生物技术通报 ›› 2021, Vol. 37 ›› Issue (11): 225-236.doi: 10.13560/j.cnki.biotech.bull.1985.2021-0288

• 研究报告 • 上一篇    下一篇

西藏沙棘根瘤及根际土壤放线菌分离及抗菌活性研究

罗亚军1(), 孙红敏2, 何宁2, 袁丽杰1(), 解云英2()   

  1. 1.华北理工大学基础医学院 河北省慢性疾病基础医学重点实验室 唐山市慢性病临床基础研究重点实验室,唐山 063210
    2.中国医学科学院药物合成生物学重点实验室 中国医学科学院&北京协和医学院医药生物技术研究所,北京 100050
  • 收稿日期:2021-03-11 出版日期:2021-11-26 发布日期:2021-12-03
  • 作者简介:罗亚军,男,硕士,研究方向:微生物的分离鉴定、抗菌活性及天然产物挖掘;E-mail: 592062203@qq.com
  • 基金资助:
    国家自然科学基金项目(81973219);中国医学科学院医学与健康科技创新工程经费(2016-I2M-2-002);河北省自然科学基金项目(H2021209027)

Isolation and Antibacterial Activity of Actinomycetes from the Nodules and Rhizosphere Soil of Hippophae rhamnoides in Tibet

LUO Ya-jun1(), SUN Hong-min2, HE Ning2, YUAN Li-jie1(), XIE Yun-ying2()   

  1. 1. School of Basic Medical Science,North China University of Science and Technology,Hebei Key Laboratory for Chronic Disease,Tangshan Key Laboratory for Preclinical and Basic Research on Chronic Disease,Tangshan 063210
    2. CAMS Key Laboratory of Synthetic Biology for Drug Innovation,Institute of Medicinal Biotechnology,Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100050
  • Received:2021-03-11 Published:2021-11-26 Online:2021-12-03

摘要:

探索西藏林芝与山南地区沙棘根瘤及其根际土壤样品中放线菌的多样性及抗菌活性,为天然产物药物研发提供新的微生物资源。采用稀释涂布法对10份样品中可培养放线菌进行分离,后经菌株16S rRNA序列测序,以邻接法(neighbor-joining method,N-J)构建系统发育树分析放线菌种属多样性;利用微孔板法对代表性菌株进行发酵培养及抗菌活性评价;进一步利用超高效液相色谱-质谱联用(UPLC-MS)考察活性菌株代谢产物情况。从10份样品中分离到112株放线菌,分属于7个目12科23个属,优势菌属为链霉菌属(Streptomyces);抗菌活性结果表明有17株放线菌至少对一种检定菌有抗菌活性,阳性抑菌率为36.9%。有16株放线菌对临床常见的耐甲氧西林金黄色葡萄球菌(methicillin-resistant Staphylococcus aureus,MRSA)有较强抑制活性,其中13株放线菌对MRSA抑制率超过阳性对照药万古霉素;另外有7株放线菌对耐碳青霉烯鲍曼不动杆菌(carbapenem-resistant Acinetobacter baumannii,CRAB)有抑制活性,其中Streptomyces sp. XZ19-363对CRAB抑菌作用最强,抑制率为91.3%,UPLC-MS分析显示,具有广谱抗菌活性的菌株在不同培养基中产生了较为丰富的次级代谢产物。西藏林芝与山南地区沙棘根瘤及其根际土壤中可培养放线菌资源丰富多样,同时具有较好的抗菌活性且代谢产物丰富。

关键词: 沙棘根瘤, 根际土壤, 放线菌, 抗菌活性

Abstract:

In order to provide new microbial resources for the development of natural product drugs,the diversity and antibacterial activity of actinomycetes from the root nodules and rhizosphere soil samples of Hippophae rhamnoides in Nyingchi and Shannan of Tibet were explored. Dilution coating method was empployed to isolate the cultivable actinomycetes from 10 samples. After the 16S rRNA sequence alignment of the strains,the neighbor-joining method(N-J)was used to construct a phylogenetic tree to analyze the species diversity of actinomycetes. The microplate method was adapted to conduct fermentation and antibacterial activity evaluation of representative strains;and ultra-performance liquid chromatography-mass spectrometry(UPLC-MS)was to investigate the metabolites of active strains. A total of 112 strains of actinomycetes belonged to 23 genera,12 families and 7 orders were isolated from 10 samples,in which Streptomyces was the dominant genus. The results of antibacterial activity showed that 17 strains of estimated actinomycetes presented antibacterial activity against at least one of the tested bacteria,and the positive inhibition rate was 36.9%. Sixteen actinomycetes showed strong inhibitory activity against common clinical methicillin-resistant Staphylococcus aureus(MRSA),13 of which had higher inhibitory rate than the positive control drug vancomycin. In addition,7 of evaluated actinomycetes demonstrated inhibitory activity against carbapenem-resistant Acinetobacter baumannii(CRAB),from which Streptomyces sp. XZ19-363 had the strongest antibacterial activity against CRAB,with an inhibition rate of 91.3%. UPLC-MS analysis revealed that the strains with broad-spectrum antibacterial activities produced relatively abundant secondary metabolites in different media. The resources of cultivable actinomycetes from H. rhamnoides nodules and rhizosphere soils in Nyingchi and Shannan regions of Tibet both are rich and diverse,and possess promising antibacterial activities with rich metabolites.

Key words: Hippophae rhamnoides nodules, rhizosphere soil, actinobacteria, antibacterial activity