He Guang-Hua, Wang Wen-Ming, Liu Guo-Qing, Hou Lei, Xiao Yue-Hua, Tang Mei, Yang Zheng-Lin, Pei Yan
Center of Biotechnology, Southwest Agricultural University, Chongqing 400716, China.
Yi Chuan Xue Bao. 2002 Sep;29(9):798-802.
F2 population derived from Shanyou63, F1 hybrid developed from the cross Zhenshan97 A/Zhenshan97B, was used in this study. Fertile bulk was constructed by polling equal amount of 15 highly fertile lines. Sterile bulk was obtained by pooling equal amount of 15 highly sterile lines. Minghui63 and Zhenshan97A, parents of Shanyou63, were analyzed with 302 pairs of SSR primers. 244 pairs of primers gave amplification products, of which 58 pairs of primers on 12 different chromosomes showed polymorphism between the two parents with polymorphic frequency up to 23.77%. Gene bulks were further assayed with the 5 pairs of primers. RM1 on chromosome 1 and RM258, RM304 on chromosome 10 was found to be polymorphic between the two gene bulks. In theory, there should be no difference detected between the two gene bulks except for the target traits governed by fertility-restoring genes. RM1, RM258 and RM304 were probably related to the restorer genes. Ten highly fertile and ten highly sterile lines were selected from F2 population of Shanyou63 to screen the gene bulks. The results showed that specific bands of Minghui63 were detected in all ten highly fertile lines while not observed in all the sterile lines. It indicated that the three SSR markers might be linked to fertility-restoring genes. Dominant lines were not selected due to their inalbility to distinguish recombinant lines from non-recombinant lines. Pure recessive lines were chosen to conduct mapping analysis. A total of 53 highly sterile lines were selected from 900 lines of Shanyou63 F2 population to estimate the genetic distance between three SSR markers and fertility-restoring genes respectively. The results demonstrated that recombination occurred in 2, 3, lines with RM1 and RM258 while no one with RM304. Using MAPMAKER/EXP 3.0, the genetic distance between RM1, RM258, RM304 and the related restorer genes were calculated as 1.9, 2.9 and 0.0 cM, respectively. It is possible that the fertility restoring gene(s) on chromosome 10 for three different types of cytoplasmic male sterility(WA, BT and HL) are of the same, or belong to a gene family.
本研究使用了由汕优63衍生而来的F2群体，汕优63是珍汕97A/珍汕97B杂交育成的F1杂种。通过对15个高育性株系等量取样构建可育混合池。通过对15个高度不育株系等量取样获得不育混合池。用302对SSR引物对汕优63的亲本明恢63和珍汕97A进行分析。244对引物产生了扩增产物，其中12条不同染色体上的58对引物在两个亲本间表现出多态性，多态频率高达23.77%。用这5对引物对基因池进行进一步检测。发现第1染色体上的RM1以及第10染色体上的RM258、RM304在两个基因池间表现出多态性。理论上，除了受育性恢复基因控制的目标性状外，两个基因池之间不应检测到差异。RM1、RM258和RM304可能与恢复基因有关。从汕优63的F2群体中挑选出10个高育性株系和10个高度不育株系对基因池进行筛选。结果表明，在所有10个高育性株系中均检测到明恢63的特异带，而在所有不育株系中均未观察到。这表明这3个SSR标记可能与育性恢复基因连锁。由于显性株系无法区分重组株系和非重组株系，因此未选用。选择纯合隐性株系进行定位分析。从汕优63 F2群体的900个株系中总共挑选出53个高度不育株系，分别估算3个SSR标记与育性恢复基因之间的遗传距离。结果表明，RM1和RM258分别有2个、3个株系发生了重组，而RM304没有。使用MAPMAKER/EXP 3.0分别计算出RM1、RM258、RM304与相关恢复基因之间的遗传距离为1.9、2.9和0.0 cM。第10染色体上针对三种不同类型细胞质雄性不育（野败型、包台型和红莲型）的育性恢复基因可能相同，或属于一个基因家族。
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