农杆菌介导Cry1Ac基因转化菊花

园艺学报 ›› 2005, Vol. 32 ›› Issue (1): 65-69.

农杆菌介导Cry1Ac基因转化菊花

蒋细旺;包满珠;吴家和;张献龙;田颖川

(¹华中农业大学园艺林学学院, 武汉430070; ² 江汉大学医学与生命科学学院, 武汉430056; ³ 华中农业大学作物遗传改良国家重点实验室, 武汉430070; ⁴中国科学院微生物研究所, 北京100080)

收稿日期:2004-06-19 修回日期:2004-09-16 出版日期:2005-02-25 发布日期:2005-02-25

Transformation of Chrysanthemum with a Synthetic Cry1Ac Gene Mediated by Agrobacteriumtum efaciens

Jiang Xiwang;Bao Manzhu;Wu Jiahe;Zhang Xianlong;Tian Yingchuan

(¹College of Horticulture and Forestry Science, Huazhong Agricultural University, Wuhan 430070, China; ²College of Medicine and Life Science, Jianghan University, Wuhan 430056, China; ³ The National Key Labrotary of Crops Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, China;⁴ Institute of Microbiology, Chinese Academy of Science, Beijing 100080, China)

Received:2004-06-19 Revised:2004-09-16 Online:2005-02-25 Published:2005-02-25

摘要/Abstract

摘要： 用农杆菌介导法, 以茎段(节间) 为外植体, 将含有苏云金芽孢杆菌(Bt) 毒蛋白Cry1Ac基因的植物遗传载体导入菊花(切花菊) 品种‘日本黄’中, 得到126株抗性植株, PCR检测及基因组DNA
的Southern杂交分析表明Cry1Ac基因已整合到菊花总DNA中, 共获得6株转基因植株。利用棉铃虫做转基因植株的盆栽和叶片平皿试验, 发现其中2株对棉铃虫具有很高抗性, 校正死亡率达90%以上; 1株对棉铃虫具有高抗性, 校正死亡率达80%以上; 3株对棉铃虫抗性较明显, 校正死亡率44.7%～60.9%。

关键词: 菊花, 苏云金芽孢杆菌, Cry1Ac基因, 抗虫性, 转基因植株, 棉铃虫, 根癌农杆菌

Abstract: A synthetic Bt (Bacillus thuringiensis) Cry1Ac gene was introduced into the genome of cut flower chrysanthemum (Chrysanthemum morifolium ) cultivar‘Japanese Yellow’in vitro with explants of internodal segmentsmediated by Agrobacterium tum efaciens. Results of PCR and Southern blot analysis indicated that Cry1Ac gene were integrated as one copy into the genome of chrysanthemum. Six transgenic lineswere obtained in total. Upon insect bioassay using Heliothis arm igera (Hubner) in the pot2culture and in petri dishes, two of the screened plants were demonstrated to be very highly resistant to the insect larvae, the corrected mortality of insect larvae was higher than 90% in five days. One individual line showed strong resistance,
the corrected mortality of insect larvae was higher than 80%. Three transformed lines showed relative strong resistance, the corrected mortality of insect larvae was ranged from 44.7% to 60.9%.

Key words: Chrysanthemum, Bt (Bacillus thuringensis), Cry1Ac gene, Insect-resistant, Transgenic plants, Heliothis armigera, Agrobacterium tumefaciens

中图分类号:

S682.1

蒋细旺;包满珠;吴家和;张献龙;田颖川. 农杆菌介导Cry1Ac基因转化菊花[J]. 园艺学报, 2005, 32(1): 65-69.

Jiang Xiwang;Bao Manzhu;Wu Jiahe;Zhang Xianlong;Tian Yingchuan. Transformation of Chrysanthemum with a Synthetic Cry1Ac Gene Mediated by Agrobacteriumtum efaciens[J]. ACTA HORTICULTURAE SINICA, 2005, 32(1): 65-69.

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