转水稻NibT基因玉米植株的获得及抗病性研究  

Transgenic Maize Plants with Rice NibT Gene and Their MDMV-Resistance

作　　者：杜建中[1] 孙毅[1] 王景雪[2] 郝曜山[1] 王亦学[1] 张丽君[1,3] 

机构地区：[1]山西省农业科学院生物技术研究中心,太原030031 [2]山西大学生命科学技术学院,太原030006 [3]山西农业大学,山西太谷030801

出　　处：《西北植物学报》2011年第5期893-901,共9页Acta Botanica Boreali-Occidentalia Sinica

基　　金：转基因生物新品种培育重大专项重点课题(2009ZX08003-017B;2009ZX08003-005B)

摘　　要：以玉米（Zea maysL.）自交系金黄96B为受体材料,供体为质粒pWM101并携带有水稻矮缩病病毒复制酶基因Nib的提前终止突变体基因NibT,采用超声波处理花粉介导植物基因转化方法将NibT-基因导入受体,经PCR检测和Southern杂交分析证实获得转基因植株,进而对T1－T3代转基因植株（株系）进行分子分析、田间抗病鉴定和农艺性状调查.逐代分子检测分析结果证明,目的基因可稳定遗传.抗病鉴定结果证明转基因植株（株系）各代抗病水平基本一致,抗病性比对照提高3级.农艺性状调查分析表明,与对照比较,转基因植株株高增加7－18cm、穗位高增高0－13 cm、穗长增加0.7－2.1 cm、穗粒数多8－35粒、百粒重增加1.1－2.6 g,转基因株系与阴性对照间、各代转基因株系相互间都差异显著（P〈0.05）;调查还发现转基因植株的株高和穗位高随着世代的增加,与对照间的差异逐代减少.研究也说明,超声波处理花粉介导植物基因转化方法是一种简捷、快速和有效的植物转化工具.In this study,maize（Zea mays L.）inbred line ＇Jinhuang 96B＇ was used as acceptor,plasmid pWM101,containing NibT gene that derived from the mutant of rice stunt disease virus replicase gene Nib,was used as donator.NibT gene was transformed into maize plants by supersonic wave pollen treating process to induce plant＇s gene transfer method.The transgenic plants were obtained through PCR and Southern blot hybridization detection.Further molecular detection for T1~T3 generation transgenic plants and disease-resistance analysis in field as well as agronomical traits investigation for them were conducted.The results of molecular detection to each generation transgenic plants showed that target gene could stably inheritance from parents to their progenies.Disease-resistance analysis by artificial MDMV severity confirmed that the disease-resistance of transgenic plants of each generation without difference,and the disease-resistance was 3 level higher than that of control plants.Also,the investigation results of agronomical traits displayed that there was a significant difference（P〈0.05） between transgenic plants and non-transformed plants in 5 traits,compared with that of control plants,the plant height,ear height,Ear length,grains/ear and 100-grain Wt.were incresed 7~18 cm,0~13 cm,0.7~2.1 cm,8~35 grains and 1.1~2.6 g,respectively.Even between each transgenic line in same generation,there was a great difference.Moreover,from T1 to T3,the plants height,ear height of transgenic line progenies tended to recover to the characters of non-transgenic plants.Above all,supersonic wave pollen treating process to induce plant＇s gene transfer method is a simple,fast and effective tool used for plant transformation.

关 键 词：玉米 NibT基因 超声波处理花粉介导植物基因转化 抗病性 玉米矮花叶病毒(MDMV) 

分 类 号：Q789[生物学—分子生物学]