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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2016, Vol. 42 Issue (1): 17-22    DOI: 10.3785/j.issn.1008-9209.2015.07.151
生物科学与技术     
2个转基因抗虫杂交棉Bt蛋白含量的时空表达特性研究
余恩, 蔡芸菲, 赵茹冰, 陈进红, 祝水金
浙江大学农业与生物技术学院农学系/浙江省作物种质资源重点实验室，杭州310058
Studies on temporal-spatial expression characters of Bt protein in two transgenic hybrid cotton
YU En, CAI Yunfei, ZHAO Rubing, CHEN Jinhong, ZHU Shuijin
(Department of Agronomy, College of Agriculture and Biotechnology, Zhejiang University/Zhejiang Key  Laboratory of Crop Germplasm, Hangzhou 310058, China)
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摘要： 采用双抗夹心酶联免疫 (enzyme linked immuno-sorbent assay,ELISA)的方法，分析了2个转苏云金杆菌（Bacillus thuringiensis,Bt）基因杂交棉（浙大13-1和浙大13-2）以及转Bt基因纯系亲本不同生育期和不同器官的Bt蛋白表达规律。结果表明，Bt基因在所检测的转基因抗虫杂交棉品种和转基因纯系亲本各器官中均有表达，但不同器官Bt蛋白含量明显不同，品种之间各器官的Bt蛋白含量也有明显差异。2个转基因抗虫杂交棉品种苗期以三叶期的子叶最高，苗期叶片次之;苗期根系和茎秆中Bt蛋白表达呈抛物线形，分别在第33天和第40天达到峰值，其中根系表达量高于茎秆。叶片中的Bt蛋白含量随着棉花生长发育进程而降低，盛花期Bt蛋白含量最低，之后又有所增强，整个生育期呈“V”字形规律。转基因抗虫杂交棉品种盛花期棉株各器官中，花药的Bt蛋白含量显著高于同期其他器官。2个杂交棉品种之间各器官的Bt蛋白含量的趋势基本相同，但浙大13-1器官中的Bt蛋白含量大多高于浙大13-2。亲本各器官中的Bt蛋白含量高于其杂交棉品种，但植株生长后期的叶片和花药中的Bt蛋白低于其杂交棉品种，二者有明显差异。
Abstract: Cotton bollworm, Helicoberpa armigera, was one of the most harmful pests in cotton-production areas all over the world. The transgenic insect resistant cotton with Bt gene from Bacillus thuringiensis was developed to control this pest in 1987. It was increased quickly in recent years, accounting for more than 93% of all cotton planted in China, and huge economic, social, and ecological benefits were obtained. The levels of Bt protein in cotton organs can directly affect the insect resistance of cotton plants. Many reports showed that the Bt cotton expressed the Bt protein at a high level in early season which provided good control of the second generation of cotton bollworm, but it was decreased in middle and late stages which lead to the decrease of the controlling efficacy to bollworm. However, few  reports had been found about the levels of Bt protein in different organs and tissues during the whole growth stages of the transgenic hybrid cotton. Whole season levels of Bt protein in two transgenic hybrid cotton, Zheda 13-1 and Zheda 13-2, and their pure line parent, Zayou 2012, were investigated  in 2014 in Hangzhou City, Zhejiang Province, located in southeast coastal region of China. Enzyme linked immuno-sorbent assay (ELISA)  method was used to quantify Bt protein. The results showed that the Bt protein could be detected in all organs and tissues in the two hybrid cultivars and their parent, however, its levels changed significantly among the different organs from time to time. During the seedling stage, cotyledon was the highest organ, following by the first true leaf. The Bt protein level in root and stem was increased at the beginning, reached the peak value on 33 and 40 days after sowing, respectively for two cultivars, then decreased steadily afterward. In general, the Bt protein in root was much higher than that in stem. In the leaves of the two hybrid cottons, the upper third leaf, Bt protein was high in the early stage, declined in mid-season but rebounded in late-season. While for their parent, it was high in early season, then declined steadily in middle and latter season. The difference might  relate to the growth habit of the hybrid cotton which kept growing in late-season for relatively long time. So it is suggested that the pesticide control might be necessary for the hybrid cotton in the mid-season of cotton growth. In addition, the study showed that Bt protein contents in the anther and stamens were much higher than those in leaves, squares, bolls, ovules, and petals. While for their parent, it was much lower in the anther and stamens than that in other organs and tissues. It should be studied further whether or not this phenomenon was resulted from the heterosis of the hybrid cotton, the high concentration of Bt protein in the anther and stamens was beneficial to the cotton bollworm control of the hybrid cotton as the cotton bollworms were more interested in the cotton reproductive organs.
出版日期: 2016-01-20
CLC:  Q 946.1  
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余恩
蔡芸菲
赵茹冰
陈进红
祝水金

引用本文:

余恩, 蔡芸菲, 赵茹冰, 陈进红, 祝水金. 2个转基因抗虫杂交棉Bt蛋白含量的时空表达特性研究[J]. 浙江大学学报(农业与生命科学版), 2016, 42(1): 17-22.

YU En, CAI Yunfei, ZHAO Rubing, CHEN Jinhong, ZHU Shuijin. Studies on temporal-spatial expression characters of Bt protein in two transgenic hybrid cotton. Journal of Zhejiang University (Agriculture and Life Sciences), 2016, 42(1): 17-22.

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http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2015.07.151    或    http://www.zjujournals.com/agr/CN/Y2016/V42/I1/17

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