三份南海岛礁珊瑚砂样品中可培养细菌多样性

doi:10.11978/2021050

热带海洋学报 ›› 2022, Vol. 41 ›› Issue (2): 149-158.doi: 10.11978/2021050CSTR: 32234.14.2021050

三份南海岛礁珊瑚砂样品中可培养细菌多样性

李存^1,^3,⁴(), 崔林青^1,^3,⁴, 杨红强², 龙丽娟^1,³(), 田新朋^1,³()

1. 中国科学院热带海洋生物资源与生态重点实验室(中国科学院南海海洋研究所), 广东广州 510301
2. 中国科学院边缘海与大洋地质重点实验室(中国科学院南海海洋研究所), 广东广州 510301
3. 南方海洋科学与工程广东省实验室(广州), 广东广州 511458
4. 中国科学院大学, 北京 100049

收稿日期:2021-04-15 修回日期:2021-05-12 出版日期:2022-03-10 发布日期:2021-05-12
通讯作者: 龙丽娟,田新朋
作者简介:李存(1995—), 男, 山东省枣庄市人, 硕士研究生, 从事海洋微生物研究。email: licun18@mails.ucas.ac.cn
基金资助:
中国科学院战略性先导科技专项(XDA19060301);南方海洋科学与工程广东省实验室(广州)人才团队引进重大专项(GML2019ZD0402);广东省“珠江人才计划”本土创新科研团队项目(2019BT02Y262)

Diversity of cultured bacteria isolated from three coral reef sediments in South China Sea

LI Cun^1,^3,⁴(), CUI Linqing^1,^3,⁴, YANG Hongqiang², LONG Lijuan^1,³(), TIAN Xinpeng^1,³()

1. Key Laboratory of Tropical Marine Bio-resources and Ecology, Chinese Academy of Sciences (South China Sea Institute of Oceanology, Chinese Academy of Sciences), Guangzhou 510301, China
2. Key Laboratory of Ocean and Marginal Sea Geology, Chinese Academy of Sciences (South China Sea Institute of Oceanology, Chinese Academy of Sciences), Guangzhou 510301, China
3. Southern Marine Science and Engineering Guangdong Laboratory (Guangzhou), Guangzhou 511458, China
4. University of Chinese Academy of Sciences, Beijing 100049, China

Received:2021-04-15 Revised:2021-05-12 Online:2022-03-10 Published:2021-05-12
Contact: LONG Lijuan,TIAN Xinpeng
Supported by:
Strategic Priority Research Program of the Chinese Academy of Sciences(XDA19060301);Key Special Project for Introduced Talents Team of Southern Marine Science and Engineering Guangdong Laboratory (Guangzhou)(GML2019ZD0402);Local Innovative and Research Teams Project of Guangdong Pearl River Talents Program(2019BT02Y262)

摘要/Abstract

摘要：

岛礁珊瑚砂环境中存在着大量未被培养和利用的微生物资源, 微生物多样性研究是了解微生物生态功能、开发和利用微生物资源的基础。文章采用多种寡营养培养基选择性分离南海岛礁珊瑚砂中的可培养细菌, 共获得纯培养细菌菌株349株, 通过16S rRNA基因序列分析, 发现它们隶属于4门(Actinobacteria、Proteobacteria、Firmicutes和Bacteroidetes)、6纲、26目、43科、73属、134种, 可培养细菌的优势类群为放线菌门, 占所有分离菌株数量的60%; 而且还发现18个16S rRNA 基因序列相似性低于97%的潜在新种。本研究使用改良优化的寡营养培养基进行分离, 较好地显示出样品中微生物的群落组成, 且获得了大量潜在的稀有新物种资源。研究结果表明, 岛礁珊瑚砂样品的可培养细菌资源十分丰富、细菌群落所涉及的生态功能完整、潜在新种比例较高, 为后期岛礁微生物资源挖掘打下了良好的基础, 也为后期开发应用积累了丰富且稀有的菌种资源。

关键词: 南海岛礁, 寡营养培养基, 可培养细菌多样性, 微生物资源

Abstract:

There are a large number of uncultured microbial resources in coral reef environments. Microbial diversity research will help us understand their ecological functions, and also help develop and utilize microbial resources. In this study, a total of 349 pure bacterial strains were obtained by selective isolation media with multiple oligotrophic components, from three samples collected from three coral reef islands in the South China Sea. Based on 16S rRNA gene sequence analysis, we found the cultivable bacteria belong to four phyla of Actinobacteria, Proteobacteria, Firmicutes and Bacteroidetes; and they spread into six class, 26 orders, 43 families, 73 genera, and 134 species. There are 18 potential new taxa with 16S rRNA gene sequence similarities less than 97%. The dominant group is Actinobacteria, which accounts for 60% of all isolates. In this study, the improved and optimized oligotrophic media were used, which showed better isolation results including the composition of the microbial community and rare or new taxa in every sample. The results showed that the cultivable microbial resources are rich and diverse in coral reef island environments of the South China Sea, and the microbial community has comprehensive functions for material and energy metabolism; and the proportion is higher to obtain potential new taxa. This research has laid a good foundation for the methods to mine microbial resources in coral reef islands; it also accumulated abundant and rare microbial resources for the deep development and application in the future.

