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内生真菌HND5菌株抗香蕉枯萎病菌相关非核糖体多肽合成酶基因簇的鉴定

引用本文：杨扬,陈奕鹏,周维,蔡吉苗,王宝,黄贵修.内生真菌HND5菌株抗香蕉枯萎病菌相关非核糖体多肽合成酶基因簇的鉴定.植物保护学报,2019,46(5):1110-1120

DOI：10.13802/j.cnki.zwbhxb.2019.2018151

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作者	单位	E-mail
杨扬	中国热带农业科学院环境与植物保护研究所, 农业部热带作物有害生物综合治理重点实验室, 海南省热带农业有害生物监测与控制重点实验室, 海南省热带作物病虫害生物防治工程技术研究中心, 海口 571101
陈奕鹏	中国热带农业科学院环境与植物保护研究所, 农业部热带作物有害生物综合治理重点实验室, 海南省热带农业有害生物监测与控制重点实验室, 海南省热带作物病虫害生物防治工程技术研究中心, 海口 571101
周维	广西壮族自治区农业科学院生物技术研究所, 香蕉抗病种质创制与病虫害防控联合实验室, 南宁 530007
蔡吉苗	中国热带农业科学院环境与植物保护研究所, 农业部热带作物有害生物综合治理重点实验室, 海南省热带农业有害生物监测与控制重点实验室, 海南省热带作物病虫害生物防治工程技术研究中心, 海口 571101
王宝	中国热带农业科学院环境与植物保护研究所, 农业部热带作物有害生物综合治理重点实验室, 海南省热带农业有害生物监测与控制重点实验室, 海南省热带作物病虫害生物防治工程技术研究中心, 海口 571101
黄贵修	中国热带农业科学院环境与植物保护研究所, 农业部热带作物有害生物综合治理重点实验室, 海南省热带农业有害生物监测与控制重点实验室, 海南省热带作物病虫害生物防治工程技术研究中心, 海口 571101	hgxiu@vip.163.com

中文摘要:为明确帚枝霉属内生真菌Sarocladium brachiariae HND5菌株具体的抑菌活性物质，在全基因组测序数据的基础上，利用生物信息学对抑菌活性物质合成基因簇进行定位，通过聚类分析和系统发育树分析对基因簇合成产物进行鉴定，并通过基因缺失突变构建及代谢组分析确定具体的抑菌活性物质。结果显示，HND5菌株共含有7个非核糖体多肽合成酶（non-ribosomal peptide synthe-tase，NRPS）基因簇，聚类分析结果表明基因簇29合成产物为噬铁素，基因簇30合成产物为类环孢霉素类抗菌多肽；系统发育树结合生物信息学结果显示基因簇30合成产物与已知环孢霉素结构差异大，为一个新型非核糖体多肽；将基因簇30 NRPS基因缺失突变后，HND5菌株丧失抑菌活性；同野生型菌株相比，基因簇30 NRPS基因缺失突变体缺失一个分子量为887.54 Da的多肽类物质。表明HND5菌株的抑菌活性物质为一个分子量为887.54 Da的新型类环孢霉素非核糖体多肽，基因簇30负责该多肽的生物合成。

中文关键词:内生真菌香蕉枯萎病菌抑菌多肽非核糖体多肽合成酶

Identification of the non-ribosomal peptide synthetase (NRPS) gene clusters involved in resistance to Fusarium oxysporum f. sp. cubense in the endophytic fungal isolate HND5

Author Name	Affiliation	E-mail
Yang Yang	Hainan Engineering Research Center for Biological Control of Tropical Crop Diseases and Insect Pests, Hainan Key Laboratory for Monitoring and Control of Tropical Agricultural Pests, Key Laboratory of Integrated Pest Management on Tropical Crops, Ministry of Agriculture Environmental and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, Hainan Province, China
Chen Yipeng	Hainan Engineering Research Center for Biological Control of Tropical Crop Diseases and Insect Pests, Hainan Key Laboratory for Monitoring and Control of Tropical Agricultural Pests, Key Laboratory of Integrated Pest Management on Tropical Crops, Ministry of Agriculture Environmental and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, Hainan Province, China
Zhou Wei	Joint Laboratory of Banana Resistant Germplasm Creation and Control of Diseases and Insect Pests, Biotechnology Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, Guangxi Zhuang Autonomous Region, China
Cai Jimiao	Hainan Engineering Research Center for Biological Control of Tropical Crop Diseases and Insect Pests, Hainan Key Laboratory for Monitoring and Control of Tropical Agricultural Pests, Key Laboratory of Integrated Pest Management on Tropical Crops, Ministry of Agriculture Environmental and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, Hainan Province, China
Wang Bao	Hainan Engineering Research Center for Biological Control of Tropical Crop Diseases and Insect Pests, Hainan Key Laboratory for Monitoring and Control of Tropical Agricultural Pests, Key Laboratory of Integrated Pest Management on Tropical Crops, Ministry of Agriculture Environmental and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, Hainan Province, China
Huang Guixiu	Hainan Engineering Research Center for Biological Control of Tropical Crop Diseases and Insect Pests, Hainan Key Laboratory for Monitoring and Control of Tropical Agricultural Pests, Key Laboratory of Integrated Pest Management on Tropical Crops, Ministry of Agriculture Environmental and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, Hainan Province, China	hgxiu@vip.163.com

Abstract:In order to identify the antifungal substances of the endophytic fungus Sarocladium brachiariae HND5 strain, bioinformatic methods were used to locate putative genes responsible for the biosynthesis of antifungal substances based on the whole genome sequence of this strain, and cluster analysis combined with phylogenic analysis was used to identify the products of gene clusters and the exact antifungal substance was located by constructing deletion mutants and analyzing metabolome. The results showed that HND5 strain had seven non-ribosomal peptide synthetase (NRPS) gene clusters; the product of cluster 29 was siderophore and the product of cluster 30 was cyclosporin-like non-ribosomal antifungal peptide based on cluster analysis. Phylogenetic analysis combined with bioinformatic analysis indicated that the product of cluster 30 was a new non-ribosomal peptide, the structure of which was very different from cyclosporin. HND5 strain lost its antifungal activity when cluster 30 NRPS gene was mutated by deletion. Compared with HND5 wild type strain, the deletion mutant of cluster 30 NRPS gene lost an 887.54 Da peptide. This study suggested that HND5 strain synthesized an 887.54 Da cyclosporin-like non-ribosomal peptide by gene cluster 30 as the antifungal substance.

keywords:endophytic fungus Fusarium oxysporum f. sp. cubense race 4 antifungal peptide non-ribosomal peptide synthetase

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