瑟伯氏棉着丝粒重复序列鉴定  

Identification of centromere repeat sequence in Gossypium thurberi

作　　者：陈晔 刘玉玲 韦洋洋 杨太有[1] 彭仁海[1,2] Chen Ye;Liu Yuling;Wei Yangyang;Yang Taiyou;Peng Renhai(College of Life Sciences,Henan NormaL University,Xinxiang 453007,China;CoLLege of BioLogy and Food Engineering,Anyang Institute of TechnoLogy,Anyang 455000,China)

机构地区：[1]河南师范大学生命科学学院,河南新乡453007 [2]安阳工学院生物与食品工程学院,河南安阳455000

出　　处：《河南师范大学学报（自然科学版）》2019年第6期103-109,共7页Journal of Henan Normal University(Natural Science Edition)

基　　金：国家重点研发计划(2018YFD0100300);河南省科技创新杰出人才(184200510009);河南省科技攻关(182102110048)

摘　　要：棉花着丝粒染色质主要由重复序列组成,包括高度重复的着丝粒反转录转座子和卫星重复序列等,但其着丝粒重复序列进化机制尚不清楚.为了深入了解着丝粒重复列的演化,利用染色质免疫共沉淀(ChIP)和中期染色体荧光原位杂交(FISH)技术对瑟伯氏棉(Gossypium thurberi,D 1)着丝粒序列进行了分析.结果表明,相应的ChIP-DNA序列属于着丝粒功能区域,发现了10个较为富集的着丝粒重复序列.中期染色体FISH定位发现,CL179和CL234在四倍体陆地棉中的A和D染色体组中都有杂交信号,在二倍体A组亚洲棉中没有检测到信号,由此可推断在形成四倍体后一些着丝粒特异的重复序列进行了扩增.重复序列CL95在瑟伯氏棉以及陆地棉部分染色体的着丝粒区域出现杂交信号,而在二倍体D组雷蒙德氏棉和A组亚洲棉的着丝粒区域并没有出现信号,可认为该序列是瑟伯氏棉特异的且在四倍体形成后被保留了下来.通过对瑟伯氏棉着丝粒序列及演化的分析能够为揭示棉属植物着丝粒重复序列及染色体组的演化关系提供重要的理论依据.The centromere chromatin of cotton is mainly composed of repetitive sequences,including highly repetitive centromere reverse transcriptional transposons and satellite repeats,etc.However,the evolutionary mechanism of the centromere repeats is still unclear.To further understand the evolution of centromere repeats,chromatin immunoprecipitation(ChIP)and metaphase chromosome fluorescence in situ hybridization(FISH)techniques were used to analyze the centromere sequences of Gossypium thurberi(D1).The results showed that the corresponding ChIP-DNA sequences belong to the functional region of centromere,and 10 relative abundant centromere repeats were found.FISH localization of metaphase chromosomes found that in the A and D sub-genomes chromosomes of tetraploid upland cotton CL179 and CL234 appeared FISH signals,but in G.arboreum of diploid A genome no signals were detected.Therefore,it could be inferred that some specific repeat sequences of centromeres were amplified after the formation of tetraploid.Repeated sequence CL95 showed hybrid signals in centromere regions of G.thurberi and part of upland cotton chromosomes,but no signals in centromere regions of G.raimondii G.arboreum,suggesting that the sequence was specific to G.thurberi and retained after tetraploid formation.The preliminary identification of G.thurberi centromere sequence and its evolutionary process provides a basis for further study of the structure and function of cotton centromere,which is helpful to reveal the evolutionary mechanism of the repeated centromere sequence.

关 键 词：瑟伯氏棉 着丝粒重复序列 染色质免疫沉淀 荧光原位杂交 

分 类 号：Q756[生物学—分子生物学]