异源四倍体毛棉细菌人工染色体文库的构建与鉴定

Construction and characterization of a bacterial artificial chromosome library for the allotetraploid Gossypium tomentosum.

作者信息

Liu F, Wang Y H, Gao H Y, Wang C Y, Zhou Z L, Cai X Y, Wang X X, Zhang Z S, Wang K B

机构信息

State Key Laboratory of Cotton Biology, China/Cotton Research Institute, Chinese Academy of Agricultural Sciences, Anyang, China.

College of Agronomy and Biotechnology, Southwest University, Chongqing, China.

出版信息

Genet Mol Res. 2015 Dec 15;14(4):16975-80. doi: 10.4238/2015.December.15.3.

DOI:10.4238/2015.December.15.3

PMID:26681044

Abstract

Gossypium tomentosum is a wild allotetraploid species with the (AD)5 genome. It is characterized by many useful traits including finer fiber fineness, drought tolerance, and Fusarium and Verticillium resistance. We constructed the first bacterial artificial chromosome library for Gossypium tomentosum. With high quality and broad coverage, this library includes 200,832 clones, with an average insert size of about 122 kb and fewer than 3% empty clones. Our library is approximately 10-fold the size of the (AD)5-genome (2400 Mb) and provides a 99.7% probability of isolating genes of interest or their sequences. Seven of eight simple sequence repeats markers that are located on five different chromosomes and linked with resistance to Verticillium wilt could amplify the 50 superpools and obtained one to five hits. This high capacity library will be an important genomic resource for classifying and analyzing the evolution of allotetraploid cotton species as well as for isolating disease-resistance and drought-tolerance genes.

摘要

海岛棉是一种具有(AD)5基因组的野生异源四倍体物种。它具有许多有用的性状，包括更细的纤维细度、耐旱性以及对镰刀菌和黄萎病的抗性。我们构建了首个海岛棉细菌人工染色体文库。该文库质量高、覆盖范围广，包含200,832个克隆，平均插入片段大小约为122 kb，空克隆少于3%。我们的文库大小约为(AD)5基因组(2400 Mb)的10倍，提供了99.7%的概率来分离感兴趣的基因或其序列。位于五条不同染色体上且与黄萎病抗性相关的八个简单序列重复标记中的七个，能够扩增50个超级池，并获得一到五个命中结果。这个高容量文库将成为分类和分析异源四倍体棉花物种进化以及分离抗病和耐旱基因的重要基因组资源。