【作者】 常利芳；

【导师】 马峙英； 王立新；

【作者基本信息】 河北农业大学 ， 作物遗传育种， 2003， 硕士

【摘要】 本研究采用花粉管通道技术分别将含有抗白粉病基因、抗条锈病基因以及高分子量谷蛋白5+10亚基基因导入优良品种北农6号和京冬8号，在保持受体品种原有优良性状的基础上，通过抗病性鉴定、分子标记鉴定及农艺性状鉴定，筛选出具有抗病、优质基因的植株和抗病性稳定遗传的优良品系或株系。结果如下： 1．在以含有高分子量谷蛋白5+10亚基的品种荔垦2号为供体、京冬8号为受体的D₁代中，通过分子标记鉴定获得了2株具有优质高分子量谷蛋白5+10亚基基因的的变异植株，变异率为3.3％。 2．在以含有高分子量谷蛋白5+10亚基品种荔垦2号、抗白粉病品系01067和抗条锈病品系Cp87-26-12的混合总DNA为供体、京冬8号为受体的组合中，通过分子标记鉴定，在D₁代筛选到2株具有优质高分子量谷蛋白5+10亚基基因的变异植株，变异率为1.4％。同时，获得4株具有抗条锈病基因的变异植株，变异率为2.8％。 3．在以含有抗白粉病基因的复壮30为供体、京冬8号为受体的组合中，通过白粉病抗性鉴定和分子标记鉴定以及农艺性状鉴定，D₁代筛选到7株农艺性状优良的抗白粉病植株，变异率7％；继而在D₂代中获得了1株保持抗性的单株，分子鉴定表明复壮30的抗性基因遗传到了D₂代，同时抗病株的农艺性状与京冬8号基本一致。 4．在以抗白粉病品种复壮30、矮杆品种农林10和抗条锈病的Cp87-26-12的混合总DNA为供体，京冬8号为受体的组合中，通过分子标记鉴定，D₁代筛选到3株具有复壮30的抗性基因变异的植株，变异率为6％；继而通过抗病性鉴定及农艺性状鉴定，在D₂代中筛选到6株抗病植株。同时，经田间条锈病抗病性和农艺性状鉴定，D₁代获得了5株抗病单株，以及2个株高发生变化的植株，并且其中一株的D₂代的株高性状已经稳定，株高大约在60～65cm。 5．在以抗白粉病品种复壮30为供体、北农6号为受体的组合中，经过分子鉴定、抗性鉴定及农艺性状鉴定，在D₃代获得200多株抗病单株，其中1个D₃代株系全部抗病且农艺性状优良，证明已得到1个抗白粉病的转基因株系；D₁代获得2个具有北农6号优良性状且白粉病抗性稳定的新品系，并且得到5个两个白粉病抗性稳定、农艺性状优良的新株系，分子鉴定表明抗性基因遗传到了D₄代。 6．在以抗白粉病品种小白冬麦为供体、北农6号为受体的组合中，通过抗性和农艺性状鉴定，D₃代获得1个白粉病抗性稳定、性状优良的株系，D₄代获得2个白粉病抗性稳定、农艺性状优良的新品系。 问时，恤过分析花粉管通道技术转基囚后代材料的坦传学农现，还得主帅11【一A论： 1 转化率高 采川花粉管通道技术转基因的转化率多在1％－10％，本研究中，在织合京冬S号／荔RZq的止代中筛选到了2株具有HMW－GSS＋10基冈标记的变异植株，变异率为3．30．仕组合京冬8号／混合mA的D代筛选到2株具有HMW－GSS＋10亚基基厂Ifd；记的变异植株，变异率为1．4％．同时，筛选到4株具有抗条锈病基冈的变异植株，艾并率为2．8％．在京冬8号／复壮30的广代筛选到7株农艺性状忧良的获得抗白粉病基冈的变异柏株，变异频率为7％；组合京冬8号／混合DNA的巳代S株获得抗一粉病基冈植株，变异顾率为6％．可见该技术的转化率比较高的。 2 后代材料变异)’泛 本研究中，后代材料获得了)“泛的变异，尤其是D．；代材料，除了抗病性变发生异外，转基因后代株系中株高、芒性、抽穗期、穗型、于粒形状以及于粒颜色阵也发0二一定的变异，其中株高变异表现最为明显，变异株的株高比受体高山20～30CCI左右。 ：5 变异性状稳定快 以北农6号为受体的转化后代中，在0．；代筛选到汕个FI粉炳抗性己经稳定的株系及多个抗性Y规律性分离的株系，D代义获得了d个抗病性稳定，农艺性状优良的品系及5个抗病性较好的株系，说明花粉管通道法的转化后代变异性状能够在3～，l代 l)J稳定。更多还原

