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Expression of a Synthesized Gene Encoding Cationic Peptide Cecropin B in Transgenic Tomato Plants Protects against Bacterial Diseases

机译:编码阳离子肽天蚕素B的合成基因在转基因番茄植株中的表达可预防细菌性疾病

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摘要

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 S50s (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 μg/ml and 0.29 μg/ml, respectively. The CB gene was then fused to the secretory signal peptide (sp) sequence from the barley α-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 (∼0.05 μg in 50 mg of leaves) were far lower than the S50 determined in vitro. CB transgenic tomatoes could therefore be a new mode of bioprotection against these two plant diseases with significant agricultural applications.
机译:从巨型蚕蛾(Hyalophora cecropia)分离出的阳离子裂解肽天蚕素B(CB)已被证明能有效消除革兰氏阴性菌和某些革兰氏阳性菌。在这项研究中,研究了化学合成的炭黑对植物病原体的影响。 CB对番茄的两种主要病原体青枯雷尔菌(Ralstonia solanacearum)和野油菜黄单胞菌(Xanthomonas campestris pv)的S50s(导致致病细菌存活的50%的肽浓度)。 vesicatoria分别为529.6μg/ ml和0.29μg/ ml。然后将CB基因与大麦α-淀粉酶基因的分泌信号肽(sp)序列融合,并将新的构建体pBI121-spCB用于番茄植物的转化。通过PCR证实了CB基因整合到番茄基因组中,并且通过Western印迹分析证实了其表达。对转基因番茄植株的体内研究表明,其对细菌枯萎病和细菌斑病具有明显的抗性。转基因番茄植株中的CB表达水平(50 mg叶片中约0.05μg)远低于体外测定的S50。因此,CB转基因番茄具有重要的农业应用前景,因此可以成为针对这两种植物病害的新型生物保护模式。

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