J Integr Plant Biol. ›› 2002, Vol. 44 ›› Issue (8): 963-970.

• Research Articles • Previous Articles     Next Articles

Obtaining High Pest-resistant Transgenic Upland Cotton Cultivars Carrying cry1Ac3 Gene Driven by Chimeric OM Promoter

CHEN Wan-Xin, XIAO Gui-Fang and ZHU Zhen   

  • Published:2002-08-21

Abstract:

Hypocotyl segments from aseptic seedlings of two important cultivars of upland cotton (Gossypium hirsutum L.) in Northwest China, “Xinluzao 1", “Jinmian 7", “Jinmian 12" and “Jihe 321" were transformed respectively by two efficient lant expression plasmids pBinMoBc and pBinoBc via Agrobacterium tumefaciens. In pBinMoBc,cry 1Ac3 gene, which encodes the Bt toxin, is under the control of chimeric OM promoter. In pBinoBc, it is undercontrol of CaMV 35S promoter. After cocultivation with Agrobacterium tumefimpfaciens LBA4404 (containing pBinMoBc or pBinoBc), kanamycin resistant selection, somatic embryos were induced and regenerated plants were obtained. Then the regenerated plantlets were grafted to untransformed stocks in greenhouse to produce descendants. The integration of cry1Ac3 gene and its expression in T2 generation of transgenic cotton plants were confirmed by Southern hybridization and Western blotting. The analyses of insect bioassay indicated that the transgenic plants of both onstructions have significant resistance to the larvae of cotton bollworm (Heliothis armigera ) and that cry1Ac3 gene driven by chimeric OM promoter could endue T2 generation cotton with high pest resistant ability, implicating that it has a profound application in genetic engineering to breed new pest resistant cotton varieties.

陆地棉栽培品种转复合启动子控制下的cry 1Ac3 基因获得高效抗虫植株
陈宛新 肖桂芳 朱 祯*

(中国科学院遗传与发育生物学研究所,北京100101)

摘要:以根癌土壤杆菌 (Agrobacterium tumefaciens)介导法分别将植物表达载体pBinMoBc和pBinoBc导入陆地棉(Gossypiumhirsutum L .)栽培品种“新陆早 1号”、“晋棉 7号”、“晋棉 12号”和“冀合 32 1”。pBinMoBc携带有高效启动子复合OM启动子控制下的cry1Ac3基因 ,pBinoBc携带有 35S启动子控制下的cry1Ac3基因。经过共培养、卡那霉素筛选抗性愈伤组织及体细胞胚的诱导 ,得到了再生植株。对T2 代的PCR、Southernblotting、ELISA检测及Westernblotting证明cry1Ac3基因已整合入受体棉花基因组并得到表达。抗虫性检测表明转基因后代对棉铃虫 (Heliothisarmigera )具有良好的抗性 ,转pBinMoBcT2代与转pBinoBcT2 代相比 ,对棉铃虫具有更快的致死速度。本研究建立了一套高效的陆地棉栽培品种转化体系 ;进一步的检测结果表明 ,复合OM启动子可以提高外源基因的表达量从而增强转基因棉的抗虫性。

关键词: 陆地棉;抗虫棉;转基因植物;根癌土壤杆菌;cry1Ac3 基因;复合启动子

通讯作者.Tel:010-64873490;E-mailzzhu @ genetics.ac.cn

Key words: upland cotton, insect resistant cotton, transgenic plant;Agrobacterium tumefaciens, cry1Ac3 gene, chimeric promoter

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