Ds Regional Mutagenesis: A Platform for Defining Gene Function in Maize

PI：Thomas P.Brutnell(博伊斯·汤普森植物研究所)Copi：Erik W.Vollbrecht(爱荷华州立大学)主要合作者：Jonathan P.Duvick(爱荷华州立大学)玉米是美国种植的最重要的粮食、饲料和生物燃料作物。它作为美国和国外的主要谷物的优势是建立在极其广泛的种质基础上和非常成功的育种努力之上的。然而，随着世界人口和能源成本以惊人的速度增长，我们面临着第二次绿色革命的需要。这将包括开发第二代作物和开发具有更强的抗旱性、耐盐性和提高氮素利用效率的新种质。这些努力的核心是需要将基因型与表型联系起来。也就是说，基因变异是如何影响表型变异的？突变分析一直是遗传学的基石，也是将基因变异与表型变异联系起来的重要工具。因此，本项目的目标是开发一种极其强大的反向遗传学资源来确定玉米的基因功能。一个超过1,500个D插入行的序列索引集合将作为该项目的基础。人们将利用D插入紧密连锁位点的倾向来产生功能缺失的基因插入，绘制基因的边界，包括启动子和增强子元件，并以QTL间隔定义基因空间。将开发一种社区驱动的、分级的“DS基因敲除”服务，以扩大DS插入突变的效用，为拥有不同资源的科学家提供服务。候选基因将从社区征集，期望每个目标基因的多个等位基因将从本地插入的DS中恢复出来。还将开发一个基于网络的公共界面，以提供关于集合中可获得的D插入的详细信息，以及用于设计纳入D插入位点偏好的反向遗传屏幕的新工具。这个项目的一个中心目标是通过创新的课程将正在进行的植物科学研究的兴奋带入高中课堂。在三年的补助金中，每年都会在博伊斯·汤普森学院举办课程发展讲习班。CDW的目标是为高中、社区大学和小型大学的生物课程开发基于探究的植物科学活动。教师将获得必要的实验和教学材料的资金，并将与博士后助理和外展协调员合作，在学年内实施这些模块。还将举办讲习班，对研究生、博士后和PI进行基因和分子技术方面的培训，以最好地利用在该项目期间开发的DS系。通过该项目产生的数据、项目更新和遗传物质链接将在http://www.plantgdb.org/prj/AcDsTagging/和http://www.maizegdb.org/上公布

PI: Thomas P. Brutnell (Boyce Thompson Institute for Plant Research)CoPI: Erik W. Vollbrecht (Iowa State University)Key Collaborator: Jonathan P. Duvick (Iowa State University)Maize is the most economically important food, feed and biofuel crop grown in the US. Its predominance as a staple grain for the US and abroad is built on an extremely broad germplasm base and upon highly successful breeding efforts. However, as world population and energy costs increase at an alarming rate, we are faced with the need for a second green revolution. This will include developing second generation crops and developing new germplasm with enhanced resistance to drought, salt tolerance and improved nitrogen use efficiency. At the core of these efforts is a need to link genotype to phenotype. That is, how does genotypic variation contribute to phenotypic variation? Mutational analysis has been the cornerstone of genetics and has been an essential tool to link genotypic variation to phenotypic variation. Thus, the goal of this project is to develop an extremely powerful reverse genetics resource to define gene function in maize. A sequence-indexed collection of over 1,500 Ds insertion lines will serve as a foundation for this project. The tendency of Ds to insert to closely linked sites will be exploited to generate loss-of-function gene insertions, map the boundaries of genes, including promoter and enhancer elements, and define the gene space in QTL intervals. A community-driven, tiered "Ds knockout" service will be developed to extend the utility of Ds insertional mutagenesis to serve scientists with varying resources. Candidate genes will be solicited from the community with the expectation that multiple alleles of each targeted gene will be recovered from local Ds insertions. A web-based public interface will also be developed to provide detailed information on the Ds insertions available in the collection as well as new tools for designing reverse genetic screens that incorporate the insertion site preferences of Ds. A central aim of this project is to bring the excitement of ongoing plant science research into high school classrooms through innovative curricula. For each of the three years of the grant, a Curriculum Development Workshop (CDW) will be held at the Boyce Thompson Institute. The goal of the CDW is to develop inquiry-based plant science activities for high school, community college and small college biology courses. Teachers will receive funding for necessary lab and teaching materials and will work with a postdoctoral associate and an outreach coordinator to implement these modules during the academic year. Workshops will also be held to train graduate students, postdocs and PI in the genetic and molecular techniques required to best exploit the Ds lines developed during this project. Data generated through this project, project updates and links to genetic materials will be posted at http://www.plantgdb.org/prj/AcDsTagging/ and http://www.maizegdb.org/