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Knockout of Diguanylate Cyclase Genes in Lysobacter enzymogenes to Improve Production of Antifungal Factor and Increase Its Application in Seed Coating

文献类型： 外文期刊

作者： Ren, Xuexiang 1 ; Ren, Shuangshuang 2 ; Xu, Gaoge 3 ; Dou, Wen 4 ; Chou, Shan-Ho 5 ; Chen, Yu 1 ; Qian, Guoliang 2 ;

作者机构： 1.Anhui Acad Agr Sci, Inst Plant Protect & Agroprod Safety, Hefei 230031, Peoples R China

2.Nanjing Agr Univ, Coll Plant Protect, Key Lab Integrated Management Crop Dis & Pests, 1 Weigang, Nanjing 210095, Jiangsu, Peoples R China

3.Jiangsu Acad Agr Sci, Inst Plant Protect, Nanjing 210014, Peoples R China

4.Nanjing Foreign Language Sch, Nanjing 210008, Peoples R China

5.Natl Chung Hsing Univ, Inst Biochem, Taichung, Taiwan

6.Natl Chung Hsing Univ, NCHU Agr Biotechnol Ctr, Taichung, Taiwan

期刊名称：CURRENT MICROBIOLOGY （ 影响因子：2.188； 五年影响因子：2.197 ）

ISSN： 0343-8651

年卷期：

页码：

收录情况： SCI

摘要： Heat-stable antifungal factor (HSAF) is a broad-spectrum antifungal antibiotic produced by the biological control agent, Lysobacter enzymogenes. In our earlier works, we have applied HSAF to effectively control wheat and pear fungal disease. However, a major bottleneck in its practical application is the low HSAF production level; therefore, boosting its production is essential for its wide application. In the past, we find that c-di-GMP, a universal bacterial second messenger, is inhibitory to HSAF production. In this work, we further identified eight active diguanylate cyclases (DGCs) responsible for c-di-GMP synthesis in Lysobacter enzymogenes via both bioinformatics and genetic analyses. We generated a strain lacking seven active DGC genes and found that this DGC-modified strain, OH11LC, produced a higher HSAF amount in a c-di-GMP concentration-dependent manner. Subsequently, by employing OH11LC as the host fermentation strain, we could even produce a much higher HSAF amount (> 200-fold). After improving the HSAF production, we further developed a technique of seed coating method with HSAF, which turned out to be effective in fighting against the maize seed-borne filamentous pathogen, Pythium gramineacola. Overall, via combining strain modification and fermentation optimization, we demonstrated a good example of translating fundamental knowledge of bacterial c-di-GMP signaling into biological control application in which we relieved the inhibitory effect of c-di-GMP on HSAF biosynthesis by deleting a bunch of potentially active L. enzymogenes DGC genes to improve HSAF yield and to expand its usage in antifungal seed coating.

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