割手密SsREMO-1基因的克隆与等位变异分析  

Cloning and Allelic Variation Analysis of SsREMO-1 Gene in Saccharum spontaneum L.

作　　者：刘洋 姚艳丽[2,3] 胡小文[2,3] 徐磊 高玉尧[2,3] 安东升 Liu Yang;Yao Yanli;Hu Xiaowen;Xu Lei;Gao Yuyao;An Dongsheng(Yunnan Key Laboratory of Sugarcane Genetic Improvement, Kaiyuan, 661699;Zhanjiang Experiment Station, Chinese Academy of Tropical Agri- cultural Sciences, Zhanjiang, 524013;Guangdong Engineering Technology Research Center for Dryland and Water Saving Agriculture, Zhanjiang, 524013)

机构地区：[1]云南省甘蔗遗传改良重点实验室,开远661699 [2]中国热带农业科学院湛江实验站,湛江524013 [3]广东省旱作节水农业工程技术研究中心,湛江524013

出　　处：《分子植物育种》2018年第7期2166-2174,共9页Molecular Plant Breeding

基　　金：云南省甘蔗遗传改良重点实验室开放课题(2015DG015-01);中国热带农业科学院基本科研业务费专项资金(17CXTD-07)共同资助

摘　　要：割手密抗逆基因资源的挖掘是甘蔗抗逆育种的重要工作任务。本研究以前期转录组测序得到的割手密REMO基因c DNA序列为探针,对Gen Bank中的EST数据库进行同源检索,发现高粱REMO基因(XM_002465954.1)与其具有极高的相似性(94%)。利用高粱REMO基因(XM_002465954.1)设计同源引物,后经PCR、分子克隆、序列分析验证成功分离了7个割手密REMO基因c DNA序列(Ss REMO-1a,Ss REMO-1b,Ss REMO-1c,Ss REMO-1d,Ss REMO-1e,Ss REMO-1f,Ss REMO-1g,Gen Bank登录号为MF095088-MF095094)。7条序列全长均为738 bp,5'非翻译区(UTR)长度78 bp,3'UTR长度为96 bp,包含一个564 bp的开放读码框,编码187个氨基酸。分析表明此蛋白序列具有REMO保守结构域。Ss REMO-1a、Ss REMO-1b、Ss REMO-1c、Ss REMO-1d、Ss REMO-1e、Ss REMO-1f、Ss REMO-1g的相似性在98.9%~99.7%之间,存在14个单核苷酸多态性(SNP)位点,4个氨基酸变异位点,蛋白相似性在98.4%~100%之间。Ss REMO-1蛋白序列与高粱、水稻、玉米等物种具有较高的同源性,可分为两个亚类。Abstract Mining o f resistance gene resources in Saccharum spontaneum is an important task for resistance breeding of sugarcane. In this paper, we blasted the EST database in GenBank using cDNA of REMO gene of Saccharum spontaneum L. obtained by transcriptome sequencing, and found that the sorghum REMO gene （XM_ 002465954.1） had high similarity （94%） with it. Homologous primers were designed through sorghum REMO gene （XM_002465954.1）, then the eDNA sequence of 7 REMO genes （SsREMO-1a, SsREMO-1b, SsREMO-1c, SsRE- MO-1d, SsREMO-1e, SsREMO-1f, SsREMO-1g, GenBank accession number MF095088-MF095094） from S. spontaneum was successfully separated by PCR, molecular cloning and sequence analysis verification. The analysis indicated that this protein contained REMO conserved domain. SsREMO-1a, SsREMO-1b, SsREMO-1c, SsRE- MO-1d, SsREMO-1e, SsREMO-1f SsREMO-1g all were 738 bp, containing 78 bp of 5＇ untranslated region （UTR）and 96 bp of 3＇ UTR, and a 564 bp open reading frame （ORF） encoding 187 amino acids sequence. The similarity of SsREMO-1a, SsREMO-1b, SsREMO-1c, SsREMO-1d, SsREMO-1e, SsREMO-1f SsREMO-1gcDNA sequen- ces was between 98.9% and 99.7%, having 14 single nucleotide polymorphisms （SNPs） and 4 amino acid mutation sites, and the protein similarity of SsREMO-1a, SsREMO-1b, SsREMO-1c, SsREMO-1d, SsREMO-1e, SsRE- MO-1f SsREMO-1g was between 98.4% and 100%, The SsREMO-1 protein sequence had high homology with sorghum, rice and maize, which could be divided into two subgroups.

关 键 词：割手密 REMO基因 基因克隆 变异分析 

分 类 号：S566.1[农业科学—作物学]