed specific promoter. Both methionine and total proteins contents of the 

transgenic seeds were significantly increased, suggesting that CGS is a rate-limiting enzyme of methionine 

synthesis in seeds, and that methionine availability limits seed protein synthesis. AtCGS expression altered 

the protein profile and enhanced the expression of several methionine rich proteins. In the transgenic 

seeds the amounts of glutathione (GSH) and cysteine, a precursor for methionine and GSH, were reduced, 

suggesting that methionine accumulates at the expense of GSH. Although these methionine -enriched, low 

GSH seeds, germinated slower than wild type, adult plants had no phenotype indicating that this effect is 

limited to seeds. The delayed germination could be reversed by application of GSH. In addition to an 

important step toward the development of high methionine food crops, this work contributes to the 

understanding of methionine metabolism, its interactions with GSH and cysteine metabolism and their roles 

in seed germination. 

GENETIC ENGINEERING OF CAROTENOIDS BIOSYNTHESIS IN PHAEODACTYLUM 

TRICORNUTUM 

Ulrike Eilers, G. Sandmann, C. Büchel 

Goethe-Universität Frankfurt, Frankfurt am Main, Germany 

E-mail: u.eilers@sciencemail.org 

Carotenoids are produced by all photosynthetic organisms and take part in light harvesting as well as in 

photoprotection. Their antioxidative character and other functions are reasons for their important role in 

the humans' nutrition. The high costs of synthetic carotenoid production give reason to find suitable hosts 

for natural biosynthesis. 

For a proof-of-principle we use the model organism Phaeodactylum tricornutum, a unicellular diatom 

occurring in salt and brackish water. Since rather little is known about the enzymes involved in carotenoid 

biosynthesis in diatoms we focus here on four endogenous genes, (Psy, Zep1- 3), coding for phytoensynthase 

and zeaxanthin-epoxidases 1-3, respectively. Psy catalyses the entry and limiting reaction of the 

carotenoids biosynthesis. Zep1-3 are coding for the enzymes responsible for the conversion of zeaxanthin 

to violaxanthin.To investigate these four putative enzymes in P. tricornutum we cloned cDNA and genomic 

DNA in order to identify their functions by complementing systems in E. coli.An interesting step for 

industrial purposes is the carotenoid over expression. Therefore we will transform P. tricornutum with an 

inducible Psy-construct to overcome the limiting entry reaction.Moreover, an additional gene (bkt), coding 

for a ketolase, is cloned to set a bypass for astaxanthin synthesis without disturbing the physiology. 

A GUIDELINE TO FAMILY-WIDE COMPARATIVE STATE-OF-THE-ART QRT-PCR ANALYSIS 

EXEMPLIFIED WITH A BRASSICACEAE CROSS-SPECIES SEED GERMINATION CASE STUDY 

Ada Linkies, Kai Graeber, Andrew Wood, Gerhard Leubner-Metzger 

Albert-Ludwigs-University Freiburg, Faculty of Biology, Plant Physiology, Freiburg, Germany 

E-mail: ada.linkies@biologie.uni-freiburg.de 

qRT-PCR is a sensitive method to quantify gene expression. Even small differences in expression profiles 

can be determined, given that the right precautions during data analysis are taken. One major concern is 

56 



the normalization of transcript data, usually done by reference genes that show stable expression in the 

analyzed tissue or treatment. It has been shown that many commonly used reference genes are not stable 

expressed in several developmental states i.e. seed germination. During seed germination huge 

transcriptional changes take place, complicating the identification of reference genes. Therefore we carried 

out a cross-species approach with Arabidopsis thaliana, Lepidium sativum and Brassica napus using seeds as 

challenging tissue to identify family-wide reference genes for Brassicaceae seed germination and 

maturation. We used transcriptome data of Lepidium sativum seed tissues and Arabidopsis thaliana whole 

seeds to select candidate reference genes and verified their stability with qRT-PCR in both species. Further 

analysis of transcriptome data from Brassica napus and Arabidopsis thaliana seeds during maturation 

confirmed that the majority of the new reference genes are also stable expressed during seed maturation. 

Our cross-species Brassicaceae case-study can therefore be used as a guideline to identify reference genes 

for other families or other demanding tissues and treatments. 

COMPUTATIONAL ANALYSIS OF GENES INVOLVED IN NITRIC OXIDE, SA AND JA 

SIGNALING IN RICE UNDER CADMIUM STRESS 

Indra Singh 1 , Neetu Singh Yadav 1 , Pragat Agrawal 1 , Kavita Shah 2 

1 Bioinformatics Division, Mahila Mahavidyalaya, Banaras Hindu University, Varanasi, India 

2 Institute of Environment & Sustainable Development, Banaras Hindu University, Varanasi, India 

E-mail: singhanjanabhu@gmail.com 

Complete genome availability of rice provides for study of candidate genes in nitric oxide, jasmonic acid and 

salicylic acid signalling pathway in plants. We examined in silico, the genes involved and the point of 

convergence of these signalling pathways under cadmium stress in rice. In NO pathway nitrate reductase 

and nitric oxide synthase are enzymes that catalyze a balanced production of NO in plants under abiotic 

stress. Retrieval of NR and NOS gene sequence from rice and detection of their homologs, gene ontology 

studies and homology modelling provided an insight into the protein products and their function in stress 

signalling pathway in rice. In SA-JA interacting pathways, 13 genes were found to be expressed that either 

activate (EDS-1, R-gene, CPR-6, PAD-4, EDS-5, SID-2, TGA-2, SFD-1, PR) or inhibit (CPR-5, NPR-1, SNI-1, SSI-2) 

the effect of abiotic stress in A. thaliana. Out of these, 7 gene sequences were obtained upon similarity 

search from O. sativa and are yet not annotated. Using various computational parameters and structural 

data, these genes were annotated and their functions predicted. Results suggest genes, involved in NO, SA 

and JA pathways are crosslinking in O. sativa having synergistic as well as antagonistic effects in rice under 

cadmium toxicity. 

PLANT PEROXISOMAL PARALOGUES OF GLYCOLATE OXIDASE POSSESS DIFFERENT 

BIOCHEMICAL PROPERTIES AND PHYSIOLOGICAL FUNCTIONS 

Anke Kuhn, Martin K.M. Engqvist, Nils Jaspert, Veronica G. Maurino 

Department of Botany, Cologne Biocenter, University of Cologne, Cologne, Germany 

E-mail: akuhn1@uni-koeln.de 

During photorespiration glycolate oxidase (GO) catalyses the oxidation of glycolate into equimolar amounts 

of glyoxylate and H2O2 in the peroxisomes. The Arabidopsis genome contains a family composed of five GO 

homologs. Two close related genes, GO1 and GO2, are highly expressed in photosynthetic tissues and 

support photorespiration. A third paralogue, GO3, showed lower expression in autotroph tissues and the 

two more distant paralogues showed high homology to human long- and medium-chain 2-hydroxyacid 

oxidases (HAOX) and are thus called HAOX1 and 2. In silico analysis of differing active site residues 

suggested different substrate specificities for the different homologues. In this work, the five Arabidopsis 

GO homologues were expressed as recombinant protein and characterized at the biochemical level. The 

results obtained together with the isolation and characterization of knock-out mutants allowed to propose 

that GOX3 would function as a lactate oxidase during recovery of hypoxia and that HAOX 1 and -2 would 

have a role principally during seed development. 

57 



CYTOKININS IN FRESHWATER AND SOIL CYANOBACTERIA AND ALGAE 

Eva Žižková 1 , Lenka Záveská Drábková 2 , Petre Dobrev 1 , Pavel Přibyl 3 , Silvia Gajdošová 1 , Klára Hoyerová 1 , 

Miroslav Kamínek 1 , Václav Motyka 1 

1 Institute of Experimental Botany AS CR, Prague, Czech Republic 

2 Institute of Botany AS CR, Průhonice, Czech Republic 

3 Institute of Botany AS CR, Třeboň, Czech Republic 

E-mail: zizkovae@ueb.cas.cz 

Cytokinins (CKs) are plant hormones regulating many aspects of growth and development. A great diversity 

of CK derivatives was identified in vascular plants while only some of them occur in evolutionary older nonvascular 

organisms. With the aim to contribute to the present knowledge of CK distribution throughout the 

plant kingdom, endogenous CK profiles were analysed in fifteen freshwater and soil species of 

Cyanobacteria, Chlorophyta and Chromophyta. 

In general, isopentenyladenine(iP)-type CKs prevailed over those of trans-zeatin (Z), cis-zeatin (cisZ) and 

dihydrozeatin (DHZ) in most taxa. Interestingly, in Cyanobacteria (Chroococcales, Nostocales and 

Oscillatoriales) methylthio-derivatives of iP represented the highest proportion of CKs. None or only trace 

amounts of CK-N- and O-glucosides were detected in all tested species in contrast to the land plants. 

Another general trait found for most of analysed species was a higher concentration of cisZ than Z. It is 

therefore possible that functions of CK-N-glucosides (i.e. deactivation or lowering of biological activity) in 

cyanobacteria and algae are, at least partially, substituted by cisZ, which is prevailing and less active form 

compared to Z. Absence of cisZRMP suggest an existence of a cisZ biosynthetic pathway different from 

that of Z. 


Supported by the Czech Science Foundation project P506/11/0774. 

CHLOROPHYLL FLUORESCENCE AND CHANGES IN STARCH CONTENT IN ROOTSTOCK OF 

ELEPIDOTE RHODODENDRON CULTIVAR 'CUNNINGHAM'S WHITE' CUTTING GRAFTS 

DURING PROPAGATION 

K. Dokane, L. Mertena, D. Megre, U. Kondratovics 

University of Latvia, Riga, Latvia 

E-mail: kristine.dokane@gmail.com 

The cutting-grafting technique is based on grafting scions onto unrooted rootstocks. This method is 

comparatively quick because during the propagation two different processes occur - graft union formation 

and adventitious rooting. 

The aim of this study was to find out, what changes in starch content and chlorophyll fluorescence occur in 

rootstock with or without apical bud during propagation of elepidote rhododendron cultivar 'Cunningham's 

White' by cutting grafts.Due to wounding stress a decrease with small fluctuations in Fv/Fm parameter in 

rootstock [with and without apical bud] leaves was observed for first 18 days of experiment. Afterwards 

the callus bridge formation was observed and Fv/Fm started to increase. In first days of experiment 

increase in starch content was observed, followed by decrease until Day 21 (with apical bud) or Day 18 

(without apical bud). These changes can be related to graft union formation. For last 9 or 12 days of 

experiment starch accumulated to be used in rooting process. 

In conclusion, the presence of apical bud probably can delay graft union formation, but further studies are 

necessary to prove this hypothesis. 

LIGHT QUALITY AFFECTS FLOWERING INDUCTION IN FRAGARIA VESCA 'HAWAII-4' 

Marja Rantanen, K. Mouhu, P. Elomaa, P. Palonen, T. Hytönen 

Department of Agricultural Sciences, University of Helsinki, Helsinki, Finland 

E-mail: marja.rantanen@helsinki.fi 

58 



We studied the effects of light quality on long day flowering Fragaria vesca 'Hawaii-4'. Seedlings were 

subjected to short day (SD) plus low intensity daylength extension by blue (B), far-red (FR), red (R), or 

incandescent light (INC). Flowering integrator gene FLOWERING LOCUS-T (FT) and floral identity gene 

FRUITFULL (FUL) were up-regulated after one week in INC and FR treatments. In contrast, we could not 

detect expression of FT and FUL in R and SD treatments during the treatment period of five weeks. In a 

further study at higher temperature, both genes were expressed similarly to previous results in R, SD and 

INC treatments while in B light both genes were expressed at low level. Furthermore, INC and FR treated 

plants flowered early and flowering response in B light was intermediate, whereas both R and SD treated 

plants flowered significantly later. The results show that the expression of FT and FUL correlates with 

flowering initiation in F.vesca 'Hawaii-4'. Strong responses to FR and R light and milder response to B light 

indicate that phytochromes are the major photoreceptors controlling flowering. Ongoing functional 

analysis of FT will confirm its role in the light regulated flowering induction. 

THYLAKOID CALCIUM SENSING RECEPTOR AS A REGULATOR OF PHOTOSYNTHETIC 

LIGHT REACTIONS 

Markus Nurmi, Saijaliisa Kangasjärvi, Eva-Mari Aro 


E-mail: mjnurm@utu.fi 

Calcium sensing receptor (CaS) is a 40 kDa phosphoprotein on the photosynthetic thylakoid membrane. It is 

phosphorylated by STN8 kinase in a C-terminal stroma-exposed motif, which predicts interactions with 14-3- 

3-proteins and with proteins containing fork-head associated domains. Analysis of mechanically 

fractionated thylakoid membranes by immunoblotting revealed differential light-intensity-dependent 

localization for CaS. In darkness and under low light conditions, CaS resides in granal membranes, whereas 

under bright light, high amounts of CaS is detected in stromal regions of the thylakoid membrane. Results 

obtained by immunoblotting of 2D Blue Native gels suggest that CaS interacts with Photosystem II (PSII). 

Comparative transcript profiling shows increased transcript levels for the PSII oxygen evolving complex 

protein PsbO2 and for two Photosystem I core proteins in knock-out cas plants. Intriguingly, publicly 

available co-expression data also points to a connection between CaS and these components. The cas 

mutant also shows a distinct up-regulation of PGRL1 encoding a key component of cyclic electron transfer. 

Furthermore, the steady-state protein level of PGRL1 follows the phosphorylation status of CaS in 

differentially light-treated leaves, suggests that these two proteins are functionally connected. Altogether, 

CaS seems to be involved in the regulation of photosynthetic light reactions in the changing environmental 

cues. 

ACCLIMATION RESPONSE OF BARLEY PLANTS GROWN UNDER LOW AND HIGH PAR TO 

SUPPLEMENTAL UV-A EXPOSURE 

Michal Štroch, Alena Kotyzová, Jakub Nezval, Zdeněk Nosek, Václav Karlický, Rostislav Páník, Vladimír 

Špunda 

Department of Physics, Faculty of Science, Ostrava University, Ostrava, Czech Republic 

E-mail: michal.stroch@osu.cz 

The dynamics of the acclimation response of barley assimilation apparatus was examined after transfer of 

plants grown under low and high PAR to conditions with the same level of PAR and supplemental UV-A 

radiation. The aim of experiment was to evaluate, how the level of PAR during growth of barley plants will 

affect the induction of regulatory and protective mechanisms of the assimilation apparatus (e.g. epidermal 

UV-shielding, utilization of absorbed light energy within photosystem II, xanthophyll cycle activity) in the 

course of UV-A exposure. Spring barley (Hordeum vulgare L. cv. Bonus) was grown under low and high PAR 

(50 and 1000 µmol m -2 s -1 ) in the absence of UV radiation for 8 days. Then the plants were exposed to UV-A 

radiation (8 W m -2 for 16 h per day) for the next 6 days. During the period of UV-A exposure we monitored 

UV-shielding efficiency, functional state of photosystem II and composition of photosynthetic pigments and 

UV-absorbing compounds. The results are discussed with regard to the effect of supplemental UV-B 

exposure of barley plants acclimated to low and high PAR. 

59 



DYNAMIC REGULATION OF LIGHT HARVESTING AND ENERGY DISTRIBUTION BY 

THESTN7 KINASE AND NPQ PROMOTES ACCLIMATION TO FLUCTUATING LIGHT 

CONDITION 

Michele Grieco, Mikko Tikkanen, Eva-Mari Aro 


E-mail: micrie@utu.fi 

Studies on the function of light-harvesting complex II (LCHII) phosphorylation for the acclimation to 

changing light conditions in plants have been focused mainly on different quality of light, in relation to state 

transitions. However, natural light conditions mainly face changes in the quantity of white light requesting 

acclimation strategies for both high and low light in short and long term periods. Here we have investigated 

the role of the thylakoid kinases STN7 and STN8 as well as the regulatory protein PsbS in dynamic 

acclimation of Arabidopsis thaliana plants when grown under short-term fluctuations in the intensity of 

white light. It is concluded that the lack of the dynamic control of excitation energy distribution to PSII and 

PSI in the absence of the STN7 kinase induces controversial commands within short time periods to initiate 

signalling cascades thus interfering a proper adjustment of photosystems stoichiometry. Indeed, the STN7 

mutants when grown under constant light intensity can compromise the lack of the STN7 kinase by 

drastically decreasing the PSII/PSI ratio. On the contrary, under fluctuating white light such acclimation 

mechanism is prohibited and the STN7 mutants show stunted growth phenotype. 

EFFECT OF INTEGRATION OF CYANOBACTERIAL PCS GENE ON THE PROTEOME OF 

CISGENICALLY TRANSFORMED ANABAENA SP. PCC 7120 

Neha Chaurasia 1,2 , Yogesh Mishra 1 , L.C.Rai 1 

1 Molecular Biology Section, Laboratory of Algal Biology, Center of Advanced Study in Botany, Banaras Hindu University, Varanasi, India 

2 Department of Biotechnology and Bioinformatics, North Eastern Hill University, Shillong, India 

E-mail: cyanoneha@gmail.com 

Cyanobacteria, the photosynthetic prokaryotes present in varied habitats are bestowed with unique ability 

to fix nitrogen and carbon dioxide simultaneously. To proliferate in nature, cyanobacteria develop a 

multitude of stress tolerant strategies. The present study provides first hand information of the impact of 

cisgenic transformation with a cyanobacterial gene phytochelatin synthase (pcs) over the proteome of the 

transformed cyanobacterium Anabaena sp. PCC 7120. This gene was first cloned in E.coli and its tolerance 

against multiple abiotic stresses was examined. Following which Anabaena sp. PCC 7120 was transformed 

using an integrative expression vector, pFPN containing the pcs gene expressed from astrong 

cyanobacterial psbA1 promoter. The transformed strain AnFPN-pcs demonstrated a better growth and 

nitrogen fixing ability over wild type when challenged with UV-B, salt, heat, copper, carbofuran and 

cadmium. The proteome of transformed strain AnFPN-pcs was compared with wild type Anabaena sp PCC 

7120 using 2DE and MALDI-TOF/ MS. Most of the proteins related to cell metabolism were upregulated in 

transformed strain as compared to the control cells expect the downregulation of transketolase and ketolacid 

reductoisomerase. Thus the stress induced protein genes hold potential for the development of 

multiple stress tolerant organisms. 

