中国农业科学Issue(6)：1095-1102,8.DOI:10.3864/j.issn.0578-1752.2015.06.06
小檗内生放线菌H21的鉴定及抑菌活性成分分析
Identification and Antimicrobial Ingredients of an Endophytic Actinomycete Strain H21 fromBerberis thunbergii
摘要
Abstract
[Objective]The objectives of this experiment are to screen bioactive endophytic actinomycete fromBerberis thunbergii, a traditional Chinese medicinal plant, study the taxonomic status and antimicrobial activity of the active strain, investigate the chemical structures and antimicrobial activity of active ingredients in the fermentation, and to provide a basis for the development of agriculturalfungicide.[Method] The endophytic actinomycete was isolated from the leaf ofB. thunbergii by using selective culture techniques, and the obtained strain was identified by using 16S rDNA sequencing. The antibacterial and antifungal activities of the fermentation broth against seven species of tested bacteria (such asRalstonia solanacearum) and plant pathogenic fungi (such asBotrytis cinerea) were tested by cylinder-plate method and mycelial growth method, respectively. The active ingredients were obtained by the techniques of macroporous resin adsorption, silica gel column chromatography and preparative reverse phase high performance liquid chromatography. Structures of the obtained compounds were elucidated by NMR and ESI-MS techniques. Minimum inhibitory concentrations (MICs) of active ingredients against tested bacteria were determined by micro-broth dilution method. Antifungal activity of H2101 againstB. cinerea was tested by the methods of mycelial growth and spore germination, respectively.[Result]Strain H21, an active species, was isolated from the plant, and it was identified asStreptomycessp. by the method of 16S rDNA sequencing. ExceptPseudomonas aeruginosa, the fermentation broth of H21 showed strong antibacterial against the tested bacteria, such asR. solanacearum,P. syringae pv. actinidiae,Bacillus subtilis,B. cereus,Staphylococcus aureus andEscherichia coli. However, the fermentation broth could only inhibit the mecelial growth ofB. cinerea. No inhibitory effect was observed against other tested fungi. Holomycin, a member of the pyrrothine class of antibiotics, as well as N-acetyl-2-(4-hydroxyphenyl)-ethylamine and cyclo (L-Leu-L-Arg), was isolated from the broth of strain H21. Holomycin exhibited broad antimicrobial activity, and the MIC values againstR. solanacearum,P. syringae pv. actinidiae,B. cereus,B. subtilis,E. coli andS. aureus were 0.156, 0.313, 0.078, 0.313, 0.156 and 0.313 µg·mL-1, respectively. Meanwhile, holomycin had strong antifungal activity againstB. cinerea, and its IC50 values against mycelial growth and spore germination were 13.56 and 7.89 µg·mL-1, respectively.[Conclusion]Streptomycessp. is a newly discovered producer of holomycin, and it has the antibacterial holomycin againstR. solanacearum andP. syringae pv. actinidiae, two species of plant pathogenic bacteria. In general, holomycin is a valuable lead compound for the development of agricultural fungicides with those acts on bacteria.关键词
小檗/内生放线菌/全霉素/抑菌活性Key words
Berberis thunbergii/endophytic actinomycete/holomycin/antimicrobial activity引用本文复制引用
魏少鹏,国政,姬志勤..小檗内生放线菌H21的鉴定及抑菌活性成分分析[J].中国农业科学,2015,(6):1095-1102,8.基金项目
国家自然科学基金（31371973，31301700）、陕西省自然科学基金（2013JQ3003）、中央高校基本科研业务费专项 （）
