细胞质雄性不育辣椒育性恢复基因特异分子标记的筛选  

IDENTIFICATION OF MOLECULAR MARKERS LINKED TO THE FERTILITY RESTORING GENE FOR THE CMS CAPSICUM ANNUUM L.

作　　者：常彩涛[1] 王春国[2] 陈成彬[2] 吴峰[1] 吴锋[1] 孙德岭[1] 

机构地区：[1]天津市科润蔬菜研究所,天津300382 [2]南开大学生命科学学院,天津300071

出　　处：《实验生物学报》2005年第3期227-232,共6页Acta Biologiae Experimentalis Sinica

基　　金：天津市自然科学基金(013614711)

摘　　要：利用集团分离分析法(Bulked segregant analysis BSA),以辣椒细胞质雄性不育系BU-12、恢复系RF-12为材料共筛选了336条RAPD引物,其中引物S418在恢复系中呈现特异性扩增,得到一条约3000bp的特异片段。回扩得到两条片段,测序表明大小为1515bp,1162bp。荧光原位杂交证实1515bp片段为恢复系特有,命名为S418_(1515)。序列分析表明S418_(1515)为一新发现的序列,Blastn序列比对同源性小于40%,tBlastx比对发现该序列与水稻2、4、7、10号染色体的几个BAC克隆上的序列高度同源。推测可能与其具有相似的编码功能,为进一步从分子水平研究辣椒育性恢复打下了坚实的基础。根据测序结果设计特异引物,将S418_(1515)转化成特异PCR标记,证明能用于候选材料的初筛。Bulked segregant analysis method was used to identify random amplified polymor- phic DNA (RAPD) markers linked to the fertility restoring gene for the cytoplasmic male sterility (CMS) capsicum annum L. Totally 336 random primers were screened on the DNA samples of restorer and sterile bulks. Primer S418 produced a special band in restorer line. It was about 3000bp, including two fragments 1515 bp and 1162 bp. Fluorescence in situ hybridization(FISH) indicated the fragment of 1515 bp only existed in restorer line. It was designed to S418_(1515). Analy- sis of the sequence indicated S418_(1515) was unknown before. The homology of blastn was less than 40%, however the homology of tBlastx indicated this sequence was high homologous with the part sequences of rice which were distributed on 2,4,7,10 chromosomes. It suggested this sequence might have the similar function with them. This result offered a good foundation to research the molecular mechanism of fertility restoration for CMS capsicum. Based on the sequence, special primers were designed to transform the RAPD marker to PCR marker. The result indicated that these primers could be used to screen the restorer lines from a large quantitive of candidate lines.

关 键 词：育性恢复基因 分子标记 辣椒 筛选 analysis 细胞质雄性不育系 荧光原位杂交 特异性扩增 BAC克隆 PCR标记 恢复系 RAPD 特异片段 序列比对 分子水平 特异引物 分析法 分析表 同源性 染色体 材料 测序 水稻 

分 类 号：S641.3[农业科学—蔬菜学]