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

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1.3 OBJECTIVES My hypothesis is that Phytophthora infestans suppresses defense responses in potato (Solanum tuberosum) by alterating the expression of defense-related genes, accumulation of secondary metabolites and secondary metabolism pathways genes. This hypothesis will be tested by i) developing a new molecular biology approach that will help to identify novel genes in the potato-P.infestans interaction, such as genes potentially controlling pathogenesis or avr genes in P. infestans, as well as those potentially involved in potato resistance or susceptibility to this pathogen, ii) studying the gene expression of the genes identified in this research, in order to identify a possible defense suppression, iii) evaluating the suppression of known potato genes by Phytophthora infestans effectors and iv) assessing the accumulation of potato secondary metabolites. The research accomplished in this thesis is presented in several chapters: Chapter 2: Identification and cloning of differentially expressed genes involved in the interaction between potato and Phytophthora infestans using a subtractive hybridization and cDNA-AFLP combinational approach (Henriquez, M.A. and Daayf F. 2010. Journal of Integrative Plant Biology 52: 453–467) Chapter 3: Cloning and characterization of a novel gene from the plastid-localized 2-c- methyl-d-erythritol 4-phosphate (doxp-mep) pathway in potato infected by phytophthora infestans. (Henriquez, M.A. and Daayf F. 2010. Ms. Ref. No.: Journal of Integrative Plant Biology /2010/021261; 24p. Submitted Sept 29, 2010). 23
Chapter 4: Phytophthora infestans effectors suppress secondary metabolism pathways in potato. Chapter 5: Alterated metabolic profile of secondary metabolites in potato leaves after inoculation with Phytophthora infestans 24
Page 1 and 2: Molecular characterization of potat
Page 3 and 4: ABSTRACT Henriquez Maria Antonia. P
Page 5 and 6: ACKNOWLEDGEMENT I would like to tha
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: (Tian et al. 2005; Tian et al. 2007
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
Page 57 and 58: Figure 2.1. Schematic representatio
Page 59 and 60: Figure 2.2. Example of TDFs in a po
Page 61 and 62: espectively. Transcripts DL32 and D
Page 63 and 64: Table 2.1. Similarities between seq
Page 65 and 66: US8 and US11, respectively. qRT-PCR
Page 67 and 68: and Schmittgen 2001) was used to ca
Page 69 and 70: Only in RB+US8, the DL21 precursor
Page 71 and 72: Constitutive gene expression has be
Page 73 and 74: iosynthesis via the diamino-pimelat
Page 75 and 76: Further validation of the subtracti
Page 77 and 78: CHAPTER 3: CLONING AND CHARACTERIZA
Page 79 and 80: Until recently, it was considered t
Page 81 and 82: 3.3.2. Cloning of DXS cDNA Using th
Page 83 and 84: 3.3.4. Bioinformatic analyses Deduc
Page 85 and 86: Table 3.1. Primers used for the syn
Page 87 and 88: StDXS1 has been submitted to the Ge
Page 89 and 90: Figure 3.2. Alignment of deduced am
Page 91 and 92:
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)
Page 191 and 192:
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
Page 201 and 202:
TCAGTCGCTATTTTAGGTGGCGATGGTAGCGTACT
Page 203 and 204:
MALCAYAFPGILNRTAVVSDSSKTAPLFSEWIHGT
Page 205 and 206:
Appendix 3 (Chapter 3) Defender Eco
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