两种油菜隐性核不育系综合利用及中国近年70份甘蓝型油菜品种（系）遗传多样性研究
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摘要
油菜是我国最主要的油料作物,具有显著的杂种优势。油菜杂种优势的利用途径主要有细胞质雄性不育(CMS)、细胞核雄性不育(GMS)、化学杀雄、自交不亲和等。其中,细胞核雄性不育因其具有不育性稳定、不受环境及胞质的影响等特点而倍受青睐,已成为油菜杂种优势利用的主要途径之一。目前,生产上广泛应用的细胞核雄性不育材料主要有两类:一是两对隐性重叠基因控制的细胞核雄性不育,此类材料不育性稳定、彻底,不受环境影响,恢复源广,缺点是找不到完全“保持系”,用于杂交种生产时需大面积拔除50%可育株,其利用受到了较大限制;二是三对隐性基因互作控制的核不育,即2对隐性重叠不育基因和1对隐性上位抑制基因互作控制的细胞核雄性不育,由于其存在上位抑制基因,可获得100%的全不育系,从而实现核不育“三系”配套制种。然而,隐性上位抑制基因对双隐性不育基因是否也有抑制作用,目前缺乏系统的试验证据。本研究将该两类材料杂交,欲将三隐性不育材料中的上位抑制基因转移到双隐性材料中,从而选育双隐性不育材料的临保系,实现双隐性核不育的“三系”配套,这样可以同时结合双隐性和三隐性核不育两大基因库,扩大不育材料、恢复材料的遗传背景,培育出优质、高产的油菜杂交种。所得研究结果如下:
在自交的131个F3株系中,39个株系表现为育性分离,92个株系表现为全可育。卡方检验,结果并不支持前人的假设,即甘蓝型油菜隐性上位基因与隐性重叠不育基因之间的互作是非专一性的假设;在测交的147个株系中,81个株系表现为育性分离,66个株系表现为全可育,没有全不育测交株系出现,卡方检验结果,虽然两种假设都不符合,但实际的结果更符合没有专一性抑制作用假设,因为测交后代没有出现全不育的组合。
本试验结果暗示前人提出的“甘蓝型油菜隐性上位基因与隐性重叠不育基因之间的互作是非专一性的假设”是错误的,即甘蓝型油菜9012AB中隐性上位抑制基因的对其不育基因的抑制作用是专一的。
本论文还对中国近年审定的70份甘蓝型油菜品种的遗传多样性进行了SSR分析。SSR分析共得到113条清晰的谱带,其中104条呈多态性,多态性比率为92%,平均每条引物扩增出4.3条带,片段大小在100-600bp。主成分分析结果显示,参试材料分布比较分散,没有因来源地域相同而明显地聚在一起。我国近年审定的冬油菜品种遗传多样性比较丰富;不同冬油菜大区品种变异以区内变异为主;参试油菜品种中,长江中游区材料组内变异最为丰富。
Rapeseed is one of the most important oilcrops worldwide, and has significant heterosis. The main approaches of utilizing heterosis of rapeseed included cytoplasmic male sterility (CMS), genic male sterility (GMS), chemical hybridizing, and self-incompatibility. Because GMS has stable sterility and isn’t influenced by environment and cytoplasm,this approach was paid more and more attentions by the rapeseed breeders. Currently, the main GMS materials used for producing hybrid seeds are divided to two categories. The first category was the GMS materials controlled by two duplicate recessive genes. This kind of material has stable, complete male sterility, wide resource of its restorers, and isn’t influenced by environment, but its disadvantage is that it hasn’t complete restorers, and when it is used to produce hybrid seeds, 50% segregating fertile plants in the double purpose line are needed to be removed, which limits its utilization in the production of hybrid seeds. The second category was the GMS materials controlled by two duplicate recessive sterile genes and one recessive epistatic suppression gene interactively. With the existence of epistatic inhibitor gene, a 100% sterile line can be obtained. Three-line systems can be obtained for hybrid seeds production for this kind of materials.The research about verification of existence of inhibition of recessive epistatic suppression gene on two duplicate recessive sterile genes hasn’t been reported currently. On the basis of previous researches, which reported that these genes controlled sterility of the above two materials are non-allelic, and the interaction of recessive epistatic suppression gene and duplicate recessive sterile genes in Brassica napus is non-specific, crosses were made between the two categories of materials in order to transfer the recessive epistatic suppression gene of the Reeessive Genic Male Sterility to the two duplicate recessive materials, and develop temporary maintainer lines for the two duplicate recessive sterile lines. The results obtained are as follows.
In the 131 F3 lines obtained from F2 fertile plants of the two categories of materials, 39 lines displayed fertility segregation, 92 lines were fertile completely.χ2 test showed that the interaction of recessive epistatic suppression gene and duplicate recessive sterile genes in B. napus wasn’t specific. In the 147 lines obtained from the testcross between F2 fertile plants of the two categories of materials and sterile plants of the two duplicate recessive sterile materials, 81 lines displayed fertility segregation, 66 lines were fertile completely. Inspected byχ2, our result doesn’t meet the two assumptions. But our result met the inference that the testcross progenies won’t have sterile completely lines if recessive epistatic suppression gene hasn’t specific inhibition on two duplicate recessive sterile genes.These results revealed that the previous assumption, which assumed that the interaction of recessive epistatic suppression gene and duplicate recessive sterile genes in B. napus is non-specific, is not correct. Our results showed that the recessive epistatic suppression gene has specific suppression on its sterile genes in B. napus.
