蚂蚁抗菌肽Ponericin W1对植物病原菌的体外抑菌活性及其转基因拟南芥的抗病性
收稿日期: 2013-04-01
修回日期: 2013-05-08
网络出版日期: 2013-08-25
基金资助
转基因生物新品种培育重大专项(编号2009ZX08009-036B)资助
Antimicrobial activities of ant Ponericin W1 against plant pathogens in vitro and the disease resistance in its transgenic Arabidopsis
Received date: 2013-04-01
Revised date: 2013-05-08
Online published: 2013-08-25
抗菌肽的抗菌谱很广, 非植物来源的抗菌肽在植物抗病基因工程方面的应用越来越受到重视。蚂蚁抗菌肽Ponericin W1可抑制革兰氏阴性、革兰氏阳性细菌和酿酒酵母(Saccharomyces cerevisiae)的生长, 但在植物抗病方面尚无报道。文章对人工合成的Ponericin W1在体外进行了植物病原菌的抑制实验, 结果表明, Ponericin W1可以抑制稻瘟病菌(Magnaporthe oryzae)、葡萄灰霉病菌(Botrytis cinerea)的菌丝生长, 可以抑制小麦赤霉病菌(Fusarium graminearum)的菌丝生长和产孢, 还可以抑制丁香假单胞杆菌番茄致病变种(Pseudomonas syringae pv. tomato)和水稻白叶枯病菌(Xanthomonasoryzae pv. oryzae)的生长。Ponericin W1的基因序列经密码子优化, 人工合成后构建至植物表达载体, 采用蘸花法由农杆菌介导转化拟南芥。在转基因植株中, 抗菌肽定位于细胞间隙。转基因植株接种病原真菌白粉病菌(Golovinomyces cichoracearum)后, 叶片上菌丝和分生孢子梗都较野生型少, 且有未萌发的孢子; 接种病原细菌丁香假单胞杆菌后, 叶片内的细菌量较野生型显著减少, 表明转基因植株的抗病性较野生型显著增强。体外的和转基因拟南芥的病原菌抗性证明, Ponericin W1在植物抗病基因工程中有良好的应用前景。
关键词： 转基因; Ponericin W1; 植物病原; 抗菌肽
陈永芳 孙朋卫 唐定中 . 蚂蚁抗菌肽Ponericin W1对植物病原菌的体外抑菌活性及其转基因拟南芥的抗病性[J]. 遗传, 2013 , 35(8) : 1023 -1029 . DOI: 10.3724/SP.J.1005.2013.01023
The antimicrobial peptides (AMPs) exhibit a broad antimicrobial spectrum. The application of AMPs from non-plant organisms attracts considerable attention in plant disease resistance engineering. Ponericin W1, isolated from the venom of ant (Pachycondyla goeldii), shows antimicrobial activities against Gram-positive bacteria, Gram-negative bacteria and the budding yeast (Saccharomyces cerevisiae); however, it is not clear whether Ponericin W1 is effective against plant pathogens. The results of this study indicated synthesized Ponericin W1 inhibited mycelial growth of Magnaporthe oryzae and Botrytis cinerea, as well as hyphal growth and spore production of Fusarium graminearum. Besides, Ponericin W1 exhibited antibacterial activities against Pseudomonas syringae pv. tomato and Xanthomonas oryzae pv. oryzae. After codon optimization, Ponericin W1 gene was constructed into plant expression vector, and transformed into Arabidopsis thaliana by floral dip method. The Ponericin W1 was located in intercellular space of the transgenic plants as expected. Compared with the wild-type plants, there were ungerminated spores and less hyphal, conidia on the leaves of transgenic plants after innoculation with the powdery mildew fungus Golovinomyces cichoracearum. After innoculation with the pathogenic bac-terium Pseudomonas syringae pv. tomato, the baceria in the leaves of transgenic plants was significantly less than the wild-type plants, indicating that the transgenic plants displayed enhanced disease resistance to pathogens. These results demonstrate a potential use of Ponericin W1 in genetic engineering for broad-spectrum plant disease resistance.
Key words： Ponericin W1; transgenic; antimicrobial peptides; plant pathogen
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