Key words: Coral reef island, oligotrophic medium, cultivable bacterial diversity, microbial resources

中图分类号:

P735.51

李存, 崔林青, 杨红强, 龙丽娟, 田新朋. 三份南海岛礁珊瑚砂样品中可培养细菌多样性[J]. 热带海洋学报, 2022, 41(2): 149-158.

LI Cun, CUI Linqing, YANG Hongqiang, LONG Lijuan, TIAN Xinpeng. Diversity of cultured bacteria isolated from three coral reef sediments in South China Sea[J]. Journal of Tropical Oceanography, 2022, 41(2): 149-158.

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培养基名称	培养基成分/(g·L^-1)
5% 2216	蛋白胨5.0, 酵母浸粉1.0, 柠檬酸铁0.1, NaCl 19.45, MgCl₂ 5.98, Na₂SO₄ 3.24, CaCl₂ 1.8, KCl 0.55, Na₂CO₃ 0.16, KBr 0.08, SrCl₂ 0.034, H₃BO₃ 0.022, Na₂O·SiO₂ 0.004, NaF 0.0024, NH₄NO₃ 0.0016, Na₂HPO₄ 0.008, 稀释20倍, 琼脂15. 0
5% AIA	酪蛋白酸钠2.0, 天冬酰胺0.1, 丙酸钠4.0, K₃PO₄ 0.5, MgSO₄ 0.1, FeSO₄ 0.001, 沸水浴1min后加入甘油5.0, 稀释20倍, 琼脂15. 0
5% R2A	胰蛋白胨0.25, 酸水解酪蛋白0.5, 酵母浸粉0.5, 可溶性淀粉0.5, K₂HPO₄ 0.3, MgSO₄ 0.1, 丙酮酸钠0.3, 蛋白胨0.25, 葡萄糖0.5, 稀释20倍, 琼脂15. 0
SN	NaNO₃ 0.75, K₂HPO₄ 0.0159, EDTA二钠0.0056, Na₂CO₃ 0.0104, VitaminB₁₂ 0.001(过滤除菌), 1×10^-6微量盐(乙酸6.25, 柠檬酸铁铵6.0, MnCl₂·4H₂O 1.4, Na₂MoO₄·2H₂O 0.39, Co(NO₃)₂·6H₂O 0.025, ZnSO₃·7H₂O 0.222, 单独灭菌), 琼脂15. 0
10%菌液琼脂	离心收集菌株Gaiella occulta F2-233^T的发酵液100mL, 过滤除菌, 加入900mL纯水, 琼脂15. 0

样品名称	Shannon指数	Simpson指数	Invsimpson指数	Chao1指数	ACE指数
Y11	2.39	0.85	6.78	53.00	42.54
BK-S	3.83	0.95	20.40	182.67	259.60
BS2-S	3.81	0.97	30.07	169.11	176.52

菌种编号	合并菌株数	最相似菌种	16S RNA序列相似性	新分类单元
SCSIO 63424	1	Arboricoccus pini	89.88	潜在新属
SCSIO 63409	3	Inquilinus limosus	92.80	潜在新属
SCSIO 63602	1	Nocardioides mesophilus	93.64	潜在新属
SCSIO 63346	1	Brevibacterium ammoniilyticum	94.61	潜在新属
SCSIO 63385	1	Gordonia terrae	94.74	潜在新属
SCSIO 63263	3	Brevibacterium casei	95.07	潜在新属
SCSIO 63650	1	Brachybacterium paraconglomeratum	95.66	潜在新属
SCSIO 63659	6	Nocardioides iriomotensis	96.00	潜在新种
SCSIO 63265	2	Haoranjiania flava	96.19	潜在新种
SCSIO 63649	1	Prauserella aidingensis	96.34	潜在新种
SCSIO 63671	1	Mesorhizobium oceanicum	96.45	潜在新种
SCSIO 63658	1	Prauserella rugosa	96.56	潜在新种
SCSIO 63505	1	Cohnella cellulosilytica	96.58	潜在新种
SCSIO 63329	1	Bacillus dafuensis	96.71	潜在新种
SCSIO 63500	1	Microbacterium oxydans	96.72	潜在新种
SCSIO 63633	1	Maricaulis virginensis	96.77	潜在新种
SCSIO 68054	1	Oceanobacillus iheyensis	97.01	潜在新种
SCSIO 63591	1	Sphingomonas aestuarii	97.02	潜在新种

三份南海岛礁珊瑚砂样品中可培养细菌多样性

Diversity of cultured bacteria isolated from three coral reef sediments in South China Sea

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