农杆菌介导的双价抗虫基因对番茄遗传转化的研究

J4 ›› 2013, Vol. 48 ›› Issue (7): 9-13.

农杆菌介导的双价抗虫基因对番茄遗传转化的研究

杨俊杰1,于惠敏1,侯丙凯2*,王兴智3

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

收稿日期:2013-01-10 发布日期:2013-12-03
通讯作者: 侯丙凯(1964- ),男,博士,教授,博士生导师,研究方向为植物分子遗传学与基因工程.Email:bkhou@sdu.edu.cn
作者简介:杨俊杰(1981- ),女,硕士,讲师,研究方向为植物细胞工程.Email:junjie198100@163.com
基金资助:
齐鲁师范学院青年教师基金资助项目(2011L1514)

Agrobacterium-mediated introduction of insect-resistant genes into tomato

YANG Jun-jie1, YU Hui-min1, HOU Bing-kai2*, WANG Xing-zhi3

1. Department of biology, Qilu Normal University, Jinan 250013, Shandong, China;
2. Key Laboratory of Plant Cell Engineering and Germplasm Innovation, Ministry of Education,
Shandong University, Jinan 250100, Shandong, China;
3. School of Life Sciences, Northeast Normal University, Changchun 130024, Jilin, China

Received:2013-01-10 Published:2013-12-03

摘要/Abstract

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。

关键词: 农杆菌;番茄;杀虫蛋白基因;遗传转化;抗虫性

Abstract:

A plant expression vector pCAMBIA3300-Pta-Cry1Ac harboring two insecticidal genes was constructed and employed in Agrobacterium-mediated tomato transformation by using cotyledons as the explants. We harvested 24 phosphinothricin(PPT0-resistant independent regenerated plantlets after co-culture, selection culture and regeneration. PCR and Southern blot hybridization analyses confirmed that the two target genes were successfully integrated into 8 lines of tomato transformants. Bioassay of transgenic tomatos using insect larvae showed that transgenic plants were conferred with increased insect-resistance compared to untransformed plants. This study laid a potential foundation for the further research on tomato insect-resistance and for the tomato genetic improvement.

Key words: Agrobacterium tumefaciens; Tomato; insecticidal gene;Genetic transformation; Insect-resistance

中图分类号:

Q943

杨俊杰1,于惠敏1,侯丙凯2*,王兴智3. 农杆菌介导的双价抗虫基因对番茄遗传转化的研究[J]. J4, 2013, 48(7): 9-13.

YANG Jun-jie1, YU Hui-min1, HOU Bing-kai2*, WANG Xing-zhi3. Agrobacterium-mediated introduction of insect-resistant genes into tomato[J]. J4, 2013, 48(7): 9-13.

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