甜椒细胞质雄性不育基因的SRAP标记  

SRAP marker linked to cytoplasmic male sterility in sweet pepper

作　　者：吴国平[1,2] 袁稳[1,2] 刘金兵[1] 刁卫平[1] 王述彬[1] 潘宝贵[1] 戈伟[1] 侯喜林[2] 

机构地区：[1]江苏省农业科学院蔬菜研究所,江苏南京210014 [2]南京农业大学园艺学院,江苏南京210095

出　　处：《江苏农业学报》2012年第5期1114-1118,共5页Jiangsu Journal of Agricultural Sciences

基　　金：国家大宗蔬菜产业技术体系淮安综合试验站项目(CARS-25-G-14);江苏省自然科学基金项目(BK2010464);江苏省农业科技自主创新基金项目[CX(10)103;CX(11)1004]

摘　　要：为了筛选出甜椒细胞质雄性不育基因的连锁相关序列扩增多态性(SRAP)标记,以甜椒细胞质雄性不育系(CMS)T6A和相应的保持系T6B为材料,利用SRAP分子标记技术,筛选与甜椒不育基因相关的特异片段。对该特异片段进行回收、克隆和测序,并用Blastn和Blastx程序在NCBI数据库中进行同源性检索和相似性比对。128对SRAP引物扩增共获得了3 844条100～1 000 bp的条带,平均每对引物组合可扩增出31条清晰的条带。检测到3个多态性位点,其中仅在不育系中扩出1条稳定的特异条带CMS T6A201。用Blastn程序进行核苷酸同源序列比较,特异片段CMS T6A201的序列与Ty3-gypsy类逆转座子的部分序列高度同源;用Blastx程序与蛋白质数据库进行同源性比对,由该片段推测的氨基酸序列与水稻假拟Ty3-gypsy逆转座子蛋白质、蒺藜苜蓿的反转录酶、水稻gag-pol蛋白质以及马铃薯的假拟gag-pol蛋白质等相似性较高。甜椒细胞质雄性不育系特异片段CMS T6A201的功能可能与逆转座子有关。To get SRAP（sequence-related amplified polymorphism） marker linked to cytoplasmic male sterility in sweet pepper,SRAP was analyzed in cytoplasmic male sterile（CMS） line T6A and its maintainer line T6B.The specific SRAP fragment was reclaimed,cloned and sequenced.DNA sequence of the specific fragment was analyzed with Blastn and Blastx in GenBank of NCBI.Total 128 pairs of SRAP primers were used.Three thousand eight hundred and forty four bands were amplified in two lines and 31 bands were amplified for an average primer combination.There were three polymorphic loci in two lines,but only one stable polymorphic fragment（named CMS T6A201） was found in CMS line.The results of Blastn and Blastx online showed that the fragment shared high homology with part of Ty3-gypsy-like retrotransposon sequence and had high similarities with partial protein sequence of Oryza sativa japonica group putative Ty3-gypsy retrotransposon protein,Medicago truncatula reverse transcriptase,Oryza sativa indica group gag-pol polyprotein and Solanum demissum putative gag-pol polyprotein,respectively.The function of this specific fragment CMS T6A201 is likely to be closely related to retrotransposon.

关 键 词：甜椒 细胞质雄性不育(CMS) SRAP标记 

分 类 号：S641.3[农业科学—蔬菜学]