小麦抗病分子标记辅助育种技术研究
摘要
本研究运用GISH、RAPD和SCAR分析方法，对抗全蚀病、黄矮病和兼抗黄矮病白粉病的小麦杂交后代材料进行分析，获得了一些对育种实践有重要意义的易位系、异附加系材料和几个可用于辅助育种的特异性分子标记。
选育抗病品种是防治全蚀病最为有效、经济和安全的方法。本研究先用基因组原位杂交(GISH)对经鉴定抗全蚀病的材料进行分析，检测出三个抗性源于簇毛麦的端体异附加系材料，再利用RAPD方法，对这三个材料和抗感病亲本进行分析，得到了两个与全蚀病抗性基因紧密相关的特异性RAPD标记。
目前，研究得较为深入的抗黄矮病材料的抗性基因Bdv2源于中间偃麦草7X。中间偃麦草与小麦杂交获得的部分双二倍体无芒中4所含的抗病基因位于中间偃麦草第二染色体组2Ai-2上，相对于Bdv2基因，其对中国流行的GPV和GAV黄矮病株系表现出更好的抗性，因此对具2Ai-2的材料进行易位系选育有重要的意义。本研究采用RAPD方法，对具2Ai-2端体和完整染色体异附加或异代换系进行分析，从随机引物中，选出两个分别位于2Ai-2染色体两臂上的分子标记，然后用这两个标记对中间偃麦草和普通小麦的杂交后代进行筛选，选出小麦抗黄矮病易位系新材料。
选育携带多个抗性基因的多基因聚合体材料具有重要的育种意义，但是传统的育种方法难以同时快速准确鉴定两个以上的抗性基因。分子标记辅助育种技术的发展，使对多基因聚合体材料的选育变得简单易行。本研究利用黄矮病抗性基因Bdv2特异的SCAR标记SCW-37，及对白粉病抗性基因Pm13和Pm21特异的标记，对54株杂交回交后代进行扩增，检测选择到Pm13+Pm21+Bdv_2抗病优质材料1株1484-16；Pm13+Bdv_2一株1484-9；Pm21+Bdv_2基因聚合株11株。
In this research , by using of genomic in situ hybridization (GISH) , random amplified polymorphism DNA (RAPD) technology and three specific SCAR molecular markers, the author analyzed wheat hybrid generations carrying the desired genes that were resistant to take-all, BYDV and/or powdery mildew, and developed some alien chromosome addition and translocation lines that were important in breeding practice, and some specific molecular markers that could be applied in marker-assisted selection.
Breeding resistant or immune cultivars was the most economical and efficient way to control wheat take-all disease. In this study, the author used GISH technology to analyze the material that was resistant to take-all in test, and got three chromosome arm addition lines that carried the resistance genes derived from the amphiploid of Triticum-Haynaldia. The above arm addition lines and their resistant and sensitive parents were analyzed by RAPD method and two specific molecular markers that were tightly linked to resistant gene were obtained.
Amount studies on BYDV resistant gene Bdv2 on 7X of Thinopyrum intermedium were reported.Another resistance gene on 2Ai-2 of chromosome group 2 of Thinopyrum intermedium from amphiploid Zhong 4 Awnless had better resistance to GPV and GAV serotype of BYDV that were prevail in China ,so it's important to develop the chromosome translocation lines of Zhong 4 Awnless. In this research, the author utilized RAPD method to analyze 2Ai-2 chromosome or arm of addition or substitution lines of Zhong 4 Awnless . Two RAPD molecular markers located on different arms of 2Ai-2 chromosome from Thinopyrum intermedium were selected and the generations of the hybrid generation derived from Zhong 4 Awnless and wheat were analyzed. Two chromosome translocation lines were screened for wheat breeding program to control BYDV.
It's important to breed varieties(lines) with multiple disease resistance genes, but it's difficult for traditional breeding ways to identify two or more resistant characteristics quickly and immediately at the same time. Marker-assisted selection technology made this issue simple and feasible. In this study, the author used a molecular marker SCW-37 that was specific for BYDV resistant gene Bdv2, and two molecular markers that were specific for powdery mildew resistant genes Pm13 and Pm21 to analyze 54 candidate plants. As a result, 1 plant pyramided Pm13+Pm21+ Bdv2 resistant genes, 1 plant pyramided Pm13+ Bdv2, 11 plants pyramided Pm21+ Bdvi, were selected for further breeding practice.
