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合成基因编码阳离子肽 Cecropin B 在转基因番茄植物中的表达可预防细菌病。

Expression of a synthesized gene encoding cationic peptide cecropin B in transgenic tomato plants protects against bacterial diseases.

机构信息

National Nano Device Laboratories, Nano Biomedical and MEMS Technology Division, Hsinchu, Taiwan.

出版信息

Appl Environ Microbiol. 2010 Feb;76(3):769-75. doi: 10.1128/AEM.00698-09. Epub 2009 Dec 4.

Abstract

The cationic lytic peptide cecropin B (CB), isolated from the giant silk moth (Hyalophora cecropia), has been shown to effectively eliminate Gram-negative and some Gram-positive bacteria. In this study, the effects of chemically synthesized CB on plant pathogens were investigated. The S(50)s (the peptide concentrations causing 50% survival of a pathogenic bacterium) of CB against two major pathogens of the tomato, Ralstonia solanacearum and Xanthomonas campestris pv. vesicatoria, were 529.6 microg/ml and 0.29 microg/ml, respectively. The CB gene was then fused to the secretory signal peptide (sp) sequence from the barley alpha-amylase gene, and the new construct, pBI121-spCB, was used for the transformation of tomato plants. Integration of the CB gene into the tomato genome was confirmed by PCR, and its expression was confirmed by Western blot analyses. In vivo studies of the transgenic tomato plant demonstrated significant resistance to bacterial wilt and bacterial spot. The levels of CB expressed in transgenic tomato plants ( approximately 0.05 microg in 50 mg of leaves) were far lower than the S(50) determined in vitro. CB transgenic tomatoes could therefore be a new mode of bioprotection against these two plant diseases with significant agricultural applications.

摘要

阳离子溶细胞肽 CB(从巨型蚕蛾(Hyalophora cecropia)中分离得到)已被证明可有效消除革兰氏阴性和一些革兰氏阳性细菌。在这项研究中,研究了化学合成的 CB 对植物病原体的影响。CB 对番茄的两种主要病原体(茄青枯病菌和番茄疮痂病菌)的 S(50)(导致病原菌存活 50%的肽浓度)分别为 529.6μg/ml 和 0.29μg/ml。然后,将 CB 基因融合到大麦α-淀粉酶基因的分泌信号肽(sp)序列中,新构建体 pBI121-spCB 用于转化番茄植物。通过 PCR 证实了 CB 基因整合到番茄基因组中,并通过 Western blot 分析证实了其表达。对转基因番茄植物的体内研究表明,对细菌性萎蔫病和细菌性斑点病具有显著的抗性。在转基因番茄植物中表达的 CB 水平(约为 50mg 叶片中的 0.05μg)远低于体外测定的 S(50)。因此,CB 转基因番茄可能成为防治这两种植物病害的一种新的生物保护模式,具有重要的农业应用价值。

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