农杆菌介导的双价抗虫基因对番茄遗传转化的研究  

Agrobacterium-mediated introduction of insect-resistant genes into tomato

作　　者：杨俊杰[1] 于惠敏[1] 侯丙凯[2] 王兴智[3] 

机构地区：[1]齐鲁师范学院生物系,山东济南250013 [2]山东大学植物细胞工程与种质创新教育部重点实验室,山东济南250100 [3]东北师范大学生命科学学院,吉林长春130024

出　　处：《山东大学学报（理学版）》2013年第7期9-13,共5页Journal of Shandong University(Natural Science)

基　　金：齐鲁师范学院青年教师基金资助项目(2011L1514)

摘　　要：采用农杆菌介导法将含有苏云金芽孢杆菌毒蛋白基因(CryIAc)与半夏凝集素抗虫基因(Pta)的高效植物表达载体pCAMBIA3300转入番茄品系Micro Tom的子叶外植体中。经过共培养、除草剂筛选和分化再生,获得了24个具有除草剂抗性的株系。再将转化后的番茄植株经过PCR检测和Southern Blot检测,确定检测后呈阳性反应的株系为8个。通过小菜蛾幼虫初步抗性试验证明,转基因株系表现出较强的抗虫性。实验结果为进一步研究番茄抗虫性和培育抗虫番茄新品种奠定了重要基础。A plant expression vector pCAMBIA3300-Pta-CrylAc harboring two insecticidal genes was constructed and employed in Agrobacterium-mediated tomato transformation by using cotyledons as the explants. We harvested 24 phos- phinothricin（ PPT0-resistant independent regenerated plantlets after co-cnllmre, selection culture and regeneration. PCR and Southern blot hybridization analyses confirmed that the two target genes were successfully integrated into 8 lines of tomato trmlsformants. Bioassay of transgenic tomatos using insect larvae showed that transgenic plants were conferred with increased insect-resistance compared to nntransformed plants. This study laid a potential foundation for the further research on tomato insect-resistance and for the tomato genetic improvement.

关 键 词：农杆菌 番茄 杀虫蛋白基因 遗传转化 抗虫性 

分 类 号：Q943[生物学—植物学]