Conservation of natural diversity of legume nodulation symbiotic genes using crop wild relatives core collection

1.植物根瘤菌遗传资源的保护：对日本冲绳县石垣岛、东帝汶、老挝和巴布亚新几内亚进行了探索，收集和保存了新的植物根瘤菌遗传资源。根瘤菌多样性分析分析了泰国Vigna寄主植物根瘤菌的多样性。结果表明，与温带和泰国大豆慢生根瘤菌相比，泰国葡萄慢生根瘤菌在Nod基因RFLP谱上具有更高的变异，并产生了多种Nod因子。通过序列分析，将泰国Vigna根瘤菌分为日本慢生根瘤菌（bryyrhizobium japonicum）和贝氏根瘤菌（b.s elkanii）两类。日本石垣岛试验田和自然生境根瘤菌16S rRNA基因测序发现根瘤菌、慢生根瘤菌和中华根瘤菌。从试验田豇豆（Vigna unguiculata）和鸽豆（Cajanus cajan）中分离出根瘤菌菌株，并对逃逸种群进行了研究。从豇豆、鸽豆、豇豆和野生豇豆种及其自然生境中分离得到了一株慢生根瘤菌。从野葡萄种群中分离到一株中华根瘤菌。这是在日本首次分离到的中华根瘤菌菌株。寄主植物多样性分析利用日本莲（Lotus japonicus） NFR1和NFR5基因的序列信息，从豇豆（Vigna angularis）中分离出Nod因子受体（NFR）基因。分离得到的Vigna NFR1-like和Vigna NFR5-like基因与荷花中对应基因的氨基酸序列同源性分别为63%和71%，且在LysM、跨膜和蛋白激酶结构上保持一致。根据azuki nfr样基因的序列信息，从其他Vigna物种中分离出同源基因。通过对不同种类野葡萄的序列比较，发现野葡萄LysM结构域的氨基酸序列存在差异，表明野葡萄属内nfr样基因存在功能分化。少

1.Conservation of plant-rhizobia genetic resourcesIshigaki island, Okinawa prefecture, Japan, East Timor, Laos and Papua New Guinea have been explored and new plant-rhizobia genetic resources have been collected and are conserved.2.Diversity analysis of rhizobiaDiversity of rhizobia strains from Thai Vigna host plants were analyzed. Thai Vigna Bradyrhizobium rhizobia were revealed to have much higher variation in RFLP profiles of Nod gene and produced diverse Nod factors compared with temperate and Thai soybean Bradyrhizobium. Based on sequence analysis, Thai Vigna rhizobia strains were classified into two groups corresponding to Bradyrhizobium japonicum and B.elkanii.16S rRNA gene sequencing of rhizobia strains collected from experimental field and natural habitat on Ishigaki island, Japan. revealed Rhizobium, Bradyrhizobium and Sinorhizobium rhizobia strains. Rhizobium strains were isolated from cowpea (Vigna unguiculata) and pigeon pea (Cajanus cajan) grown in experimental fields an … More d an escaped population of siratro (Macroptilium atropurpureum). Bradyrhizobium strains were isolated from cowpea, pigeon pea, siratro and wild Vigna species in experimental fields and their natural habitat. Sinorhizobium strains were isolated from wild populations of Vigna marina. This is the first record of Sinorhizobium strains isolated in Japan.3.Diversity analysis of host plantsPutative Nod factor, receptor (NFR) genes have been isolated from azuki bean (Vigna angularis) using the sequence information of NFR1 and NFR5 genes of Lotus japonicus. Isolated Vigna NFR1-like and Vigna NFR5-like genes showed 63% and 71% amino acid sequence homology to counter part genes in Lotus and conserved the same structure of LysM, trans membrane and protein kinase. Based on the sequence information of azuki NFR-like genes, homologous genes have been isolated from other Vigna species. Comparison of the sequence of various Vigna species revealed amino acid sequence variation in LysM domain indicating the functional differentiation of NFR-like genes within the genus Vigna. Less