Disease cycle of potato late blight - MSpace at the University of ...

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this pathogen. Using the subtractive hybridization (SH)/cDNA-AFLP combinational approach, DOXP-MEP pathway genes were identified for first time in potato and its regulation in response to the oomycete P. infestans. The DOXP-MEP pathway, also called the non-mevalonate route is an alternative terpenoids' biosynthetic route that was discovered first in eubacteria and soon after in photosynthetic organisms such as higher plants, algae as well as in cyanobacteria. In addition, potato plants treated with glucans extracted from P. infestans, the elicitor eicosapentanoic acid (EPA) and P. infestans isolates were analyzed to study the accumulation of phenolic compounds and expression level of genes from the phenylpropanoid, mevalonate and DOXP-MEP pathway. IV
ACKNOWLEDGEMENT I would like to thank my supervisor Dr. Fouad Daayf for giving me the opportunity to work in his laboratory. I am grateful to my committee members Dr. Claudio Stasolla, Dr. Michele Piercey-Normore and Dr. Lakhdar Lamari (rip) for their helpful advice and discussion. I also want to thank the Crop Technology Centre at the University of Manitoba, Drs. G. Li and D. Levin for the use of lab facilities, as well as Drs. G.M. Ballance and M. Tahir for reviewing a manuscript from my thesis. I appreciate all the past and present members at the Crop Technology Centre for creating an enjoyable team environment. My thanks also go to all the members of Plant Science Department who have helped me one way or the other and members from Dr. Daayf’s Lab and my friends in the Animal Science Department. I would like to give my special thanks to Lorne Adam and Carlo Carere for technical assistance. My appreciation is also extended to Dr. Adriana Andreu who supplied research material for my project and Erika Wolski for her collaboration. Financial support from University of Manitoba Graduate Fellowship and a research grant from The Natural Sciences and Engineering Research Council of Canada (NSERC) to F. Daayf are also appreciated. I thank my beloved husband Oscar Molina for his love, collaboration, understanding and support to me. I would not have been able to go this far without him. I wish to express my deep gratitude to my parents, brothers and sisters for their love, support and encouragement. V
Page 1 and 2: Molecular characterization of potat
Page 3: ABSTRACT Henriquez Maria Antonia. P
Page 7 and 8: CHAPTER 2: IDENTIFICATION AND CLONI
Page 9 and 10: CHAPTER 4: PHYTOPHTHORA INFESTANS E
Page 11 and 12: LIST OF TABLES Table 2.1. Similarit
Page 13 and 14: Figure 4.1. Effect of EPA elicitor
Page 15 and 16: SH: subtractive hybridization TDFs:
Page 17 and 18: FORWARD This thesis has been writte
Page 19 and 20: the use of moderately resistant var
Page 21 and 22: subtropical and temperate environme
Page 23 and 24: FALL Disease Disease cycle cycle of
Page 25 and 26: plant intercellular space (apoplast
Page 27 and 28: signaling cascades, alteration in g
Page 29 and 30: The recent research target in late
Page 31 and 32: (lignin), UV protection (quercetin,
Page 33 and 34: 1.2.5.4.2 Terpenoids pathways Terpe
Page 35 and 36: Isopentenyl diphosphate (IPP) is th
Page 37 and 38: hemiterpenes C5 (1 isoprene unit),
Page 39 and 40: (Tian et al. 2005; Tian et al. 2007
Page 41 and 42: Chapter 4: Phytophthora infestans e
Page 43 and 44: slightly up-regulated in response t
Page 45 and 46: The objective of the current study
Page 47 and 48: days growth in Pea Broth Medium at
Page 49 and 50: 2.3.5. SH- Second Strand synthesis
Page 51 and 52: Second Strand for digestion. Howeve
Page 53 and 54: CAGCTACTTGGGAGGCTGAG-3’ and DL81-
Page 55 and 56:
2.4. RESULTS 2.4.1. Inoculation res
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Figure 2.1. Schematic representatio
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Figure 2.2. Example of TDFs in a po
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espectively. Transcripts DL32 and D
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Table 2.1. Similarities between seq
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US8 and US11, respectively. qRT-PCR
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and Schmittgen 2001) was used to ca
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Only in RB+US8, the DL21 precursor
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Constitutive gene expression has be
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iosynthesis via the diamino-pimelat
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Further validation of the subtracti
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CHAPTER 3: CLONING AND CHARACTERIZA
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Until recently, it was considered t
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3.3.2. Cloning of DXS cDNA Using th
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3.3.4. Bioinformatic analyses Deduc
