The National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
BMC Plant Biol. 2014 Mar 17;14:65. doi: 10.1186/1471-2229-14-65.
Grasspea (Lathyrus sativus L., 2n = 14), a member of the family Leguminosae, holds great agronomic potential as grain and forage legume crop in the arid areas for its superb resilience to abiotic stresses such as drought, flood and salinity. The crop could not make much progress through conventional breeding in the past, and there are hardly any detailed molecular biology studies due to paucity of reliable molecular markers representative of the entire genome.
Using the 454 FLX Titanium pyrosequencing technique, 651,827 simple sequence repeat (SSR) loci were identified and 50,144 nonredundant primer pairs were successfully designed, of which 288 were randomly selected for validation among 23 L. sativus and one L. cicera accessions of diverse provenance. 74 were polymorphic, 70 monomorphic, and 144 with no PCR product. The number of observed alleles ranged from two to five, the observed heterozygosity from 0 to 0.9545, and Shannon's information index ranged from 0.1013 to 1.0980, respectively. The dendrogram constructed by using unweighted pair group method with arithmetic mean (UPGMA) based on Nei's genetic distance, showed obvious distinctions and understandable relationships among the 24 accessions.
The large number of SSR primer pairs developed in this study would make a significant contribution to genomics enabled improvement of grasspea.
作为豆科的一员,兵豆(Lathyrus sativus L.,2n=14)是一种在干旱地区具有巨大农业潜力的粮食和饲料作物,因为它对干旱、洪水和盐度等非生物胁迫具有极强的适应能力。过去,该作物通过常规育种进展不大,而且由于缺乏代表整个基因组的可靠分子标记,几乎没有任何详细的分子生物学研究。
利用 454 FLX Titanium 焦磷酸测序技术,鉴定出 651827 个简单重复序列(SSR)位点,并成功设计了 50144 对非冗余引物对,其中 23 个兵豆和 1 个不同起源的野豌豆属(L. cicera)品系随机选择了 288 对进行验证。其中 74 对具有多态性,70 对为单态性,144 对无 PCR 产物。观察到的等位基因数从 2 到 5 不等,观察到的杂合度从 0 到 0.9545,Shannon 信息指数从 0.1013 到 1.0980 不等。基于 Nei 遗传距离的不加权对组平均法(UPGMA)构建的聚类树状图,在 24 个品系之间表现出明显的区别和可理解的关系。
本研究开发的大量 SSR 引物对将对兵豆的基因组学改良做出重要贡献。