FUNCTION OF FERREDOXIN-NADP + -OXIDOREDUCTASE (FNR) LEAF ISOFORMS IN 


Nina Lehtimäki, Minna Lintala, Yagut Allahverdiyeva-Rinne, Eva-Mari Aro, Paula Mulo 

Molecular Plant Biology, Department of Biochemistry and Food Chemistry, University of Turku, Turku, Finland 

E-mail: nkleht@utu.fi 

60 



In light reactions of photosynthesis electrons are transferred in a linear fashion from water to NADP + , which 

is reduced to NADPH by ferredoxin-NADP + -oxidoreductase (FNR). In Arabidopsis thaliana two distinct 

genes, At5g66190 and At1g20020, encode the chloroplast targeted isoforms, FNR1 and FNR2. Both exist as 

membrane bound and soluble forms. We have investigated physiological roles of FNR using single and 

double knock-out mutants. Single mutant plants were highly viable suggesting that both isoforms are 

functional and participate in linear and cyclic electron transfer. Nonetheless, FNR1 isoform is needed for the 

membrane binding of FNR2, which suggests FNR heterodimer formation. Viability of fnr1 mutants having 

only soluble FNR proves that soluble form is photosynthetically active. The fnr1xfnr2 (F1 generation) plants 

were slow-growing due to downregulated photosynthetic capacity. High excitation pressure in the 

fnr1xfnr2 plants points to over-reduction of electron transfer chain, which was partly discharged via 

induction of cyclic electron transfer. Scarcity of FNR led to induction of oxidative stress, which induced 

protective mechanisms in mutant plants. The fnr1 fnr2 plants (F2 generation) lacking both FNR isoforms 

survived only under heterotrophic conditions. Their photosynthetic machinery was extremely 

downregulated, but yet their leaves contained a few deformed plastids. 

MITOCHONDRIAL D-2-HYDROXYGLUTARATE DEHYDROGENASE PARTICIPATES IN AN 

ALTERNATIVE PATHWAY OF RESPIRATION AND LINKS PHOTORESPIRATION AND THE 

TRICARBOXYLIC ACID CYCLE 

Anke Kuhn, Martin K.M. Engqvist, Judith Wienstroer, Andreas P.M. Weber, Veronica G. Maurino 

Institute for Developmental- and Molecular Biology of Plants, Heinrich-Heine-University, Düsseldorf, Germany 

E-mail: Veronica.Maurino@uni-duesseldorf.de 

D-2-hydroxyglutarate (D-2HG) is a dicarboxylic acid with the hydroxy group in the alpha-carbon. Locus 

At4g36400 encodes a mitochondrial D-2HG dehydrogenase (D-2HGDH) which specifically converts D-2HG 

into into 2-oxoglutarate (2-OG). In wild-type plants the activity of D-2HGDH increases gradually during 

developmental and dark-induced senescence while D-2HG accumulates to very high levels under these 

conditions in loss-of-function mutants. Gene coexpression analysis indicated that D-2HGDH is in the same 

network as several genes involved in the mobilization of alternate substrates from proteolysis and/or lipid 

degradation, the electron transfer protein (ETF) and the ETF-ubiquinone oxidoreductase (ETFQO). This and 

the fact that AtD-2HGDH does not effectively use cytochrome c nor NAD(P) as electron acceptors indicate 

that AtD-2HGDH may donate electrones to the mitochondrial electron transport chain through ETF/ETFQO. 

Moreover, a metabolic characterization indicated that D-2HG originates during the catabolism of lysine. On 

the other hand, D-2HG accumulates to high levels in the photorespiratory mutant lacking serinehydroxymethyl-transferase 

(shm1) after transfer from high CO2 atmosphere to normal air conditions only in 

the light. This was accompanied by a huge accumulation of 2-OG and glycine. Probably, in vivo the activity of 

AtD-2HGDH is inhibited by these metabolites as both inhibited the activity of the recombinant enzyme. 

CA(II)-BINDING PROTEINS, CCAP1 AND CCAP2, ARE INDUCED BY A LONG DARK PERIOD 

IN ARABIDOPSIS THALIANA 

Yuya Ouchi 1 , Akira Nagatani 2 , Masayoshi Maeshima 1 

1 Nagoya University, Nagoya, Japan 

2 Kyoto University, Kyoto, Japan 

E-mail: ouchi.yuya@g.mbox.nagoya-u.ac.jp 

We found novel Ca(II)-binding proteins in A. thaliana: namely, CCaP1 (cytoplasmic Ca(II)-binding protein) 

and CCaP2. These proteins have no common motif found in other proteins or enzymes. CCaP proteins are 

localized in the cytoplasm when expressed as GFP-fusion proteins. The promoter-GUS analysis revealed that 

CCaP1 was predominantly expressed in petioles and CCaP2 in roots. When plants were grown in the dark for 

>20 h, the mRNA levels of CCaP1 and CCaP2 were increased approximately 10-fold of that of the daytime 

level. These high level expression were decreased to the daytime level by light illumination for


of CCaP2 expression in the dark, we cut shoots off and quantified the mRNA. There was no induction of 

CCaP2 by dark, suggesting that induction of CCaP2 expression needs the signal from shoot. Moreover, we 

will report the gene-expression profiles in ccap1 and ccap2 mutant lines. 

FUNCTION OF CDT1 PROTEINS IN GAMETOPHYTE DEVELOPMENT AND S-PHASE 

CHECKPOINT 

Séverine Domenichini 1 , Moussa Benhamed 1 , Gert de Jaeger 2 , Sophie Blanchet 1 , Lieven de Veylder 2 , Catherine 

Bergounioux 1 , Cécile Raynaud 1 

1 Institute of Plant Biology, Université Paris XI, Paris, France 

2 VIB, Ghent University, Ghent, Belgium 

E-mail: cecile.raynaud@u-psud.fr 

CDT1 proteins are involved in S-phase initiation in all eukaryotes: it is a sub-unit of the pre-replication 

complex (pre-RC) that allows the firing of replication origins. Previous work has established that this 

function is conserved in Arabidopsis, although AtCDT1a appears to have a dual role in cell cycle regulation 

and chloroplast division. Here we show that unlike other sub-units of the pre-RC, AtCDT1a is strictly required 

for female gametophyte development. Male gametophyte development is not completely arrested in cdt1a 

hemizygous mutants, but some pollen grains die before anther dehiscence. By contrast, cdt1b null mutants 

have no phenotype, but loss of cdt1b further reduces pollen viability in cdt1a hemizygous plants. 

Furthermore, we show that AtCDT1 proteins could be involved in the perception or repair of double-strand 

breaks. Indeed, both AtCDT1a and AtCDT1b interact with DNA-polymerase epsilon, which functions both to 

replicate the leading strand during S-phase, and in the S-phase check-point. Furthermore, CDT1-RNAi plants 

are more sensitive to gamma-irradiation than the wild type, and accumulate more double-strand breaks. 

Taken together, our results indicate that CDT1 function is crucial to genome stability not only because of its 

role in S-phase onset, but also via a role in the monitoring of DNA integrity. 

CONIFERYL ALCOHOL AND POTASSIUM IODIDE AFFECT THE CELL VIABILITY OF NON- 

LIGNIFYING NICOTIANA TABACUM BY-2 CELLS 

Enni Väisänen 1 , Teemu Teeri 2 , Kurt Fagerstedt 1 , Anna Kärkönen 2,3 

1 Department of Biosciences, Division of Plant Biology, University of Helsinki, Helsinki, Finland 

2 Department of Agricultural Sciences, University of Helsinki, Helsinki, Finland 

3 MTT Agrifood Research Finland, Jokioiken, Finland 

E-mail: enni.vaisanen@helsinki.fi 

In lignifying cells monolignols are produced in the cytosol, deposited into the cell wall and polymerized to 

lignin. It has been suggested that monolignol alcohols are toxic, but the experimental evidence to support 

this claim is hard to find. We have tested monolignol toxicity by treating non-lignifying Nicotiana tabacum 

BY-2 cells in liquid culture with 50 µM - 2 mM monolignol coniferyl alcohol (CA) with or without 5 mM 

potassium iodide (KI) supplementation, which prevents lignification by catalyzing H2O2 removal. It was 

observed that CA treatment alone rarely affected cell viability. In contrast, KI treatment greatly reduced cell 

viability. In combined treatments with KI and 2 mM CA cell viability was lower than with KI only. These 

results suggest that the hypothesis on monolignol toxicity is plausible. However, in combined treatments 

with KI and 500 µM CA the cell viability was higher than with KI only. Similar effects on growth were 

observed in preliminary experiments, where N. benthamiana seeds were germinated in liquid cultures 

supplemented with KI and CA. The unexpected effects of KI and KI + CA raise questions about the 

biochemical reactions that take place during the treatments. 

CHARACTERIZATION OF A GLYCOSIDE HYDROLASE WITH RESPECT TO THE 

BIOSYNTHESIS OF ARABINOGALACTAN PROTEINS 

Eva Knoch 1 , Henriette L. Petersen 1 , William Willats 1 , Satoshi Kaneko 2 , Henrik V. Scheller 3 , Naomi Geshi 3 

1 Department of Plant Biology and Biotechnology, Faculty of Life Sciences, University of Copenhagen, Frederiksberg C, Denmark 

62 



2 Biological Function Division, National Food Research Insitute, Tsukuba, Ibaraki, Japan 

3 Joint BioEnergy Institute, Feedstocks Division, Lawrence Berkeley National Laboratory, Emeryville, California, USA 

E-mail: evaknoch@life.ku.dk 

CAZy glycoside hydrolase family 17 is one of the plant dominant GH families, and includes glucan 1,3-βglucosidase, 

glucan endo-1,3-β-glucanase and lichenase. In Arabidopsis there are 51 genes in this family and 

in silico analysis suggests that several members are highly coexpressed with a glycosyltransferase from 

CAZy family 31 involved in arabinogalactan protein (AGP) biosynthesis. We are interested in the relationship 

between these enzymes with respect to AGP biosynthesis. We chose the glycosyl hydrolase (GH17) which 

shows the highest coexpression profile for further investigation. 

This GH17 is expressed in the embryo and shoot apex in Arabidopsis. The protein sequence consists of an Nterminal 

signal sequence, a GH17 catalytic domain, CBM43 domain, and a site for GPI-anchoring on the Cterminus. 

The protein sequence between the catalytic domain and the CBM domain also contains SPSPSSSP 

sequence which is a potential motif for attachment of arabinogalactan side chains. We are interested to see 

whether this GH17 is an acceptor-peptide for type II arabinogalactan attachment and/or it is involved in the 

modification of AGP on the plasma membrane. We are investigating these possible functions by analysis of 

Arabidopsis knockout mutants, and hydrolase assays using recombinant protein expressed in 

N.benthamiana and E.coli. 

DUAL-TARGETING OF A EUKARYOTIC TRANSCRIPTION FACTOR, AT2G44940 

Lan Yin, Kirsten Krause 

Department of Arctic and Marine Biology,University of Tromsø, Tromsø, Norway 

E-mail: lyi000@uit.no 

In recent years, a growing number of proteins have been shown to be localized in both nuclear and plastid 

compartments [1]. Since such proteins might be involved in chloroplast-nuclear crosstalk,an in silico-based 

screening of transcription factors from Arabidopsis and rice was carried out to find the putative proteins 

that are both targeted to plastids and nuclear [2] (Schwacke R. et al., 2007). Transcription factor At2g44940 

(TF1) is one of the putative proteins that are dual-targeted to both chloroplast and nucleus. Transient 

transformation of tobacco protoplasts showed indeed that a GFP fusion protein is targeted to the nucleus 

and the chloroplasts in the same cell. Higher resolution analysis revealed, moreover, that the TF1 protein 

was co-localized with a plastid DNA marker. SDS-PAGE and Western Blot results indicated that precursor TF1 

overexpressed in E. coli was a protein around 40kDa, its mature protein was around 35kDa, both being 

bigger than the predicted MW 32kDa and 27kDa. A protein of 35kD was detected in the chloroplast stroma 

as well as in isolated nuclei, indicating that a processed form might prevail in both cell compartments. 

In addition, the semi-quantitative RT-PCR of the TF1 transcript indicated that the expression of TF1 was only 

moderately expressed in darkness but increased significantly after transfer to light. The significance of this 

will be further investigated. 

Reference 

[1] Krause and Krupinska, Trends Plant Sci. 2009 

[2] Schwacke et al., Mol. Genet. Genomics2007 

INFLUENCE OF SALINITY ON ANTIOXIDANT ACTIVITY IN CANOLA (BRASSICA NAPUS L.) 

CULTIVARS 

E. Shahbazi, A. Arzani and G. Saeidi 


E-mail: es_shahbazi@yahoo.com 

The effects of salt stress on germination, seedling growth parameters (root and shoot lengths, root and 

shoot fresh and dry weights) and activity of antioxidant enzymes in leaves of six cultivars of canola (Brassica 

napus L.) were investigated. Two F1 hybrids (Hyola401, Hyola330) and four open pollinated cultivars 

(Zarfam, Okapi, RGs003 and Sarigol) were used in this study. Seeds were germinated under various level of 

salinity 0, 50, 100, 150 and 200 mM NaCl concentrations. Results showed that an increase in NaCl 

63 



concentrations progressively inhibited seed germination. Hyola401 showed the greatest germination 

percentage at all salinity levels. Seedling growth parameters were affected by salt stress particularly at 150 

mM and 200mM. Leaf antioxidant activities of SOD, APX and GR were increased by salinity increment up to 

150mM while they were decreased in 200 mM NaCl concentration. Although constitutive levels of activity of 

antioxidative enzymes were almost the same among canola cultivars, Hyola401 induced antioxidant enzyme 

activities more efficiently when subjected to NaCl treatment. Among the tested cultivars, F1 hybrid 

'Hyola401' could be considered as salt tolerance as possessing superior germination percentage, seedling 

growth parameters, antioxidant activities under salinity stress. On the other hand, F1 hybrid 'Hyola330' 

performed inferior in those aspects and was the most susceptible cultivars to salinity stress. 

THE ROLE OF RHOMBOID PROTEASES IN ARABIDOPSIS 

Elinor P. Thompson 1 , Beverley J. Glover 2 

1 University of Greenwich, London, United Kingdom 

2 University of Cambridge, Cambridge, United Kingdom 

E-mail: te30@gre.ac.uk 

Rhomboid proteins constitute a family of membrane-bound serine proteases, which are found in nearly 

every genome so far sequenced. Both eukaryotic and prokaroytic rhomboids mediate communication or 

interactions between cells (eg, during infection) by releasing or activating membrane-anchored substrate 

proteins. Plant rhomboids are not yet well characterised. In Arabidopsis, mutation of the rhomboid 'KOM' 

results in malformed pollen; enzymatic activity of another has been shown in vitro. Investigating several 

members of the multigene family in Arabidopsis, we found that other rhomboids besides KOM affected 

development of floral organs. Indeed, several rhomboids' promoter-GUS constructs yield pollen- and seedlocalised 

staining. Other family members are expressed more widely, their transcripts being found in many 

tissues, and a few Arabidopsis rhomboid genes contain predicted organelle-targeting presequences. GFPtagging 

of a chloroplast rhomboid suggests it is situated in the organelle's outer membrane; the null 

mutant of this protein shows both fertility and photosynthetic defects. We discuss our findings in relation 

to the different subtypes of rhomboids, presence of transcript splice-variants, and evolution of rhomboids' 

roles in plants and other organisms. 

CYTOSOLIC NADP-ISOCITRATE DEHYDROGENASE (ICDH) IS DIFFERENTIALLY 

REGULATED BY REACTIVE NITROGEN SPECIES (RNS) IN PHOTOSYNTHETIC AND NON- 

PHOTOSYNTHETIC ARABIDOPSIS ORGANS 

M. Leterrier 1 , J.B. Barroso 2 , R. Valderrama 2 , J.C. Begara-Morales 2 , B. Sanchez-Calvo 2 , M. Chaki 1 , J.M. Palma 1 , 

F.J. Corpas 1 

1 Departamento de Bioquímica, Biología Celular y Molecular de Plantas, Estación Experimental del Zaidín, CSIC, Granada, Spain 

2 Grupo de Señalización Molecular y Sistemas Antioxidantes en Plantas, Unidad Asociada al CSIC (EEZ), Departamento de Bioquímica y Biología 

Molecular, Universidad de Jaén, Jaén, Spain 

E-mail: josemanuel.palma@eez.csic.es 

NADPH is an essential component in the cellular homeostasis and its regeneration is critical for reductive 

biosynthesis and detoxification processes. One of the most important NADPH cell sources is the enzyme 

NADP-isocitrate dehydrogenase (ICDH) which concomitantly converts isocitrate into alpha-ketoglutarate. 

Arabidopsis contains three genes coding for cytosolic, mitochondrial/chloroplastic and peroxisomal NADP- 

ICDH isoenzymes. The cytosolic isoenzyme is the most abundant representing more that 90% of the total 

activity. 

Biochemical comparative analyses revealed that the kinetic parameters (Km and Vmax) of cytosolic NADP- 

ICDH were different in photosynthetic and non-photosynthetic organs of Arabidopsis plants. Moreover, the 

activity showed a differential regulation in response to the treatment with reducing agents, nitric oxide 

(NO), and oxidants such as peroxynitrite and hydrogen peroxide (H2O2). Thus, H2O2 did not affect the NADP- 

ICDH activity in any organ; however, reduced glutathione inhibited the activity in leaves but not in roots. On 

the other hand, S-nitrosoglutathione (an NO donor) provoked inhibition of NADP-ICDH in both organs and 

similar behaviour was found with peroxynitrite. Taken together, these data indicate that cytosolic NADP- 

64 



ICDH seems to be greatly modulated by reactive nitrogen species (RNS) in both organs, whereas this 

isoenzyme is only sensitive to redox and oxidative changes in Arabidopsis leaves. 

