Transgenic maize lines expressing a cry1C* gene are resistant to insect pests
Transgenic maize lines expressing a cry1C* gene are resistant to insect pests
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DOI：
10.1007/s11105-013-0663-3
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发表时间：
2014-04-01
影响因子：
2.1
通讯作者：
Qiu, Fazhan
中科院分区：
文献类型：
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作者：
Du, Dengxiang;Geng, Cunjuan;Qiu, Fazhan
玉米象是玉米生产上最严重的害虫之一。Cry 1C蛋白是苏云金芽孢杆菌（Bacillus thuringiensis，Bt）的一组蛋白，对昆虫的肠道具有毒性。Cry 1C蛋白的过表达已经导致几种作物对鳞翅目害虫的抗性增加。在本研究中，合成cry 1C * 基因，以前在水稻中进行了测试，引入玉米Hi-II基因型通过基因枪介导的转化。共获得9个独立的推定愈伤组织，87个转基因植株根据聚合酶链反应（PCR）分析cry 1C * 阳性。通过PCR分析、田间鉴定和基因组southern印迹分析，进一步证实了3株高抗虫转基因植株ZmKc-2-3、ZmKc-3-2和ZmKc-3-5。田间和室内昆虫生物测定结果表明，3个转基因株系的后代在整个生育期对鳞翅目害虫均表现出显著的抗性。通过RT-PCR和ELISA检测，证实了cry 1C * 基因在转基因植株中的稳定整合和表达。将转基因株系ZmKc-2-3与优良自交系郑58杂交获得杂种。转cry 1C * 基因玉米杂交种和回交后代的抗虫性差异较小，表明转cry 1C * 基因玉米可用于抗虫杂交种的选育，并可作为Bt玉米的抗虫源。
The lepidopteran Ostrinia furnacalis is one of the most serious pests of maize production. The Cry1C proteins are group of Bacillus thuringiensis (Bt) proteins that are toxic to the intestine of insects. Overexpression of Cry1C protein has led to increased resistance to lepidopteran pests in several crops. In the present study, the synthetic cry1C* gene that was previously tested in rice was introduced into maize Hi-II genotypes via biolistic gun-mediated transformation. A total of nine independent putative callus were obtained and 87 transgenic plants were positive with cry1C* according to polymerase chain reaction (PCR) analysis. Three highly insect-resistant transgenic plants, ZmKc-2-3, ZmKc-3-2, and ZmKc-3-5, were further confirmed by PCR analysis, field assessment, and genomic southern blotting in the T-3 generations. Insect bioassays were conducted in both the field and the laboratory, and showed that progeny of the three transgenic lines were significantly resistant to lepidopteran maize pests during the whole development and growth period. The stable integration and expression of the cry1C* in the three transgenic plants' progeny were confirmed by reverse transcription-PCR (RT-PCR) and enzyme-linked immuno sorbent assay (ELISA) methods. Hybrids were produced by crossing transgenic line ZmKc-2-3 with the elite inbred line Zheng58. There was small variation among the hybrids and backcross offspring, indicating that these cry1C* transgenic lines can be used to produce insect-resistant hybrids and served as insect-resistant sources for the development of Bt maize.