| 引用本文: | 刘敏,车文学,边伟杰,甘禧霖,赵怀宝.八门湾红树林土壤放线菌多样性及抗病原菌活性分析.海洋与湖沼,2022,53(2):352-363. |
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| 八门湾红树林土壤放线菌多样性及抗病原菌活性分析 |
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刘敏, 车文学, 边伟杰, 甘禧霖, 赵怀宝
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海南热带海洋学院 海南省现代化海洋牧场工程研究中心 海南三亚 572022
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| 摘要: |
| 红树林土壤中蕴藏着丰富的放线菌资源,是新菌种的重要来源。为获得更多新的放线菌资源以用于开发新的生物活性化合物,了解放线菌的多样性是非常重要和必要的。研究采用扩增子高通量测序和传统培养法对八门湾红树林土壤放线菌进行了非培养和培养水平多样性研究,同时对其抗病原菌活性进行了分析。对于非培养水平多样性研究,与细菌通用引物相比,利用放线菌相关引物可以提高放线菌丰度的检测水平,其百分比含量提高2.47倍;可以检测到放线菌门更多的目、科和属;对于在目水平上的放线菌类群组成来说,Acidimicrobiales、Corynebacteriales、Gaiellales、Kineosporiales、Solirubrobacterales是优势类群,但是在用两对不同引物得到的结果中,其百分含量差异较大。放线菌相关引物更适合环境样品中放线菌多样性的分析。对于培养水平多样性研究,分离到256株放线菌,属于7个目、9个科、14个属,Shannon-Wiener多样性指数为1.32,Streptomyces(42.58%)和Micromonospora(42.19%)是优势属;与模式菌株的相似性小于98.5%的有13株,其中菌株HA161004和HA161010与Amycolatopsis thermoflava N1165T相似性分别为95.56%和96.80%,是潜在新种。病原微生物拮抗活性测定表明,来自9个属的92株代表性菌株具有抗病原菌活性,其中对Colletotrichum gloeosporioides、Fusarium oxysporum、Escherichia coli、Staphylococcus aureus和Vibrio neocaledonicus有拮抗活性的菌株数量分别为71、25、6、9和19株。研究为选择更合适的引物以便更客观地反映环境中放线菌多样性提供了数据支持,同时也为后续红树林放线菌资源收集、活性评价和开发利用提供菌种保障。 |
| 关键词: 放线菌 多样性 抗菌活性 红树林 |
| DOI：10.11693/hyhz20210800176 |
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| 基金项目:海南热带海洋学院引进人才科研启动项目,RHDRC201901号;海南省重点计划项目,ZDYF2020190号;海南省基础与应用基础研究计划(自然科学领域)高层次人才项目,2019RC241号。 |
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| DIVERSITY AND ANTIMICROBIAL ACTIVITY OF ACTINOBACTERIA IN THE SOIL OF THE BAMENWAN MANGROVE IN HAINAN, CHINA |
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LIU Min, CHE Wen-Xue, BIAN Wei-Jie, GAN Xi-Lin, ZHAO Huai-Bao
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Modern Marine Ranching Engineering Research Center of Hainan, Hainan Tropical Ocean University, Sanya 572022, China
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| Abstract: |
| Mangrove is recognized a rich source of new Actinobacteria strains. To obtain more Actinobacteria for development of new bioactive compounds, it is important and necessary to understand their diversity. The diversity at culture-independent and culture-dependent level of Actinobacteria in the Bamenwan mangrove soil (Hainan, South China) was investigated by using amplicon high-throughput sequencing and traditional culture methods. The antimicrobial activities of representative isolates were characterized. For the diversity at culture-independent level, compared with bacterial universal primers, the detection level of actinobacterial abundance can be improved using Actinobacteria-related primers by 2.47 times in percentage. More orders, families, and genera of actinomycetes can be detected. Acidimicrobiales, Corynebacteriales, Gaiellales, Kineosporiales, Solirubrobacterales are the dominant orders, but their percentage contents are quite different in the results obtained by two pairs of different primers. Actinobacteria-related primers are more suitable for the analysis of actinobacterial diversity in environmental samples. For the diversity at culture-dependent level, 256 strains of actinobacterial strains were isolated, being affiliated to 7 orders, 9 families, and 14 genera. The Shannon-Wiener diversity index was 1.32. Streptomyces (42.58%) and Micromonospora (42.19%) were the dominant genera. The similarities of 13 strains with the type-strain were below 98.5%. Among them, strains HA161004 and HA161010 were the closest to Amycolatopsis thermoflava N1165T with similarities of 95.56% and 96.80%, respectively, indicating that both were potentially novel strains of species. The determination of antagonistic activity of pathogenic microorganisms showed that 92 representative strains from 9 genera had antimicrobial activity, among which 71, 25, 6, 9, and 19 strains had antagonistic activity against Colletotrichum gloeosporoides, Fusarium oxysporum, Escherichia coli, Staphylococcus aureus,and Vibrio neocaldonicus, respectively. This study provides data support for selecting more appropriate primers for the actinobacterial diversity, and supplies strain resources for the collection, activity evaluation, and utilization of Actinobacteria in mangrove. |
| Key words: Actinobacteria diversity antimicrobial activity mangrove |
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