DYNAMICS OF PLANT HORMONES DURING DEVELOPMENT OF WHEAT CARYOPSES 

P. Dobrev 1 , J. Albrechtová 2 , M. Trčková, V. Motyka 1 , M. Kamínek 1 

1 Institute of Experimental Botany, AS CR, Prague, Czech Republic 

2 Department of Plant Physiology, Faculty of Science, Charles University, Prague, Czech Republic 

3 Crop Research Institute, Prague, Czech Republic 

E-mail: kaminek@ueb.cas.cz 

Discovery of cytokinin (CK) accumulation in young seeds shortly after pollination has attracted attention to 

the potential role of plant hormones in control of seed development. In addition to CKs, transient increases 

in the contents of indole-3-acetic acid (IAA) and its conjugates as well as of abscisic acid (ABA) were found 

during the latter anatomically distinguished phases of seed development. 

Using advanced HPLC/MS we determined the dynamics of concentrations of CKs, IAA, ABA and some of 

their metabolites in developing wheat grains and related them to the changes in grain anatomy (coenocyte 

mitosis, endosperm cellularization, development of starchy endosperm and grain filling). Results indicate 

that CK/auxin ratio, that is evidently important for initiation of specific anatomical events, is 

enhanced/adjusted not only by biosynthesis or degradation of a particular hormone (as CKs shortly after 

anthesis) but also by inactivation of the hormonal counterpart (auxin) due to its conjugation. Results will be 

discussed with respect to our recent detection of CK biosynthesis in wheat grains shortly after anthesis 

using stable isotope non-specific labeling following in vivo feeding with 2H2O. 

CLONING AND CHARACTERIZATION OF TWO SERK GENES TO INVESTIGATE THE 

MOLECULAR BASIS OF ORGANOGENESIS AND SOMATIC EMBRYOGENESIS IN CYCLAMEN 

PERSICUM 

M. Savona 1,3 , R. Mattioli 2 , S. Nigro 2 , G. Falasca 1 , F. Della Rovere 1 , P. Costantino 2 , S. De Vries 4 , B. Ruffoni 3 , M. 

Trovato 2 , M.M. Altamura 1 

1 “Sapienza” University of Rome, Dept. of Biology and Biotechnology, Rome, Italy 

2 “Sapienza” University of Rome, Dept. of Environmental Biology, Rome, Italy 

3 C.R.A.-Research Unit for Floriculture and Ornamental Species, Sanremo, Italy 

4 Wageningen University, Laboratory of Biochemistry, Wageningen, the Netherlands 

E-mail: maurizio.trovato@uniroma1.it 

Somatic embryogenesis and organogenesis in vitro are essential for propagation, particularly for those 

species, such as Cyclamen spp., which can be only propagated in vitro. However, the genetic basis of the 

specification of the respective stem cells remains obscure. Since SOMATIC EMBRYOGENESIS RECEPTOR- 

LIKE KINASE (SERK) genes are involved in somatic embryogenesis as well as, in some species, in 

organogenesis, we searched for AtSERK homologs, in in vitro culture of Cyclamen persicum immature 

ovules. By a combination of 5'- and 3' Race approaches on RT-PCR templates, and of genome walking and 

IP-PCR strategies on genomic DNA templates, we eventually identified full cDNA and genomic sequences of 

two SERK-like genes named CpSERK1 (GenBank accession no. JF511659), and CpSERK2 (GenBank accession 

no.GU189408), as well as a partial cDNA fragment similar to AtSERK3 (GenBank accession no. EF661828). As 

in vitro culture of Cyclamen persicum immature ovules provide, in the same genotype and hormonal 

conditions, either lines forming organs and/or embryos or recalcitrant calli, this study aims to use CpSERK1 

and CpSERK2 as tools to understand the molecular basis of stem cell formation and maintenance in somatic 

embryogenesis and organogenesis in vitro. 

PLANT PROTEINS CONTAINING LEUCINE-RICH REPEAT (LRR) WITH NON-LRR, ISLANDS 

INTERRUPTING LRRS 

Norio Matsushima, Tomoko Mikami, Hiroki Miyashita 

65 


Sapporo Medical University, Sapporo, Japan 

E-mail: matusima@sapmed.ac.jp 


LRRs (leucine rich repeats) are present in over 50,000 proteins. Non-LRR, island regions (IRs) interrupting 

LRRs are widely distributed. Here we describe LRR@IR proteins from various plant species, identified by a 

method to identify IRs (LRR@IRpred). A great number of LRR-RLKs and LRR-RLPs, that contain a single 

transmembrane-spanning region, have LRRs intersected by a single IR in which the number of repeat units 

in the preceding LRR block (N1) is greater than the number of the following block (N2); N1 " N2. Examples 

include; the A. thaliana TMK1 homologs have 13 LRRs with N1 = 10 and N2 = 3; and A. thaliana BRI1-related 

proteins have 22 LRRs with N1 = 17 and N2 = 5. The homologs of A. thaliana BRI1, A. thaliana RPP27, and 

tomato Cf have very variable N1, while N2 is highly conservative; N1 = 14 - 33 and N2 = 4. The IRs in the 

homologs of TMK1, BRI1 and BRI1-related proteins contains cysteine clusters that likely form a cap structure. 

The rule of N1 " N2 would play a novel, significant role in ligand-interaction and/or dimerization of the LRR- 

RLKs and the LRR-RLPs. The structure and evolution of plant LRR@IR proteins also are discussed. 

PROTEOMICS OF PEPPER (CAPSICUM ANNUUN L.) FRUITS DURING RIPENING 

Paz Alvarez 1 , Ana Jiménez 2 , Mounira Chaki 1 , Daniel Bonilla-Valverde 1 , María J. Campos 1 , Luis A. del Río 1 , 

Francisca Sevilla 2 , Francisco J. Corpas 1 , José M. Palma 1 

1 Estacion Experimental del Zaidin, CSIC, Granada, Spain 

2 Centro de Edafología y Biología Aplicada del Segura, CSIC, Murcia, Spain 

E-mail: paz.alvarez@eez.csic.es 

Pepper (Capsicum annuum L.) is one of the most consumable worldwide vegetable and its fruits are rich in 

vitamins C, A and calcium. In pepper the main ripening visible feature is the shift from green to red fruits 

due to the conversion of chloroplasts into chromoplasts. Besides, during fruit ripening other important 

metabolic changes have been reported such as taste alteration, emission of volatile organic compounds, 

destruction of chlorophyll, synthesis of new pigments and pectins, synthesis of new proteins and 

degradation of former ones, changes in total soluble reducing equivalents, and alteration of organelles' 

metabolism (chloroplasts, peroxisomes and mitochondria), among others. In spite of that all those 

processes have been reported, the information of the events which take place at molecular level is still 

scarce. Proteomics has been recently proposed as a very useful tool to the understanding of fruit ripening 

and development [1]. Thus, the analysis of the proteome of both green and red pepper fruits has been 

accomplished in this work. Whole fruits have been studied, but also the sub-proteome of both peroxisomes 

and mitochondria from both kinds of fruits have been investigated. 


Financed by the Ministry of Science and Innovation (AGL2002-00834), Spain 

References 

[1] Palma et al., J. Proteomics2011 

STIMULATION OF ROOT GROWTH INDUCED BY ALUMINUM IN QUERCUS SERRATA 

THUNB. IS RELATED TO ACTIVITY OF NITRATE REDUCTASE AND MAINTENANCE OF IAA 

CONCENTRATION IN ROOTS 

Rie Tomioka, Mikiko Kojima, Hitoshi Sakakibara, Chisato Takenaka, Masayoshi Maeshima, Takafumi Tezuka 

Nagoya University, Nagoya, Japan 

E-mail: tomiokar@agr.nagoya-u.ac.jp 

Aluminum is the most abundant metal in the earth's crust. Excess Al 3+ released by soil acidification in soil 

solution is toxic to many cultivated plant species, but it has been reported to stimulate plant growth in 

some crop and tree species in certain concentration of Al 3+ . In this study, we investigated the mechanism of 

Al-induced root development, focusing on the change in NR activity, and indole-3-acetic acid (IAA) and 

cytokinins concentration in roots of Q. serrata seedlings. NR activity in Al-treated roots was increased 3 days 

after treatment. To clarify the process of Al-induced root development, roots were treated for 1 h with Al or 

66 



Ca. In Al-treated roots, NR activity was increased and IAA concentration was maintained at the same level 

as pretreatment, and indole-3-acetyl-L-aspartic acid (IAAsp), which is a metabolic intermediate of IAA 

degradation, was not detected in roots. In Ca-treated roots, NR activity increased, but IAA concentration 

decreased as IAAsp increased. Thus, the maintenance of IAA concentration in Al-treated roots seemed to 

suppress the process of IAA decomposition. Al treatment increased the length and number of second 

lateral roots but Ca treatment did not. We concluded that root development induced by Al was related to 

NR activity and maintenance of IAA concentration in roots. 

PHYSIOLOGICAL AND GENETIC CHARACTERIZATION OF NOVEL INDOLE-3-BUTYRIC ACID 

RESISTANT MUTANTS 

Taiki Hanzawa 1 , Gloria Muday 2 , Abidur Rahman 1 

1 Cryobiofrontier Research Center, Iwate University, Morioka, Japan 

2 Wake Forest University, Winston-Salem, USA 

E-mail. a2511008@iwate-u.ac.jp 

Although two predominant forms of endogenous auxins, IAA, and IBA are produced by the plant, the 

majority of the research to date has been focused on IAA. The conversion of IBA to IAA via beta-oxidation 

predicts that IBA may act solely via IAA pathways. However, IBA has been shown to have more potent 

activity than IAA in some physiological responses. Further, it has been recently reported that the loss of IAA 

uptake carrier or efflux carrier does not affect the intracellular IBA transport. These results suggest that IBA 

transport and signaling pathways may differ from those used by IAA. However, the molecular mechanism 

of IBA specific transport or signaling pathways remains elusive. In the present study, we tried to identify 

IBA specific signaling by screening for novel IBA-resistant mutants. From this screen we recovered three 

mutant lines, which show only a specific resistance to IBA induced root growth inhibition and lateral root 

formation, but respond to other growth regulators, including IAA, exactly as wild type. All three mutants 

showed reduced transport activity for IBA, but not IAA, indicating that the mutated genes may regulate the 

IBA specific transport process. 

THE ROLE OF GDSL-LIPASES IN ARABIDOPSIS PETAL DEVELOPMENT AND FUNCTION 

Tamar Rosilio-Brami, Michele Zaccai, Moriyah Zik 

Life Sciences Department, Ben-Gurion University of the Negev, Beer Sheva, Israel 

E-mail: tamarosilio@gmail.com 

The GDSL-lipase gene family encodes for hydrolytic enzymes named after a unique sequence of amino 

acids, the GDSL motif found near the N-terminus of the protein. GDSL-lipases have a flexible active site that 

enables broad substrate specificity and facilitates different enzymatic activities, including lipase, esterase 

and protease activities. Several microarray studies, aimed at identifying genes involved in the development 

of floral organs, demonstrated that members of this gene family are significantly and differentially 

expressed in these organs, particularly in petals. Thus far the in planta function of only a few GDSL-lipases 

have been demonstrated. In order to elucidate GDSL-lipases unique and common roles in petals, we 

initiated a detailed study of the expression patterns of six Arabidopsis GDSL-lipase genes which are 

abundantly expressed in petals. Possible functions of these genes in planta can be drawn from the close 

correlation between their spatial and temporal expression and physiological events that take place 

throughout flower development. The expression of the six studied genes is mainly associated with 

physiological events related to anthesis and senescence that involve lipid metabolism processes, plant 

defense pathways and cuticule metabolism. In parallel to the expression pattern analysis, we are 

conducting phenotypic analyses of petal GDSL-lipases mutant lines. 

THE USE OF THE 7B-1 AND CRY1-1 MUTANTS UNRAVELED A SPECIFIC INVOLVEMENT OF 

IP DURING DE-ETIOLATION IN TOMATO (SOLANUM LYCOPERSICUM L.) 

67 



Véronique Bergougnoux 1 , David Zalabák 2 , Hana Pospíšilová, Michaela Jandová 3 , Ondřej Novák 1 , Martin 

Fellner 1 

1 

Palacky University in Olomouc and IEB ASCR, Laboratory of Growth Regulators, Olomouc, Czech Republic 

2 

Centre of the Region Haná for Biotechnological and Agricultural Research, Department of Molecular Biology, Faculty of Science, Palacky 

University, Olomouc, Czech Republic 

3 

Department of Botany, Palacky University in Olomouc, Faculty of Science, Olomouc, Czech Republic 

E-mail: v.bergougnoux @yahoo.fr 

As sessile organisms, plants have evolved sophisticated mechanisms to adapt to environmental conditions. 

Light is one of the most important factor influencing plant growth and development all through their life 

cycle. One of the well known light-regulated processes is de-etiolation, i.e. the switch from 

skotomorphogenesis to photomorphogenesis. The hormones cytokinins (CKs) play an important role 

during de-etiolation as high concentration of CKs in the culture medium induced photomorphogenesis in 

dark-grown seedlings. Using two tomato mutants deficient in blue light (BL)-induced responses and their 

corresponding wild-types (WT), de-etiolation in tomato was investigated. Under BL, the two mutants had 

longer hypocotyls than their corresponding WT. This was correlated with longer epidermal cells, higher 

proportion of epidermal cells with high ploidy and lower osmotic pressure in hypocotyl. The changes in 

endogenous CK were measured after exposure to BL and the cytokinin iP was found to be specifically 

involved in the BL-induced de-etiolation. We revealed also that BL modulates expression of genes encoding 

proteins involved in the iP metabolism. To our knowledge, this study unravels for the first time the specific 

role of iP in BL-induced de-etiolation. 


This work was supported by grant MSM6198959215 (MšMT) and P501/10/0785 

INTER-ORGANELLAR H2O2 SIGNALING IN ARABIDOPSIS 

Altynai Adilbayeva, Jodi Maple, Sigrun Reumann, Simon Geir Møller 


E-mail:altynai.adilbayeva@uis.no 

Reactive oxygen species (ROS) such as hydrogen peroxide (H2O2) are highly toxic molecules that are daily 

produced as by-products of normal metabolism and at increased rate under biotic and abiotic stress 

conditions. Apart from their toxic function, ROS also play essential roles as second messengers in several 

signal transduction cascades, ultimately inducing the expression of defence and adaptation genes and 

allowing organisms to tolerate adverse conditions. The ROS signaling network is highly complex and 

dynamic due to compartmentalization of both ROS production and scavenging at multiple sites. We aim at 

studying cellular H2O2 dynamics by an integrative approach considering several major cell compartments. 

Proteinaceous ROS sensors and spectral variants of green fluorescent protein are targeted to different cell 

compartments (e.g., chloroplasts, mitochondria, peroxisomes) to create multiply labeled Arabidopsis cell 

lines for integrated, temporal and spatial high-resolution analysis of H2O2 dynamics by confocal microscopy 

(Nikon A1R). ROS homeostasis will be perturbed by various means (e.g., abiotic stress) in wild-type plants 

and gain- and loss-of-function mutants of antioxidative enzymes and major signal transduction 

components. Cutting edge imaging technologies will be applied to comprehensively dissect abiotic H2O2 

signaling and accurate monitoring of H2O2 production, diffusion and scavenging by a non-invasive method. 

NIGHT-TIME PR:PFR RATIO AND DAYTIME STOMATAL OZONE UPTAKE 

Ane V. Vollsnes 1 , Aud B. Eriksen 1 , Cecilia M. Futsaether 2 , Ole Mathis Opstad Kruse 2 

1 University of Oslo, Oslo, Norway 

2 Norwegian University of Life Sciences, Ås, Norway 

E-mail: a.v.vollsnes@bio.uio.no 

In Trifolium subterraneum, oxidative stress due to ozone has been shown to result in more severe visible 

foliar injuries when phytochrome is predominantely in the far-red absorbing conformation during night 

than when it is in the red absorbing conformation. Phytochrome may be involved in stomata opening in the 

68 



morning, which could lead to differences in subsequent ozone uptake and consequently differences in 

visible foliar injuries. This hypothesis was tested by studying leaf temperature measurements obtained noninvasively 

through thermography. Leaf temperature is strongly dependent on transpiration, which in turn 

depends on stomata opening. Plants kept in darkness during night were compared to plants kept in dim farred, 

red or white light during night. Half the plants were exposed to ozone on three consecutive days. 

The main finding was that morning leaf temperatures were the same regardless of night-time illumination. 

Thus, all plants had the same potential for ozone uptake before ozone exposure started. On the other 

hand, evening leaf temperatures, shortly after the end of an ozone exposure period, were affected by an 

interaction between ozone exposure and night light conditions. 

MITOGEN-ACTIVATED PROTEIN KINASE ACTIVITY IN ROBINIA PSEUDOACACIA LEAFLETS 

D. Vidal, Q. Wang, E. Simón 

Plant Physiology Department, Faculty of Biology, University of Barcelona, Barcelona, Spain 

E-mail: dvidal@ub.edu 

Mitogen-activated protein kinases (MAPKs) are involved in the signal transduction of plant responses to 

environmental stimuli. Protein phosphorylation by MAPKs is typically organized in signalling cascades. It is 

estimated that about 10 % of all plant kinases (PKs) are involved in MAPK pathways. We previously reported 

MAPK activity, affected by red (R) and far-red (FR) light, in cucumber cotyledons. Here we examine 

whether R. pseudoacacia leaflets show MAPK activity. By performing immunodetection assays with 

polyclonal antibodies directed against mammalian ERK1/2 and in-gel phosphorylation assays, a MAPK-like 

activity was detected in Robinia leaflet extracts. Immunoprecipitation with antibodies anti-pERK (Tyr 204), 

anti-pERK1/2 and anti-ERK1/2 was used to purify putative MAPKs from the extracts. Two proteins (of about 

53 y 50 kDa) were recognised by the antibodies, indicating that they had immunological characteristics of 

MAPKs. Both proteins were also recognised in the immunoblotting assays and detected by the 

chemiluminescent system. The in-gel phosphorylation assays using SDS-PAGE polymerized with myelin basic 

protein (MBP) as exogenous substrate and [γ-32P] ATP, as phosphate donor, showed that the proteins of 

53 kDa and 50 kDa had MAPK-like activity, although the PK activity of the smaller protein was only scarcely 

detected. 

