摘要
9012AB,是一个被广泛应用于甘蓝型油菜杂种优势利用的隐性上位细胞核雄性不育两型系。遗传分析表明,其育性由两对隐性不育基因ms3、ms4和一对隐性上位基因rf控制。由于育性受3对基因控制并且基因之间存在互作,依靠传统的杂交、测交等表型鉴定方法,选出性状优良的不育系和临保系将是一项繁琐低效的工作。但利用与上位抑制基因(rf)和不育基因(ms3和ms4)连锁的分子标记进行分子标记辅助育种,就能取到事半功倍的效果。
本研究以隐性上位核不育材料9012A(ms3ms3ms4ms4RfRf)为供体亲本,高产,高含油量的浙双72(ms3ms3ms4ms4rfrf)为受体亲本,利用回交结合分子标记辅助选择的方法,选育出浙双72遗传背景的纯合不育系和同型临保系,为隐性核不育杂种优势利用提供新的优良材料。主要实验结果如下:
1.利用9012A与浙双72衍生的分离群体进行遗传分析,结果表明浙双72的基因型为ms3ms3ms4ms4rfrf。为了加快遗传背景的回复率,我们针对F2代群体中24株不育株(ms3ms3ms4ms4Rfrf或者ms3ms3ms4ms4RfRf)进行背景选择。聚类分析得到5株背景与浙双72相近的单株,与浙双72回交得到FzBC1。
2.利用与Rf基因连锁的SCAR标记WSC1从235株F2BC1群体(Ms3ms3ms4ms4Rfrf或者Ms3ms3ms4ms4rfrf)中选出124株基因型为Ms3ms3ms4ms4Rfrf的单株,采用AFLP标记进行背景选择,选出与浙双72背景最接近的5株单株,与浙双72回交得到F2BC2代。
3.利用与ms3连锁的BSR7和BSR67,与Rf连锁的WSC1和IBP5-3对F2BC2群体进行前景选择,挑出23株基因型为Ms3ms3ms4ms4Rfrf的单株,结合田间长势,选择3株与浙双72回交,得到F2BC3。
4.利用上述连锁标记对F2BC3群体进行前景选择,获得96株基因型为Ms3ms3ms4ms4Rfrf的单株。再利用SSR标记进行背景选择,最终得到5株背景与浙双72最为接近的单株,自交得到F2BC3F2。
A recessive epistatic genic male sterility (REGMS) two-type line,9012AB, has been used for rapeseed hybrid seed production. The male sterility of 9012AB is controlled by two recessive sterile genes ms3、ms4 with one recessive suppressor gene (rf). As the sterility is determined by three recessive genes and the existence of gene interaction, the breeding procedure for REGMS lines and temporary maintainer lines is very complicated, and the identification of the different genotypes with expected traits in backcross or selfed generations is laborious and time-consuming with traditional breeding methodology. However, the breeding process can be accelerated through MAS, which makes it easier to identify different genotypes in the segregating generations by selection of molecular markers tightly linked to the genes (ms3, ms4 and rf).
In the present study, we transferred the male sterile genes of the 9012A line into high-yield and high oil contents variety ZS72 through mlecular marker assisted backcross breeding and develop a noval REGMS line and their homologous temporary maintainer line. Our study developed a new material for the utilization of heterosis. The main experiment results are as follows:
1. Genetic analysis of the population derived from 9012A and ZheShuang 72 showed that the genetype of ZheShuang 72 is Ms3Ms3ms4ms4rfrf.23 sterile plants were selected (ms3ms3ms4ms4Rfrf or ms3ms3ms4ms4RfRf) in the F2 populations for background selection using AFLP markers, and 5 plants with high recovery rates were seclected to backcross with Zheshuang 72.
2. Using SCAR marker WSC1 tihtly linked to Rf,124 individuals with Ms3ms3ms4ms4Rfrf were selected out from the F2BC1 population of 235 plants (Ms3ms3ms4ms4Rfrf or Ms3ms3ms4ms4rfrf. According to the background selection results of 124 plants, we selected out 5 individuals with the closest Zheshuang 72 background for next backcross. As a result, the F2BC2 population was gained.
3. Molecular markers linked to ms3 (BSR7 and BSR67) and rf(WSC1 and IBP5-3) were used to screen the F2BC2 population, and 23 individuals with genotype of Ms3ms3ms4ms4Rfrf were obtained. Based on the agronomic traits, we selected 3 plants to backcross to ZheShuang 72, resulting in F2BC3 population.
4. Through foreground selection, we selected 96 plants with genetpye of Ms3ms3ms4ms4Rfrf in the F2BC3 population. Through background selection using SSR markers,5 individuals with the closest Zheshuang 72 were self-crossed, and then F2BC3F2 was gained.
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