二倍体野生棉与四倍体栽培棉基于GISH的遗传关系分析

doi:10.11963/issn.1002-7807.201701011

棉花学报 ›› 2017, Vol. 29 ›› Issue (1): 99-107.doi: 10.11963/issn.1002-7807.201701011

二倍体野生棉与四倍体栽培棉基于GISH的遗传关系分析

肖水平^1,²(),陈宜¹,柯兴盛¹,王玉红²,王春英²,杨磊¹,刘新稳¹,孙亮庆¹,杨绍群¹,王坤波^2,^*(),刘方^2,^*()

1. 江西省棉花研究所/国家棉花产业技术体系鄱阳湖综合试验站,江西九江332105
2. 中国农业科学院棉花研究所/棉花生物学国家重点实验室,河南安阳 455000

收稿日期:2015-11-18 出版日期:2017-01-15 发布日期:2017-01-15
通讯作者: 王坤波,刘方 E-mail:shuipingxiao@163.com;wkbcri@163.com;liufcri@163.com
作者简介:肖水平(1984―)，男，硕士，助研，shuipingxiao@163.com
基金资助:
国家科技支撑计划——经济作物种质资源发掘与创新利用(2013BAD01B03);国家现代农业产业技术体系——棉花产业技术体系(CARS-18-36);国家科技重大专项——转基因生物新品种培育子课题（2014ZX08005002- 009）;江西省科技支撑计划项目（20141BBF60011）

Genomic in situ Hybridization Analysis of the Genetic Relationship between Wild-Type Diploid and Cultivated Allotetraploid Gossypium Species

Xiao Shuiping^1,²(), Chen Yi¹, Ke Xingsheng¹, Wang Yuhong², Wang Chunying², Yang Lei¹, Liu Xinwen¹, Sun Liangqing¹, Yang Shaoqun¹, Wang Kunbo^2,^*(), Liu Fang^2,^*()

1. Cotton Research Institute of Jiangxi Province / Poyang Lake Cotton Experiment Station, China Agriculture Research System, Jiujiang, Jiangxi 332105, China
2. Institute of Cotton Research of Chinese Academy of Agricultural Sciences / State Key Laboratory of Cotton Biology, Anyang, Henan 455000, China

Received:2015-11-18 Online:2017-01-15 Published:2017-01-15
Contact: Kunbo Wang,Fang Liu E-mail:shuipingxiao@163.com;wkbcri@163.com;liufcri@163.com

摘要/Abstract

摘要： 目的研究二倍体野生棉与四倍体栽培棉间的遗传亲缘关系,进一步探索各棉种间的起源与进化。方法以5个二倍体基因组的代表种B₁（异常棉）、C₁（斯特提棉）、E₂（索马里棉）、F₁（长萼棉）以及G₁（比克氏棉）的基因组DNA（gDNA）为探针,以2个四倍体栽培种（陆地棉中棉所16、海岛棉新海7号）有丝分裂中期染色体为靶DNA,进行了基因组原位杂交（Genomic in situ hybridization, GISH）分析。结果以B₁、E₂和F₁ gDNA为探针时,杂交信号主要分布在2个栽培种较长的13对A亚组染色体上;各产生3对较强的GISH-NOR信号,其中1对分布在较长的A亚组上,2对分布在较短的D亚组上,其GISH-NOR信号强度与分布情况与以D基因组棉种为探针时相似。说明二倍体B、E、F基因组与四倍体棉A亚基因组具有较高的同源性,亲缘关系更近。这一点与它们的地理分布情况相符;而它们基因组中的45S rDNA重复序列与二倍体D基因组的45S rDNA重复序列同源性较高。C₁和G₁中以gDNA为探针时,杂交信号分布在2个栽培种全部26对染色体上,无法区分开A或D亚组染色体,都有3对较强的GISH-NOR信号。这一现象与D基因组拟似棉（D₆）gDNA为探针的GISH相似,表明二倍体C和G基因组与四倍体棉的A和D亚基因组均具有较高的同源性,或者C和G基因组同时含有A基因组(或其他非洲棉基因组)和D基因组成分,进一步证实了其基因组成分的杂合性;而它们基因组中的45S rDNA重复序列同属D基因组类型。结论这些发现可为棉花杂交育种和棉属起源与演化研究提供有用信息。

关键词: 棉属; 二倍体基因组; 四倍体栽培棉; 基因组原位杂交; 遗传亲缘关系

Abstract:

[Objective] This study investigated the genetic relationship between wild-type diploid and cultivated allotetraploid cotton species, and explored the origin and evolution of each species. [Method] Genomic in situ hybridization (GISH) was performed on chromosomes of Gossypium hirsutum var. CCRI 16 and G. barbadense var. Xinhai 7 as representative species of wild-type diploid cotton B, E, and F genome species using G. anomalum, G. somalense, and G. longicalyx genomic DNA as probes. [Results] The fluorescent signals were mainly distributed on A sub-genome chromosomes and were accompanied by three pairs of strong GISH-nucleolar organizer regions (NORs), one pair on the A sub-genome chromosomes and two pairs on the D sub-genome chromosomes. GISH-NOR signal intensities and distribution were similar for all diploid D genome gDNA probes. These results indicate that the G. anomalum, G. somalense, and G. longicalyx genome species have a closer genetic relationship with the tetraploid A sub-genome, which is consistent with their geographic distribution, while their 45S rDNA repeat sequences show higher homology with those of the D genome. Using G. sturtianum (C1) and G. bickii (G₁) gDNA as probes, fluorescent signals were distributed on all 26 pairs of chromosomes, the A and D sub-genomes were unable to be discriminated. This phenomenon was also observed using the D₆ genome species as a probe when three pairs of strong GISH-NOR signals were also detected. These findings suggest that the C and G genomes are highly homologous with the A and D sub-genomes of allotetraploid cotton species, or that they both contain A and D genome components leading to genome heterozygosity. However, their 45S rDNA repeat sequences appear to belong to the D genome type. [Conclusion] These findings provide useful information for cotton hybrid breeding and researches on the origin and evolution of Gossypium species.

Key words: Gossypium; diploid cotton species; allotetraploid cotton species; genomic in situ hybridization; genetic relationship

肖水平, 陈宜, 柯兴盛, 王玉红, 王春英, 杨磊, 刘新稳, 孙亮庆, 杨绍群, 王坤波, 刘方. 二倍体野生棉与四倍体栽培棉基于GISH的遗传关系分析[J]. 棉花学报, 2017, 29(1): 99-107.

Xiao Shuiping, Chen Yi, Ke Xingsheng, Wang Yuhong, Wang Chunying, Yang Lei, Liu Xinwen, Sun Liangqing, Yang Shaoqun, Wang Kunbo, Liu Fang. Genomic in situ Hybridization Analysis of the Genetic Relationship between Wild-Type Diploid and Cultivated Allotetraploid Gossypium Species[J]. Cotton Science, 2017, 29(1): 99-107.

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二倍体野生棉与四倍体栽培棉基于GISH的遗传关系分析

Genomic in situ Hybridization Analysis of the Genetic Relationship between Wild-Type Diploid and Cultivated Allotetraploid Gossypium Species

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