REDOX INSENSITIVE2, A NOVEL CHLOROPLAST PROTEIN LINKING REDOX REGULATION 

OF PHANG EXPRESSION TO PEP ACTIVITY IN THE CHLOROPLASTS 

Peter Kindgren, Juan de Dios Barajas López, Dmitry Kremnev, Aurora Piñas Fernández, Christian Tellgren- 

Roth, Ian Small, Åsa Strand 

Umeå Plant Science Centre, Umeå, Sweden 

E-mail: dmitry.kremnev@plantphys.umu.se 

The photosynthetic apparatus is composed of proteins encoded in nuclear and chloroplastic genomes and 

their activities are coordinated through intracellular signaling. The plastids produce multiple retrograde 

signals throughout development and in response to changes in the environment. These signals regulate the 

expression of photosynthesis-associated nuclear genes (PhANGs). Using forward genetics we identified 

REDOX INSENSITIVE2 (RIN2), a novel chloroplast component involved in redox-mediated retrograde 

signaling. The allelic mutants rin2-1 and rin2-2 demonstrated a mis-regulation of PhANG expression in 

response to excess light and inhibition of photosynthetic electron transport. As a consequence of the misregulation 

of PhANGs, the rin2 mutants displayed a high irradiance-sensitive phenotype with significant 

photoinactivation of PSII. RIN2 is localized to the nucleoids and plastid transcriptome analyses 

demonstrated that RIN2 is required for full expression of genes transcribed by the plastid-encoded RNA 

Polymerase (PEP). A direct role for PEP activity in redox-mediated retrograde signaling. Taken together, our 

results indicate that RIN2 is part of the PEP machinery and that the PEP complex responds to 

photosynthetic electron transport and generates a retrograde signal enabling the plant to synchronize the 

expression of photosynthetic genes from both the nuclear and plastidic genomes. 

69 



INVOLVEMENT OF CGMP IN THE NITRIC OXIDE EFFECT ON THE PHYTOCHROME- 

MEDIATED NYCTINASTIC CLOSURE OF ALBIZIA LOPHANTHA LEAFLETS 

S. Chellik, C. Bergareche, L. Moysset, E. Simón 

Plant Physiology Department, Faculty of Biology, University of Barcelona, Barcelona, Spain 

E-mail: esimon@ub.edu 

Albizia lophantha leaflets show both rhythmic and nyctinastic movements, from a horizontally extended 

position in daylight to a folded position at night. These movements are under phytochrome control, 

through a low fluence response. Leaflet movements depend on the curvature of a specialized motor organ, 

the pulvinus, located at the base of leaflets. Pulvinar curvature is caused by turgor changes in both extensor 

and flexor pulvinar motor cells, which in turn are driven by K + and Cl - ionic fluxes. Nitric oxide (NO) is a 

cellular signalling molecule which affects the activity of ionic channels. Previous data indicate that NO 

inhibits nyctinastic closure and this inhibition it is more apparent after red-light irradiation. Here we 

examine whether the NO effect is associated with changes in cGMP, by testing inhibitors of guanylate 

cyclase (ODQ and Ly85.583) and phosphodiesterase (sildenafil) as well as an analogue of cGMP (8-BrcGMP). 

Exogenous donors of NO inhibited nyctinastic closure, but simultaneous application of ODQ 

cancelled this inhibitory effect. ODQ (25-200 µM) and Ly85.583 (50 µM) enhanced nyctinastic closure. 

Sildenafil and 8-Br-cGMP inhibited nyctinastic closure. All these data implicate cGMP is involved in the NO 

effect on phytochrome-mediated nyctinastic closure. 

PROTEIN PHOSPHATSE 2A REGULATORY SUBUNITS ARE ESSENTIAL FOR METABOLISM 

AND PLANT DEVELOPMENT 

Behzad Heidari, Polina Matre, Else Müller Jonassen, Dugassa Nemie-Feyissa, Christian Meyer, Odd Arne 

Rognli, Simon G. Møller, Cathrine Lillo 

Center for Organelle Research, University of Stavanger, Stavanger, Norway 

E-mail: behzad.hidary@uis.no, cathrine.lillo@uis.no 

Canonical protein phosphatases 2A (PP2A) are trimeric protein complexes present in all eukaryotes cells, 

and known to be involved in regulation of cell cycle, hormone signaling and stress response. PP2A consists 

of a catalytic (C), scaffolding (A) and regulatory (B) subunit. The B subunits are divided into three (nonrelated) 

groups B55, B' and B''. The B subunits are generally thought to confer substrate specificity and 

cellular localization of the PP2A complex, hence providing for the specific functions of different PP2As. 

Bimolecular fluorescence complementation (BiFC) was used to identify PP2A regulatory subunits 

interacting with nitrate reductase (NR), and the two B55 (α and β) subunits were found to be positive. 

Dosage test of B55 effect on NR activation revealed that B55 promoted activation of NR. Interestingly, the 

homozygous double mutant (b55αβ) appeared to be lethal, which shows that the B55 group has essential 

functions that cannot be replaced by other regulatory B subunits. The B'α subfamily consists of two 

members B'α and B'β. A double mutant b'αβ null in B'α, but 1% of wild type B'β transcript levels showed a 

striking phenotype with poor seed set, pointing also to the B'α subfamily as being necessary for survival of 

the plant. 

THE ROLE OF NONSPECIFIC PHOSPHOLIPASE C IN PLANT STRESS RESPONSES 

Daniela Kocourkova, Premysl Pejchar, Zuzana Krckova, Olga Valentova, Jan Martinec 

Institute of Experimental Botany, ASCR, Prague, Czech Republic 

E-mail: martinec@ueb.cas.cz 

Phospholipid signalling is one of the key cellular regulatory mechanisms in plants in which phospholipases 

play a central role. A novel member of plant phospholipid signalling, nonspecific-phospholipases C (NPC) 

has been recognized only recently. Molecular, cellular and functional characterization of the small NPC gene 

family is still in its infancy. The gene family in Arabidopsis consists of six genes NPC1-NPC6. Among them, 

only NPC4 and NPC5 were heterologously expressed and partially characterized. Almost nothing is known 

about mechanism of NPC activation and downstream processes. 

70 



Here we summarize our recent results concerning a role of NPC in responses of plants to abiotic stresses 

mainly to aluminium and salt. Aluminium inhibited NPC activity in both tobacco BY-2 cells and pollen tubes. 

The inhibition was time and concentration dependent. Aluminium arrested growth of pollen tubes was 

rescued by diacylglycerol (DAG) treatment. Treatment with phosphatidic acid was ineffective. Salt stress 

caused rapid, concentration dependent and transient upregulation of NPC4 expression in Arabidopsis 

roots. NPC4prom::GUS experiments revealed root tip localization of NPC4 expression. The in situ activity of 

NPC increased after salt treatment in Arabidopsis seedlings in time and concentration dependent manner 

as well. The expression of ABA related genes in salt treated NPC4 knockout plant was impaired. The 

possible role of DAG will be discussed. 

IMPORTANCE OF PHOSPHORYLATION SITES IN SAMDC UORF PEPTIDE FOR 

MAINTAINING THE HOMEOSTATIC POLYAMINE LEVELS BY THE REGULATION OF 

DEGRADATION OF ITS OWN MRNA TRANSCRIPT AND PROTEIN 

Yu Jin Choi, Ky Young Park 

Sunchon National University, Jeollanam-do, South-Korea 


S-Adenosylmethionine decarboxylase (SAMDC), a key enzyme for polyamines biosynthesis, was tightly 

regulated for homeostatic levels by translational inhibition of its own protein. The degradation of 

downstream GUS mRNA in transgenic tobacco plants containing point-mutation of phosphorylated small 

uORF peptide at Ser10 (P10), Ser17 (P17), Ser28 (P28) and Ser54 (P54), respectively, were about 1.8-fold 

delayed than WT uORF plants. These data suggest that phosphorylation of small uORF peptide is an 

essential component for maintaining the transcript level of the downstream ORF. Also, we showed that 

degradation rate of GUS and SAMDC protein after treatment with protein synthesis inhibitor cycloheximide 

and specific proteasome inhibitor MG115 was accelerated in the presence of small uORF peptide. These 

results implied that small uORF peptide might effectively act as a functional regulator for its related ORF in 

cis in the manner of proteasome-dependent degradation. However, these effects of SAMDC uORF protein 

did not occurred on endogenous activity of ADC, which was other enzyme for polyamine biosynthesis and 

contained its own ADC uORF sequence in 5' UTR. Therefore, SAMDC uORF has a sequence-specific function 

of transcriptional and/or translational inhibitor for its own SAMDC protein not only in cis but also in trans. 

CHARACTERIZATION OF CYSTEINE-RICH RLKS IN ARABIDOPSIS THALIANA 

Niina Idänheimo 1 , Adrien Gauthier, Mikael Brosché 1,2 , Hannes Kollist 2 , Jaakko Kangasjärvi 1 , Michael 

Wrzaczek 1 

1 University of Helsinki, Helsinki, Finland 

2 University of Tartu, Tartu, Estonia 

E-mail: niina.idanheimo@helsinki.fi 

The production of reactive oxygen species (ROS) is a common response to many stresses in plants. The 

production and the action of ROS are critical to the stress recognition and the coordination of the plant's 

response to those stresses. Plants can suffer from oxidative stress when ROS production exceeds 

antioxidizing capacity and can ultimately lead to cell death by ROS toxicity. In contrast to ROS as toxic 

molecules, ROS is also a signalling molecule and important regulator of plant development and controls 

processes such as root development, pollen tube growth and cell expansion. 

ROS are sensed via yet unidentified mechanisms. We have found several Cysteine-rich RLKs (CRKs), which 

are transcriptionally regulated in response to ROS. The members of the CRK subfamily could act as ROS 

“receptors” by sensing ROS through redox modifications of the extracellular domain which could activate 

receptor complex and lead to signal transduction and response. Phenotypic analysis of the crk knock-out 

and the overexpression plants under various conditions have provided more information about the 

biological relevance of these proteins and supported our hypothesis. We have started physical and 

molecular characterization of these proteins to analyze their role in ROS perception and subsequent signal 

transduction. 

71 



THE ROLE OF F-BOX-PROTEINS IN STRESS SIGNALING IN ARABIDOPSIS THALIANA 

Maria Piisilä, Mehmet Ali Keçeli, Günter Brader, Tapio Palva 

Department of Biological and Environmental Sciences, University of Helsinki, Helsinki, Finland 

E-mail: maria.piisila@helsinki.fi 

Induced defenses and stress responses play a major role in plant disease resistance and are regulated by a 

network of interconnected signal transduction pathways with the plant hormones ethylene (ET), jasmonic 

acid (JA) and salicylic acid (SA) as crucial mediators. These specific hormone-mediated signaling cascades 

trigger distinct sets of stress-related genes leading to tolerance or resistance. F-box proteins (roughly 700 

in Arabidopsis) are important components in these stress responses. They act as members of SCF protein 

complexes, which target substrate proteins for modification and degradation and thereby allow plants to 

respond rapidly to environmental stress factors. They are involved in various processes including plant 

metabolism, hormonal responses as well as responses to environmental stresses. The objective of this 

study is to identify and characterize specific F-Box proteins involved in stress signaling and components 

interacting with these F-box proteins. To elucidate the function of F-box proteins belonging to subfamily C3 

in the regulation of stress responses, we employ a reverse genetic screening approach and characterize T- 

DNA mutant lines of this subfamily of F-box proteins for their performance under abiotic (cold, ozone, 

drought) and biotic (Erwinia carotovora, Botrytis cinerea, Alternaria brassicicola) stresses. Mutants with 

altered stress response phenotypes will be subjected to microarray analysis to elucidate the response 

pathways a given F-box protein is involved in. So far, more than 50 F-Box mutant lines were screened with 

the pathogens Erwinia carotovora and Botrytis cinerea and cold, ozone, salt and different hormones. We 

have found many candidates which are showing clear phenotype to pathogen (Erwinia), ozone, and 

hormone (ABA) treatments. Our research is focusing on these confirmed candidates. 


This work is supported by the Academy of Finland Center of Excellence program. 

EMBRYO-SPECIFIC PROTEINS IN CYCLAMEN PERSICUM ANALYZED WITH 2-D DIGE 

Ragnhild Lyngved, Jenny Renaut, Jean-Francois Hausman, Tor-Henning Iversen, Anne Kathrine Hvoslef-Eide 

Sør-Trøndelag University College, Trondheim, Norway 

E-mail: ragnhild.lyngved@hist.no 

Somatic embryogenesis can be used to produce artificial seeds of Cyclamen persicum, one of the most 

important ornamental plants for the European market, both as a potted plant in northern Europe and a 

bedding plant in the cool winters in southern Europe. The aim of this study was to obtain new insights into 

the molecular biology of somatic embryogenesis, which in turn can be useful for the improvement of tissue 

culture methodology. Total proteins were characterized from two isogenic cell lines of Cyclamen persicum, 

one that was embryogenic and one that never has shown any embryogenic capacity. The extracted 

proteins were separated by two-dimensional differential gel electrophoresis (2-D DIGE) and selected 

proteins were treated using the ETTAN Dalt Spot Handling Workstation. Protein identification was 

performed using MALDI-TOF-MS. More than 1200 Cyclamen proteins were detected; 943 proteins were 

common to both lines. The different protein patterns of the embryogenic and non-embryogenic cell lines 

were obvious: One hundred eight proteins were more abundant in the embryogenic cells, and 97 proteins in 

the non-embryogenic cells. Among the differentially expressed proteins, 128 were identified. MALDI-TOF- 

MS analysis enabled 27 spots to be proposed as candidates for embryo-specific proteins, as they were 

unique to the embryogenic cell line. The proteins identified are involved in a variety of cellular processes, 

including cell proliferation, protein processing, signal transduction, stress response, metabolism, and 

energy state, but the majority are involved in protein processing and metabolism. The main functions of the 

putative embryo-specific proteins have been discussed in proportion to their role in the somatic 

embryogenesis process. 

STUDYING THE MECHANISMS UNDERLYING SUCROSE-DEPENDENT GENE EXPRESSION 

IN A. THALIANA CELL CULTURES 

72 



Sabine Kunz, Edouard Pesquet, Leszek A. Kleczkowski 

Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, Sweden 

E-mail: sabine.kunz@plantphys.umu.se 

The ability of a plant to cope with different environmental factors depends on its energy state. In plants, 

the energy available for each cell underlies a continuous fluctuation, which is influenced by carbon 

assimilation and respiration. The response of a plant to available carbohydrates (CH), the main source of 

energy, is partly coordinated through cell signaling by specific changes in gene expression. These are 

determined by the interaction of transcription factors (TF) with cis-regulatory DNA elements in promoters 

of responsive genes. Here we describe experimental approaches to verify the functionality of the three 

insilico predicted cis-elements SUC/ROS 3, 5 and 6, which are likely to be involved in sucrose-dependent cellsignaling. 

The use of A. thaliana cell cultures, grown on xylose as carbon source, allows us to distinguish the 

response of the cells to different CH signals. This set-up, applied to cells transformed with promoterreporter 

gene constructs that bear the cis-elements in both the forward and reverse orientation in a native 

as well as synthetic environment, is a valuable tool to determine the involvement of these cis-elements in 

the different transcriptional responses in the adaptation to changes of available CH. 

ENDOGENOUS REACTIVE OXYGEN SPECIES (ROS) STATUS AND CELL DEATH IN NITRIC 

OXIDE (NO) MUTANTS UNDER COPPER EXCESS 

Zsuzsanna Kolbert, Andrea Pető, Nóra Lehotai, László Erdei 

Department of Plant Biology, Faculty of Science and Informatics, University of Szeged, Hungary 

E-mail: kolzsu@bio.u-szeged.hu 

Enhanced ROS production is associated with abiotic stresses, and NO as a general plant signal plays a role in 

cell death induction. The possible relationship between ROS and NO was investigated in copper-treated 

wild type (WT), NO-overproducer (nox1) and -deficient mutant (nia1nia2, nia1nia2noa1-2) Arabidopsis by 

microscopic methods. Seven-day-old plants were grown in MS medium supplemented with 0, 5, 25 and 50 

µM CuSO4. In the elongation zone (EZ) of wild type roots the levels of NO and superoxide anion (O2 .- ) 

decreased, peroxynitrite (ONOO - ) enhanced as the effect of Cu 2+ exposure, which suggests the reaction 

between NO and O2 .- to produce ONOO - . The level of ROS was also increased by Cu 2+ , which can be partly 

responsible for oxidative cell death. In roots of NO-overproducer and -deficient plants the copper-induced 

alterations in the ROS metabolism and the degree of cell death was more pronounced compared to WT. 

Based on our results it can be stated, that under Cu 2+ excess NO acts as an antioxidant by eliminating O2 .- 

and yielding the less toxic ONOO - . The copper-induced cell death is strictly regulated by endogenous NO 

status: NO content in the root being over or under the optimal level intensifies cell death process. 

PLASMA MEMBRANE AQUAPORIN PIP2;3 IS RELATED TO HEAT STRESS RESPONSE IN 


Ayako Tsuchihira 1 , Yuko Hanba 2 , Masayoshi Maeshima 1 

1 Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan 

2 Kyoto Institute of Technology, Kyoto, Japan 

E-mail: tsuchihira.ayako@f.mbox.nagoya-u.ac.jp 

Plants have a large protein family of aquaporin, which facilitate membrane transport of water and other 

small molecules. Arabidopsis thaliana contains 35 aquaporins divided into four subfamilies: PIP, TIP, NIP and 

SIP. Plasma membrane aquaporins (PIPs) play a key role of intercellular water transport. PIPs have been 

reported to response to abiotic stresses. We focused on the physiological function of PIPs under high 

temperature conditions. 

