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Cloning and Functional Validation of Pathogen Responsive Promoter from Rice

KrishiKosh

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Title Cloning and Functional Validation of Pathogen Responsive Promoter from Rice
Ph.D.
 
Creator JOSHITHA VIJAYAN
 
Contributor Sharma, T. R.
 
Subject genes, planting, rice, biological phenomena, enzymes, pathogens, proteins, transcription, diseases, fungi
 
Description Cloning and Functional Validation of Pathogen-Responsive Promoter from Rice
ABSTRACT
Rice blast caused by Magnaporthe oryzae is one of the most destructive diseases causing
extensive yield loss in rice crop throughout the world. The present study deals with
identification of pathogen stress responsive genes in susceptible indica rice cultivar HR
12 challenged with blast pathogen M. oryzae and cloning and characterization of
pathogen responsive promoter of CYP76M7 gene. Transcript profiling using the
Affymetrix 57 K Rice GeneChip revealed a total of 72 differentially expressed genes at
initial interaction stage of 6 hours post inoculation (hpi) with M. oryzae isolate Mo-si-63
which were significantly altered after pathogen infection. Molecular function enrichment
analysis suggested that the differentially regulated genes were mainly related to protein
degradation and modification, cell signalling and biotic stress mechanism. Majority of
the genes of protein degradation and modification, transport, signalling and transcription
factors were repressed at the initial stages of interaction whereas hormonal signalling,
cell wall defense and transcription factors belonging to the WRKY and Trihelix family
genes were up regulated. Further, a comprehensive transcriptional profiling experiment
at four time points viz. 12, 24, 48 and 72 hpi with M. oryzae isolate Mo-ei-11 on rice
cultivar HR 12 revealed differential regulation of 178 to 3862 transcripts at various time
points. GO enrichment analysis revealed that most of the differentially expressed genes
were involved in transcription, protein metabolism, signal transduction, stress, secondary
metabolism and hormone signalling. Our analysis has identified an inventory of genes
with altered expression regulated by M. oryzae infection. The data extend the current
understanding of rice defense mechanism during compatible interaction with pathogen
and serve as a genomic resource for robustly selected, differentially expressed genes for
further investigation. A CYP76M7 gene which showed consistently high upregulation
from 24 hpi after pathogen challenge was also functionally validated. Promoter
sequences spanning -222 bp to -520 bp was found to confer Magnaporthe inducibility to
CYP76M7 promoter. This promoter can be used to develop two component system for
development of rice lines with durable blast resistance to M. oryzae.
 
Date 2016-03-07T15:52:23Z
2016-03-07T15:52:23Z
2013
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/64889
 
Language en_US
 
Format application/pdf
 
Publisher IARI, NATIONAL RESEARCH CENTRE ON PLANT BIOTECHNOLOGY, NEW DELHI