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

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
3.1. Abstract…………………………………………………….…………... 60
3.2. Introduction………………………………………………….………… 61
3.3. Materials and Methods…………………………………….……………
3.3.1. Plant material and treatments Phytophthora infestans strains
63
and inoculation …………………………………………….……… 63
3.3.2. Cloning of DXS cDNA …………………………………………… 64
3.3.3. Southern blotting …………………………………………………. 65
3.3.4. Bioinformatic analyses ……………………………………………… 66
3.3.5. qRT-PCR and data analyses ………………………………………… 66
3.3.6. Statistical analysis………………………………………………… 67
3.4. Results………………………………………………………………….. 68
3.4.1. Inoculation results ………………………………………………… 68
3.4.2. Cloning and sequence analysis …………………………………….. 69
3.4.3. Southern blot …………………………………………………….... 72
3.4.4. StDXS1 is a member of Class I plant DXS ……………………… 73
3.4.5. StDXS1 expression under pathogen inoculation in potato ……... 75
3.4.6. Transcriptional hierarchy of genes in the MEP Pathway ………..... 76
3.5. Discussion……………………………………………………………… 79
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