Expression of 13 PIP genes was quantified by qRT-PCR in A. thaliana. Among them, only PIP2;3 transcript 

level was transiently increased when the temperature raised to 36°C . The increased level of mRNA was 

rapidly decreased within 2 hr even at high temperatures. PIP2;3 promoter-GUS fusions revealed the tissuespecific 

expression under high temperature. After heat stress, PIP2;3 was predominantly expressed in the 

73 



veins of leaves and the vascular tissue of roots. We estimate that this transient alteration of PIP2;3 is tightly 

related to initial response of heat stress and the increased PIP2;3 functions as water channel in order to 

regulate the water conditions in plants at high temperature. 

THE LIGHT-DEPENDENT INDUCTION OF LEAF SENESCENCE 

Bastiaan Brouwer, Olivier Keech, Agnieszka Ziolkowska, Per Gardeström 

Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, Sweden 

E-mail: bastiaan.brouwer@plantphys.umu.se 

Often, leaves have to deal with reduced light-conditions, generally resulting from shading by neighboring 

plants. As a common response, they can relocate nutrients from shaded to developing leaves through a 

process called leaf senescence. Shade- and dark-induced leaf senescence are thought to be initiated by 

either the relative increase of far-red light on phytochromes or the light dropping below the light 

compensation point. However, the regulatory mechanisms by which these factors induce leaf senescence 

are still unclear. 

We investigated this matter by shading individual attached leaves and analyzing the resulting processes by 

physiological measurements, transmission electron microscopy and the expression of senescenceassociated 

genes. The results showed that mild shading led to photosynthetic acclimation within the 

shaded leaf, whereas strong shading led to leaf senescence when the light intensity was lower than the 

acclimated light compensation point. Furthermore, using phytochrome null-mutants, we showed that 

despite an increase in chlorophyll-degradation upon shading, these mutants did not increase the expression 

of senescence-associated genes. Altogether, our results suggest that the induction of shade-induced leaf 

senescence mainly depends on the efficiency of carbon fixation rather than on a phytochrome-dependent 

signaling pathway. 

OZONE SENSITIVITY OF THE MOST IMPORTANT INDIAN CROPS AND TREE SPECIES 

Elina Oksanen, Vivek Pandey, Sari Kontunen-Soppela, Sarita Keski-Saari 

University of Eastern Finland, Joensuu, Kuopio and Savonlinna, Finland 

E-mail: Elina.Oksanen@uef.fi 

Indo-Gangetic Plains (IGP) is the bread-basket of rapidly developing India, where intense agriculture, landuse 

changes, industrialization, urbanization and population growth lead to increasing emissions of 

precursors for greenhouse gases, such as ozone. It has indicated that surface ozone levels are much above 

critial levels at most places in the Indian region. Ozone caused losses in gross primary production in India is 

expected to be roughly 20-30% by 2100. However, species-level information about ozone sensitivity is very 

scarce. 

In this development project we screen the genetic variation of ozone sensitivity of the most important 

Indian crop and tree species (rice, wheat, pea, mustard, teak) addressing tolerance mechanisms. We utilize 

different and complementary methodological approaches: laboratory experiments in Finland and free-air 

approaches and natural field sites (low- and high-ozone regions) in IGP area in India. The plants will be 

studied e.g. for growth, gas exchange, ozone uptake, changes in metabolome and protein profiles, and key 

genes and enzymes. Metabolome assays from grains (e.g. nutrients and amino acid profiles) will be linked 

to sensitivity screening. The results from plant responses and ozone monitoring data will be integrated for 

ozone risk assessment, which is useful for agriculture managers and planners, and ultimately farmers. 

VARIATION IN HIGH-LIGHT STRESS TOLERANCE BETWEEN ARABIDOPSIS ECOTYPES 

Espen Granum, Maria Luisa Pérez Bueno, Matilde Barón 

Department of Biochemistry and Molecular and Cellular Biology of Plants, Estación Experimental del Zaidín, CSIC, Granada, Spain 

E-mail: e.granum@csic.es 

74 



Photosynthetic responses and tolerances to short- and long-term high-light stress were investigated in 

seven Arabidopsis ecotypes collected from different environments, including high altitude, high salinity, dry 

and sunny regions. Plants grown under standard conditions (light intensity 150 µmol m -2 s -1 ) were compared 

to plants grown at different periods of high light (300-1000 µmol m -2 s -1 ). The different ecotypes showed 

different degrees of change in morphology, with Col-0 displaying the strongest stress symptoms under high 

light. Analyses by multicolour fluorescence imaging (blue, green, red and far-red) indicated enhanced stress 

tolerances in Pf-0, Po-0 and Ws-0, whereas Col-0 and C24 were the most sensitive. Chlorophyll fluorescence 

kinetics showed that quantum yield is strongly stimulated by high light in Cvi-0 and Nok-2, and nonphotochemical 

quenching indicates better high-light adaptation in Cvi-0 and C24. Photosynthetic 

performances in the different ecotypes were also evaluated by gas exchange analysis, and photosynthetic 

pigments and anthocyanins determined by spectrophotometry. Further work is underway to resolve the 

mechanisms of stress tolerance in Arabidopsis by comparative proteomics as well as metabolic and 

transcriptional analyses. 


This work was supported by grants MCINN-FEDER/AGL2008-00214 and CVI 03475 (Junta de Andalucía). 

PEROXISOME-MEDIATED MECHANISMS OF ABIOTIC STRESS TOLERANCE 

Gopal Chowdhary 1, 2 , Sigrun Reumann 2 

1 KIIT School of Biotechnology, Campus XI, KIIT University, Bhubaneswar, Orissa, India 


E-mail: gopal.chowdhary@uis.no 

Present alterations in environmental scenario have led to a serious concern about the loss of plant 

productivity due to extreme abiotic stress conditions, which mainly include drought, salt, metal toxicity 

high and low temperature. The tremendous amount of losses caused a thrust on high end research to make 

plants tolerant to these stress conditions. Many plant species evolved effective, endogenous mechanisms 

to cope with abiotic stress conditions. Our group focuses of stress adaptation mechanisms in the model 

plant Arabidopsis that involve peroxisome functions. Over the past years, our lab has established the 

Arabidopsis peroxisome proteome database. Novel plant peroxisomal proteins have been identified on a 

large scale by experimental proteome analyses and bioinformatics-based protein prediction models. These 

proteins of largely unknown function have been subjected to a 2nd order screen for genes that are highly 

inducible by abiotic stress conditions in microarray-based transcriptome studies using the Genevestigator 

database. Selected candidate genes were subcloned and validated as peroxisome targeted by in vivo 

subcellular targeting analysis. Many proteins have been validated as peroxisomal, including, for instance, a 

calcium binding annexin homolog, an alanine:glyoxylate aminotransferase homolog, and a polyamine 

oxidase. Stress-inducible gene expression is presently being validated by real-time PCR. The predicted 

functions and mechanisms underlying abiotic stress resistance are being predicted by bioinformatics tools 

such as co-expression analysis and experimentally investigated by loss- and gain-of-function mutants. To 

translate the knowledge from model Arabidopsis to important crop plants, Oryza orthologs are being 

analyzed in parallel. 

THE EFFECT OF DIFFERENT RADIATION CONDITIONS ON THE QUALITATIVE AND 

QUANTITATIVE COMPOSITION OF UNBOUND PHENOLIC COMPOUNDS IN BARLEY 

LEAVES EXAMINED BY UV-VIS ABSORPTION SPECTROPHOTOMETRY AND HPLC-DAD. 

Jakub Nezval 1 , Jiří Kalina 2 , Michal štroch 1 , Vladimír špunda 1 

1 Department of Physics, Faculty of Science, University of Ostrava, Ostrava, Czech Republic 

2 Department of Chemistry, Faculty of Science, University of Ostrava, Ostrava, Czech Republic 

E-mail: nezval.jakub@gmail.com 

In this work we focused on the role of phenolic compounds (PC) in the plant protection against deleterious 

effects of the ultraviolet radiation (UV-R). These compounds are able to absorb UV-R before it can reach 

sensitive biomolecules (UV-shielding mechanism) or they can inactivate harmful reactive compounds, which 

are produced in cells by UV-R. We tried to elucidate interaction between UV-R and photosynthetic active 

75 



radiation (PAR) in the induction of PC accumulation. As a model plant we used spring barley (Hordeum 

vulgare L. cv. Bonus) cultivated in 4 different radiation conditions. Two groups of plants were cultivated in 

the absence of UV-R under the high (1000 µmol m -2 s -1 ) and low intensities (50 µmol m -2 s -1 ) of PAR. Other 

two groups were cultivated in aforesaid PAR conditions, but they were treated by higher doses of UV-B 

(280-320 nm). Important part of this work is the standardization of spectrophotometric and HPLC-DAD 

methods, which allow us to determine the qualitative and quantitative composition of unbound leaf PC. We 

made summarization of obtained parameters (absorption spectra, retention times, calibration equations). 

The results of the HPLC analysis of leaf extracts show that response of specific PC to various levels of UV-B 

and PAR differs significantly. 

EFFECT OF DIFFERENT ANION CHANNEL INHIBITORS ON THE GERMINATION OF THE 7B- 

1, A TOMATO (SOLANUM LYCOPERSICUM L.)MUTANT DEFECTIVE IN BL SIGNALING. 

Jan Humplík 1,2 , Jana Balarynová 2 , Veronique Bergougnoux 2 , Martin Fellner 2 

1 

Centre of the Region Haná for Biotechnological and Agricultural Research Department of Growth Regulators, Palacky University, Olomouc, 

Czech Republic 

2 

Palacky University and Institute of Experimental Botany, Academy of Science of the Czech Republic, Olomouc, Czech Republic 

E-mail: jfhumplik@gmail.com 

The tomato mutant 7B-1 was previously described as being more tolerant than corresponding wild-type 

(WT) to various abiotic stresses in a blue-light dependent manner. In order to understand the role of light 

during germination, 7B-1 and WT seeds germinated on the basal MS medium in different light conditions. 

Under continuous blue-light (BL), mutant seeds germinated better than seeds of the WT. The involvement 

of anion channels in germination was assessed by adding anion channel blockers 9-AC, niflumic acid or DIDS 

to the culture medium. Under BL, niflumic acid strongly inhibited germination of 7B-1 seeds whereas 

germination of WT seeds was less affected. Inhibitors 9-AC and DIDS had no essential effect on germination 

of 7B-1 seeds. Differently in WT, 9-AC inhibited seed germination additionally to BL, whereas DIDS abolished 

the inhibitory effect of BL. Preliminary data from expression analysis of ESTs homologous to selected 

Arabidopsis channels suggest that during seed germination under BL, their expression is affected by the 7B- 

1 mutation. Our results suggest an important role of anion channels and 7B-1 gene in the process of 

imbibition/germination under BL. 


The work was supported by Ministry of Education of the Czech Republic (grants no. ME10020, MSM6198959215). 

CPD PHOTOLYASE REPAIRS ULTRAVIOLET-B-INDUCED CPDS IN RICE CHLOROPLAST AND 

MITOCHONDRIAL DNA 

Masaaki Takahashi, Mika Teranishi, Hiroyuki Ishida, Junji Kawasaki, Atsuko Takeuchi, Jun Hidema 

Graduate School of Life Sciences, Tohoku University, Sendai, Japan 

E-mail: j-hidema@ige.tohoku.ac.jp 

Ultraviolet-B (UVB) (280 to 320 nm) radiation in sunlight suppresses photosynthesis and protein 

biosynthesis, thereby decreasing growth and productivity. A principal cause of such UVB-induced growth 

inhibition in plants is the major UVB-induced DNA lesion, cyclobutane pyrimidine dimers (CPDs). UVB 

radiation induces the formation of CPDs in nuclear, chloroplast, and mitochondrial DNA. Repair of CPDs is 

therefore essential for plant survival while exposed to UVB-containing sunlight. Nuclear repair of the UVBinduced 

CPDs by CPD photolyase is well known, but repair processes in plant chloroplasts and mitochondria 

are not understood. Here, we report the photoreactivation of CPDs in chloroplast and mitochondrial DNA in 

rice. Biochemical and subcellular localization analyses using rice strains with different levels of CPD 

photolyase activity and transgenic rice strains showed that full-length CPD photolyase is encoded by a 

single gene, not a splice variant, and is expressed and targeted not only to nuclei but also to chloroplasts 

and mitochondria. Thus, a single DNA repair enzyme CPD photolyase is “triple targeted” in rice cells. The 

results indicate that rice may have evolved a CPD photolyase that functions in all three organelles that 

contain DNA to protect cells from the harmful effects of UVB radiation. 

76 



OZONE-INDUCED CHANGES IN CAROTENOIDS AND CHLOROPHYLLS IN THREE POPULUS 

CLONES 

Sarita Keski-Saari, Jennifer Dumont, Markku Keinänen, Sari Kontunen-Soppela, Elina Oksanen, Didier Le 

Thiec 


E-mail: sarita.keski-saari@uef.fi 

Ozone is a phytotoxic air pollutant causing oxidative stress. We studied the effect of ozone on carotenoids, 

chlorophylls and polyisoprenoid alcohols in three euramerican poplar clones (Populus deltoides x Populus 

nigra : 'Carpaccio', 'Cima' and 'Robusta'). Poplars originating from cuttings were grown for 6 weeks and 

exposed to ozone in fumigation chambers (120 ppb each day for 13h). Leaf samples were collected 2, 4, 11, 

15 and 17 days after the start of ozone treatment. Chemical analyses were made by HPLC-APCI-MS. The 

clones clearly differed in growth rate and ozone responses. 'Carpaccio' had the highest above-ground 

biomass, and it was also the most ozone tolerant genotype. Greatest growth reductions by ozone were 

measured for 'Cima ', while 'Robusta' showed the greatest number of visible injuries. Ozone and control 

samples were clearly separate in PCA based on analysed chemical profiles, so that ozone treatment 

explained 50% of the variance. Ozone reduced the concentrations of chlorophylls a and b, β-carotenes, 

luteins, neoxanthin and some unidentified isoprenoid derivatives in all clones. The difference between the 

ozone and control treatments was the strongest on the last two sampling dates. The genotypes differed 

from each other, but genotype explained only 12% of the variation. 

THE FIRST BURST OF ROS IN STRESS-INDUCED BIPHASIC GENERATION OF ROS AND 

ETHYLENE IS AN IMPORTANT DETERMINANT FOR STRESS TOLERANCE 

Na Ri Ji, Myung Hee Nam, Ky Young Park 

Sunchon National University, Jeollanam-do, South Korea 


The biphasic generation of ROS and ethylene, which was peaked twice in a first transient phase and a 

second massive phase was occurred after treatment of abiotic/biotic stresses. The activation of both 

members of NADPH oxidase, RbohD and RbohF, was observed at both phases of ROS production, with 

much more rapid and abundant increase of RbohD transcripts compared to that of RbohF in first phase. The 

treatment of ACC was induced more specifically the expression of RbohD in first phase of ROS production. 

Also, the treatments of AVG and NBD, which are inhibitors of ethylene biosynthesis and ethylene signaling, 

respectively, were significantly reduced ROS accumulation during phase I. The accumulation of ROS and cell 

damage, which was significantly decreased after abiotic stresses in transgenic plants, in which ethylene 

biosynthetic pathway or signaling was impaired. This means that after onset of stress treatment the first 

peak of endogenous ROS accumulation was followed by ethylene production during phase I, and then ROS 

and ethylene biosynthesis were occurred for a larger and more prolonged response with necrosis during 

phase II. Therefore, the first burst of ROS is more responsible for making a role as a signaling molecule 

during stress-induced response. 

PHOTOSYNTHETIC RESPONSES OF 3 BIRCH SPECIES TO ELEVATED CO2 CONCENTRATION 

WITH FACE SYSTEM IN NORTHERN JAPAN 

Makoto Watanabe, Qiaozhi Mao, Eka Novriyanti, Hirotaka Ito, Kentaro Takagi, Kaichiro Sasa, Takayoshi 

Koike 

Hokkaido University, Sapporo, Japan 

E-mail: nab0602@for.agr.hokudai.ac.jp 

The increasing CO2 concentration ([CO2]) is one of the most critical environmental changes. Since CO2 is the 

primary substrate for photosynthesis, the increased [CO2] will act as a fertilizer. However, a decrease in 

photosynthetic activity, called 'down-regulation of photosynthesis', is occasionally observed. In general, the 

77 



responses to elevated [CO2] are different between species. We examined photosynthetic responses of 3 

kinds of birches to elevated CO2 under 2 soil types. Two-year-old seedlings of Betula ermanii, B. 

maximowicziana and B. platyphylla var. japonica were grown on fertile brown forest soil or infertile volcanic ash 

soil and exposed to 500 µmol CO2 mol -1 in a free air CO2 enrichment system located in northern Japan for 

one growing season of 2010. The exposure to elevated [CO2] increased light-saturated net photosynthetic 

rates at growth [CO2] in the leaves of B. ermanii and B platyphylla, whereas no photosynthetic enhancement 

was observed in B. maximowicziana. The decrease in maximum rates of carboxylation and electron 

transport due to the decreasing leaf N content was a factor of down-regulation of photosynthesis in B. 

maximowicziana under elevated [CO2]. Our results suggest that there is a difference of photosynthetic 

response to elevated [CO2] even within the same genus of Betula. 

DEHYDRIN IN BUCKWHEAT SEEDS 

Michiko Momma 

National Food Research Institute, Tsukubi, Ibaraki, Japan 

E-mail: michiko@affrc.go.jp 

Dehydrin is a class of LEA proteins that are expressed in the late stage of seed maturation in response to 

water stress and formation of abscisic acid. While involvement of dehydrin in the desiccation tolerance of 

plants has been extensively studied, little is clarified about the property and function of dehydrin proteins 

in seeds. We have shown protective activity of dehydrin proteins in soybean and rice seeds to the 

freezes/thaw denaturation of enzyme proteins. Studies also suggested that the accumulation of dehydrin in 

peanut relate to human allergenic response. In this study, the presence of dehydrin proteins in buckwheat 

was indicated for the first time, using immunoblotting for antibody against its highly conserved lysine-rich 

motif. A major (20 kDa) and a minor (16 kDa) bands were found in buckwheat flours, noodles and boiled 

water used for noodle cooking. The recognition of the major dehydrin protein (20 kDa) was difficult in the 

SDS-PAGE analysis, since its molecular weight was very close to that of legumin, most abundant storage 

protein of buckwheat. The minor dehydrin-like protein at 16 kDa showed moderate resistance to pepsindigestion. 

