昆虫学报 ›› 2022, Vol. 65 ›› Issue (11): 1444-1451.doi: 10.16380/j.kcxb.2022.11.005

• 研究论文 • 上一篇    下一篇

美洲大蠊成虫肠道抗细菌共生放线菌的筛选及鉴定

殷鹏凯, 王明明, 杨自忠, 杨大松*, 杨银河*   

  1. (大理大学药学院, 云南省昆虫生物医药研发重点实验室, 云南大理 671000)
  • 出版日期:2022-11-20 发布日期:2022-12-02

Screening and identification of symbiotic actinomycetes with antibacterial activity from the gut of adult Periplaneta americana (Blattodea: Blattidae)

YIN Peng-Kai, WANG Ming-Ming, YANG Zi-Zhong, YANG Da-Song*, YANG Yin-He*   

  1. (Yunnan Provincial Key Laboratory of Entomological Biopharmaceutical R&D, College of Pharmacy, Dali University, Dali, Yunnan 671000, China) 
  • Online:2022-11-20 Published:2022-12-02

摘要: 【目的】鉴于野外美洲大蠊Periplaneta americana对恶劣环境适应性强,本研究旨在从云南省大理州的野外美洲大蠊成虫肠道中分离、筛选出抗细菌活性放线菌,为抗生素开发提供菌种资源。【方法】采用涂布平板法和平板划线法对美洲大蠊成虫肠道放线菌进行分离;以金黄色葡萄球菌Staphylococcus aureus、耐甲氧西林金黄色葡萄球菌、铜绿假单胞菌Pseudomonas aeruginosa、粪肠球菌Enterococcus faecalis、大肠杆菌Escherichia coli和鼠伤寒沙门氏菌Salmonella typhimurium共6种人体病原细菌为指示菌株,采用牛津杯法对分离自这些放线菌的次生代谢产物进行抗菌活性测定;通过形态学特征和16S rRNA基因序列分析,对具有广谱和明显抗菌活性的放线菌进行鉴定,并经16SrRNA基因序列的BlAST同源性比对及系统发育分析确定它们的分类地位。【结果】从美洲大蠊成虫肠道共分离获得41株放线菌。抗菌活性测定结果表明,34株(82.9%)放线菌对至少1种指示病原细菌具有抑制作用,其中有7株对3种以上病原细菌具有抑制作用,9株表现出明显的抗菌活性。根据系统发育分析结果,这些放线菌均被鉴定为链霉菌属Streptomyces spp.。【结论】野外美洲大蠊成虫肠道含有丰富的抗细菌活性的放线菌资源,为后续挖掘新型抗生素提供重要的微生物资源。

关键词: 美洲大蠊, 肠道放线菌, 抗细菌活性, 16S rRNA, 分子鉴定

Abstract: 【Aim】 In view of the strong adaptability of Periplaneta americana in harsh environments, this study aims to isolate and screen actinomycetes with antibacterial activity from the gut of P. americana adults in the wild in Dali prefecture of Yunnan province so as to provide strain resources for the development of antibiotics. 【Methods】 The gut actinomycetes of P. americana adults were isolated by coating plate method and plate marking method, and six strains of common human pathogenic bacteria, Staphylococcus aureus, methicillin-resistant S. aureus, Pseudomonas aeruginosa, Enterococcus faecalis, Escherichia coli and Salmonella typhimurium, were used as the indicator strains. The antibacterial activities of the secondary metabolites extracted from these actinomycetes were determined by the Oxford cup method. The actinomycetes with broad spectrum and obvious antibacterial activities were identified according to their morphological features as well as 16S rRNA gene sequence analysis. Their taxonomic statuses were confirmed by the BLAST analysis of their 16S rRNA gene sequences and phylogenetic analysis. 【Results】 Totally, 41 actinomycetes strains were isolated from the gut of P. americana adults. The results of antibacterial activity test showed that 34 strains (82.9%) of actinomycetes had inhibitory activities against at least one indicator pathogenic bacterium. Among them, seven strains had inhibitory activities against more than three pathogenic bacteria, and nine strains had obvious antibacterial activities. Based on the results of phylogenetic analysis, these actinomycetes were identified as Streptomyces spp. 【Conclusion】 The gut of P. americana adults in the wild contains rich actinomycete resources with antibacterial activity, which provides an important microbial source for further exploration of new antibiotics.

Key words: Periplaneta americana, gut actinomycetes, antibacterial activity, 16S rRNA, molecular identification