摘要
昆虫肠道共生放线菌作为一种特境放线菌,因与土壤中的放线菌生存环境的不同,研究其种类和抗菌活性具有重要意义。将金黄色葡萄球菌和耐甲氧西林金黄色葡萄球菌(Methicillin-resistant Staphylococcus aureus,MRSA)这两种致病菌作为指示细菌,对分离自美洲大蠊Periplaneta americana肠道的159株内生放线菌使用牛津杯法实验,分别对其进行体外抑菌活性筛选,随后利用16S rRNA PCR、BLAST同源性对比和构建系统发育树相结合的方法对菌株进行分子生物学鉴定,对关键酶基因非核糖体多肽合成酶(Nonribosomal peptide synthetase,NRPS)基因、多聚酮合酶(Polyketone synthase,PKS)基因和卤化酶基因FADH2进行初步筛选。结果表明49株放线菌能够不同程度的抑制金黄色葡萄球菌和MRSA,有11株放线菌对两种致病菌均具有不同程度的抑制作用。49株放线菌中链霉菌属有36株(73.47%),属于优势菌属;其余的13株放线菌分别属于戈登氏菌属、微杆菌属、短状杆菌属、分枝杆菌属、无色杆菌属和小单孢菌属。琼脂糖凝胶电泳结果显示,11株放线菌中有6株放线菌均检测到关键酶基因PKS I、NRPS和卤化酶基因FADH2。本研究表明美洲大蠊肠道中含有具有抗菌活性的放线菌资源。
As a special actinomycete,intestinal symbiotic actinomycetes are different from the actinomycetes in the soil.Therefore,it is of great significance to study their types and antibacterial activities.Anti-Staphylococcus aureus and MRSA activities of 159 actinomycetes strains of Periplaneta americana were screened using the Oxford cup and confrontation co-culture method in vitro while using 2 species of human pathogenic bacteria of S.aureus and MRSA as the research object.Molecular biological identification of strains was carried out by 16S rRNA PCR,Blast homology comparison and phylogenetic tree construction.The result showed 49 endophytic actinomycetes had different degrees of anti-Staphylococcus aureus and MRSA activity,and 11 strains had antagonistic effects on both bacteria.Among the 49 strains of Actinomycetes,36 strains(73.47%)belong to the genus Streptomyces,which belongs to the dominant genus;the other 13 strains were Gordon s,Microbacteria,Brevibacterium,Mycobacterium,Achromobacte r and Micromono-spora.The results of agarose gel electrophoresis showed that the key enzyme genes PKS I、NRPS and FADH2-dependent halogenase were all detected in the 6 strains.This study showed that there were rich actinomycetes with antibacterial activity on the guts of P.americana.
作者
蒋诗林
叶宇歆
沈娟
朱家勇
金小宝
JIANG Shi-Lin;YE Yu-Xin;SHEN Juan;ZHU Jia-Yong;JIN Xiao-Bao(College of Life Science and Biopharmaceutical,Guangdong Key Laboratory of Pharmaceutical Bioactive Substances,Guangdong Pharmaceutical University,Guangzhou 510006,China)
出处
《环境昆虫学报》
CSCD
北大核心
2021年第2期453-461,共9页
Journal of Environmental Entomology
基金
广东省公益研究与能力建设项目(2017A020211008)
广东省普通高校基础研究与应用基础项目(2018KZDXM041)。
关键词
美洲大蠊
放线菌
抗金黄色葡萄球菌
抗耐甲氧西林金黄色葡萄球菌
分类鉴定
Periplaneta americana
Actinomycetes
anti-Staphylococcus aureus
anti-methicill-in-resistant Staphylococcus aureus
identification