Results suggested that buckwheat and its product contain dehydrin-like proteins, which 

associated with pepsin resistance. 

EFFECT OF PHOSPHORYLATION ON FUNCTION OF RICE CPD PHOTOLYASE 

Mika Teranishi, Kentaro Nakamura, Haruya Furukawa, Masaaki Takahashi, Jun Hidema 

Graduate School of Life Sciences, Tohoku University, Sendai, Tōhoku, Japan 

E-mail: tera@ige.tohoku.ac.jp 

Ultraviolet-B (UVB) radiation induces photodamage in DNA, including the formation of cyclobutane 

pyrimidine dimers (CPDs). CPDs are repaired by CPD photolyase that monomerizes dimers by using 

blue/UVA light as the energy source. In rice, CPD photolyase is a crucial factor for determining UVB 

sensitivity. We previously demonstrated that the native rice CPD photolyase was phosphorylated [1]. There 

is no report that CPD photolyase is phosphorylated in vivo in organism other than rice plant. First, we 

analyzed the phosphorylation status of CPD photolyase in several poaceous species. It was found that the 

phosphorylation of CPD photolyase was not a common event among plant species, wheat CPD photolyase 

was phosphorylated, whereas barley and maize CPD photolyase were not. We also found that CPD 

photolyase localized to chloroplasts, mitochondria and nuclei [2]. Mitochondria showed a high proportion 

of non-phosphorylated CPD photolyase, whereas nuclei and chloroplasts had a relatively high proportion of 

the phosphorylated form. These results suggested that phosphorylation of CPD photolyase was involved in 

its translocation to organelles. In this congress, we discuss about the significance of phosphorylation on 

CPD photolyase in rice plants. 

References 

[1] Teranishi et al., Plant Physiol. 2008 

[2] Takahashi et al., Plant J. 2011 

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INCREASING PROTEIN QUALITY/QUANTITY AND ZN CONTENT OF BARLEY GRAIN 

Mohammad Nasir Uddin 1 , Mette Lange 3 , Jan K. Schjørring 2 , Preben Bach Holm 1 , Eva Vincze 2 

1 University of Aarhus, Aarhus, Denmark 

2 University of Copenhagen, Copenhagen, Denmark 

3 University of Aalborg, Aalborg, Denmark 

E-mail: MohammadNasir.Uddin@agrsci.dk 

Atmospheric CO2 enrichment may lead to increased yields of cereal species but there will be a “dilution” of 

other nutrients like minerals and proteins in the grain. The nutritional short-comings can not be met solely 

by using higher amounts of nitrogen and mineral fertilizer since this would increase nitrogen and mineral 

loads into the environment. Therefore, alternative approaches embracing creation of transgenic lines or 

exploitation of the natural variation are needed to increase grain Zn content. 

In our experiments, transgenic barley lines with a reduced C-hordein content showed not only an altered 

amino acid composition but also a 2-3 fold increase in the grain Zn concentration. From bioinformatic 

studies we have identified protein families which potentially may bind more Zn. The expression levels of the 

selected gene/protein families are currently being assessed by quantitative PCR. We also have a collection 

of close to 300 barley cultivars with high grain protein content and a substantial genetic variation in grain 

Zn concentration. We will use them to further explore the potential link between protein quality/quantity 

and Zn content under elevated atmospheric CO2 level. 

ANALYSIS OF CARBON ION BEAM INDUCED UVB TOLERANT AND SENSITIVE RICE 

MUTANTS 

Nao Takano 1 , Yuko Takahashi 1 , Misturu Yamamoto 1 , Mika Teranishi 1 , Yoshihiro Hase 2 , Ayako Sakamoto 2 , 

Atsushi Tanaka 2 , Jun Hidema 1 

1 Graduate School of Life Sciences, Tohoku University, Sendai, Tōhoku, Japan 

2 Radiation-Applied Biology Division, Quantum Beam Science Directorate, Japan Atomic Energy Agency, Ibaraki Prefecture, Tokaimura, Japan 

E-mail:n-takano@ige.tohoku.ac.jp 

UVB radiation can damage plant growth and development. To develop the factors related to UVB 

resistance, Japanese rice cultivar “Sasanishiki” was exposed to carbon ion beam (320 MeV: 12C6+, 80 Gy), 

which have been used to generate new mutants varieties, and two UVB tolerant and three UVB sensitive 

mutants were isolated. We focused on UVB tolerant mutant utr-319 (UV Tolerant Rice-319) and UVB 

sensitive mutant usr-1 (UV Sensitive Rice-1), because the levels of UV-absorbing compounds or CPD 

photorepair activity, which has been thought as factors related UVB tolerance of each mutant wasn't 

affected compared to wild-type. First, to estimate the mutated regions of each mutant, we performed 

Array Comparative Genomic Hybridization, and then, genomic DNA sequence analysis using BAC clone of 

each mutant. As a result, we found that approximately 40 kb on chromosome 7 containing 2 genes 

(AK111251, AK071492: function of each gene product has not been unknown) was deleted in utr-319. In usr-1 

mutant, we found that approximately 53 kb on chromosome 7 containing 2 genes (AK073834; encodes 

thioesterase domain, AK121807; encodes Rpb6) was deleted. In this meeting, we introduce the characters 

of utr-319 and usr-1, and discuss about relationships between deleted genes and UVB tolerance or 

sensitivity. 

THE INTERACTION BETWEEN DEVELOPMENT AND STRESS IN THE LOW TEMPERATURE 

METABOLOME OF ARABIDOPSIS THALIANA 

Paulina Stachula, M. Lundmark, L. Hendrickson, E. Pesquet, C. Külheim, A. Vlčková, H. Stenlund, T. Moritz, 

Å. Strand, V. Hurry 

Umeå Plant Science Centre, Umeå, Sweden 

E-mail: paulina.stachula@plantphys.umu.se 

79 



In an attempt to discriminate between a low temperature stress versus cold acclimation responses we 

contrasted the anatomical, physiological and metabolic profile changes of Arabidopsis thaliana plants 

grown under different temperature regimes. We contrasted the characteristics of 4-week old 23°Cgrown 

plants that were subsequently exposed to 5°C and allowed to develop new mature leaves (CD) to plants 

that germinated and spent their life cycle in 5°C (CG). For most parameters measured, the CG phenotype 

exhibited intermediate characteristics between WG and CD plants. This result was accompanied by 

improved photosynthetic capacity and daily photosynthetic carbon gain of CG relative to CD plants when 

considering primary resource investment per leaf. The difference between cold stress and development 

(CD) versus cold growth (CG) was further demonstrated by the fact that the CD and CG leaves had equal 

freezing tolerance yet metabolically the CG plants were more similar to WG leaves. Thus, these experiments 

demonstrate that if we are to develop an more informative and integrative approach to understand abiotic 

stress acclimation responses it is important to find experimental systems where we can begin to filter the 

large number of changes in gene expression, protein and metabolite abundance that occur in response to 

the imposed stress, but are not acclimative, in order to determine those, that are a direct response leading 

to acclimation. 

GROWTH AND METABOLIC ACCLIMATION OF BETULA PENDULA TO DIFFERENT NIGHT- 

TIME TEMPERATURES 

Sari Kontunen-Soppela, Maarit Mäenpää, Vladimir Ossipov, Markku Keinänen, Matti Rousi, Elina Oksanen 


E-mail: sari.kontunen-soppela@uef.fi 

The climatic change with increasing average air temperature, but without a change in photoperiod, poses 

plants to a new combination of light and temperature. Night temperatures in northern latitudes are 

expected to increase more than the day temperatures. 

Three genotypes of silver birch (B. pendula) were exposed to same daytime temperature (22°C), but to five 

different night-time temperatures (6, 10, 14, 18, 22°C) in growth chambers. Photoperiod was 14h full light 

with gradually increasing morning (2h) and decreasing evening (2h) light. Gas exchange, growth and the 

concentrations of ca. 280 metabolites were measured. 

Warmer night temperatures increased dark-respiration, but no effects were seen in daytime CO2 

assimilation. In contrast, stomatal conductance increased at 22°C during daytime, but no changes were 

observed at night. The height growth of birches was greater at warmer temperatures; total biomass was, 

however, not affected. Genotype was the strongest factor for determining metabolic composition of the 

plants. Genotypes had both unique and shared metabolic responses to night temperatures. The responses 

common for all genotypes were connected to growth and the developmental state of plants. Changes in 

growth and metabolite concentrations were non-linear along the temperature gradient and the sensitivity 

to react and the critical temperatures were genotype-specific. 

TRANSCRIPTIONAL REGULATION OF THE COLD-INDUCIBLE DREB1 GENES IN 

ARABIDOPSIS 

Satoshi Kidokoro, Kyonoshin Maruyama, Nobutaka Mitsuda, Masaru Ohme-Takagi, Kazuo Shinozaki, 

Kazuko Yamaguchi-Shinozaki 

Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan 

E-mail: akido@mail.ecc.u-tokyo.ac.jp 

Arabidopsis transcription factors DREB1s/CBFs specifically interact with a cis-acting element DRE/CRT/LTRE 

involved in the expression of low-temperature-, drought- and high-salinity-inducible genes. Expression of 

the DREB1 genes is kept at very low level and regulated by circadian control, but induced rapidly and 

significantly by low temperature. In this study, we analyzed the molecular mechanisms of DREB1 gene 

expression under both low temperature and circadian control conditions. 

Using a GUS reporter gene fused to a DREB1C promoter, we identified a region including cis-acting elements 

involved in regulation of gene expression under low temperature and circadian control conditions. We 

isolated cDNAs encoding the transcriptional factors, PIF7 and CAMTA2, which specifically bind to the region 

80 



by using the yeast one-hybrid screening. We showed that PIF7 functions as a transcriptional repressor for 

DREB1 expression under circadian control. In contrast, CAMTA proteins activate expression of the GUS 

reporter gene driven by the DREB1 promoters in transient experiments using Arabidopsis protoplasts. We 

are analyzing interaction between PIF7 and CAMTA proteins by using the BiFC assay. 

THE TISSUE-SPECIFIC GLYCINE BETAINE ACCUMULATION IN BARLEY PLANTS - HOW DO 

BARLEY PLANTS UTILIZE THE GLYCINE BETAINE? 

Takashi Fujiwara, Shiro Mitsuya, Tasuku Hattori, Tetsuko Takabe 

Nagoya University, Chikusa-ku, Nagoya, Japan 

E-mail: takafuji@nuagr1.agr.nagoya-u.ac.jp 

The accumulation of glycine betaine (GB) is one of the adaptive strategies to adverse salt stress conditions. 

It has been demonstrated that some plants accumulate GB in response to salt stress and various analyses of 

enzymes that catalyze the GB synthesis were performed. However, it remains unknown where the plants 

accumulate GB at plant level. Thus, it would be difficult to clarify the role of GB in plants. 

Since GB is not degraded in plants, accumulation patterns of GB at plant levels are regulated by the 

synthesis and transport. In this report, we have investigated the distribution of GB at organ level, tissue 

localization of BADH protein that catalyzes the last step in the GB synthesis and that of HvProT2 mRNA, 

betaine transporter in barley plants. GB was increased by salt treatment and accumulated more in young 

leaves than in older leaves. BADH proteins were expressed around the xylem vessels of leaves, and in the 

pericycle cells of roots. Moreover, HvProT2 mRNA was expressed in the mestome sheath and lateral root 

cap cells. These results indicate that barley plants accumulate GB at vascular tissues under salt stress 

conditions. We discuss the possible function of GB in barley plants. 

EVALUATION OF SUGARBEET MONOGERM O-TYPE LINES FOR SALINITY TOLERANCE AT 

VEGETATIVE STAGE 

Zahra Abbasi, Ahmad Arzani and Mohammad Mehdi Majidi 


E-mail: z.abbasi@ag.iut.ac.ir 

Sugar beet is a salt tolerant crop species that is cultivated in the Iranian areas where salinity can be a 

serious threat. Screening of genotypes for salinity tolearance is difficult in field due to heterogeneity of 

physical and chemical properties of soil. To evaluate the salinity tolerance of 21 sugarbeet monogerm Otypes 

lines a greenhouse experiment was conducted using a split plot design with 3 replications. Two levels 

of control (2 dS m-1) and salinity (16 dS m-1) were used as main plots and 21 lines were allocated to subplots. 

In each plastic pot (18cm diameter) 24 monogerm seeds were sown in perlit bed and a drop irrigation 

system was implemented. Germination percentage, germination rate, seedling survival percentage, 

chlorophyl content, dry weight of shoots and roots after 8 weeks of the treatments were evaluated.The 

results of analysis of variance showed the significant effects of salinity on the tested traits. Sugar beet 

genotypes differed significantly (P


Arabidopsis RNA polymerase II (RNAPII) C-terminal domain (CTD) phosphatases regulate stress-responsive 

gene expression and plant development via the dephosphorylation of serine (Ser) residues of the CTD. 

Some of these phosphatases (CTD phosphatase-like 1 (CPL1) to CPL3) negatively regulate ABA and stress 

responses. 

We isolated AtCPL5, a cDNA encoding a protein containing two CTD phosphatase domains (CPDs). To 

characterize AtCPL5, we analyzed the gene expression patterns and subcellular protein localization, 

investigated various phenotypes of AtCPL5-overexpressors and knockout mutants involved in ABA and 

drought responses, performed microarray and RNA hybridization analyses using AtCPL5-overexpressors, 

and assessed the CTD phosphatase activities of the purified AtCPL5 and each CPD of the protein. 

Transcripts of the nucleus-localized AtCPL5 were induced by ABA and drought. AtCPL5-overexpressors 

exhibited ABA-hypersensitive phenotypes (increased inhibition of seed germination, seedling growth, and 

stomatal aperture), lower transpiration rates upon dehydration, and enhanced drought tolerance, while the 

knockout mutants showed weak ABA hyposensitivity. AtCPL5 overexpression changed the expression of 

numerous genes, including those involved in ABA-mediated responses. In contrast to Ser-5-specific 

phosphatase activity of the negative stress response regulators, purified AtCPL5 and each CPD of the 

protein specifically dephosphorylated Ser-2 in RNAPII CTD. We conclude that AtCPL5 is a unique CPL family 

protein that positively regulates ABA-mediated development and drought responses in Arabidopsis. 

CALLUS INDUCTION AND FATTY ACID PROFILES OF JATROPHA CURCAS INDUCED CALLI 

ChongSiang Tee 1 , ThenSoong Siow 1 , Adelin Ting Su Yien 2 

1 Department of Biological Science, Faculty of Science, Universiti Tunku Abdul Rahman, Jalan Universiti, Bandar Barat, Kampar, Perak, Malaysia 

2 School of Science, Monash University, Jalan Lagoon Selatan, Bandar Sunway, Selangor Darul Ehsan, Malaysia. 

E-mail: teecs@utar.edu.my 

Jatropha curcas, a biodiesel production plant, is intensively cultivated in many countries. The main 

objectives of this study were to induce callus from various types of J. curcas explants and study the fatty 

acid content of the induced calli. In this study, different types of auxins at various concentrations were used 

to induce callus. In addition, fatty acid compositions and total oil content in the induced calli were also 

investigated. It was observed that 4 µM of picloram, 4 µM of dicamba and 2 µM of 2,4-D, cotyledon and 

petioles explants, respectively. For leaf explants, 2,4-D was used in combination with cytokinin to induce 

callus. Besides, embryogenic calli induced from cotyledon explants using MS medium containing 4 µM 

dicamba. The fatty acid analysis revealed that fatty acid compositions in the induced calli were similar to 

that of seed kernels particularly the fatty acid profile of embryogenic calli. Generally, induced calli had lower 

oil content than seed kernels. Among all types of induced calli studied, embryogenic calli had higher total oil 

content than non- embryogenic calli but significantly lower than the seed kernels. 

NEW PERSPECTIVE IN PLANT VITAMIN D PRODUCTION 

Daniele Silvestro 1 , Christina Fredslund 1 , Rie Bak Jäpelt 2 , Poul Erik Jensen 1 

1 Department of Plant Biology and Biotechnology, Faculty of Life Sciences, University of Copenhagen, Copenhagen, Denmark 

2 Division of Food Chemistry, National Food Institute, Technical University of Denmark, Kongens Lyngby, Denmark 

E-mail: dasi@life.ku.dk 

The presence of vitamin D compounds has been established in a few plant species and could represent an 

alternative source of vitamin D. Interestingly, the metabolic pathway leading to the formation of vitamin D 

in plants is basically unknown. The enzyme Δ5,7-sterol-Δ7-reductase (7DHCR) is known to catalyze the 

conversion of 7DHC into cholesterol in vertebrates while in plants a similar enzyme (DWARF5) converts the 

Δ5,7-sitosterol (7DHS) into sitosterol. The structure similarity should make possible the conversion of 7DHS 

into vitamin D (vitamin D5) by exposure to UV B light. 

To verify if this conversion is occurring in plants A. thaliana dwarf5 mutants have been exposed to UV B 

light. This mutant is accumulating Δ5,7-sterols (mainly 7DHS) due to a mutation in the Δ7-reductase and it 

displays a dwarf phenotype due to the down regulation of the brassinosteroid biosynthesis. 

Moreover two cDNA sequences highly homologous to A. thaliana DWARF5 were isolated from S. 

lycopersicum. The activity of the codified enzymes was assessed by enzymatic assay via expression studies 

in yeast and plant. The results on yeast made by GC-MS sterol characterization of the transformants 

82 



showed the accumulation of new compounds due to the Δ5,7-sterolΔ7-reductase activity on ergosterol and 

its Δ5,7-sterol precursor. 

