利用SSR标记定位明恢63的2对恢复基因

选取珍汕97A和明恢63杂交组合的F2高可育和高不育单株构建基因池，利用302对SSR引物对其进行了多态性分析。结果表明，位于第1染色体上的RM1和位于第10染色体上的RM258，RM304在亲本，基因池间表现多态性，用F2单株验证证明它们与野败型恢复基因连锁，完全不育株分析表明，与恢复基因间的遗传距离分别为1.9,2.9和0.0cM，野败型，红莲型，BT型3种不育胞质恢复基因在第10染色体上可能为同一基因或家族成员。

Mapping of Two Fertility-restoring Gene for WA Cytoplasmic Male Sterility in Minghui 63 Using SSR Markers

F 2 population derived from Shanyou63,F 1 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 F 2 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 F 2 population to estimate the genetic distance between three SSR markers and fertility restoring genes respectively. The results demonstrated thatrecombination 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 0.0cM ,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.