Construction and characterization of a bacterial artificial chromosome library for Gossypium mustelinum.

Yuling Liu,Shulin Zhang,Baohong Zhang,Zhen Liu,Quanwei Lu,Xinpeng Wen,Kunbo Wang,Renhai Peng,Fang Liu,Yangyang Wei,David D Fang

doi:10.1371/journal.pone.0196847

Yuling Liu, Shulin Zhang + Show 9 more

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https://doi.org/10.1371/journal.pone.0196847

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Abstract

A bacterial artificial chromosome (BAC) library for G. mustelinum Miers ex G. Watt (AD4) was constructed. Intact nuclei from G. mustelinum (AD4) were used to isolate high molecular weight DNA, which was partially cleaved with Hind III and cloned into pSMART BAC (Hind III) vectors. The BAC library consisted of 208,182 clones arrayed in 542 384-microtiter plates, with an average insert size of 121.72 kb ranging from 100 to 150 kb. About 2% of the clones did not contain inserts. Based on an estimated genome size of 2372 Mb for G. mustelinum, the BAC library was estimated to have a total coverage of 10.50 × genome equivalents. The high capacity library of G. mustelinum will serve as a giant gene resource for map-based cloning of quantitative trait loci or genes associated with important agronomic traits or resistance to Verticillium wilt, physical mapping and comparative genome analysis.

Highlights

Gossypium genus comprises more than 50 species, including eight diploid groups and six tetraploid species [1,2,3]
Phylogenetic and phenetic ananlyses demonstrate that G. mustelinum had been isolated as one branch of the earliest split following allopolyploid formation, and is genetically farthest from G. hirsutum and other tetraploid species [4, 5, 7]
We firstly report the construction of a high capacity library in G. mustelinum, a wild tetraploid species, which is far from G. hirsutum in phylogeny, and has many excellent traits, including finer fibers and Verticillium resistance, which will help to save cotton germplasm resources, and provide important genome resources for cultivated cotton breeding programs about genetic diversity

Summary

Introduction

Gossypium genus comprises more than 50 species, including eight diploid groups and six tetraploid species [1,2,3]. Among the six tetraploid species, G. hirsutum L. The remaining tetraploid species are all wild species, and have much narrower distributions: G. tomentosum Nuttall ex Seemann (AD3) in the Hawaiian Islands, and G. mustelinum Miers ex Watt (AD4) in NE Brazil, G. darwinii Watt (AD5) in the Galapagos Islands[4], except for G. ekmanianum Wittmack (AD6), whose distribution needs to be further determined[3, 5]. Because of long-term natural selection under the distrubution environment, these wild species have rich genetic diversity and contain many excellent genes which can be used in cotton breeding, such as drought resistance, disease resistance, cold resistance, and the potential properties of fine fibers [6]. Phylogenetic and phenetic ananlyses demonstrate that G. mustelinum had been isolated as one branch of the earliest split following allopolyploid formation, and is genetically farthest from G. hirsutum and other tetraploid species [4, 5, 7]. Most likely there have some new genetic loci that can be used for genetic improvement of upland cotton, which is helpful to solve the serious bottleneck of genetic

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