Book of Abstracts - Geyseco

P - Posters
legume grains whose proteins have a low level of methionine.
Therefore, the At D-CGS was seed-specific expressed in soybean
seeds. The level of soluble methionine increased up to seven-fold,
while the methionine that was incorporated into the protein seeds
increased up to 18-fold. The phenotype and germination rate did
not alter in the transgenic seeds compared to the wild-type seeds.
These results have shown new ways of elevating methionine content
in seeds, thus enhancing their nutritional qualities.
P05-024: AN IMPROVED PROTOCOL FOR TRANSFOR-
MATION OF ANTIRRHINUM MAJUS
Manchado Rojo, M.* - Portero Martinez, M.A. - Weiss, J. - Egea-
Cortines, M.
Universidad Politécnica De Cartagena
*Corresponding author e-mail: maria.manchado@upct.es
Genetic transformation is a cornerstone to obtain information
of gene functions. We have developed an improved protocol for
transformation and regeneration of Antirrhinum majus, obtaining
a highly reproducible method that has yielded up to a high efficiency,
close to 10% (the final results will be discussed in the congress).
Several aspects affect transformation efficiency. We tested
two lines, 165E and Vilmorin Nain and two different explants,
leaf discs and hypocotyls from seedling of two and four weeks.
As a proof of concept we transformed A. majus with a pHellsgate12
construct expressing RNAi of the homeotic gene Deficiens.
Two week old hypocotyls explants from the line Vilmorin Nain
had the highest transformed rate. Putative transformants were
tested by PCR using NPTII primers and by their phenotypes.
The resulting plants showed classic phenotypes corresponding to
hypomorphic alleles of Def, which included sepaloid petals and
sterile stamens and fell somewhere in strength between deficiens
chlorantha and deficiens nicotianoides.
P05-025: MAKING A TRANSGENIC RICE WITHOUT SE-
LECTION
Jung, H.* - Bang, S.W. – Kim, Y.S. – Kim, J.K.
School of Biotechnology and Environmental Engineering, Myongji
University
*Corresponding author e-mail: harin0723@gmail.com
Over the past several years, consumer and environmental groups
have expressed concern about the use of antibiotic-and herbicideresistance
genes against ecological and food safety perspective.
Although no scientific basis has been established for these concerns,
generating marker-free plants would certainly contribute
to the public acceptace of transgenic crops. Here we present
a technology that allows us to make transgenic plants without
using selectable markers. This technology relies on an efficient
Agrobacterium-mediated transformation method and PCR-based
selection of transformants. After co-cultivated, transformed cells
were allowed to regenerate on MS medium without any antibiotics
or herbicides. In about 2-3 weeks after regeneration, a leaf
disc of regenerated plants from one callus and was punched and
pooled in one tube. We performed PCR with the leaf discs for
the presence of transgene in the pools. We then select the positive
pools and punched a leaf disc of individual rice plants in the
positive pools. Second PCR from individual plants of the PCRpositive
pools led us to identify transgenic plants. This series of
process was referred to as the Clean T-DNA technology. Such
transgenic rice plants are marker-free and further analyzed after
grown in a paddy field. Thus, our Clean T-DNA technology may
provide marker-free transgenic rice plants that can be used directly
for commercial purpose
P05-026: USSING OF TRANSGENOSIS FOR INDUCTION
OF VIRUS RESISTANCE IN PE
Hanacek, P.¹* - Rohrer, M.¹ - Reinöhl, V.¹ - Prochazka, S.¹ - Safarova,
D.² - Navratil, M.² - Horacek,
J.³ - Svabova, L.³ - Smykal, P.³ - Griga, M.³
¹Mendel University in Brno
²Palacky University Olomouc
³Agritec, Research, Breeding & Services,Ltd.
*Corresponding author e-mail: hanacek@mendelu.cz
Viral diseases are one of the serious problems that can cause
great economical loss during pea cultivation. In the Czech Republic
Pea enation mosaic virus (PEMV) and Pea seed-borne mosaic
virus (PSbMV) were the most frequently detected viruses in
samples collected from inspected pea fields. Integrated plant protection
is a modern trend against pest and diseases, and includes
also creation of resistant varieties. Breeding for resistance could
be used, but the process is lengthy and sometimes the resistance
sources are not available.
This goal can be achieved utilizing biotechnological tools, especially
transgenosis. Here we present results of testing of this
alternative approach – the use of genetic modification to achieve
the induction of resistance to viral diseases by inverted repeats
post-transcriptional gene silencing (IR-PTGS). For both PEMV
and PSbMV we cloned sense and antisense CP cDNA between
the 35S promoter and CAMV terminator that result in hairpin
(hp)RNA. This dsRNA is processed by a dsRNase resulting in
21-23 nt siRNAs (small interfering RNA). Incorporation of the
siRNAs into a nuclease complex that degrades ssRNA in a sequence
specific manner induces viral resistance. Preparation and
testing of plasmid vectors as well as results of testing of transgenic
pea plants will be presented. This research was supported by a
project of the Czech Ministry of Agriculture QI91A229.
P05-027: PHYTOREMEDIATION OF STABLE CESIUM
AND LEAD FROM SOLUTIONS BY CHENOPODIUM
ALBUM
Moogouei, R.¹*- Borghei, M.² - Arjmandi, R.¹ - Vosoughi, M.²
¹Department of Environmental Management, Faculty of Environment
and Energy, Science and Research Branch, Islamic Azad
University, Tehran, Iran
²Sharif University of Technology
*Corresponding author e-mail: r_moogoui@iau-tnb.ac.ir
Chenopodium album plants were tested for their potential to remediate
stable cesium and lead from solutions in 15 d. Hydroponically
grown Plants were exposed to CsCl and Pb(C 2
H 3
O 2
) 2
solutions at three different concentrations (0.6, 2 and 5 mg l -1 ). When
plants were incubated in CsCl solutions 68.08 ± 2.12%, 39.66 ±
3.48% and 56.37 ± 1.90% cesium was found to be remediated
after 15d, respectively. Moreover more than 99% lead was removed
from the Pb(C 2
H 3
O 2
) 2
solution in all three concentrations
after 15d. When both CsCl and Pb(C 2
H 3
O 2
) 2
were supplemented
together to the solution, 14.53 ± 1.62%, 47.25 ± 0.96% and 48.01
± 1.43% cesium and 71.22 ± 0.25%, 94.31 ± 0.24% and 98.40 ±
0.05% lead were removed after 15d. The present study suggests
that hydroponically grown Chenopodium album could be used as
a potential candidate plant for phytoremediation Cesium and lead
from solutions, however plants were found to be more efficient
for the remediation of lead than cesium.
P05-028: THE DEVELOPMENT OF TRANSGENIC FLAX
WITH ENHANCED HEAVY METALS BINDING CAPABI-
LITY
Vrbová, M.¹* - Horacek, J.¹ - Vetrovcova, M.¹ - Macek, T.² - Griga,
M.¹
¹AGRITEC Ltd., Plant Biotechnology Department, Šumperk,
Czech Republic
²Joint Laboratory of IOCB and ICT Prague, Institute of Organic
Chemistry and
Biochemistry CAS, Prague, and Department of Biochemi
*Corresponding author e-mail: vrbova@agritec.cz
Flax is an industrial crop utilized mainly for technical purposes
and for this reason is a good candidate for phytoextraction of
P