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油菜Pol-CMS Rfp候选基因区域SSR标记开发与应用

Development of SSR Markers Associated with Polima CMS Fertility Restorer in Brasscia napus L.and It’s Applicaton
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摘要 波里马细胞质雄性不育系(Pol-CMS)的育性受环境温度影响较大,在杂交制种过程中易自交结实产生假杂种,从而影响杂交种的纯度质量,在生产上对母本假杂种的检测是杂交种纯度鉴定的关键。本研究利用已公布的Pol-CMS候选恢复基因Rfp编码序列,通过在白菜型油菜和甘蓝型油菜基因组进行同源序列比对,然后基于Rfp候选基因区域同源序列开发恢复基因的特异SSR标记,并应用于油菜杂交种纯度鉴定研究。研究结果表明,利用已公布Pol-CMS恢复基因1953 bp编码序列获取Rfp在基因组所在区段,在该区域及上、下游15K区域共获得32个SSR位点;其中,1个位点在供试的Pol-CMS不育系和恢复系间具有多态性,该标记为共显性标记,在不育系中的扩增产物大小为162 bp,在恢复系中为184 bp;与不育系相比,恢复系的扩增序列存在一个15 bp和一个7 bp的片段插入;在‘沣油737’的F_(2)分离群体中,该标记与育性恢复性状呈共分离遗传;利用该标记对‘沣油737’、‘沣油320’和‘沣油958’杂交制种样品的纯度鉴定结果与田间育性鉴定结果一致,表明本研究开发的SSR标记为Rfp基因紧密连锁的共显性标记,在Pol-CMS杂交品种的纯度鉴定应用中,可有效检测出混杂的父母本假杂种。本研究结果可为油菜杂交品种纯度鉴定与遗传多样性分析提供技术参考。 The Polima system of cytoplasmic male sterility(Pol-CMS)is widely used for hybrid seed production,but with high probability of occurrence of maternal false hybrid due to the fertility of Polima CMS(Pol-CMS)line is greatly affected by the ambient temperature,therefore,the detection of maternal false hybrids is the key to purity identification.The present study,the SSR markers were designed based on the flanking sequences of Pol-CMS restorer gene Rfp,and application of SSR markers in hybrid seed purity test of rapeseed.The main results were as follows:the sequences of Rfp were subjected to basic local alignment search tool queries against the Brasscia napus genome to determine chromosome positions,and 32 SSR markers were developed based the upstream and downstream sequences of Rfp.Among the 32 SSR markers,one codominant marker was polymorphic between Pol-CMS sterile line and Pol-CMS restorer line,the PCR product length was 162 bp in Pol-CMS sterile lines,and184 bp in restorer lines.Based on the PCR sequence,two insert fragments(15 bp and 7 bp)were identified in Pol-CMS restorer lines.The SSR marker co-segregated with plant fertility of individuals in the‘Fengyou737’F_(2)population.The purity of‘Fengyou737’,‘Fengyou320’and‘Fengyou958’were identified by means of by means of PCR-SSR markers and field-planting,and the results of two ways were highly consistent,indicated that the codominant SSR marker tightly linked to Rfp,and the marker can detect paternal and maternal false hybrids.The present findings will provide technical assistance for hybrid seed purity test and genetic diversity analysis in rapeseed.
作者 王同华 李宝 郭一鸣 张瑶婷 刘新红 范连益 曲亮 邓力超 李莓 Wang Tonghua;Li Bao;Guo Yiming;Zhang Yaoting;Liu Xinhong;Fan Lianyi;Qu liang;Deng Lichao;Li Mei(Hunan Hybrid Rape Engineering Technology Research Center,Crop Research Institute of Hunan Province,Changsha,410125)
出处 《分子植物育种》 CAS 北大核心 2022年第4期1206-1213,共8页 Molecular Plant Breeding
基金 湖南省农业科学院科技创新项目(2019YC02) 国家油菜产业技术体系项目(CARS-12)共同资助。
关键词 油菜(Brassica napus L.) Pol-CMS 恢复基因 SSR Brassica napus L. Pol CMS Restorer gene SSR
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