Yi Bin, Chen Yuning, Lei Shaolin, Tu Jinxing, Fu Tingdong
National Key Laboratory of Crop Genetic Improvement, National Sub-center of Rapeseed Improvement in Wuhan, Huazhong Agricultural University, Wuhan 430070, China.
Theor Appl Genet. 2006 Aug;113(4):643-50. doi: 10.1007/s00122-006-0328-9. Epub 2006 Jun 28.
A recessive genic male sterility (RGMS) system, S45 AB, has been developed from spontaneous mutation in Brassica napus canola variety Oro, and is being used for hybrid cultivar development in China. The male sterility of S45 was controlled by two duplicated recessive genes, named as Bnms1 and Bnms2. In this study, a NIL (near-isogenic line) population from the sib-mating of S45 AB was developed and used for the fine mapping of the Bnms1 gene, in which the recessive allele was homozygous at the second locus. AFLP technology combined with BSA (bulked segregant analysis) was used. From a survey of 2,560 primer combinations (+3/+3 selective bases), seven AFLP markers linked closely to the target gene were identified, of which four were successfully converted to sequence characterized amplified region (SCAR) markers. For further analysis, a population of 1,974 individuals was used to map the Bnms1 gene. On the fine map, Bnms1 gene was flanked by two SCAR markers, SC1 and SC7, with genetic distance of 0.1 cM and 0.3 cM, respectively. SC1 was subsequently mapped on linkage group N7 using doubled-haploid mapping populations derived from the crosses Tapidor x Ningyou7 and DH 821 x DHBao 604, available at IMSORB, UK, and our laboratory, respectively. Linkage of an SSR marker, Na12A02, with the Bnms1 gene further confirmed its location on linkage group N7. Na12A02, 2.6 cM away from Bnms1, was a co-dominant marker. These molecular markers developed from this research will facilitate the marker-assisted selection of male sterile lines and the fine map lays a solid foundation for map-based cloning of the Bnms1 gene.
一种隐性核雄性不育(RGMS)系统S45 AB,是从甘蓝型油菜品种奥罗的自发突变中培育出来的,目前正在中国用于杂交品种的培育。S45的雄性不育由两个重复的隐性基因控制,命名为Bnms1和Bnms2。在本研究中,通过S45 AB的同胞交配构建了一个近等基因系(NIL)群体,并用于Bnms1基因的精细定位,其中隐性等位基因在第二个位点是纯合的。采用AFLP技术结合BSA(混合分离群体分析法)。通过对2560个引物组合(+3/+3选择性碱基)的筛选,鉴定出7个与目标基因紧密连锁的AFLP标记,其中4个成功转化为序列特征性扩增区域(SCAR)标记。为了进一步分析,利用一个1974个个体的群体对Bnms1基因进行定位。在精细图谱上,Bnms1基因两侧分别有两个SCAR标记SC1和SC7,遗传距离分别为0.1 cM和0.3 cM。随后,利用分别来自英国IMSORB和我们实验室的Tapidor×Ningyou7以及DH 821×DHBao 604杂交产生的双单倍体定位群体,将SC1定位到连锁群N7上。一个SSR标记Na12A02与Bnms1基因的连锁进一步证实了其在连锁群N7上的位置。Na12A02与Bnms1的距离为2.6 cM,是一个共显性标记。本研究开发的这些分子标记将有助于雄性不育系的标记辅助选择,精细图谱为基于图谱的Bnms1基因克隆奠定了坚实基础。