Engineering Pseudomonas protegens Pf-5 to improve its antifungal activity and nitrogen fixation
Engineering Pseudomonas protegens Pf-5 to improve its antifungal activity and nitrogen fixation
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对保护假单胞菌 Pfâ5 进行工程改造以提高其抗真菌活性和固氮作用
DOI：
10.1111/1751-7915.13335
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发表时间：
2020-01-01
影响因子：
5.7
通讯作者：
Zhang, Youming
中科院分区：
文献类型：
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作者：
Jing, Xiaoshu;Cui, Qingwen;Zhang, Youming
在农业生产中，可持续性是目前最重要的问题之一。植物促生根瘤菌的基因改造为利用天然细菌作为微生物接种剂提供了一条新的途径。在本研究中，对根定殖菌株假单胞菌蛋白Pf-5进行了基因改造，使其成为具有生物固氮活性的生物防治剂和生物肥料。遗传失活的retS促进了2,4-二乙酰间苯三酚的产生，这有助于增强抗真菌活性。然后，将带有stutzeri假单胞菌DSM4166原生启动子的整个氮酶岛导入到ret突变株中进行表达。通过共聚焦激光扫描显微镜评估的根定植模式证实，gfp标记的细菌主要位于根表面和表皮根细胞之间的连接处。此外，在病原菌和限氮双重处理条件下，接种重组retS -nif突变株的幼苗鲜重比对照有所增加。总之，这项研究创新地开发了一种生态友好的农药替代品，将大大有利于全球植物生产。
In agricultural production, sustainability is currently one of the most significant concerns. The genetic modification of plant growth-promoting rhizobacteria may provide a novel way to use natural bacteria as microbial inoculants. In this study, the root-colonizing strain Pseudomonas protegens Pf-5 was genetically modified to act as a biocontrol agent and biofertilizer with biological nitrogen fixation activity. Genetic inactivation of retS enhanced the production of 2,4-diacetylphloroglucinol, which contributed for the enhanced antifungal activity. Then, the entire nitrogenase island with native promoter from Pseudomonas stutzeri DSM4166 was introduced into a retS mutant strain for expression. Root colonization patterns assessed via confocal laser scanning microscopy confirmed that GFP-tagged bacterial were mainly located on root surfaces and at the junctions between epidermal root cells. Moreover, under pathogen and N-limited double treatment conditions, the fresh weights of seedlings inoculated with the recombinant retS mutant-nif strain were increased compared with those of the control. In conclusion, this study has innovatively developed an eco-friendly alternative to the agrochemicals that will benefit global plant production significantly.