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Table 3.1. Primers used for the syn
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StDXS1 has been submitted to the Ge
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Figure 3.2. Alignment of deduced am
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Figure 3.3. Phylogenetic tree of DX
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StDXS1 transcripts in RB+US8 were i
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Fold induction 7 6 5 4 3 2 1 1 0 2
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tentative consensus-TC of Solanum t
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Hevea brasiliensis (HbDXS) two DXS
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This work opens up new perspectives
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4.2 INTRODUCTION Plant pathogens ha
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catalyze the first steps in the bra
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type A2) (kindly provided by Dr. Ad
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synthesis pool from each treatment,
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using a Stratagene Mx3005p cycler,
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with the glucan fraction and then i
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4.4.2. Analysis of variance Data co
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4.4.3 Gene expression analysis in t
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Relative Expression 1.6 1.4 1.2 1 0
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strains D-03 (lineage US11, A1 mati
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Table 4.6. RT-PCR analysis of the r
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There was a down-regulation of StDX
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Figure 4.6. Transcript profiles of
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cultivar Defender (DF+GL+US8). Expl
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(Ac-MVA) pathway, participate in es
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4.5.2.2 Russet Burbank defense resp
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Russet Burbank. With the exception
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CHAPTER 5: ALTERED METABOLIC PROFIL
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pathogen interactions, the cell wal
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at 0, 12, 72, and 120 hours post in
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5.4 RESULTS 5.4.1 Disease developme
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Finally, an early stage of the inte
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Figure 5.3. Chromatograpic profiles
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µg Catechin /g FW µg Catechin /g
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levels of flavonoid (P3) in RB+US11
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Terpenoid (T1) was detected in Russ
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control plant, would be associated
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Flavonoids possess a wide range of
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explanation of such a specific supp
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diaminopimelate aminotransferase wh
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the DOXP-MEP pathway, we detected c
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• Sesquiterpene cyclase (SC) gene
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good disease control strategy. For
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REFERENCES Abramovitch R, Kim Y, Ch
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Chappell J (1995) Biochemistry and
Page 173 and 174:
Daayf F, Platt HW, Peters RD (2000)
Page 175 and 176:
Enyedi AJ, Yalpani N, Silverman P,
Page 177 and 178:
Haas BJ, Kamoun S, Zody MC, Jiang R
Page 179 and 180:
Kamoun S (2005) A catalogue of the
Page 181 and 182:
McGarvey DJ, Croteau R (1995) Terpe
Page 183 and 184:
Pieterse C, de Wit P, Govers F (199
Page 185 and 186:
Sels J, Mathys J, De Coninck BMA, C
Page 187 and 188:
van Loon LC, Rep M, Pieterse CMJ (2
Page 189 and 190:
Yao KN, Deluca V, Brisson N (1995)
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CGCGGCGCGGCGGCCGGGTCATTTGCCCAGCTTTC
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Sequence GGCATGAAATGCATCGGGTGGAGTAA
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TCCCTTGGGCATGGTCCCCTCAGTTATACTTCGTG
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Sequence CTGAGGCGAGTGGCAGAAACTAATAC
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EST#: DL127 Sequence CCTGGGTGGGGCCG
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TCAGTCGCTATTTTAGGTGGCGATGGTAGCGTACT
Page 203 and 204:
MALCAYAFPGILNRTAVVSDSSKTAPLFSEWIHGT
Page 205 and 206:
Appendix 3 (Chapter 3) Defender Eco
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