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Functional Analysis Identifies Glycine Max Genes Involved in Defense to Heterodera GlycinesMatsye, Prachi D 17 August 2013 (has links)
The infection of plants by Heterodera glycines, commonly known as soybean cyst nematode (SCN), is a serious agricultural problem of worldwide extent. Meanwhile, it provides an excellent experimental model to study basic aspects of how cells function, in particular, during biotic challenge. Heterodera glycines challenges plant cells by initiating, developing and sustaining an interaction that results in the formation of a nurse cell from which the nematode derives nourishment. The presented experiments examine (1) how a cell can be de-differentiated and reprogrammed to perform a much different biological role and (2) how a cell’s immune responses can be engaged or suppressed to accomplish that goal. The observation of alpha soluble N-ethylmaleimide-sensitive factor attachment protein (alpha-SNAP) expression, its location within the rhg1 locus and known involvement in the vesicular transport machinery relating to defense made it a strong candidate for further functional analysis. Functional studies demonstrated that overexpression of alpha-SNAP in the susceptible G. max[Williams 82/PI 518671] genotype that lacks its expression results in the partial suppression of H. glycines infection. This indicated that the vesicles could be delivering cargo to the site of infection to engage a defense response. High levels of expression of a cell wall modifying gene called xyloglucan endotransglycosylase also occur during defense. XTHs associate with vesicles, act in the apoplast outside of the cell, and have a well-known function in cell wall restructuring. These observations indicated that alterations in the cell wall composition of nurse cells could be important for the successful defense response. Overexpression of a G. max xyloglucan endotransglycosylase (Gm-XTH) in the susceptible G. max[Williams 82/PI 518671] genotype resulted in a significant negative effect on H. glycines as well as R. reniformis parasitism. The results, including preliminary experiments on components of the vesicle transport system, identify a potent mechanism employed by plants to defend themselves from two types of plant-parasitic nematodes.
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Functional Genomic Studies of Soybean Defenses against Pests and Soybean Meal ImprovementLin, Jingyu (Lynn) 01 December 2011 (has links)
Soybean [Glycine max (L.) Merr.] is an important crop worldwide. It has been widely consumed for protein, oil and other soy products. To develop soybean cultivars with greater resistance against pests and improved meal quality, it is important to elucidate the molecular bases of these traits. This dissertation aims to investigate the biochemical and biological functions of soybean genes from four gene families, which are hypothesized to be associated with soybean defense against pests and soybean meal quality. There are three specific objectives in this dissertation. The first one is to determine the function of components in the salicylic acid (SA) signaling pathway in soybean resistance against soybean cyst nematode (Heterodera glycines, SCN). The second one is to determine whether insect herbivory induce the emission of volatiles from soybean, and if so, how these volatiles are biosynthesized. The third objective is to identify and characterize soybean mannanase genes that can be used for the improvement of soybean meal quality. The soybean genome has been fully sequenced, which provides opportunities for cross-species comparison of gene families of interest and identification of candidate genes in soybean. The cloned cDNAs of putative genes were expressed in Escherichia coli to produce recombinant enzymes. Through biochemical assays, these proteins were proved to be soybean salicylic acid methyltransferase (GmSAMT1), methyl salicylate esterase (GmSABP2-1), α[alpha]-farnesene synthase (GmTPS1) and E-β[beta]-caryophyllene synthase (GmTPS2), and endo-β[beta]-mannanase (GmMAN1). Through a transgenic hairy root system harboring overexpression of GmSAMT1 and GmSABP2-1, both of these two genes were evaluated for their biological function related to resistance against SCN. The results showed that the over-expression of GmSAMT1 and GmSABP2-1 in the susceptible soybean background lead to enhanced resistance against SCN. Among four putative soybean mannanase genes, one gene was cloned and characterized. GmMAN1 showed the endo-β[beta]-mannanase hydrolyse activity and can hydrolyze cell walls isolated from soybean seeds. In summary, using comparative and functional genomics, a number of genes involved in soybean defense and meal quality were isolated and characterized. This study provides novel knowledge and molecular tools for the genetic improvement of soybean for enhanced resistance and improved meal quality.
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