Institute for Plant Breeding, Genetics and Genomics/Center for Applied Genetic Technologies, University of Georgia, Athens, Georgia 30602, USA.
Plant Physiol. 2011 Oct;157(2):552-62. doi: 10.1104/pp.111.181206. Epub 2011 Aug 15.
Insertional mutagenesis of legume genomes such as soybean (Glycine max) should aid in identifying genes responsible for key traits such as nitrogen fixation and seed quality. The relatively low throughput of soybean transformation necessitates the use of a transposon-tagging strategy where a single transformation event will produce many mutations over a number of generations. However, existing transposon-tagging tools being used in legumes are of limited utility because of restricted transposition (Ac/Ds: soybean) or the requirement for tissue culture activation (Tnt1: Medicago truncatula). A recently discovered transposable element from rice (Oryza sativa), mPing, and the genes required for its mobilization, were transferred to soybean to determine if it will be an improvement over the other available transposon-tagging tools. Stable transformation events in soybean were tested for mPing transposition. Analysis of mPing excision at early and late embryo developmental stages revealed increased excision during late development in most transgenic lines, suggesting that transposition is developmentally regulated. Transgenic lines that produced heritable mPing insertions were identified, with the plants from the highest activity line producing at least one new insertion per generation. Analysis of the mPing insertion sites in the soybean genome revealed that features displayed in rice were retained including transposition to unlinked sites and a preference for insertion within 2.5 kb of a gene. Taken together these findings indicate that mPing has the characteristics necessary for an effective transposon-tagging resource.
豆科植物(如大豆)基因组的插入突变应该有助于鉴定负责固氮和种子质量等关键性状的基因。大豆转化的相对低通量需要使用转座子标记策略,其中单个转化事件将在多个世代中产生许多突变。然而,由于转座限制(Ac/Ds:大豆)或需要组织培养激活(Tnt1:紫花苜蓿),目前在豆科植物中使用的转座子标记工具的用途有限。最近从水稻(Oryza sativa)中发现的一个可移动元件 mPing 及其移动所需的基因被转移到大豆中,以确定它是否会优于其他可用的转座子标记工具。对大豆中的稳定转化事件进行了 mPing 转座的测试。在早期和晚期胚胎发育阶段对 mPing 切除的分析表明,大多数转基因系在晚期发育过程中切除增加,这表明转座是受发育调控的。鉴定出产生可遗传 mPing 插入的转基因系,其中活性最高的系的植物每代至少产生一个新的插入。对大豆基因组中 mPing 插入位点的分析表明,保留了在水稻中显示的特征,包括转座到非连锁位点以及在基因 2.5kb 内插入的偏好。总之,这些发现表明 mPing 具有作为有效转座子标记资源的必要特征。