中国农业科学Issue(20)：4064-4076,13.DOI:10.3864/j.issn.0578-1752.2015.20.008
芽孢杆菌的抑菌活性与其产脂肽类抗生素的相关性
The Antifungal Activities of Bacillus spp. and Its Relationship with Lipopeptide Antibiotics Produced by Bacillus spp
摘要
Abstract
Objective] The objective of this study is to clear the inhibition effect of 55Bacillus spp. strains to pathogenic fungi,Rhizoctonia cerealis,Fusarium graminearum andFusarium oxysporumf. sp. niveum, and the qualitative and quantitative relationship with lipopeptide antibiotics produced by the Bacillus strains.[Method]Inhibition rates of the 55Bacillus spp. strains and their crude lipopeptides extracts against three pathogenic fungi were tested, respectively, by dual culture on plate. Three pairs of primers designed based on the known lipopeptide genes were used to amplify the corresponding genes,srfA, ituAandfenA, from the genome of the 55 strains. The lipopeptides antibiotics were identified by high performance liquid chromatography (HPLC) and mass spectrum (MS) techniques.[Result] The 55 strains ofBacillus spp. and their crude lipopeptides extracts had certain biological activities to the tested pathogenic fungi. The crude lipopeptides extracts of 16.5% strains had higher antifungal activities than the Bacillus strainsin vivo, the antifungal activities against the pathogenic fungi of crude lipopeptides extracts of 55.8% strains were weaker than living cells of the strains in vivo, while the antifungal activities of 27.7% strains did not change basically. The results of PCR amplification showed that among the 55Bacillu strains, the genomes of 89.1%Bacillusstrains containedsrfA, 87.3% strains containedituA, only 56.4% strains containedfenA. Among the 55Bacillusstrains, 53Bacillus strains were identified with surfactins in the crude lipopeptides extracts by the MS analysis, 48 strains were identified with iturins, only Bs916 (B. subtilis916) was identified having fenfycins in the crude lipopeptides extracts. Those strains that were not identified having the three kinds of lipopeptides had weaker or no antifungal activity. There were 48 strains which were found to have iturins all identified with bacillomycin D, only 2 strains were identified with bacillomycin L, 12 strains were identified with bacillomycin F, iturin A or mycosubtilin. Among the 55Bacillus strains, 53 strains were detected with suefactins by HPLC, 51 strains were detected with iturins, only Bs916 strain was detected with fengycins and 2 strains were not detected with the 3 kinds of lipopeptide antibiotics. According to the time and type of chromatographic peak, iturins were divided into 4 types, named as types 1, 2, 3, and 4. The antifungal activities intensity of 4 types of iturins were type 1>type 2>type 3>type 4. There were two unknown lipopeptides (named as Unknowns: Unknown1, 2 ) which were speculated as new lipopeptides were detected with MS analysis in 3 strains JCC-1, and JCC-9 and JCC-18.[Conclusion] Lipopeptides antibiotics produced byBacillus spp. were the main composition against plant pathogenic fungi and had diversity characteristics. The structure changes and secretion of iturins had certain relevance to antifungal activities. There were some unknown new resisted resources in lipopeptides compounds from Bacillus spp..关键词
芽孢杆菌/抗真菌活性/脂肽类抗生素/相关性Key words
Bacillus spp./antifungal activity/lipopeptides/correlation引用本文复制引用
向亚萍,陈志谊,罗楚平,周华飞,刘永锋..芽孢杆菌的抑菌活性与其产脂肽类抗生素的相关性[J].中国农业科学,2015,(20):4064-4076,13.基金项目
国家高技术研究发展计划（“863”计划）（2011AA10A201）、江苏省农业科学院科技自主创新资金（CX（14）2128）、国家公益性行业（农业）科研专项 （）
