转基因玉米株系纯合系选育及其农艺性状表现

Selection Breeding of Transgenic Maize Lines and Their Performance of Agronomic Characters

目的:以携带水稻矮缩病毒(RDV)运动蛋白缺陷型(MP-)基因的转基因玉米种子T0为试验材料,通过分子分析、抗病鉴定及农艺性状筛选得到转基因纯合株系。方法:首先对转化种子进行潮霉素抗性筛选,其后对各代转基因材料进行PCR检测、农艺性状调查和抗病鉴定。结果:从645粒T0转化种子得到抗潮霉素植株246株,即T1转基因植株。T1、T2、T3、T4代材料的PCR检测阳性率分别为56.9%、83.9%、94.6%和99.8%,证明RDVMP-基因已被导入玉米自交系中,且目的基因可以稳定遗传到转基因植株及其后代。田间人工接种抗病鉴定结果表明,经过连续筛选,转基因后代植株(系)的抗病性不断提高,T1、T2、T3和T4代中的高抗病材料分别占总材料数的8.9%、31.5%、70.7%和100%;所选纯合系连续2年的发病率均为0,抗病性比相应对照株系提高4级。农艺性状调查结果表明,转基因株系的株高比对照株系高15.0～33.4cm,穗位高提高13.4～20.2cm,;穗长增加2.0～3.8cm,穗粒数增加10.2～22.8粒。结论:根据分子检测、田间抗病鉴定及农艺性状鉴定结果,选育到96C0502、96C0507和96C0513等抗矮花叶病转基因玉米纯合株系。

Objective： In order to obtain transgenic pure lines, the transgenic maize carrying RDV MP- gene inbred lines seeds To with RDV MP- gene in them were subjected to further studies and selection. Methods： First, the To seeds were subjected to hygromycin-serening, then, the PCR detection, the investigation of disease-resistance and their agronomic characters performance of transgenic line T1 to T4 generation were conducted. Results： From putative 645 To seeds, 246 hygromycin resistant plants（T1 plants） were obtained. PCR amplification test to T,, T2, T3 and T4 transgenic plants（lines） dis- played that the ratio of positive results to total plants（lines） were 56.9%, 83.9%, 94.6% and 99.8%, respectively, which suggested that RDV MP- gene was introduced into transgenic maize plants and could be stablely inherited. MDMV resis- tance evaluation to each generation plants （lines） by artificial inoculation showed that the disease resistance of transgenic plants（lines） was enhanced gradually, the ratio of plant（lines） number of highly disease-resistance in T1 to T4 generation to total plants（lines） number were 8.9%, 31.5%, 70.7% and 100%, respectively. Compare with control, the average disease in- cidence of pure transgenie lines obtained by selection breeding were 0 during 2 years, their disease-resistant activities en- hanced 4 degrees. Compared with control plants, the plants height increased by 15.0~33.4 cm, the ear height enhanced 13.4-20.2 era, ear length increased 2.0-3.8 cm, kernels/ear increased 10.2-22.8. Conclusion： The transgenic MDMV resistant maize pure lines 96C0502, 96C0507 and 96C0513 were obtained.