A Novel Approach to Identify Plant Genes Involved in Agrobacterium-mediated Transformation

项目摘要：标题：鉴定参与农杆菌介导的植物转化的植物基因(本建议)首席研究员：Kirankumar Mysore建议：0445799农杆菌是一种土壤细菌，可引起植物的冠瘤病。根癌农杆菌通过将其T-DNA转移到植物细胞核并整合到植物染色体中来遗传转化植物。农杆菌介导的植物转化是获得转基因植株最常用和最受欢迎的方法。尽管农杆菌介导的植物转化技术已经取得了重大进展，但利用农杆菌对玉米和大豆等重要作物进行遗传转化仍然不容易。更好地了解这种独特的跨界DNA在农杆菌和植物之间的转移将有助于提高作物的转化效率，并有助于预防葡萄等重要经济作物的冠瘤病。PI提出了一种新的遗传方法来鉴定农杆菌介导的植物转化过程中涉及的植物基因。这种方法使用一种被称为病毒诱导的基因沉默的技术来瞬时敲除或减少植物基因的表达。与目前用来确定农杆菌介导的转化所涉及的植物因子的方法相比，这种方法非常快速和稳健。这种方法不仅将识别参与农杆菌介导的转化的新的植物基因，而且还将鉴定已被推测参与植物转化的基因。更广泛的影响：拟议研究的结果将有助于提高目前顽抗农杆菌介导的转化的植物的转化效率，并培育出抗冠瘤病的转基因植物。这可能会对美国农业产生重大的经济影响。PI参与了为高中教师提供扩展教育的既定计划，以及促进强化学习和实践经验的大型暑期实习生计划。拟议的项目将大大有助于继续和加强SE方案，并将为一名博士后研究员和两名暑期学生实习生提供高度技术性和多学科方法的培训。

PROJECT ABSTRACT:Title: Identification of plant genes involved in Agrobacterium-mediated plant transformation (this proposal)Principal Investigator: Kirankumar Mysore Proposal: 0445799Agrobacterium tumefaciens is a soil bacterium that can cause crown gall disease in plants. A. tumefaciens genetically transforms plants by transferring its T-DNA into the plant cell nucleus and integrating into the plant chromosome. Agrobacterium-mediated plant transformation is the most common and preferred method to produce transgenic plants. In spite of significant progress in Agrobacterium-mediated plant transformation techniques, it is still not easy to genetically transform several important crops like corn and soybean using A. tumefaciens. A better understanding of this unique inter-kingdom DNA transfer between Agrobacterium and plants will help improve the crop transformation efficiency and to prevent crown gall disease on economically important fruit crops like grapes. The PI has proposed a novel genetic approach to identify the plant genes that are involved during Agrobacterium-mediated plant transformation. This approach uses a technique called virus-induced gene silencing to transiently knockout or to reduce the expression of a plant gene. This approach is very fast and robust compared to currently used approaches to identify plant factors involved in Agrobacterium-mediated transformation. This approach will not only identify novel plant genes involved in Agrobacterium-mediated transformation but will also characterize the genes that have been speculated to be involved in plant transformation. Broader Impacts: Results from the proposed research will help to improve the transformation efficiency of plants that are currently recalcitrant to Agrobacterium-mediated transformation and also to generate transgenic plants that will be resistant to crown gall disease. This can produce a significant economic impact on United States agriculture. The PI participates in established programs that provide extended education to high school teachers and a large summer intern program that promote enhanced learning and hands-on experience. The Proposed project will contribute significantly to continuation and enhancement of the se programs and also will provide training for one postdoctoral fellow and two summer student interns in highly technical and multidisciplinary approaches.