摘要
利用农杆菌介导的子叶节遗传转化体系,将Cry1Ac/Ab融合抗虫基因表达载体导入栽培大豆品种GP03-8-23中,通过Cry1Ac/Ab融合基因在大豆中的表达,获得了抗虫转基因大豆新材料。试验共转化子叶节外植体1 540个,再生转化苗经PCR、Southern杂交和Bar试纸条检测,获得85株阳性转基因植株,平均转化率为5.47%。利用实时荧光定量PCR技术(qRT-PCR)检测了目标基因在转基因大豆不同株系及不同组织中的表达量。结果表明:不同转基因株系表达量存在显著差异,目标基因在大豆根、茎、叶、子粒中均有表达,在叶中的表达量最高,根中的表达量较高,茎和子粒中的表达量最低。经室内抗食叶性害虫离体鉴定,获得抗虫性较好的T1代转Cry1Ac/Ab基因大豆材料,其中1份表现为高抗,抗虫性显著优于对照,可为培育抗虫大豆新品系提供新的种质材料。
Insect-resistant CrylAc/Ab fusion expression vector was transformed into soybean cuhivar GP03-8-23 by Agrobacterium tumefaciens infection of cotyledonary nodes, and transgenic soybean lines containing CrylAc/Ab fusion genes showed resistance to insects. At the beginning, 1 540 soy- bean cotyledonary node explants were transformed. Through molecular examination of the transgenic soybean plants, 85 positive transgenic plants were obtained, and transformation rate was 5.47%. The results of Quantitative Real Time PCR （qRT- PCR） showed that the expression levels of CrylAc/Ab fusion genes were the highest in the leaves, higher in the roots, and high in stems and seeds. The resistance of one T1 transgenic soybean line to soybean pod borer was significantly higher than the control, which will provide new germplasm materials for breeding new soybean varieties with insect-resistant characteristics.
作者
钱雪艳
苏颖
姚瑶
李启云
郭东全
董英山
QIAN Xueyan;SU Ying;YAO Yao;LI Qiyun;GUO Dongquan;DONG Yingshan(Institute of Agricultural Biotechnology, Jilin Academy of Agricultural Sciences, Changchun 130033, China)
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2017年第5期527-533,共7页
Journal of Jilin Agricultural University
基金
农业部转基因生物新品种培育重大专项(2016ZX08004-004)
关键词
大豆
抗虫基因
遗传转化
苏云金芽孢杆菌
soybean
insect-resistant gene
genetic transformation
Bacillus thuringiensis
