Agrobacterium-transformed rice plants expressing synthetic cryIA(b) and cryIA(c) genes are highly toxic to striped stem borer and yellow stem borer
Agrobacterium-transformed rice plants expressing synthetic cryIA(b) and cryIA(c) genes are highly toxic to striped stem borer and yellow stem borer
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DOI：
10.1073/pnas.95.6.2767
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发表时间：
1998-03-17
影响因子：
11.1
通讯作者：
Altosaar, I
中科院分区：
文献类型：
--
作者：
Cheng, XY;Sardana, R;Altosaar, I
经过农杆菌介导的转化后，超过 500 个独立选择的潮霉素抗性愈伤组织再生出 9 个品系的 2,600 多个转基因水稻植株。使用来自苏云金芽孢杆菌的两种合成的cryIA(b)和cryIA(c)编码序列以及分别编码潮霉素磷酸转移酶和β-葡萄糖醛酸酶的hph和gus基因的完全修饰（植物密码子优化）版本转化植物。这些序列置于玉米泛素启动子、CaMV35S 启动子和芸苔属 Bp10 基因启动子的控制下，以实现鳞翅目特异性 δ-内毒素的高组织特异性表达。这些基因的整合、表达和遗传在 R-0 和 R-1 代中通过 Southern、Northern 和 Western 分析以及其他技术得到了证实。在 R-0 植物中检测到高水平的 CryIA(b) 和 CryIA(c) 蛋白积累（高达 3% 的可溶性蛋白）。 R-1转基因植物的生物测定表明，转基因植物对两种主要的水稻害虫：二化螟（Chile二化螟）和二化螟（Scirpophaga incertulas）具有高毒性，在侵染后5天内死亡率达97-100%，从而为转基因水稻植物提供了有效抗虫的潜力。
Over 2,600 transgenic rice plants in nine strains were regenerated from >500 independently selected hygromycin-resistant calli after Agrobacterium-mediated transformation. The plants were transformed with fully modified (plant codon optimized) versions of two synthetic cryIA(b) and cryIA(c) coding sequences from Bacillus thuringiensis as well as the hph and gus genes, coding for hygromycin phosphotransferase and beta-glucuronidase, respectively. These sequences were placed under control of the maize ubiquitin promoter, the CaMV35S promoter, and the Brassica Bp10 gene promoter to achieve high and tissue-specific expression of the lepidopteran-specific delta-endotoxins. The integration, expression, and inheritance of these genes were demonstrated in R-0 and R-1 generations by Southern, Northern, and Western analyses and by other techniques. Accumulation of high levels (up to 3% of soluble proteins) of CryIA(b) and CryIA(c) proteins was detected in R-0 plants. Bioassays with R-1 transgenic plants indicated that the transgenic plants were highly toxic to two major rice insect pests, striped stem borer (Chile suppressalis) and yellow stem borer (Scirpophaga incertulas), with mortalities of 97-100% within 5 days after infestation, thus offering a potential for effective insect resistance in transgenic rice plants.