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拓展用于印度绿豆(Vigna radiata L. Wilczek)种质多态性研究的微卫星标记库。

Expanding the repertoire of microsatellite markers for polymorphism studies in Indian accessions of mung bean (Vigna radiata L. Wilczek).

作者信息

Shrivastava Divya, Verma Priyanka, Bhatia Sabhyata

机构信息

National Institute of Plant Genome Research, Post Box No. 10531, Aruna Asaf Ali Marg, New Delhi, 110067, India.

出版信息

Mol Biol Rep. 2014 Sep;41(9):5669-80. doi: 10.1007/s11033-014-3436-7. Epub 2014 Jun 10.

Abstract

Limited availability of validated, polymorphic microsatellite markers in mung bean (Vigna radiata), an important food legume of India, has been a major hurdle towards its improvement and higher yield. The present study was undertaken in order to develop a new set of microsatellite markers and utilize them for the analysis of genetic diversity within mung bean accessions from India. A GA/CT enriched library was constructed from V. radiata which resulted in 1,250 putative recombinant clones of which 850 were sequenced. SSR motifs were identified and their flanking sequences were utilized to design 328 SSR primer pairs. Of these, 48 SSR markers were employed for assessing genetic diversity among 76 mung bean accessions from various geographical locations in India. Two hundred and thirty four alleles with an average of 4.85 alleles per locus were detected at 48 loci. The polymorphic information content (PIC) per locus varied from 0.1 to 0.88 (average: 0.49 per locus). The observed and expected heterozygosities ranged from 0.40 to 0.95 and 0.40 to 0.81 respectively. Based on Jaccard's similarity matrix, a dendrogram was constructed using the unweighted pair-group method with arithmetic averages (UPGMA) analysis which revealed that one accession from Bundi, Rajasthan was clustered out separately while remaining accessions were grouped into two major clusters. The markers generated in this study will help in expanding the repertoire of the available SSR markers thereby facilitating analysis of genetic diversity, molecular mapping and ultimately broadening the scope for genetic improvement of this legume.

摘要

绿豆(Vigna radiata)是印度一种重要的食用豆类,经过验证的多态性微卫星标记数量有限,这一直是其改良和提高产量的主要障碍。开展本研究是为了开发一套新的微卫星标记,并利用它们分析来自印度的绿豆种质资源的遗传多样性。从绿豆构建了一个富含GA/CT的文库,得到1250个推定的重组克隆,其中850个进行了测序。鉴定了SSR基序,并利用其侧翼序列设计了328对SSR引物对。其中,48个SSR标记用于评估来自印度不同地理位置的76份绿豆种质资源的遗传多样性。在48个位点检测到234个等位基因,每个位点平均有4.85个等位基因。每个位点的多态信息含量(PIC)从0.1到0.88不等(平均每个位点为0.49)。观察到的杂合度和预期杂合度分别在0.40至0.95和0.40至0.81之间。基于杰卡德相似性矩阵,采用非加权组平均法(UPGMA)分析构建了一个聚类图,结果显示来自拉贾斯坦邦邦迪的一份种质单独聚类,其余种质分为两个主要类群。本研究中产生的标记将有助于扩大可用SSR标记的范围,从而便于分析遗传多样性、进行分子图谱绘制,并最终拓宽这种豆类遗传改良的范围。

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