TEMPERATURE DEPENDENCE OF RUTIN-FE(II) BINDING CONSTANTS DETERMINED BY 

ISOTHERMAL TITRATION CALORIMETRY 

Ingunn W. Jolma, Cathrine Lillo, Peter Ruoff 

Center for Organelle Research, University of Stavanger, Stavanger, Norway 

E-mail: ingunn.w.jolma@uis.no 

Flavonoids are an important class of plant secondary metabolites, which not only set colors on fruits and 

vegetables, but also play important roles as antioxidants and are considered to promote human health. 

Flavonoid glycosides, such as rutin (quercetin-3-rutinoside) can also bind cations, such as Fe(II), effectively 

reducing the Fenton reaction which would otherwise form reactive oxygen species (ROS). This ability to 

bind iron ions might also play an important role in modulating iron homeostasis. 

The binding between quercetins and a variety of cations, especially Fe(II) was the subject of several studies 

indicating that the stability constant was quite large. Using isothermal titration calorimetry (ITC), we 

investigated the stability constant between rutin and Fe(II) at pH 7.0 at various temperatures in the range 

between 20-40 degrees Celsius. The results indicate that Fe(II) strongly binds rutin, and that the binding 

constant decreases with increasing temperature. Stoichiometries indicate that Fe(II) has the possibility to 

bind up to two molecules of rutin, which is favoured at low temperatures, while at higher temperatures the 

equilibrium is shifted in direction to a 1:1 Fe(II)-rutin complex. The ITC experiments showed that the 

formation of the Fe(II)-rutin complexes are associated with large exothermic enthalpies. 

IMPACT OF GLUCOSE 1-PHOSPHATE POOLS ON STARCH METABOLISM IN HIGHER 

PLANTS 

Joerg Fettke 

University of Potsdam, Potsdam, Germany 

E-mail: fettke@uni-potsdam.de 

For several glucosyl transfer reactions glucose 1-phosphate is an essential metabolite that acts either 

immediately as glucosyl donor or as a substrate for the formation of the more general donors, ADPglucose 

and UDPglucose. New data revealed that a glucose 1-phosphate transport over both the plasma- and 

plastidial membrane is possible [1,2]. The glucose 1-phosphate uptake by the cells and the import into intact 

plastids is highly specific for the anomeric position of the phosphate ester as glucose 6-phosphate does not 

substitute the carbon 1 ester. Glucose 1-phosphate is taken up by both autotrophic and heterotrophic cells 

(such as mesophyll protoplasts and potato tuber parenchyma cells). Following uptake, glucose 1-phosphate 

is in part metabolized in the cytosol [3] but the majority of the glucose 1-phosphate is imported into the 

plastids and subsequently enters the plastidial path(s) of starch biosynthesis. The import into plastids as 

well as the conversion of glucose 1-phosphate to starch has been characterized by both in situ and in vitro 

experiments. 

References 

[1]Fettke et al. New Phytol. 2010 

[2]Fettke et al. Plant Physiol. 2011 

[3]Fettke et al. Plant Physiol. 2008 

EFFECT OF LIGHT QUALITY ON FLAVONOID BIOSYNTHESIS IN BILBERRY (VACCINIUM 

MYRTILLUS) 

Laura Zoratti, Marko Suokas, Marian Sarala, Hely Häggman, Laura Jaakola 

Department of Biology, University of Oulu, Oulu, Finland 

E-mail: Laura.Zoratti@oulu.fi 

83 



Flavonoids are plant secondary metabolites well known for their high antioxidant activity, which is an 

important property for plants´ interaction and adaptation with the surrounding environment. As well, they 

contribute to the organoleptic quality of plant products, like fruits, and are of major interest in human 

nutrition and health. 

Bilberry is a characteristic berry species of boreal forests and naturally contains high amounts of flavonoids, 

in particular anthocyanins. In bilberry, the synthesis and accumulation of these compounds is genetically, 

developmentally, and also environmentally regulated. 

Generally, our bilberry research is focused on regulation of flavonoid biosynthesis related to adaptation. In 

particular, we are studying the role of light quality on the production of flavonoids. Results show 

considerable differences in the expression of structural and regulatory flavonoid biosynthesis genes, as well 

as the photoreceptor genes PHYTOCHROME B and CRYPTOCHROME, in response to different light 

treatments. 

CHANGING THE BIOMASS COMPOSITION OF BRACHYPODIUM DISTACHYON -MORE 

FRUCTANS FOR A MORE EFFICIENT BIOMASS CONVERSION 

Maria Lundmark, Helle Kildal Mogensen, Tom Hamborg Nielsen 

Department of Plant Biology and Biotechnology, University of Copenhagen, Copenhagen, Denmark 

E-mail: mlun@life.ku.dk 

Fructans are non-structural polysaccharides consisting of either β -2,1- or β -2,6-linked linear and branched 

fructose chains that act as a carbohydrate reserves as an alternative or in addition to starch. Fructans are 

commonly found in many of the important biofuel crops such as Switchgrass, Perennial ryegrass and 

Miscanthus. 

Plant biomass is the largest available resource for the production of renewable liquid fuels. However, the 

conversion of lignocelluloic biomass into biofuel is difficult due to the recalcitrance of these structures and 

the efficiency in terms of yield and cost is still too poor for it to be feasible at a commercial level. 

We intend to shift the carbon flux of the photosynthetic cells of Brachypodium distachyon towards more 

soluble carbohydrates by overexpressing the enzymes of the fructan biosynthetic pathway. The shift in 

biomass composition will allow for more efficient degradation. 

We have identified three putative fructosyl transferases from Brachypodium. These have been cloned and 

we are waiting for the first generation of the transgenic plants. 

Alongside, we have also cloned genes encoding fructan exohydrolases in Brachypodium and an 

endohydrolase from Bacillus subtilis, in order to obtain new enzymes suitable for degrading fructans stored 

within the leaves. 

CLONING, CHARACTERIZATION AND EXPRESSION PROFILES OF TWO UDP- 

GLUCOSYLTRANSFERASES, UGT85K4 AND UGT85K5, RESPONSIBLE FOR THE LAST STEP 

IN THE CYANOGENIC GLUCOSIDE BIOSYNTHETIC PATHWAY IN CASSAVA 

Rubini Kannangara 1 , Mohammed Saddik Motawia 1 , Natascha Kristine Krahl Hansen 1 , Suzanne Michelle 

Paquette 2 , Birger Lindberg Møller 1 , Kirsten Jørgensen 1 

1 

Department of Plant Biology and Biotechnology, Villum Foundation Research Centre “Pro-Active Plants”, UNIK Center for Synthetic Biology, 

University of Copenhagen, Copenhagen, Denmark. 

2 

Department of Biological Structure, University of Washington, Seattle, WA, USA 

E-mail: natascha@life.ku.dk 

Cyanogenic glucosides are amino acid-derived bioactive natural products biosynthesized by two 

cytochrome P450 enzymes and an UDP-glucosyl transferase. Cassava (Manihot esculenta) biosynthesizes 

the two cyanogenic glucosides, linamarin and lotaustralin, derived from L-valine and L-isoleucine 

respectively. In this study cDNAs encoding two UGT paralogs (assigned UGT85K4 and UGT85K5) have been 

identified and isolated from cassava. The two paralogs show 96 % identity and belong to a family containing 

UGTs involved in cyanogenic glucoside biosynthesis in Prunus dulcis and Sorghum bicolor. The two UGT 

paralogs were recombinantly expressed in Escherichia coli and biochemically characterized. UGT85K4 and 

84 



UGT85K5 were able to glucosylate acetone cyanohydrin and 2-hydroxy-2methylbutyronitrile forming 

linamarin and lotaustralin, and furthermore displays broad in vitro substrate specificity as shown by their 

ability to glucosylate other hydroxynitriles, some flavonoids and simple alcohols. Immunolocalization 

studies of UGT85K4 and UGT85K5 indicated co-occurrence with the enzymes catalyzing the first two steps 

in the cyanogenic glucoside biosynthesis, CYP71E7 paralogs and CYP79D1/D2, in mesophyll and xylem 

parenchyma cells in the first unfolded leaf of cassava. Furthermore In situ PCR showed that UGT85K4 and 

UGT85K5 are co-expressed with CYP79D1 and both CYP71E7 paralogs in the cortex, phloem and xylem 

parenchyma and in specific cells in the endodermis of the petiole of the first unfolded leaf. 

PINOSYLVIN BIOSYNTHESIS IN SCOTS PINE (PINUS SYLVESTRIS) 

Tanja Paasela, Kean-Jin Lim, Teemu H. Teeri 


E-mail: tanja.paasela@helsinki.fi 

The stilbene pinosylvin is a phenolic compound that is synthesised in Scots pine under both developmental 

and stress induced control. In heartwood of adult trees pinosylvin concentration correlates with decay 

resistance; trees that have the highest amount of stilbenes in their heartwood are the most resistant ones 

[1]. Our aim is to find out what regulates pinosylvin formation in adult trees when heartwood is developing 

and in young seedlings in stress conditions. Do these two processes have common genetic determinants, 

i.e. are they regulated by converging signal transduction pathways? Finding genetic causes for the variation 

in heartwood stilbenes would make early selection of seedlings with DNA markers possible. 

UV-C light activates transcription of pinosylvin synthase gene (PST-1) within hours after treatment in sixweek-old 

seedlings. Protein translation inhibitor cycloheximide doesn't have an effect on transcriptional 

activation of PST-1 under UV stress which infers that specific transcription factors are already present in 

uninduced tissues. Wounding activates PST-1 transcription in xylem of five-year-old seedlings. Transcription 

is perceivable 24 hours after wounding. Additionally, osmotic stress induces transcription of PST-1 in 

seedlings. 

References 

[1] Venäläinen et al., Ann.Forest Sci. 2003 

METABOLOMICS REVEALS A SIGNIFICANT METABOLIC SHIFT IN ARABIDOPSIS PLANTS 

GROWN IN THE FIELD AND CLIMATE CHAMBERS 

Yogesh Mishra 1,2 , Hanna Johansson Jänkänpää 1 , Christiane Funk 2 , Wolfgang Peter Schröder 1,2 , Stefan 

Jansson 1 

1 Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, Umeå, Sweden 

2 Umeå Plant Science Centre, Department of Chemistry, Umeå University, Umeå, Sweden 

Email: stefan.jansson@plantphys.umu.se, yogesh.mishra@plantphys.umu.se 

Plants are most consummate and sophisticated system in world, with tremendous capacity to adjust them 

self against the natural variation, called as phenotypic plasticity. Therefore, it is utmost import to 

understand their adaptation capacity in natural condition (where they actually developed) rather in growth 

chamber. In previous study (Mishra et al, unpublished) we find that Arabidopsis growing in field are 

significantly differ from growth chamber in terms of leaf morphology and photo system components. The 

metabolome provides a tool for understanding the function of gene even if that gene has minimal or no 

effect on phenotype. Therefore, to achieve exclusive insights in above differences into the metabolic level, 

we compare the metabolite profiles of leaves of wild type Arabidopsis thaliana (Col) growing under 

constant laboratory conditions and field. Sugar and sugar derivatives fructose, sucrose, glucose, galactose 

and rafinose registered ten fold accumulations in climate chamber over filed grown plants. However, the 

levels of aminoacids display large increase in field grown plants. Several intermediates of TCA cycle 

including succinate, fumarate and malate three fold higher in growth chamber compared to field grown 

plants. The above changes in metabolites suggest a metabolic shift has occurred. This finding provides new 

insight into the mechanism of plant adaptation at metabolomic level, and highlights the role of known 

protectants under natural conditions. 

85 



STEM HYDRAULIC CONDUCTANCE IS INFLUENCED BY LIGHT - EXPERIMENTAL EVIDENCE 

FROM SILVER BIRCH (BETULA PENDULA) 

Eele Õunapuu, Arne Sellin 

Department of Botany, Institute of Ecology and Earth Sciences, University of Tartu, Tartu, Estonia 

E-mail: ounapuu@ut.ee 

Stem hydraulic conductance (Kstem) has previously been shown to depend mainly on the anatomical 

features of xylem conduits and the possibility of rapid flow control in xylem has not been considered. Here 

we report evidence for short-term changes in Kstem for silver birch (Betula pendula) related to light 

conditions. First we sampled shoots cut from lower (shade shoots) and upper (sun shoots) thirds of 

naturally growing ~25-year-old forest trees. Before hydraulic measurements the shoots were exposed to 

photosynthetic photon flux density (PPFD) of 70, 140, 330 or 610 µmol m -2 s -1 for 7 h. Both canopy position 

(long-term effect) and incident PPFD (short-term effect) had a significant impact on Kstem. Sun shoots 

exhibited consistently higher Kstem compared with shade shoots. For both canopy positions maximum 

values of Kstem were recorded at PPFD of 330 µmol m -2 s -1 . In a second experiment conducted in 4-year-old 

saplings growing in an experimental plantation, Kstem as well as potassium ion concentration ([K + ]) in 

xylem sap varied considerably along the canopy vertical profile, both increasing from bottom to top in 

accord with the light availability gradient in the canopy; there existed a strong relationship between Kstem 

and [K + ]. These results suggest that Kstem is dynamic on a short time scale and that potassium is involved 

in the regulation of Kstem in relation to light availability. 

FRUCTAN REGULATION IN TIMOTHY (PHLEUM PRATENSE L.) AS AFFECTED BY STAGES 

OF DEVELOPMENT AND BY WILTING 

M. Ould Baba Ahmed 1,2 , A. Morvan-Bertrand 1 , Ml. Decau 1 , C. Lafreniere 2 , Mp Prud'homme 1 , P. Drouin 2 

1 Ecophysiologie Végétale, Agronomie et nutritions NCS, Esplanade de la Paix, Université de Caen, Caen cedex, France 

2 Unité de Recherche et de Développement en Agroalimentaire en Abitibi-Témiscamingue, UQAT, Rouyn-Noranda, Québec, Canada 

E-mail: Marouf.Ould-Ahmed@uqat.ca 

In temperate grasses chloroplastic starch accounts for about 15% of non-structural carbohydrate (NSC). The 

predominant NSCs are vacuolar sucrose and fructans. Fructans are known to have a role in physiological 

responses to low temperatures and regrowth after defoliation. Fructan degradation during silage 

contributes to forage preservation and studies suggest that plant fructan exohydrolases (FEHs) are partially 

responsible for that degradation. However, fructan breakdown by plant enzymes in harvested tissues as 

well as the availability of soluble sugars during storage are little documented. Here, FEH activity and NSC 

profiles, including starch, were evaluated during development (vegetative growth, stem elongation, 

heading and anthesis) and during 24 hours of wilting after cutting for the last two stages in leaves and 

stems of timothy. This study shows that whereas starch content decreases, fructan content increases with 

growth being maximal at anthesis in N non-limiting conditions. After cutting, fructans content was relatively 

stable and FEH activity low, possibly due to desiccation. Besides, three putative FEH coding genes were 

isolated from cDNA library on these same tissues. Expression analysis of the putative FEH transcript levels 

and functional characterization by heterologous expression in Pichia pastoris of the corresponding genes 

are in progress to complete this study. 

MICROTUBULE (+)-END-ASSOCIATED PROTEIN FROM SOLANACEAE INTERACTING WITH 

POTYVIRAL HELPER COMPONENT PROTEINASE 

Tuuli H. Haikonen, Minna-LiisaRajamäki, Jari P. T. Valkonen 


E-mail: Tuuli.Haikonen@helsinki.fi 

86 



Microtubule (+)-end-associated protein SPR2/TOR1 from Arabidopsis is necessary for cortical microtubule 

array dynamics in cell elongation. Potato homolog of SPR2, designated as HIP2, interacts with Helper 

component proteinase of Potato virus A (HCpro of PVA). PVA belongs to the genus Potyvirus (family 

Potyviridae), the largest group of plant viruses. PotyviralHCpro is a multifunctional protein involved in virus 

movement within host and suppression of the fundamental antiviral defence system, RNA silencing. The 

aim of this study is to characterize potato HIP2 and explain function of the interaction between viral HCpro 

and microtubules in infected plants. 

Potato HIP2 complemented twisting phenotype of Arabidopsis spr2-mutant, confirming functional 

homology between HIP2 and SPR2. Transiently expressed HIP2 localized to microtubules in planta. In yeast 

two-hybrid system (YTHS) and bi-molecular fluorescence complementation (BiFC) analyses, HIP2 interacted 

with HCpro in yeast and plant cells, respectively. Importantly, the interaction took place during PVAinfection 

in leaves and aligned to cortical microtubules. HCpro domain required for HIP2-interaction was 

mapped in YTHS. Mutagenesis of the virus at this putative HIP2-interaction domain reduced virus 

accumulation in systemically infected leaves of potato. These data suggest the microtubulus-interaction via 

HIP2 might be beneficial for PVA. 

THE ROLE OF PEROXISOME-TARGETED NDR1/HIN1 LIKE (NHL) PROTEINS IN PATHOGEN 

DEFENCE 

Amr Kataya, Chimuka Mwaanga, Sigrun Reumann 


E-mail: amr.kataya@uis.no 

Peroxisomes are subcellular organelles that are traditionally known to be involved in processes like 

photorespiration, fatty acid β-oxidation and detoxification of reactive oxygen species. However, recent 

evidence indicates an important role of leaf peroxisomes also in defence against pathogens and herbivores. 

By proteome analyses and protein targeting prediction, we identified candidate defense-related proteins 

from the model plant Arabidopsis thaliana that are potentially targeted to peroxisomes. In particular, 

several yet unknown homologs of Arabidopsis NDR1 and tobacco HIN1 have been predicted. The full-length 

Arabidopsis cDNAs were fused to the reporter protein, enhanced yellow fluorescent protein (EYFP), and 

expressed transiently in plant cells. Peroxisome targeting was difficult to show in onion epidermal cells but 

convincingly demonstrated in tobacco protoplasts. The predicted peptides were validated as functional 

peroxisome targeting signals type 1 (PTS1). Real-time PCR has been established to investigate gene 

induction by defence hormones, elicitors, and bacterial pathogens. The NHL genes of interest are induced 

by salicylic acid but not jasmonic acid to different degree. All homologs were induced upon infection by 

virulent Pseudomonas syringaepv. Tomato DC3000, while only one homolog was induced by an avirulent 

strain carrying avrRpt2. The same gene was induced in wild-type plants by the bacterial elicitor, FLG22, but 

remained unaffected in plants carrying mutations in the flagellin receptor gene FLS2, suggesting that the 

defence protein is involved in basal PAMP triggered immunity (PTI). In homozygous knock-out mutants, 

significant differences in bacterial growth rates were observed compared to wild-type plants upon infection 

with Pst DC3000 (avrRpt2). Taken together, the data indicate for the first time that NDR1/HIN1 homologs 

are located in peroxisomes and play important roles in plant innate immunity. 

