thesis

Functional analysis of soybean PR-10 genes

Abstract

Soybean (Glycine max) is the most important oilseed source and one of the most important crops worldwide. In 2014, 249 million metric tons were produced across the world. Even though the worldwide production is increasing every year, scientists, farmers and companies are still struggling to match production increases, to increase in world population growth. The challenge is not simply to produce more, but also to produce more on the same area of land. A main limit to increasing crop yields is the variety of diseases, mainly bacterial and fungal. It is estimated that around 15% of the crop production every year is lost due to biological threats. Understanding how soybean responds in defense to pathogens at a molecular level would help produce innovated seeds and plants that can better withstand biological attacks. This research was designed to characterize a soybean gene family that responds to multiple pathogens within a few hours of infection, the PR-10 gene family. We identified six members of the PR-10 gene family based on expression patterns from in-house microarray studies. Gene-specific PCR primers were designed to clone full-length cDNA of selected PR-10 genes. The cDNA were sequence verified and transferred into an Agrobacterium overexpression vector, expression controlled by the CaMV35S constitutive promoter. Four PR-10 family members were transformed into Arabidopsis thaliana, and alterations in defense responses were monitored in the PR-10 transformants. Two RNAi constructs were made for future transformation into soybean, to silence these genes and ascertain their function in soybean defense. Additionally, the response of a PR-10 promoter was assayed by studying GFP expression controlled by a PR-10 promoter versus the constitutive promoter GmUbi. These research results increase our understanding of PR-10 function and verify the effectiveness of PR-10 in defense response to pathogen infection, which could potentially lead to the development of markers that are associated with pathogen resistance, and also provide genetic material for basic research and possible development of transgenics with enhanced resistance

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