【Abstract】 In order to obtain choicely breeding resource several wheat varieties genomic DNA which include different resistant disease and dwarf gene and HMW-GS 5+10 gene are respectively introduced into the popularize wheat varieties Beinong #6 and Jingdong #8, which need to be improved through pollen tube pathway. Some superior plants and lines with fine qualities are selected through resistant and molecular and agricultural identifying. The main results are as follows:1. Two plants with HWM-GS 5+10 gene are selected from the D, generation. Liken #2 with quality gene as the donor and the acceptor is Jingdong #8. The ratio is about 3.3%.2. In the combination base on the donor composed of mixed genomic DNA of three different varieties contained Liken #2 with HMW-GS5+10 gene and O1067 with Pml3 gene and Cp87-26-12 with Yrl7 gene and the acceptor also is Jingdong #8, two plants which contained HMW-GS 5+10 gene were selected via resistant and molecular and agricultural test. The ratio is 1.4%. And four plants with resistant stripe rust gene are also singled out via identifications. The ratio is 2.8%.3. In the combination of Fuzhuang 30# as the donor and Jingdong #8 as the acceptor, seven plants with resistant gene of powdery mildew and perfect agricultural character are singled out of D1 progeny through molecular identifying and agricultural identifying. The ratio is 7%. Further selecting in D2 generation one plant with resistant Pm gene and agricultural test is the same as molecular identifying.4. In the combination of the mixed genomic DNA as the donor composed of Fuzhuang 30# with resistant Pm gene, Nonglin 10# with dwarf gene and Cp87-26-12 with Yr17 gene and Jingdong 8# as the acceptor, there are three plants are singled out of DI generation with molecular variance and six resistance plants are selected from D2 generation via several forms identifying. Similarly, five plants are picked out with its variance of resistance of stripe rust disease in DI generation and two plants height fall.5. In the combination of Fuzhuang 30# as the donor and Beinong #6 as the acceptor, over 200 plants are obtained in D3 progenies via resistant and molecular and agricultural identifying, among these plants one line with perfect agricultural appearance and steadily resistance, which can be ascertained as a new line. In the D4 progenies, two lines with steadily resistance and similar appearance to Beinong 6# are singled out via resistant test and five lines with Pm resistant gene but intervenient appearance of parents.6. In the combination of Xiaobaidongmai as the donor and Beinong #6 as the acceptor, one line with perfect agricultural appearance and steadily resistance, which can be ascertained as a new line are obtained in D3 progenies. In the D4 progenies, two new lines with steadily resistance and similar appearance as Beinong 6# are singled out via resistant identifying and agricultural test.At the same time some conclusions by analyzing the genetic behavior of transgenic progeny as followings.1. The transform ratio is highIn this study, in the combination Jingdong #8/Liken 2#, two plants with HWM-GS 5+10 gene are selected from the D| generation. The ratio is about 3.3%. In the combination Jingdong #8/mixed genomic DNA of three different varieties contained Liken #2 with HMW-GS5+10 gene and O1067 with Pml3 gene and Cp87-26-12 with Yr17 gene, two plants which contained HMW-GS 5+10 gene were selected via molecular. The ratio is 1.4%. At the same time, four plants with resistant stripe rust gene also single out via identifications. The ratio is 2.8%. In the combination of Jingdong #8/Fuzhuang 30#, seven plants with Pm gene and perfect agricultural character are singled out of D, progeny through molecular and agricultural identifying. The ratio is 7%. In the combination of the mixed genomic DNA as the donor composed of Fuzhuang 30# with resistant Pm gene, Nonglin 10# with dwarf gene and Cp87-26-12 with Yrl7 gene and Jingdong 8# as the acceptor, there are three plants are singled out of DI generation w更多还原