热带作物学报2024，Vol.45Issue(7)：1498-1510,13.DOI:10.3969/j.issn.1000-2561.2024.07.022
基于高通量测序和纯培养技术解析海南东寨港红树植物老鼠簕根际放线菌多样性及其潜在的功能
Analysis of Rhizospheric Actinomycetes Diversity and Potential Func-tions of Mangrove Acanthus ilicifolius in Dongzhaigang of Hainan Province Based on High-throughput Sequencing and Pure Culture
摘要
Abstract
Rhizosphere is a dynamic interface where soil,plant roots and microorganisms exchange substances.Rhizospheric actinomycetes are an important component of root actinomycetes and one of the main sources of bioactive substances.In this study,five different rhizospheric soil samples of a mangrove plant,Acanthus ilicifolius,were col-lected from the Dongzhaigang Mangrove Nature Reserve of Hainan province.A combination of high-throughput se-quencing and traditional pure culture strategies were used to explore the diversity of the rhizospheric actinomycetes of A.ilicifolius and the potential to produce bioactive substances in the samples.The V3 to V4 regions of 16S rRNA gene in the rhizospheric samples of A.ilicifolius were sequenced and analyzed by Illumina Novaseq 6000 sequencing platform.A total of 32 phyla(including 25 effectively identified phyla)were annotated,of which Proteobacteria is the dominant phyla.α-diversity analysis showed that the rhizospheric bacterial community of A.ilicifolius was very abundant.The microbial communities of actinomycetes were analyzed,and 33 validated genera,four tentative genera and 28 uncul-tured new genera were noted.KEGG functional prediction analysis in different levels showed that rhizospheric actino-mycetes were mainly involved in the biosynthesis of secondary metabolites including various antibiotics.Meanwhile,actinomycetes were isolated by the traditional pure culture method.After isolation,purification,morphological charac-teristics grouping and rearrangement,the diversity of actinomycetes of mangrove A.ilicifolius was analyzed by PCR amplification,sequencing and comparison of 16S rRNA gene sequences.The antimicrobial activity was screened by the agar diffusion method.PCR-based gene screening technology was used to detect the possible antibiotic biosynthesis genes of the representative strains of PKS-I,PKS-Ⅱ and NRPS.A total of 33 representative strains of rhizospheric ac-tinomycetes were obtained from four genera,of which Streptomyces was the dominant genus,and 2 genera could not be detected by high-throughput sequencing method.The positive rates of NRPS and PKS-Ⅱ genes of the representative strains were very high(more than 90％).24.24％of rhizospheric actinomycetes had the activity against at least one or more indicator microorgnisms.In this study,high-throughput sequencing and traditional pure culture methods were used to show that rhizospheric actinomycetes of a mangrove plant A.ilicifolius from Dongzhaigang had a very rich actino-mycetic diversity.The rhizospheric actinomyces of A.ilicifolius has good potential to synthesize secondary metabolites,which could provide rich microbial resources for the screening of bioactive compounds,and also provide an important basis for the development of subsequent new drugs.关键词
老鼠簕/根际/放线菌多样性/高通量测序/纯培养Key words
Acanthus ilicifolius/rhizosphere/actinomycetic diversity/high-throughput sequencing/pure culture分类
生物科学引用本文复制引用
刘顺卿,何小稳,吴金燕,易国辉,陈芊芊,谢欣宜,隋金蕾..基于高通量测序和纯培养技术解析海南东寨港红树植物老鼠簕根际放线菌多样性及其潜在的功能[J].热带作物学报,2024,45(7):1498-1510,13.基金项目
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(No.2019RC216) （自然科学领域）
海南医学院引进人才科研启动基金项目(No.XRC190006) （No.XRC190006）
海南医学院(国家级)大学生创新创业训练计划项目(No.202311810008). （国家级）