选育抗病品种是防治全蚀病最为有效、经济和安全的方法。本研究先用基因组原位杂交(GISH)对经鉴定抗全蚀病的材料进行分析，检测出三个抗性源于簇毛麦的端体异附加系材料，再利用RAPD方法，对这三个材料和抗感病亲本进行分析，得到了两个与全蚀病抗性基因紧密相关的特异性RAPD标记。
目前，研究得较为深入的抗黄矮病材料的抗性基因Bdv2源于中间偃麦草7X。中间偃麦草与小麦杂交获得的部分双二倍体无芒中4所含的抗病基因位于中间偃麦草第二染色体组2Ai-2上，相对于Bdv2基因，其对中国流行的GPV和GAV黄矮病株系表现出更好的抗性，因此对具2Ai-2的材料进行易位系选育有重要的意义。本研究采用RAPD方法，对具2Ai-2端体和完整染色体异附加或异代换系进行分析，从随机引物中，选出两个分别位于2Ai-2染色体两臂上的分子标记，然后用这两个标记对中间偃麦草和普通小麦的杂交后代进行筛选，选出小麦抗黄矮病易位系新材料。
选育携带多个抗性基因的多基因聚合体材料具有重要的育种意义，但是传统的育种方法难以同时快速准确鉴定两个以上的抗性基因。分子标记辅助育种技术的发展，使对多基因聚合体材料的选育变得简单易行。本研究利用黄矮病抗性基因Bdv2特异的SCAR标记SCW-37，及对白粉病抗性基因Pm13和Pm21特异的标记，对54株杂交回交后代进行扩增，检测选择到Pm13+Pm21+Bdv_2抗病优质材料1株1484-16；Pm13+Bdv_2一株1484-9；Pm21+Bdv_2基因聚合株11株。
In this research , by using of genomic in situ hybridization (GISH) , random amplified polymorphism DNA (RAPD) technology and three specific SCAR molecular markers, the author analyzed wheat hybrid generations carrying the desired genes that were resistant to take-all, BYDV and/or powdery mildew, and developed some alien chromosome addition and translocation lines that were important in breeding practice, and some specific molecular markers that could be applied in marker-assisted selection.
Breeding resistant or immune cultivars was the most economical and efficient way to control wheat take-all disease. In this study, the author used GISH technology to analyze the material that was resistant to take-all in test, and got three chromosome arm addition lines that carried the resistance genes derived from the amphiploid of Triticum-Haynaldia. The above arm addition lines and their resistant and sensitive parents were analyzed by RAPD method and two specific molecular markers that were tightly linked to resistant gene were obtained.
Amount studies on BYDV resistant gene Bdv2 on 7X of Thinopyrum intermedium were reported.Another resistance gene on 2Ai-2 of chromosome group 2 of Thinopyrum intermedium from amphiploid Zhong 4 Awnless had better resistance to GPV and GAV serotype of BYDV that were prevail in China ,so it's important to develop the chromosome translocation lines of Zhong 4 Awnless. In this research, the author utilized RAPD method to analyze 2Ai-2 chromosome or arm of addition or substitution lines of Zhong 4 Awnless . Two RAPD molecular markers located on different arms of 2Ai-2 chromosome from Thinopyrum intermedium were selected and the generations of the hybrid generation derived from Zhong 4 Awnless and wheat were analyzed. Two chromosome translocation lines were screened for wheat breeding program to control BYDV.
It's important to breed varieties(lines) with multiple disease resistance genes, but it's difficult for traditional breeding ways to identify two or more resistant characteristics quickly and immediately at the same time. Marker-assisted selection technology made this issue simple and feasible. In this study, the author used a molecular marker SCW-37 that was specific for BYDV resistant gene Bdv2, and two molecular markers that were specific for powdery mildew resistant genes Pm13 and Pm21 to analyze 54 candidate plants. As a result, 1 plant pyramided Pm13+Pm21+ Bdv2 resistant genes, 1 plant pyramided Pm13+ Bdv2, 11 plants pyramided Pm21+ Bdvi, were selected for further breeding practice.
引文
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