摘要
大豆疫霉根腐病是大豆生产上的主要病害之一,在世界主要大豆产区均有发生。利用抗病品种是防治大豆疫霉根腐病最经济、有效的方法。但由于大豆疫霉菌具有高度的变异,大豆品种的抗性容易被克服,给抗病育种带来了极大的挑战。因此,必须不断地发掘新的抗病基因,以确保抗病育种工作持续、有效的开展。
豫豆25号是当期黄淮地区广泛栽培的优良大豆品种,对大豆疫霉菌具有广谱抗性。本研究通过以豫豆25为父本分别与豫豆21和早熟18进行杂交构建抗性分离群体,分析豫豆25对大豆疫霉根腐病的抗性遗传,应用分离群体分组分析法结合微卫星标记进行分析,对豫豆25中的抗病疫霉根腐病基因进行分子标记和定位。得到了如下结果:
1.豫豆25对大豆疫霉菌的抗性由一个显性单基因控制,将该基因命名为RpsYD25。
2.微卫星标记与遗传连锁性分析发现,豫豆21×豫豆25组合衍生的F2:3家系中,位于N连锁群上的五个标记sat_208、satt530、sat_084、satt125和sat_236与豫豆25中抗疫霉根腐病基因RpsYD25具有连锁性,遗传距离分别是:9.9cM、6.3 cM、7.6 cM、10.2 cM和30 cM,遗传作图表明,标记与基因之间的位置关系为sat_208- satt530- RpsYD25- sat_084- satt125- sat_236;在早熟18×豫豆25组合衍生的F2:3家系中,位于N连锁群上的五个标记satt125、sat_275、sat_266、satt660和GMABAB与抗病基因RpsYD25具有连锁性,遗传距离分别是:7.9 cM、7.8 cM、24.7 cM、38.8 cM和45.1 cM,标记与基因之间在连锁图上的位置关系是satt125- RpsYD25-sat_275- sat_266- satt660-GMABAB。
3. RpsYD25作图在N连锁群上抗疫霉根腐病基因Rps1座位附近。选择Rps1k基因序列中SSR设计引物对豫豆25、豫豆21和早熟18进行扩增,SSR标记satt1k6在豫豆25和豫豆21间具有多态性。用satt1k6鉴定豫豆21×豫豆25组合衍生的F2:3家系的基因,连锁分析该标记与基因RpsYD25连锁。将satt1k6与其他5个与RpsYD25连锁的SSR标记sat_208、satt530、sat_084、satt125、sat_236一起作图,satt1k6与基因RpsYD25的遗传距离是25.7cM,这一结果表明,RpsYD25与Rps1连锁,至于是否为Rps1座位上的一个新等位基因,还是一个新的基因还需要进一步研究。
Phytophthora root rot is one of the most important diseases on soybean production and it has occurred throughout the world. Application of the resistant cultivars is the most economic and effective way to control the disease. It brings a large challenge for the resistant breeding due to the high degree variability of Phytophthora. So we must discover novel resistance genes to meet the need for resistance breeding.
Yudou 25 is an elite soybean cultivar growing Hhuanghuai region now and has a broad resistance spectrum to Phytophthora root rot. The objectives of this study were to analysis resistance genetic to Phytophthora root rot and molecularly tagged and mapped the resistance gene on Yudou25, resistance identification of Phytophthora to Yudou25、Yudou21、Zaoshu18 and F2:3 progenies of the cross Yudou21/Yudou25 and Zaoshu18/Yudou25; genetic analysis with the F2:3 progenies from two cross lines; polymorphism analysis by microsatellite markers and bulked segregation analysis(BSA), and to locate the major resistance gene .The main results are as follows:
1. Inoculated parents and F2:3 populations of the cross Yudou21/Yudou25 and to isentify resistance to Phytophthora root rot in glasshouse at seedling stage, the result showed the resistance to Phytophthora root rot on Yudou25 was controlled by a single gene. The gene is temporarily named as RpsYD25.
2. Using polymorphic analysis of microsatellite markers and genetic linkage analysis, in F2:3 population deriverd from the cross of Yudou21/Yudou25, 5 SSR marker including sat_208, satt530, sat_084, satt125 and sat_236 on soybean molecular linkage group N were found linked to RpsYD25 with genetic distance of 9.9 cM, 6.3 cM, 7.6 cM, 10.2 cM and 30 cM respectively. The order of the resistant gene and the markers was sat_208- satt530- RpsYD25- sat_084- satt125- sat_236; in F2:3 population of the cross Zaoshu18/Yudou25, 5 SSR marker including satt125, sat_275, sat_266, satt660 and GMABAB on MLG N were found linked to RpsYD25 with genetic distance of 7.9 cM, 7.8 cM, 24.7 cM, 38.8cM and 45.1cM respectively. The order of the resistant gene and the markers was satt125- RpsYD25-sat_275- sat_266- satt660-GMABAB.
3. Because RpsYD25 was mapped on MLG N near to Rps1 locus, the genetic relationship of RpsYD25 and Rps1 was detected by using the selected SSR markers contained into the Rps1k allele sequence. A SSR marker satt1k6 into Rps1k allele was found to be linked to RpsYD25 with a genetic distance of 19.4 cM in F2:3 population from the crosses Yudou21×Yudou25. Therefore, the Phytophthora resistance gene RpsYD25 in cv. Yudou25 could be a novel allele at Rps1 locus, or novel gene.
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