大豆三系的选育及恢复基因的SSR初步定位研究

以栽培大豆ZD8319为母本，分别与SG01、JX03和PI004杂交，再用父本回交5次，选育出阜CMS1A、阜CMS2A和阜CMS3A三个高度不育的不育系。正反交结果表明，这3个不育系为质核互作不育系（CMS），花粉为典败型不育，且不育性稳定。与3个不育系广泛测交，筛选与不育系配合力高、育性恢复力强的强优势组合，从中鉴定出恢复度高的恢复系5个（蒙-06，YC04，阜84-5-4，ZY9010，Z9001），使栽培大豆质核互作不育系三系配套。通过测交发现，阜CMS1A、阜CMS2A比阜CMS3A育性易被恢复。用这2个不育系和5个恢复系的核基因组DNA为模板，进行简单重复序列（simple sequence repeat, SSR）分析，从60对核心引物中筛选出在2个不育系和5个恢复系间有特异带的SSR引物，分别为Satt143、Satt168、Satt441。表明这3个SSR标记是与恢复基因有关的3个等位位点。并把恢复基因初步定位于2个分离群体上的6个连锁群上。

Selection of Three Lines and Localization of the Restorer Genes in Soybean Using SSR Markers

Three soybean male sterile lines, FuCMS1A(ZD8319SG01BC5F1),FuCMS2A(ZD8319JX03BC5F1),FuCMS3A(ZD8319PI004BC5F1) were developed through six years crosses and backcrosses using ZD8319 as cytoplasm donor and SG01, PI004, JX03 as nuclear donors. Through reciprocal crosses, it proved that the three male sterile lines were cytoplasmicnuclear male sterile lines. All the BC5F1 plants were male sterile and the average percentage of the sterile pollen grains was more than 98%. The results of parallel crosses suggested that the female of the CMS lines was fertile and its male sterility was stable. The percentage of sterile pollen grains of ZD8319, SG01, JX03, PI004 and their F1, BC1F1, BC1F5 were calculated. The results showed that their sterility rates were different. Nuclear donors SG01, JX03 were similar in genetics, their F1's male sterility rates were high, and their BC1 and BC5's sterility rates were similar. Comparing with F1, their sterility rates were higher, but only slightly. On the contrary, the F1's sterility rate between ZD8319 and PI004 was low. BC1 and BC5's sterility rates increased largely in a gradual manner. These results suggested that the nuclear male sterile genes of SG01, JX03 were dominant, and genes of PI004 are partial dominant, and its male sterility was controlled by multiple quantitative genes. Crossing with restorer lines, FuCMS1A, FuCMS2A's fertility was restored more easily than FuCMS3A's and five Rlines were selected,i.e. Meng06, YC04, Fu8454, ZY9010, and Z9001. So, soybean three lines was developed completely and successfully. Using soybean microsatellite or simple sequence repeat (SSR) method, 60 pairs of SSR core primer were screened between sterile lines and Rlines. There are 3 pairs of primer producing polymorphic bands. Satt143, Satt168 and Satt441 have produced special band in five Rlines and two CMSlines. Satt441 and Satt143 generated a 290bp, 500bp bands in all the five Rlines respectively. The three primers locate on 6 linkage groups. We deduce that there are three Rf loci, which are distributed in the six linkage groups.