In the second part, genetic diversity of 70 B. napus varieties of approvalled during recent years in China was analyzed with SSR. In total, 113 fragments were detected with size ranged from 100 to 600 bp and 104 were polymorphism, the polymorphism rate was 92%. On average, each primer combination generated 4.3 polymorphic fragments. Principal analysis revealed that all varieties scattered well in bi-plot, and accessions from the same geographic origin did not group together. There existed considerable variance among recently registered B. napus cultivars in China. The components of variation within populations was dominant over that among populations. Varieties in middle of Yangtze valley possessed the richest genetic variation in the present investigation.
在自交的131个F3株系中,39个株系表现为育性分离,92个株系表现为全可育。卡方检验,结果并不支持前人的假设,即甘蓝型油菜隐性上位基因与隐性重叠不育基因之间的互作是非专一性的假设;在测交的147个株系中,81个株系表现为育性分离,66个株系表现为全可育,没有全不育测交株系出现,卡方检验结果,虽然两种假设都不符合,但实际的结果更符合没有专一性抑制作用假设,因为测交后代没有出现全不育的组合。
本试验结果暗示前人提出的“甘蓝型油菜隐性上位基因与隐性重叠不育基因之间的互作是非专一性的假设”是错误的,即甘蓝型油菜9012AB中隐性上位抑制基因的对其不育基因的抑制作用是专一的。
本论文还对中国近年审定的70份甘蓝型油菜品种的遗传多样性进行了SSR分析。SSR分析共得到113条清晰的谱带,其中104条呈多态性,多态性比率为92%,平均每条引物扩增出4.3条带,片段大小在100-600bp。主成分分析结果显示,参试材料分布比较分散,没有因来源地域相同而明显地聚在一起。我国近年审定的冬油菜品种遗传多样性比较丰富;不同冬油菜大区品种变异以区内变异为主;参试油菜品种中,长江中游区材料组内变异最为丰富。
Rapeseed is one of the most important oilcrops worldwide, and has significant heterosis. The main approaches of utilizing heterosis of rapeseed included cytoplasmic male sterility (CMS), genic male sterility (GMS), chemical hybridizing, and self-incompatibility. Because GMS has stable sterility and isn’t influenced by environment and cytoplasm,this approach was paid more and more attentions by the rapeseed breeders. Currently, the main GMS materials used for producing hybrid seeds are divided to two categories. The first category was the GMS materials controlled by two duplicate recessive genes. This kind of material has stable, complete male sterility, wide resource of its restorers, and isn’t influenced by environment, but its disadvantage is that it hasn’t complete restorers, and when it is used to produce hybrid seeds, 50% segregating fertile plants in the double purpose line are needed to be removed, which limits its utilization in the production of hybrid seeds. The second category was the GMS materials controlled by two duplicate recessive sterile genes and one recessive epistatic suppression gene interactively. With the existence of epistatic inhibitor gene, a 100% sterile line can be obtained. Three-line systems can be obtained for hybrid seeds production for this kind of materials.The research about verification of existence of inhibition of recessive epistatic suppression gene on two duplicate recessive sterile genes hasn’t been reported currently. On the basis of previous researches, which reported that these genes controlled sterility of the above two materials are non-allelic, and the interaction of recessive epistatic suppression gene and duplicate recessive sterile genes in Brassica napus is non-specific, crosses were made between the two categories of materials in order to transfer the recessive epistatic suppression gene of the Reeessive Genic Male Sterility to the two duplicate recessive materials, and develop temporary maintainer lines for the two duplicate recessive sterile lines. The results obtained are as follows.
In the 131 F3 lines obtained from F2 fertile plants of the two categories of materials, 39 lines displayed fertility segregation, 92 lines were fertile completely.χ2 test showed that the interaction of recessive epistatic suppression gene and duplicate recessive sterile genes in B. napus wasn’t specific. In the 147 lines obtained from the testcross between F2 fertile plants of the two categories of materials and sterile plants of the two duplicate recessive sterile materials, 81 lines displayed fertility segregation, 66 lines were fertile completely. Inspected byχ2, our result doesn’t meet the two assumptions. But our result met the inference that the testcross progenies won’t have sterile completely lines if recessive epistatic suppression gene hasn’t specific inhibition on two duplicate recessive sterile genes.These results revealed that the previous assumption, which assumed that the interaction of recessive epistatic suppression gene and duplicate recessive sterile genes in B. napus is non-specific, is not correct. Our results showed that the recessive epistatic suppression gene has specific suppression on its sterile genes in B. napus.
In the second part, genetic diversity of 70 B. napus varieties of approvalled during recent years in China was analyzed with SSR. In total, 113 fragments were detected with size ranged from 100 to 600 bp and 104 were polymorphism, the polymorphism rate was 92%. On average, each primer combination generated 4.3 polymorphic fragments. Principal analysis revealed that all varieties scattered well in bi-plot, and accessions from the same geographic origin did not group together. There existed considerable variance among recently registered B. napus cultivars in China. The components of variation within populations was dominant over that among populations. Varieties in middle of Yangtze valley possessed the richest genetic variation in the present investigation.
引文
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