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. |
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Creator |
JOSHITHA VIJAYAN
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Contributor |
Sharma, T. R.
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Subject |
genes, planting, rice, biological phenomena, enzymes, pathogens, proteins, transcription, diseases, fungi
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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. |
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Date |
2016-03-07T15:52:23Z
2016-03-07T15:52:23Z 2013 |
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Type |
Thesis
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Identifier |
http://krishikosh.egranth.ac.in/handle/1/64889
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Language |
en_US
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Format |
application/pdf
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Publisher |
IARI, NATIONAL RESEARCH CENTRE ON PLANT BIOTECHNOLOGY, NEW DELHI
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