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United States Department of Agriculture

Agricultural Research Service

Title: Identification and functional analysis of candidate Sclerotinia defense genes in soybean

Authors
item Calla, Bernarda - UNIV OF ILLINOIS
item Simmonds, Daina - AGRI/AGRI-FOODS CANADA
item Clough, Steven

Submitted to: ARS Sclerotinia Initiative Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: December 31, 2007
Publication Date: January 23, 2008
Citation: Calla, B., Simmonds, D., Clough, S.J. 2008. Identification and functional analysis of candidate Sclerotinia defense genes in soybean [abstract]. ARS Sclerotinia Initiative Annual Meeting. p. 12.

Interpretive Summary: We have conducted a series of microarray studies that enabled us to identify genes that are significantly differentially expressed in soybean plants in response to Sclerotinia sclerotiorum. We are expanding these studies to include effects of oxalic acid, a major virulence factor of S. sclerotiorum. To assist with the identification of key defense genes, we assigned genes into functional categories based on the annotation of their closest sequence match in public databases and we clustered the genes across multiple experiments. Candidate defense genes will be further characterized by quantitative real-time RT-PCR to verify that the correct gene was identified in the microarray experiments. Promising genes will be functionally characterized by obtaining knockouts of these genes in soybean and/or Arabidopsis thaliana. For Arabidopsis we will obtain the T-DNA insertion mutants of the genes that are of interest and that have high sequence identity with a soybean gene. To obtain knockouts in soybean we will use a viral induced gene silencing system and/or generate stable transgenics utilizing RNAi constructs. Additionally, over expression of candidate defense genes will be studied in Arabidopsis and/or soybean. Promising candidate defense genes will be mapped to see if they map to known QTLs related to defense to S. sclerotiorum.

Technical Abstract: We have conducted a series of microarray studies that enabled us to identify genes that are significantly differentially expressed in soybean plants in response to Sclerotinia sclerotiorum. We are expanding these studies to include effects of oxalic acid, a major virulence factor of S. sclerotiorum. To assist with the identification of key defense genes, we assigned genes into functional categories based on the annotation of their closest sequence match in public databases and we clustered the genes across multiple experiments. Candidate defense genes will be further characterized by quantitative real-time RT-PCR to verify that the correct gene was identified in the microarray experiments. Promising genes will be functionally characterized by obtaining knockouts of these genes in soybean and/or Arabidopsis thaliana. For Arabidopsis we will obtain the T-DNA insertion mutants of the genes that are of interest and that have high sequence identity with a soybean gene. To obtain knockouts in soybean we will use a viral induced gene silencing system and/or generate stable transgenics utilizing RNAi constructs. Additionally, overexpression of candidate defense genes will be studied in Arabidopsis and/or soybean. Promising candidate defense genes will be mapped to see if they map to known QTLs related to defense to S. sclerotiorum.

Last Modified: 4/20/2014
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