Dissection of Nodule and Lateral Root Development in the Model Legume Lotus japonicus

Gresshoff9808625T-DNA插入突变技术将用于鉴定在根瘤和/或侧根发育中以细胞或组织特异性方式表达并控制根瘤和/或侧根发育的植物基因。使用T-DNA边界附近具有无启动子B-谷氨酸酶(uidA，gus)报告基因的双元载体。通过农杆菌转化方案将产生多达2500个转基因株系，并筛选GUS表达和突变表型。最有希望的标记品系和突变体将通过分离分析和表达模式来表征。将分离出cDNA，并用于确定负责所需标记和突变的基因组序列。还将进行其他类型的表征。将使用模式豆莲日本莲花来研究导致结核形成的遗传和分子机制。所使用的方法将涉及对携带无启动子的uidA示踪基因的T-DNA载体进行插入突变。关于根瘤和侧根发育相关的工作假说也将通过插入突变方法进行研究。该项目的成功将大大增加对豆科植物根瘤形成的深入了解，这是我们所知的生命的核心过程

Gresshoff9808625T-DNA insertional mutagenesis will be used to identify plant genes that are expressed in a cell or tissue-specific manner in nodule and/or lateral root development and that control nodule and/or lateral root development. Binary vectors with a promoterless B-gluronidase (uidA, gus) reporter gene adjacent to the T-DNA border are used. Up to 2,500 transgenic lines will be generated by Agrobacterium transformation protocols and screened for GUS expression and mutant phenotypes. The most promising marker lines and mutants will be characterized by segregation analysis and expression patterns. cDNAs will be isolated and used to determine the genomic sequences responsible for the desired markers and mutations. Other types of characterizations will also be done.Genetic and molecular mechanisms responsible for nodule formation will be investigated using the model legure Lotus japonicus. The approach used will involve using insertional mutagenesis with a T-DNA vector carrying a promoterless uidA tracer gene. The working hypothesis that nodule and lateral root development are related will be investigated also by the insertional mutagenesis approach. The success of this project will add significantly to insight into legume nodule formation, a process central to life as we know it