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

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DOXP-MEP pathway D-glyceraldehyde 3-phosphate + DXS 1-deoxy-D-xylulose-5- DXR 2-C-methyl-D-erythritol 4- MCT 4-diphosphocytidyl-2-C-methyl-D- CMK 4-diphosphocytidyl-2-C-methyl-D-erythritol 2- MDS 2-C-methyl-D-erythritol 2, 4- HDS 1-hydroxy-2-methyl-2-(E)-butenyl 4- HDR Dimethylallyl-PP (DMAPP) Gibberellins Carotenoids ABA Phylloquinon Plastoquinone Isopentenyl-PP (IPP) Chlorophylls Tocopherols Figure 1.4. Schematic representation of the DOXP-MEP pathway. DXS: 1-deoxy-D- xylulose 5-phosphate synthase, DXR: 1-deoxy-D-xylulose 5-phosphate reductoisomerase, MCT: 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase, CMK: 4-(cytidine 50-diphospho)-2-Cmethyl-D-erythritol kinase, MDS: 2-C-methyl-D- erythritol 2,4-cyclodiphosphate synthase , HDS: (E)-4-hydroxy-3-methylbut-2-enyl diphosphate synthase 19
hemiterpenes C5 (1 isoprene unit), monoterpenes C10 (2 isoprene units), sesquiterpenes C15 (3 isoprene units), diterpenes C20 (4 isoprene units), triterpenes C30 (6 isoprene units), tetraterpenes C40 (8 isoprene units) (Dubey et al. 2003). In the mevalonate (Ac-MVA) pathway cytokinins, sesquiterpenes, triterpenes (sterols) and the polyterpene ubiquinone are synthesized. In the DOXP-MEP pathway the isoprene, monoterpenes, diterpenes (phytol) included the side chain of chlorophyll, tocopherol and gibberellin are synthesized, as well as the tetraterpenes (carotenoids) include the side chain of ABA and the polyterpene plastoquinone (Lichtenthaler 2000; Rodriguez-Concepcion and Boronat 2002; Eisenreich et al. 2004; Phillips et al. 2008). 1.2.6. Defense suppression by pathogens Plants activate defense mechanisms that help prevent invasion and microorganisms spread in response to pathogen attack. Plants produce signals, structural barriers, hypersensitive cell death and inhibitors of pathogen growth. On the other hand, only after overcoming the host defenses, can pathogenic microorganisms benefit from the host nutrients by degradation of host high molecular weight constituents and absorvance of low molecular weight metabolites. For these purposes, pathogens produce enzymes, toxins and suppressors (Agrios 2005). Suppressors are inhibitory molecules that suppress the expression of defense responses in the host plant. They allow some pathogens to interrupt or evade the host defense responses. As a result, the host cells become susceptible. However, experimental validation of this assumption has been scarce (Shiraishi et al. 1994). 20
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: Isopentenyl diphosphate (IPP) is th
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
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
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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
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Daayf F, Platt HW, Peters RD (2000)
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Enyedi AJ, Yalpani N, Silverman P,
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Haas BJ, Kamoun S, Zody MC, Jiang R
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Kamoun S (2005) A catalogue of the
Page 181 and 182:
McGarvey DJ, Croteau R (1995) Terpe
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Pieterse C, de Wit P, Govers F (199
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Sels J, Mathys J, De Coninck BMA, C
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van Loon LC, Rep M, Pieterse CMJ (2
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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
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MALCAYAFPGILNRTAVVSDSSKTAPLFSEWIHGT
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Appendix 3 (Chapter 3) Defender Eco
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