Microdissection of the A_h01 chromosome in upland cotton and microcloning of resistance gene anologs from the single chromosome.

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS ACS Applied Bio Materials Pub Date : 2017-05-18 eCollection Date: 2017-01-01 DOI:10.1186/s41065-017-0035-3

Xinchuan Cao, Yuling Liu, Zhen Liu, Fang Liu, Yalei Wu, Zhongli Zhou, Xiaoyan Cai, Xingxing Wang, Zhenmei Zhang, Yuhong Wang, Zhimin Luo, Renhai Peng, Kunbo Wang

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引用次数: 1

Abstract

Background: Chromosome microdissection is one of the most important techniques in molecular cytogenetic research. Cotton (Gossypium Linnaeus, 1753) is the main natural fiber crop in the world. The resistance gene analog (RGA) cloning after its single chromosome microdissection can greatly promote cotton genome research and breeding.

Results: Using the linker adaptor PCR (LA-PCR) with the primers of rice disease-resistance homologues, three nucleotide sequences PS016 (KU051681), PS054 (KU051682), and PS157 (KU051680) were obtained from the chromosome A_h01 of upland cotton (cv. TM-1). The Blast results showed that the three sequences are the nucleotide binding site-leucine rich repeat (NBS-LRR) type RGAs. Clustering results indicated that they are homologous to these published RGAs. Thus, the three RGAs can definitely be confirmed as NBS-LRR class of RGAs in upland cotton.

Conclusions: Using single chromosome microdissection technique, DNA libraries containing cotton RGAs were obtained. This technique can promote cotton gene cloning, marker development and even the improvement of cotton genome research and breeding.

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陆地棉Ah01染色体的显微解剖及单染色体抗性基因类似物的微克隆。

背景:染色体显微解剖是分子细胞遗传学研究的重要技术之一。棉花(Gossypium Linnaeus, 1753)是世界上主要的天然纤维作物。抗性基因类似物(RGA)单染色体显微解剖后的克隆，对棉花基因组的研究和育种具有重要的促进作用。结果:利用水稻抗病同源物引物的连接子PCR (LA-PCR)技术，从陆地棉(cv.) Ah01染色体上获得了PS016 (KU051681)、PS054 (KU051682)和PS157 (KU051680) 3个核苷酸序列。TM-1)。Blast结果表明，这三个序列均为核苷酸结合位点-亮氨酸富重复序列(NBS-LRR)型RGAs。聚类结果表明它们与这些已发表的RGAs同源。因此，这3种RGAs可以确定为陆地棉中NBS-LRR类RGAs。结论:利用单染色体显微解剖技术，可获得含棉花RGAs的DNA文库。该技术可促进棉花基因克隆、标记开发乃至棉花基因组研究和育种的改进。

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