Regional Mutagenesis Utilizing Activator (Ac) in Maize

利用玉米激活剂 (Ac) 进行区域诱变

• 批准号: 0076892
• 负责人: Thomas Brutnell
• 金额: $ 108.04万
• 依托单位: Boyce Thompson Institute Plant Research
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0076892&HistoricalAwards=false
• 关键词: Regional; Mutagenesis; Utilizing; Activator; Ac

Transposable element Activator (Ac) often moves to linked sites, thus limiting its use in non-targeted mutagenesis programs. To overcome this limitation, Ac will be distributed at approximately 10 cM intervals throughout the maize genome. Each of the approximately 200 non-transgenic 'anchor Ac (aAc) lines' in this near isogenic set will harbor a single active Ac element at a unique chromosomal locus. By utilizing an aAc line(s) that maps within 10 cM of any mapped gene or EST for directed tagging, these lines will provide an extremely powerful tool for gene isolation and functional analysis. In addition to facilitating the cloning of genes, the inbred nature of the population and low copy number of Ac elements in the genome will permit identification of subtle phenotypic changes in agronomically important traits such as plant height and flowering time. An equally important feature of this system is that multiple Ac-induced alleles can be generated from unstable Ac insertions to produce an allelic series. Thus, the generation of these aAc lines will provide an essential complement to the gene tagging programs currently being developed. This program will not only allow for the identification of novel mutant phenotypes, but will also provide a uniform genetic background for examining epistatic relationships and establishing genetic hierarchies. An important consideration in developing this system for general use, will be to determine the frequencies with which genes can be tagged relative to the position of the aAc. To address this issue, approximately 10,000 new transpositions will be selected from aAc's that have been distributed at roughly 10 cM intervals along chromosome 1S. Plants will be self-pollinated to generate segregating F2 populations that will be screened in sand benches and in the field for mutant phenotypes. Data from these studies will be used to develop a generalized strategy for directed and regional mutagenesis programs. An essential component of this program, is the development of the Emerson Summer Genetics Program to be established at the Boyce Thompson Institute and Cornell. This program will provide researchers from the US and abroad with the opportunity to utilize the Ac anchor lines while helping to generate the approximately 10,000 F1 and F2 families described above. Access to field sites is particularly important for researchers who are interested in initiating a maize genetics program and for those without access to field sites at their home institutions. To help bridge the gap between research and education, undergraduate students, high school teachers and a select group of high school seniors will be integrated into this program. Thus, teachers and students will spend up to 3 weeks in the field as a team with senior researchers from the US and abroad. The interactions and discussions that develop in the field and have long been part of the maize genetics tradition will help foster collaborations, increase public understanding of science and promote research in this agronomically important cereal. Deliverables:All maize seed stocks are propagated, genotyped and deposited as generated at:Maize Genetics Cooperation Stock Centerc/o Dr. Marty Sachs ([e-mail])To view a list of Ac stocks currently available see:http://www.agron.missouri.edu/cgi-bin/cprop/stock_list.cgi#TKand for updates on the mapping progress see:http://bti.cornell.edu/Brutnell_lab2/Projects/Tagging/BMGG_pro_tagging.html

转座因子激活子（Ac）经常移动到连接位点，从而限制了其在非靶向诱变程序中的使用。为了克服这一限制，Ac将以大约10 cM的间隔分布在整个玉米基因组中。在该近等基因组中的大约200个非转基因“锚Ac（aAc）系”中的每一个将在独特的染色体基因座处具有单个活性Ac元件。通过利用定位在任何定位基因或EST的10 cM内的aAc系用于定向标记，这些系将为基因分离和功能分析提供极其强大的工具。除了便于基因克隆外，群体的近交性质和基因组中Ac元件的低拷贝数将允许鉴定农艺学上重要的性状如株高和开花时间的细微表型变化。该系统的一个同样重要的特征是，可以从不稳定的Ac插入产生多个Ac诱导的等位基因，以产生等位基因系列。因此，这些aAc系的产生将为目前正在开发的基因标记程序提供必要的补充。该程序不仅可以识别新的突变表型，而且还可以为检查上位关系和建立遗传层次提供统一的遗传背景。在开发该系统用于一般用途时的一个重要考虑是确定相对于aAc的位置可以标记基因的频率。为了解决这个问题，将从沿1 S染色体沿着以大约10 cM间隔分布的aAc中选择大约10，000个新的易位。植物将自花授粉，以产生分离的F2群体，将在砂台和田间筛选突变表型。这些研究的数据将用于开发定向和区域诱变程序的通用策略。该计划的一个重要组成部分是在博伊斯汤普森研究所和康奈尔大学建立艾默生夏季遗传学计划。该计划将为来自美国和国外的研究人员提供利用Ac锚系的机会，同时帮助产生上述约10，000个F1和F2家族。对于那些有兴趣启动玉米遗传学计划的研究人员和那些无法在其家乡机构进入实地研究的研究人员来说，进入实地研究尤为重要。为了帮助弥合研究和教育之间的差距，本科生，高中教师和高中毕业生的选择组将被整合到这个计划。因此，教师和学生将与来自美国和国外的高级研究人员一起在该领域度过长达3周的时间。在该领域发展的互动和讨论，长期以来一直是玉米遗传学传统的一部分，将有助于促进合作，提高公众对科学的理解，并促进这一农业重要谷物的研究。所有的玉米种子库都是繁殖、基因分型和储存的，如产生于：玉米遗传合作库存中心c/o博士Marty Sachs（[e-mail]）要查看目前可用的Ac库存列表，请参见：http://www.agron.missouri.edu/cgi-bin/cprop/stock_list.cgi#TKand有关映射进度的更新http://bti.cornell.edu/Brutnell_lab2/Projects/Tagging/BMGG_pro_tagging.html