簇毛麦与普通小麦杂种后代贮藏蛋白和小麦品系抗白粉病基因的研究
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摘要
小麦白粉病是影响小麦生产的世界性病害,在我国的危害日益严重。发掘、筛选和研究新的抗白粉病基因并应用于育种中,具有重要的学术意义和经济价值。同时,在培育小麦新品种过程中,大量使用一些骨干亲本,使得小麦品质大大降低。因此,在选育抗病高产品种的同时,应把高品质作为育种的目标之一。小麦异缘神、属含有许多与丰产、优质、抗病、抗逆等相关的基因,是小麦遗传改良的巨大基因库。
本研究对簇毛麦与绵阳11杂种后代的151个稳定品系贮藏蛋白进行了遗传分析,旨在对以后的进一步选育提供依据。并对通过远缘杂交获得的小麦品系″101-3″含有的1个未知抗白粉病基因进行了白粉病田间抗性鉴定和RAPD标记分析,旨在建立抗白粉病基因RAPD分子标记,为进一步克隆抗白粉病基因、进行分子标记辅助育种提供理论依据。主要研究结果如下:
1采用SDS-PAGE法和APAGE法分析151份品系的高分子量麦谷蛋白亚基(HMW-GS)的组成和醇溶蛋白遗传多样性,结果表明:(1)品系共出现18种HMW-GS组合,Glu-A1、Glu-B1、Glu-D1三个位点上分别检测到2、5、6种不同的亚基类型。Glu-B1位点出现20、7+9、7+8+9、7等新类型;Glu-D1位点出现2+12、5+10+12等五种新的类型。优质亚基组合1,7+8,5+10的比例为21.19%。(2)醇溶蛋白APAGE共分离出差异谱带51条,126种谱带组合。α、β、γ、ω区分别检测出11、8、15和17条谱带,表明Gli-1变异最丰富,且四个区均有簇毛麦的谱带出现。90%品系的谱带数变幅在20～30之间,平均为23.93条。聚类分析表明,后代品系与亲本之间存在较大变异。
2对品系101-3与中国春杂交F2代进行田间抗病性鉴定及RAPD分子标记,结果表明:(1)杂交组合101-3//中国春的F2分离群体进行田间抗病性鉴定结果显示:抗、感分离比例符合3:1,抗病性由显性单基因控制。(2)用340条RAPD引物对F2代抗感分离群体及抗、感病亲本进行标记筛选,结果发现,有316条能扩增出清晰的谱带,分子量范围在0.2—2.5kb之间。引物E9的扩增结果表现出特异性,经连锁分析,E9与抗病基因连锁,遗传距离为9.6cM。
Quality of wheat had been gradually reduced by using same parental genotypes in breeding. Therefore,high-quality was of importance with high yield in production.Wheat powdery mildew was a damaging and strongly epidemic disease of wheat wordwide.It had important academic and economic significances to create and screen and study the new powdery mildew resistance genes and to utilize these genes in breeding programs.Many alien species related to Triticum aestivum contains favorable genes and possess many useful characteristics as genetics resources for wheat genetic improvement.
Genetic analysis of wheat storage proteins in 151 stable lines from progenies of common wheat Mianyang 11(MY11) with Dasypyrum villosum(L.) Candary were studied and disease-risistant determination and RAPD analysed for powdery mildew resistance gene in a line 101-3 derived from the progenies of MY 11 with D.villosum to establish RAPD markers related to powdery mildew resistance gene were carried out.The main results were list follows:
1 SDS-PAGE and APAGE analysis were used to analyse the variations of high molecular weight glutenin subunit and gliadin compositions in 151 lines from the progenies of MY11×D. villosum.The results showed that:(1) there were 18 different HMW-GS composition types in 151 lines.2,5 and 6 types of HMW -GS types were found in Glu-A1,Glu-B1 and Glu-D1 respectively. At Glu-B1 loci,20,7+9,7 were new subunit types.At Glu-D1 loci,there were five new subunit types.The rate of the composition of high molecular weight glutenin subunits with 1,7+8 and 5+ 10 was 21.19%;(2) 51 different gliadin bands and 126 patterns were found.There were 11,8,15, 17 variation binds in the zones ofα、β、γandωrespectively,which showed the loci of Gli-1 possessed more extensive variation.90%of studied lines had 20 to 30 bands with average of 23.93 bands.Cluster analysis showed that there were abundant variations at gliadin loci between tasted lines and their parents.
