药用昆虫桂花蝉共生放线菌的分离培养及抗菌活性研究OACSTPCD
Study on the isolation,culture and antibacterial activity of actinomyces derived from Chinese herbal medicine Kirkaldyia deyrolli and whole genome sequencing of a Gordonia strain
目的 探讨药用昆虫桂花蝉肠道共生放线菌的多样性,为抗菌药物开发提供新的微生物资源.方法 采用平板稀释法对桂花蝉共生放线菌进行分离和纯化;采用牛津杯法测定菌株抗菌活性;通过外观观察、扫描电镜和16S rDNA序列分析进行分类鉴定;对具有良好抗菌活性的菌株进行Nanopore第3代测序;利用Antismash、Prism、Circos、Rodigal、eggNOG、GO和KEGG等工具对基因组数据进行注释分析.结果 采用6种培养基从桂花蝉肠道共分离获得34株不同放线菌,其中链霉菌属(Streptomyces)32株、戈登氏菌属(Gordonia)1株和冢村氏菌属(Tsukamurella)1株,菌株GHC 11可能为1株新的链霉菌种,73.5%的菌株对至少1种病原菌具有拮抗作用.菌株GHC 54具有显著的抗S.aureus和MRSA活性,通过形态学和分子生物学方法鉴定为Gordonia terrae.菌株GHC 54基因组大小为5 346 952 bp,包含1个5 210 251 bp环状染色体和1个136 601 bp质粒,共编码4 828个基因,GC含量为67.91%,其中含有50个tRNA基因、9个rRNA基因.编码基因分别进行eggNOG、GO、KEGG、Nr、Pfam、Swiss-prot和TrEMBL功能注释,共注释到4 776个蛋白.菌株GHC 54中含有14个生物合成基因簇,包含6个NRPS、2个Terpene、1个Ectoine、1个Arylpolyene、1个Redox-cofactor、1个NAPAA、1个Ripp-like和1个PKS基因簇,85.7%的基因簇同源性小于20%,具有较高的新颖性.结论 桂花蝉共生放线菌具有良好的抗菌活性,菌株GHC 54具有合成新颖抗菌化合物的潜力.本研究将为抗菌药物的开发提供新的微生物资源.
Object To explore the diversity of symbiotic actinomycetes in the gut of the medicinal insect Kirkaldyia deyrolli,and provide new microbial resources for the development of antimicrobial drugs.Methods Isolation and purification of symbiotic actinomycetes from the gut of the Kirkaldyia deyrolli were carried out by the plate dilution method;Antibacterial activity of the strains was determined by the Oxford cup method.Classification and identification were performed by visual observation,scanning electron microscopy,and 16S rDNA sequence analysis.Strains exhibiting significant antibacterial activity were subjected to Nanopore third-generation sequencing,Genome data were annotated and analyzed by tools such as Antismash,Prism,Circos,Prodigal,eggNOG,GO,and KEGG.Results 34 strains of various actinomycetes were isolated from the gut of the Kirkaldyia deyrolli by six different culture media,including 32 strains belonging to the Streptomyces genus,1 strain to the Gordonia genus,and 1 strain to the Tsukamurella genus.Strain GHC 11 was likely a new species of Streptomyces.73.5%of the isolates were antagonistic to at least one pathogen.Strain GHC 54 exhibited significant antibacterial activity against S.aureus and MRSA.Based on morphological and molecular identification,strain GHC 54 was confirmed to belong to Gordonia terrae.The genome of strain GHC 54 had a size of 5 346 952 bp,including a circular chromosome of 5 210 251 bp and a plasmid of 136,601 bp,encoding a total of 4 828 genes with a GC content of 67.91%,which included 50 tRNA genes and 9 rRNA genes.Functional annotation of the encoded genes with eggNOG,GO,KEGG,Nr,Pfam,Swiss-prot,and TrEMBL identified a total of 4 776 annotated proteins.Strain GHC 54 contained 14 biosynthetic gene clusters,including 6 NRPS,2 Terpene,1 Ectoine,1 Arylpolyene,1 Redox-cofactor,1 NAPAA,1 Ripp-like,and 1 PKS gene clusters.The homogeneity of 85.7%of the gene clusters was less than 20%,indicating a high level of novelty.Conclusion The symbiotic actinomycetes associated with the Kirkaldyia deyrolli have good antibacterial activity,with strain GHC 54 showing potential for synthesizing novel antimicrobial compounds.This study will contribute new microbial resources for the development of antimicrobial drugs.
苏雅琳;廖婧阳;欧健鹏;刘文彬
广东药科大学基础医学院,广东广州 510006||广东药科大学生命科学与生物制药学院,广东广州 510006广东药科大学基础医学院,广东广州 510006||广东省生物活性药物研究重点实验室,广东广州 510006
生物学
桂花蝉共生放线菌戈登氏菌全基因组测序
Kirkaldyia deyrollisymbiotic actinomycesGordoniawhole genome sequencing
《广东药科大学学报》 2024 (002)
7-15 / 9
广东省大学生创新创业训练计划项目(202210573021);广州市基础与应用基础研究项目(202201010357);广东省医学科研基金(A2022069)
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