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United States Department of Agriculture

Agricultural Research Service

Research Project: Development of Protocols for Efficient Soybean and Legume Transformation Using Engineered Viral Vectors

Location: Corn Insects and Crop Genetics Research

Project Number: 5030-21000-062-02
Project Type: Specific Cooperative Agreement

Start Date: Aug 16, 2010
End Date: Sep 30, 2014

Objective:
Develop virus-induced gene silencing (VIGS) constructs to knock out putative disease resistance genes in soybean, in order to test and establish gene function. Also test and develop protocols to precisely modify genes in soybean or other legume species, using a combination of VIGS and zinc finger nuclease (ZFN) technologies.

Approach:
The cooperator will work with the ADODR and soybean researchers in the CICGRU on two tasks that use specialized knowledge and technology related to virus-induced gene silencing (VIGS) and use of viruses for transient gene over expression. First, assemble gene constructs (genes, promoters, and appropriate vector and selection sites) to substantially decrease or knock out gene expression of targeted genes. VIGS technologies used in the cooperator lab enable gene knockdown in transient assays (assays used within a single plant). Second, test and develop protocols to extend VIGS technologies to produce persistent, multi-generation gene knockout. Specifically, develop constructs containing zinc finger nucleases (which can be designed to precisely and permanently modify DNA), and deliver these to plant germ-line cells using modified VIGS vectors. The Soybean Mosaic Virus (SMV) and Bean Pod Mosaic Virus (BPMV) have been modified in the cooperator’s lab to express payload genes in the host plant. SMV additionally has the potential to express the ZFN in the seed and embryo, and the virus can be removed in a subsequent cross. The cooperator will first test for gene expression of a SMV-carried reporter gene in germ line cells and in a second generation in soybean, and then will test for expression and function of BPMV and SMV constructs that carry ZFNs.

Last Modified: 10/30/2014
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