2 The analyses of disease-risistant determined and RAPD using the parents and F2 progeny between Chinese Spring×101-3 indicated that(1) the segregation of risistant and susceptible plants for powdery mildew in F2 progeny was goodness fitted with 3:1,and it was controlled by a single dominant gene.(2) 316 of 340 RAPD primer(92.9%) producced the DNA fragments ranging from 0.2kb-2.5kb.Polymorphisms between resistant and suscepitable plants were observed using the primer SBSE09.SBSE09 was proved to link to the resistance gene with genetic distance of 9.6 cM.
本研究对簇毛麦与绵阳11杂种后代的151个稳定品系贮藏蛋白进行了遗传分析,旨在对以后的进一步选育提供依据。并对通过远缘杂交获得的小麦品系″101-3″含有的1个未知抗白粉病基因进行了白粉病田间抗性鉴定和RAPD标记分析,旨在建立抗白粉病基因RAPD分子标记,为进一步克隆抗白粉病基因、进行分子标记辅助育种提供理论依据。主要研究结果如下:
1采用SDS-PAGE法和APAGE法分析151份品系的高分子量麦谷蛋白亚基(HMW-GS)的组成和醇溶蛋白遗传多样性,结果表明:(1)品系共出现18种HMW-GS组合,Glu-A1、Glu-B1、Glu-D1三个位点上分别检测到2、5、6种不同的亚基类型。Glu-B1位点出现20、7+9、7+8+9、7等新类型;Glu-D1位点出现2+12、5+10+12等五种新的类型。优质亚基组合1,7+8,5+10的比例为21.19%。(2)醇溶蛋白APAGE共分离出差异谱带51条,126种谱带组合。α、β、γ、ω区分别检测出11、8、15和17条谱带,表明Gli-1变异最丰富,且四个区均有簇毛麦的谱带出现。90%品系的谱带数变幅在20～30之间,平均为23.93条。聚类分析表明,后代品系与亲本之间存在较大变异。
2对品系101-3与中国春杂交F2代进行田间抗病性鉴定及RAPD分子标记,结果表明:(1)杂交组合101-3//中国春的F2分离群体进行田间抗病性鉴定结果显示:抗、感分离比例符合3:1,抗病性由显性单基因控制。(2)用340条RAPD引物对F2代抗感分离群体及抗、感病亲本进行标记筛选,结果发现,有316条能扩增出清晰的谱带,分子量范围在0.2—2.5kb之间。引物E9的扩增结果表现出特异性,经连锁分析,E9与抗病基因连锁,遗传距离为9.6cM。
Quality of wheat had been gradually reduced by using same parental genotypes in breeding. Therefore,high-quality was of importance with high yield in production.Wheat powdery mildew was a damaging and strongly epidemic disease of wheat wordwide.It had important academic and economic significances to create and screen and study the new powdery mildew resistance genes and to utilize these genes in breeding programs.Many alien species related to Triticum aestivum contains favorable genes and possess many useful characteristics as genetics resources for wheat genetic improvement.
Genetic analysis of wheat storage proteins in 151 stable lines from progenies of common wheat Mianyang 11(MY11) with Dasypyrum villosum(L.) Candary were studied and disease-risistant determination and RAPD analysed for powdery mildew resistance gene in a line 101-3 derived from the progenies of MY 11 with D.villosum to establish RAPD markers related to powdery mildew resistance gene were carried out.The main results were list follows:
1 SDS-PAGE and APAGE analysis were used to analyse the variations of high molecular weight glutenin subunit and gliadin compositions in 151 lines from the progenies of MY11×D. villosum.The results showed that:(1) there were 18 different HMW-GS composition types in 151 lines.2,5 and 6 types of HMW -GS types were found in Glu-A1,Glu-B1 and Glu-D1 respectively. At Glu-B1 loci,20,7+9,7 were new subunit types.At Glu-D1 loci,there were five new subunit types.The rate of the composition of high molecular weight glutenin subunits with 1,7+8 and 5+ 10 was 21.19%;(2) 51 different gliadin bands and 126 patterns were found.There were 11,8,15, 17 variation binds in the zones ofα、β、γandωrespectively,which showed the loci of Gli-1 possessed more extensive variation.90%of studied lines had 20 to 30 bands with average of 23.93 bands.Cluster analysis showed that there were abundant variations at gliadin loci between tasted lines and their parents.
2 The analyses of disease-risistant determined and RAPD using the parents and F2 progeny between Chinese Spring×101-3 indicated that(1) the segregation of risistant and susceptible plants for powdery mildew in F2 progeny was goodness fitted with 3:1,and it was controlled by a single dominant gene.(2) 316 of 340 RAPD primer(92.9%) producced the DNA fragments ranging from 0.2kb-2.5kb.Polymorphisms between resistant and suscepitable plants were observed using the primer SBSE09.SBSE09 was proved to link to the resistance gene with genetic distance of 9.6 cM.
引文
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