龙梦,樊慧敏,夏洪丽,汪志文,喻大鹏,夏立群,鲁义善

• 1:广东海洋大学深圳研究院/广东省水生动物健康评估工程技术研究中心/深圳市海水经济动物种苗健康评价公共技术服务平台
• 2:广东海洋大学水产学院/广东省水产动物病害防控与健康养殖重点实验室

摘要(Abstract)：

【目的】研究广东深圳红树林沉积物可培养细菌多样性，筛选潜在生物活性菌株及水产病原菌拮抗菌。【方法】采用稀释涂布法分离纯化深圳红树林沉积物中可培养细菌，基于16S rRNA序列分析细菌物种多样性，通过PCR扩增聚酮合酶基因、非核糖体肽合成酶基因检测菌株产次级代谢产物潜力，并采用点种法筛选水产病原菌拮抗菌。【结果】获得177株细菌，隶属于4门20属27种，芽孢杆菌属（Bacillus）是优势菌属，6株细菌为潜在新物种，55株细菌的基因组DNA扩增出至少1种生物合成基因，其中有7株细菌对水产病原菌有拮抗活性。【结论】深圳红树林沉积物可培养细菌资源丰富，有应用于活性次级代谢产物发掘和水产病害绿色防控的巨大潜力。

关键词(KeyWords)： 红树林沉积物;细菌多样性;拮抗活性;水产病原菌;生物合成基因

Abstract:

Keywords:

基金项目(Foundation): 深圳市科技计划项目（20180507183240459）;; 深圳市大鹏新区科技创新和产业发展专项资金资助项目（PT202101-24）

