抗玉米丝黑穗病转基因株系的分子检测及抗病株系选育  

Molecular Check and Measure of Transgenic Maize Lines Resistancing to Maize Head Smut and Breeding Selection for Transgenic Lines Resistancing to Disease

作　　者：杜建中 孙毅 郝曜山 刘俊玲 

机构地区：[1]山西省农业生物技术研究中心,山西太原030031 [2]清徐县农业局,山西清徐030400

出　　处：《华北农学报》2006年第5期99-104,共6页Acta Agriculturae Boreali-Sinica

基　　金：国家"863"计划项目(2001AA212131)

摘　　要：采用花粉介导方法将质粒pGLII-RC-1导入玉米自交系海92—1中，获得了Tn种子128粒，种子经潮霉素抗性筛选得到抗潮霉素植株16株。对抗性植株分离的统计分析表明，多数情况下几丁质酶基因以单拷贝形式导入转基因植株，后代分离比约为3：1。抗性苗移栽到大田后，5—6叶期取样进行PCR扩增、PCR Southem blot杂交分析，对T1的分析结果表明，几丁质酶基因已导入转基因植物细胞中。对T2，L植株的检测结果显示，目的基因已整合到转化植株基因组中并可随植株世代稳定遗传。田间抗病鉴定结果表明，转基因植株及后代的玉米丝黑穗病发病率明显比对照降低。根据分子检测及田间抗病鉴定结果，得到GH05024，GH050282个抗丝黑穗病的纯合转基因株系。In this study 128 transgenic maize seeds that were obtained from transgenic plants transformed by pollenmediated approach on maize（Zea mays L.） inbred line Hal 92-1,the vector was plasmid pGL II-RC-1 .The result of seeds selected by hygromycin-screening had proved that 16 plants was hygromycin-resistant plants. Statistics analysis for the hygromycin-resistant plants suggested that single copy of the chintinase gene was transferred into the cell of majority of TO transgenic plants, segregation ratio of the progeny of transgenic plants for hygromycin resistant and susceptible was about 3： 1, P〈 0.01 .The results of detecting to T1 transgenic plants by PCR amplification and PCR Southern blot hybridization showed that chitinase gene was transferred into the cells of transgenic plants . The same methods detected to T2 and T3, their results indicated that chitinase gene was integrated into the genome of transgenic plants and stable inheritance in transgenic plants and their progeny. The results of Bio-assay for head smut-resistance activity indicated that the disease incidence of transgenic plants and their progeny was lower than that of control plants. Based on the results above, GH05024 and GH05028 . 2 transgenic maize lines that resisted to maize head smut were obtanined by breeding selection.

关 键 词：玉米 几丁质酶基因 转基因 丝黑穗病 选育 

分 类 号：S435.131.42[农业科学—农业昆虫与害虫防治]