CHITOSAN AFFECTS GUARD CELL PHOTOSYNTHESIS AND MEMBRANE TRANSPORT 

Attila Ördög 1 , Barnabás Wodala 1 , Éva Hideg 2 , Ferhan Ayaydin 2 , Zsuzsanna Deák 2 , Ferenc Horvath 1 

1 Department of Plant Biology, University of Szeged, Szeged, Hungary 

2 Institute of Plant Biology, Biological Research Center, Hungarian Academy of Sciences, Szeged, Hungary 

E-mail: horvathf@bio.u-szeged.hu 

Guard cells (GC) respond to the presence of microbes by narrowing stomatal pores following perception of 

microbe-associated molecular patterns, such as chitosan (CHT).It has been shown that CHT inhibits the blue 

light-induced stomatal opening and can trigger stomatal closure. These movements are related to the H + - 

ATPase activity in the GC plasma membrane, as it affects the transport of osmotically active solutes, such as 

the passive movement of K + via different sets of potassium channels, the activity of Cl - /H + symporters and 

87 



anion channels. The ATP for proton pumping is supplied mostly from mitochondrial respiration; however, a 

partial inhibition with DCMU implies a role of GC photosynthetic electron transport in the ATP supply. 

In order to investigate whether CHT affects the photosynthetic ATP production of GCs, the lightdependence 

of the photosynthetic electron transport rate of individual GCs was assayed. Vicia faba leaves 

were treated with CHT and chlorophyll fluorescence parameters of GCs were measured by Microscopy-PAM 

chlorophyll fluorometer.In addition, to test the possible effect of CHT on the activity of ion channels, Vicia 

GC protoplasts were investigated by patch clamp technique. 


This work was supported by the Hungarian Scientific Research Fund (Grant. no. OTKA K 81471). 

DEMONSTRATION OF DIFFERENTIALLY REGULATED PROTEINS DURING THE 

INTERACTION BETWEEN PHYTOPHTHORA INFESTANS AND A TRANSGENIC POTATO 

EXPRESSING RPI-BLB1 FROM SOLANUM BULBOCASTANUM 

B. Colignon 1,2 , M. Raes 2 , E. Delaive 2 , M Dieu 2 , B .Watillon 1 , S. Mauro 1 

1 Département Sciences du Vivant, Centre wallon de Recherches agronomiques, Gembloux, Belgium 

2 Unité de Recherche en Biologie Cellulaire, Facultés Universitaires Notre-Dame de la Paix, Namur, Belgium 

E-mail: b.colignon@cra.wallonie.be 

Solanum tuberosum (cv Desirée) is susceptible to the attack of Phythophthora infestans Mont de bary. The 

Rpi-blb1 gene from Solanum Bulbocastanum, encoding a cytoplasmic NBS-LRR protein which recognizes 

Phytophthora effectors in the “gene for gene” model, confers resistance to the pathogen. We made a 

dynamic total leaf proteome comparison of the wild type and transgenic potato cultivars using a 2D DIGE 

strategy coupled to MS identification. Proteomic data revealed 33 and 21 proteins to be differentially 

expressed in transgenic and wild type challenged leaves respectively. 

Using principal component analysis we could clearly separate challenged and healthy leaf tissues. By using 

a systems biology approach, we could map some of the identified proteins into pathways involved in the 

interaction with Phytophthora infestans. 

SIGNIFICANCE OF FIRST BURST OF ROS AND ETHYLENE AS SIGNALING MOLECULES IN 

PATHOGEN-INDUCED THEIR BIPHASIC GENERATION 

Soo Jin Wi, Ky Young Park 

Department of Biology, Sunchon National University, Jeollanam-do, South Korea 


We investigated the relationship between ROS and ethylene in response to biotic stress with the fungal 

pathogen using transgenic tobacco plants, in which ethylene biosynthesis or signaling or ROS production 

was impaired. It was observed that wild-type plants exhibited a gene-specific expression of NtACS 

members of ACC synthase, which were regulated in a time-dependent manner. Pathogen-induced 

expression of NADPH oxidase, RbohD and RbohF, and ROS accumulation at phase I were significantly 

suppressed in transgenic plants, in which ethylene biosynthesis and signaling were impaired. Biphasic 

ethylene production was also inhibited, especially at 1h, in stress-tolerant transgenic plants with impairment 

of RbohD and RbohF expression. The growth of Phytophthora, determined by expression of its 18s rRNA, 

was significantly reduced in transgenic plants with the impairment of ROS generation and ethylene 

biosynthesis and signaling pathway. To determine the physiological function of first burst of ethylene and 

ROS, we generated an RNAi-mediated silencing tobacco line targeted at NtACS4 under the control of the 

DEX-inducible promoter. NtACS4i lines, which were more tolerant compared to wild-type, significantly 

resulted in the blockage of massive production of ROS and ethylene in second phase. We will investigate 

more physiological significance in the interaction between ROS and ethylene in both phases. 

DEVELOPMENT OF A SCREENING METHOD FOR R GENE FUNCTION IN WHEAT 

Remy Kronbak, Chang Yin, Mogens S. Hovmøller, Preben B. Holm, Per L. Gregersen 

88 



Research Centre Flakkebjerg, Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark 

E-mail: Remy.Kronbak@agrsci.dk 

In Arabidopsis specifically mutated resistance genes (R genes) act dominant negatively conferring 

susceptibility, when introduced into plants resistant to the considered pathogen due to the presence of the 

wildtype R gene. Based on this, a strategy to match R genes to specific pathogens has been designed in 

wheat. Proof-of-concept will be established in the wheat cultivar ‘Bobwhite S-26’ using the wheat yellow 

rust system. The yellow rust R gene Yr10 will be transformed into ‘Bobwhite S-26’, and transgenic plants will 

be tested for achieved resistance against an appropriate avirulent P. striiformis isolate harboring the 

AvrYr10 gene. Plants transformed with both the native and the mutated Yr10 forms will prove the concept if 

they show susceptibility in this pathogen test. For the screening strategy, a ‘Bobwhite S-26’-specific library 

of expressed NB-LRR type R genes will be generated from deep transcriptome sequencing. Transgenic 

Bobwhite lines with mutated forms of R genes will be inoculated with selected avirulent isolates of P. 

striiformis. Susceptibility will indicate that a mutated form of the R gene that confers resistance to a 

particular isolate is present in the transgenic plant, and in this way the R gene can be identified and 

exploited in breeding for disease resistance. 

IS JASMONATE-REGULATED DEFENCE EFFICIENT AGAINST BIRD CHERRY-OAT APHID 

(RHOPALOSIPHUM PADI)? 

Sara Merhabi, Inger Åhman, Lisa Beste, Lisbeth Jonsson 

Botany Department, Stockholm University, Stockholm, Sweden 

E-mail: mehrabi@botan.su.se 

The bird cherry -oat aphid (Rhopalosiphum padi) is a cereal pest worldwide. The aphid causes no visible 

symptoms but reduces the yield at heavy infestations and is a vector of plant viruses. An accession of wild 

barley (Hordeum vulgare ssp. spontaneum) exhibited antibiosis against R. padi and was crossed with 

modern barley cultivars. Further breeding has resulted in a number of doubled-haploid breeding lines with 

partial resistance to R. padi. It has been suggested that efficient plant defence against phloem-feeding 

insects is regulated by jasmonate but that the insects are able to suppress or avoid this defence [1]. In 

microarray studies with two R. padi-resistant and two susceptible barley genotypes, methyl jasmonate 

(MeJA)-responsive genes did not appear as a category that was differentially regulated by aphids. Some 

MeJA-responsive gene sequences did however exhibit constitutive strong expression specifically in the two 

aphid-resistant genotypes. This prompted further investigation. We now report the constitutive gene 

expression of a selection of MeJA-regulated genes in a number of barley genotypes with known levels of 

aphid resistance. They represent the parents used in the crosses between wild barley and modern cultivars, 

their offspring and offspring from first and second generation of back-crosses. 

References 

[1] Walling, Plant Physiol. 2008 

UPTAKE AND TRANSLOCATION OF XENOBIOTICS IN CEREALS AND TURNIP RAPESEEDS 

Trine Eggen, Cathrine Lillo 

Bioforsk Vest Særheim, Klepp Stasjon. Norway 

E-mail: Trine.Eggen@bioforsk.no 

Uptake and translocation of xenobiotics from soil to seeds of barley (Hordeum vulgare cv. Edel), wheat 

(Triticumaestivum cv. Bjarne), oat (Avena sativa cv. Berlinda) and turnip rape (Brassica rapa, var. oleifera L. 

cv. Valo, Brassica napus cv. Sheik) were investigated. The studied compounds were emerging organic 

compounds with potential transfer from environment to terrestrial human food chain via application of 

sewage sludge on agricultural soil. The plant uptake study was performed as pot experiment in greenhouse 

with agricultural soil (0.7 % organic carbon) spiked with xenobiotics. High variations of uptake and 

translocation of different xenobiotics, and between plant species were observed; even bioaccumulation in 

seeds were measured. Expressed as a seed concentration factor 

89 


SCF = 


values in the range of 1-18 was measured. 

90 


A B S T R A C T B O O K – L I S T O F P A R T I C I P A N T S 

91 


List of Participants


PARTICIPANTS COUNTRY PAGES 

ADILBAYEVA A Norway 68 

ALAVAREZ DE MORALES P. Spain 66 

AMIR R. Israel 56 

ANDERSEN J. Norway - 

AWAD J. Germany - 

BARÓN M. Spain 51, 74 

BARAH P. Norway 5, 18 

BERGOUGNOUX V. Czech Republic 68, 76 

BONES A.M. Norway 3, 8, 18 

BROUWER B. Sweden 74 

BUENO M.L.P. Spain 8, 51, 74 

BUSHELL M. United Kingdom 3, 4, 10 

CHAURASIA N. India 60 

CHENYI G.G. Finland 54 

CHOWDHARY G. Norway 75 

COLIGNON B. Belgium 88 

DE JAEGER I. Belgium 62 

DEES M.W. Norway - 

DO Y.-Y. Taiwan 15 

DOKANE K. Latvia 58 

DRENGSTIG T. Norway 30 

DROUIN P. Canada 86 

ECKEY C. Germany - 

EGGEN T. Norway 89 

EICHACKER L. Norway 7 

EIDEM L.E. Norway - 

EILERS U. Germany 56 

ELLIS J. USA 7 

ERGONÅ. Norway - 

FAGERSTEDT K. Finland 8, 47, 62 

FETTKE J. Germany 83 

FISCHER K. Norway 3, 6 

FRANKLIN K. United Kingdom 3, 5, 27 

FREYMARK G. The Netherlands - 

FUJIWARA T. Japan 81 

GARGANO D. Norway - 

GRANUM E. Spain 74 

GRIECO M. Finland 60 

HAIKONEN T. Finland 86 

HALL M. Sweden 24 

HANZAWA T. Japan 67 

HEIDARI B. Norway 70 

HEIEN E. Norway 3 

HEMPEL J.J. Norway - 

HENNIG L. Sweden 92 

HIDEMA J. Japan 19, 78, 79 

HOLM M. Sweden 6, 37 

HORVÁTH F. Hungary 87 

92 



HUANG P.-L. Taiwan 4, 15 

HUMPLÍK J. Czech Republic 76 

HURRY V. Sweden 79 

IDÄNHEIMO N. Finland 71 

JARL-SUNESSON C. Sweden - 

JENSEN P.E. Denmark 3, 4, 5, 13, 82 

JOLMA I.W. Norway 30, 83 

JONSSON L. Sweden 7, 89 

KAKKASSERY L. Norway - 

KAMINEK M. Czech Republic 58, 65 

KANAI M. Japan 7, 42 

KANGASJÄRVI S. Finland 8, 39, 50, 59 

KANGASJÄRVI J. Finland 71 

KATAYA A.R.A. Norway 17, 87 

KEEGSTRA K. USA 3, 7, 40 

KESKI-SAARI S. Finland 74, 77 

KIDOKORO S. Japan 42, 80 

KIM M. South Korea 81 

KNOCH E. Denmark 62 

KOIKE T. Japan 77 

KOLBERT Z. Hungary 93 

KONERT G. Finland 6, 39, 50 

KONTUNEN-SOPPELA S. Finland 74, 77, 80 

KRAHL HANSEN N. Denmark 84 

KREMNEV D. Sweden 69 

KRONBAK R. Denmark 88 

KUHN A. Germany 57, 61 

KUNZ S. Sweden 73 

KÄRKÖNEN A. Finland 7, 45, 47, 54, 62 

LEHTIMÄKI N. Finland 60 

LELIVELT C. The Netherlands - 

LILLO C. Norway 70, 83, 89 

LINKIES A. Germany 38, 56 

LINTALA M. Finland 5, 23, 60 

LUNDMARK M. Denmark 79, 84 

LYNGVED R. Norway 72 

MAESHIMA M. Japan 61, 66, 73 

MAIER A. Germany 5, 21 

MAPLE J. Norway 3, 68 

MARTINEC J. Czech Republic 70 

MATSUSHIMA N. Japan 65 

MAURINO V. Germany 21, 57, 61 

MEHRABI S. Sweden 89 

MEIR S. Israel 8, 36, 48 

MISHRA Y. Sweden 60, 85 

MOMMA M. Japan 55, 78 

MORK-JANSSON A.E. Norway - 

MÜLLER JONASSEN E. Norway 70 

MØLLER I.M. Denmark - 

MØLLER S. Norway 2, 3, 4, 6, 68, 70 

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NAM M.H. South Korea 55, 77 

NEMIE-FEYISSA D. Norway 70 

NEZVAL J. Czech Republic 59, 75 

NIELSEN T.H. Denmark 54, 84 

NIWA Y. Japan 6, 29 

NUMATA T. Japan - 

NURMI M. Finland 59 

OKSANEN E. Finland 74, 77, 80 

OUCHI Y. Japan 61 

OULD AHMED M. Canada 86 

ÕUNAPUU E. Estonia 86 

PACHEBAT J. United Kingdom 5, 20 

PALMA J.M. Spain 64, 66 

PALMGREN M. Denmark - 

PARK K.Y. South Korea 4, 14, 71, 77, 88 

PARKER COUPLAND J. Germany 38, 49 

PAUL S. Norway - 

PAWLOWSKI K. Sweden 3, 6 

PEHKONEN T. Finland 45 

PELISSIER COMBESCURE H. Denmark 54 

PHILOSOPH-HADAS S. Israel 6, 36, 48 

PIISILÄ M. Finland 72 

PISTON D. Norway - 

PODLIPNA R. Czech Republic 4, 16 

PAASELA T. Finland 85 

RAHMAN A. Japan 6, 31, 67 

RANTANEN M. Finland 58 

RATKE C. Sweden 6, 35 

RAYNAUD C. France 62 

REUMANN S. Norway 3, 5, 7, 17, 32, 34, 68, 75, 87 

RINNE P. Norway - 

RINTAMÄKI E. Finland 6, 33 

ROCHAIX J.-D. Switzerland 3, 4, 12 

ROSILIO-BRAMI T. Israel 67 

RUOFF P. Norway 4, 6, 8, 30, 83 

SARMIENTO A. France - 

SCHERES B. The Netherlands 3, 5, 25 

SCHINKEL H. Germany - 

SICNER M. Czech Republic - 

SILVESTRO D. Denmark 82 

SIMÓN E. Spain 69, 70 

SINGH I. India 57 

SOLL J. Germany 3, 5, 23, 26 

SONI P. Norway 6, 34 

STACHULA P. Sweden 79 

STECCANELLA V. Denmark - 

ŠTROCH M. Czech Republic 59, 75 

TAKANO N. Japan 79 

TEE C.S. Malaysia 82 

TERANISHI M. Japan 76, 78, 79 

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THOMPSON E. UK 64 

THORSEN K. Norway - 

TILLBERG E. Sweden - 

TILLBERG J.-E. Sweden - 

TYUTEREVA E. Russian Federation 5, 22 

TOMIOKA R. Japan 66 

TROVATO M. Italy 65 

TSUCHIHIRA A. Japan 73 

TUOMINEN H. Sweden 8, 46 

UDDIN M.N. Denmark 79 

VALENTOVA O. Czech Republic 70 

VAN DER SCHOOT C. Norway - 

VAN DER ZEEUW E. The Netherlands - 

VIDAL D. Spain 69 

VILLAR J. Norway - 

VOEGELE A. Germany 6, 38 

VOITSEKHOVSKAJA O. Russian Federation 6, 22, 32 

VOLLSNES A.V. Norway 68 

VON BOTHMER R. Sweden 3, 7, 41 

VON DER LEHR N. Sweden - 

VÄISÄNEN E. Finland 47, 62 

WATANABE M. Japan 77 

WENDELL M. Norway - 

WILLIAMS M. United Kingdom 7 

WONG H.L. Malaysia 8, 52 

YAMAGUCHI-SHINOZAKI K. Japan 42, 80 

YIN L. Norway 63 

ZEEMAN S. Switzerland 3, 6, 28 

ZIMMERMANN T. Germany 7, 43 

ZIZKOVA E. Czech Republic 58 

ZORATTI L. Finland 83 

ÖRDÖG A. Hungary 87 

AASER K. Norway - 

95 


Sponsors 

onsors 

ors 

A B S T R A C T B O O K – S P O N S O R S 

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ABSTRACT BOOK COMPILATION AND DESIGN: 

D M I T R I Y S H E V E L A 

E V E N H E I E N 

K R I S T I N A A S E R

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