作者(Author): 龙梦,樊慧敏,夏洪丽,汪志文,喻大鹏,夏立群,鲁义善

参考文献(References)：

• [1] XU J. Bioactive natural products derived from mangroveassociated microbes[J]. RSC Advances, 2015, 5(2):841-892.
• [2]赵雅慧,张舒琳,吴家法,等.山口红树林根际土壤可培养细菌多样性及其活性筛选[J].海洋学报, 2018, 40(8):138-151.
• [3]李菲,李喆,胡文进,等.广西茅尾海红树林土壤放线菌多样性及功能酶活性研究[J].海洋学报, 2021, 43(2):88-97.
• [4]孟昊,薛智权,唐杰,等.深圳福田红树林土壤可培养微生物和土壤酶活性研究[J].广东农业科学, 2013, 40(1):53-56.
• [5] YANG Q, TAM N F Y, WONG Y S, et al. Potential use of mangroves as constructed wetland for municipal sewage treatment in Futian, Shenzhen, China[J]. Marine Pollution Bulletin, 2008, 57(6/7/8/9/10/11/12):735-743.
• [6] CHAI M W, SHEN X X, LI R L, et al. The risk assessment of heavy metals in Futian mangrove forest sediment in Shenzhen Bay(South China)based on SEM-AVS analysis[J]. Marine Pollution Bulletin, 2015, 97(1/2):431-439.
• [7]陈昌福,周鑫军.浅谈21世纪人类面临的危机与应对微生物耐药问题[J].当代水产, 2021, 46(1):76-77; 79.
• [8] MOFFITT M C, NEILAN B A. Evolutionary affiliations within the superfamily of ketosynthases reflect complex pathway associations[J]. Journal of Molecular Evolution,2003, 56(4):446-457.
• [9]李菲,苏芯莹,李喆,等.广西北部湾海域表层海水细菌多样性及其潜在生物合成基因研究[J].广西科学院学报,2022, 38(1):76-87.
• [10] ZHAO J, YANG N, ZENG R Y. Phylogenetic analysis of type I polyketide synthase and nonribosomal peptide synthetase genes in Antarctic sediment[J]. Extremophiles,2008, 12(1):97-105.
• [11] JOHNSON R, VOISEY C, JOHNSON L, et al. Distribution of NRPS gene families within the Neotyphodium/Epichlo?complex[J]. Fungal Genetics and Biology, 2007, 44(11):1180-1190.
• [12] EHRENREICH I M, WATERBURY J B, WEBB E A.Distribution and diversity of natural product genes in marine and freshwater cyanobacterial cultures and genomes[J].Applied and Environmental Microbiology, 2005, 71(11):7401-7413.
• [13] SILVA C S P, GENUáRIO D B, VAZ M G M V, et al.Phylogeny of culturable cyanobacteria from Brazilian mangroves[J]. Systematic and Applied Microbiology,2014, 37(2):100-112.
• [14] LI Y M, LI Y M, LI Q, et al. Biosynthetic and antimicrobial potential of Actinobacteria isolated from bulrush rhizospheres habitat in Zhalong Wetland, China[J]. Archives of Microbiology, 2018, 200(5):695-705.
• [15] ALMEIDA E, DIAS T V, FERRAZ G, et al. Culturable bacteria from two Portuguese salterns:diversity and bioactive potential[J]. Antonie Van Leeuwenhoek, 2020, 113(4):459-475.
• [16]周双清,黄小龙,黄东益,等. Chelex-100快速提取放线菌DNA作为PCR扩增模板[J].生物技术通报, 2010(2):123-125.
• [17] WALSH P S, METZGER D A, HIGUCHI R. Chelex 100as a medium for simple extraction of DNA for PCR-based typing from forensic material[J]. BioTechniques, 1991, 10(4):506-513.
• [18] STACKEBRANDT E, EBERS J. Taxonomic parameters revisited:tarnished gold standards[J]. Microbiology Today,2006, 33:152-155.
• [19] ZHANG Y Y, DONG J D, YANG B, et al. Bacterial community structure of mangrove sediments in relation to environmental variables accessed by 16S rRNA gene-denaturing gradient gel electrophoresis fingerprinting[J]. Scientia Marina, 2009, 73(3):487-498.
• [20] DIAS A C F, ANDREOTE F D, DINI-ANDREOTE F, et al.Diversity and biotechnological potential of culturable bacteria from Brazilian mangrove sediment[J]. World Journal of Microbiology and Biotechnology, 2009, 25(7):1305-1311.
• [21]李菲,李喆,覃仙玲,等.山口红树林土壤芽孢杆菌多样性及纤维素酶活性筛选研究[J].广西科学, 2021, 28(5):473-481.
• [22]林鹏,张瑜斌,邓爱英,等.九龙江口红树林土壤微生物的类群及抗菌活性[J].海洋学报, 2005, 27(3):133-141.
• [23] SHOME R, SHOME B, MANDAL A, et al. Bacterial flora in mangroves of Andaman-Part I:Isolation, identification and antibiogram studies[J]. Indian Journal of Geo-Marine Sciences, 1995, 24:97-98.
• [24] DE LIMA PROCóPIO R E, DA SILVA I R, MARTINS M K, et al. Antibiotics produced by Streptomyces[J]. The Brazilian Journal of Infectious Diseases, 2012, 16(5):466-471.
• [25] BAJ J. Taxonomy of the genus Paracoccus[J]. Acta Microbiologica Polonica, 2000, 49(3/4):185-200.
• [26] ZHOU M Y, WANG G L, LI D, et al. Diversity of both the cultivable protease-producing bacteria and bacterial extracellular proteases in the coastal sediments of King George Island, Antarctica[J]. PLoS One, 2013, 8(11):e79668.
• [27] ZHANG W Y, YUAN Y, SU D Q, et al. Gallaecimonas mangrovi sp. nov., a novel bacterium isolated from mangrove sediment[J]. Antonie van Leeuwenhoek, 2018, 111(10):1855-1862.
• [28] SUBINA N S, NAIR S, GONSALVES M J. Spongeassociated bacterium, Yangia pacifica:a potential candidate for bioremediation of azo dyes[J]. Current Science, 2017,113(5):965.
• [29] MOH T H, LAU N S, FURUSAWA G, et al. Complete genome sequence of Microbulbifer sp CCB-MM1, a halophile isolated from Matang Mangrove Forest, Malaysia[J].Standards in Genomic Sciences, 2017, 12(1):36.
• [30] HUANG H Q, MO K L, HU Y H, et al. Microbulbifer harenosus sp. nov., an alginate-degrading bacterium isolated from coastal sand[J]. International Journal of Systematic and Evolutionary Microbiology, 2020, 70(3):1639-1643.
• [31]王岳坤,洪葵.红树林土壤因子对土壤微生物数量的影响[J].热带作物学报, 2005, 26(3):109-114.
• [32] GONZALEZ-ACOSTA B, BASHAN Y, HERNANDEZSAAVEDRA N Y, et al. Seasonal seawater temperature as the major determinant for populations of culturable bacteria in the sediments of an intact mangrove in an arid region[J]. FEMS Microbiology Ecology, 2006, 55(2):311-321.
• [33] FIRA D, DIMKI?I, BERI?T, et al. Biological control of plant pathogens by Bacillus species[J]. Journal of Biotechnology, 2018, 285:44-55.
• [34] JE?EWSKA-FR?CKOWIAK J, SEROCZY?SKA K,BANASZCZYK J, et al. The promises and risks of probiotic Bacillus species[J]. Acta Biochimica Polonica, 2018, 65(4):509-519.
• [35]张静,陈红莲,鲍俊杰,等.水产养殖中嗜水气单胞菌拮抗菌的研究进展[J].江苏农业科学, 2020, 48(17):21-33.