Khai岛和Pathiu岛珊瑚礁沉积物细菌多样性及细菌粗提物延缓秀丽隐杆线虫衰老活性研究*

doi:10.11978/2019126

热带海洋学报 ›› 2020, Vol. 39 ›› Issue (5): 19-29.doi: 10.11978/2019126CSTR: 32234.14.2019126

Khai岛和Pathiu岛珊瑚礁沉积物细菌多样性及细菌粗提物延缓秀丽隐杆线虫衰老活性研究*

韩敏敏¹(), 李蜜¹, 刘昕明², 刘永宏³, 龙超³, 钟振国¹, 易湘茜¹, 高程海¹()

1.广西中医药大学海洋药物研究院, 广西中医药科学实验中心, 药学院, 广西南宁 530200
2.广西壮族自治区海洋研究院, 广西南宁 530022
3.中国科学院南海海洋研究所热带海洋生物资源与生态重点实验室, 广东广州 510301

收稿日期:2019-12-05 修回日期:2020-03-03 出版日期:2020-09-10 发布日期:2020-03-05
通讯作者: 高程海
作者简介:韩敏敏(1993—), 女, 山东省济宁市人, 硕士研究生, 主要从事海洋药物基础与产品开发研究。E-mail: 2846748658@qq. com
基金资助:
国家自然科学基金项目(41566004);国家自然科学基金项目(21662006);广西中医药大学岐黄工程高层次人才团队培育项目(2018006);广西中医药大学博士科研启动基金项目(2017BS039);广西中医药大学海洋药物研究院团队科研专项经费项目(2018ZD005-A06)

Studies on bacterial diversity in coral reef sediments in Khai Island and Pathiu Island and bacterial crude extract retards aging activity of *Caenorhabditis elegans**

HAN Minmin¹(), LI Mi¹, LIU Xinming², LIU Yonghong³, LONG Chao³, ZHONG Zhenguo¹, YI Xiangxi¹, GAO Chenghai¹()

1. Institute of Marine Drugs, Guangxi Scientific Research Center of Traditonal Chinese Medicine, Faculty of Pharmacy Guangxi University of Chinese Medicine, Nanning 530200, China
2. Guangxi Academy of Oceanography, Nanning 530022, China
3. Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China

Received:2019-12-05 Revised:2020-03-03 Online:2020-09-10 Published:2020-03-05
Contact: Chenghai GAO
Supported by:
National Natural Science Foundation(41566004);National Natural Science Foundation(21662006);Development Program of High-level Talent Team under Qihuang Project of Guangxi University of Chinese Medicine(2018006);Doctoral Research Foundation of Guangxi University of Traditional Chinese Medicine(2017BS039);Scientific Research Foundation of Institute of Marine Drugs, GUCM(2018ZD005-A06)

摘要/Abstract

摘要：

在Khai岛和Pathiu岛珊瑚礁生长海域采集2份珊瑚礁沉积物样品, 将样品预处理后采用6种分离培养基进行分离培养, 获得活性海洋细菌菌株, 通过PCR扩增和16s rRNA基因序列测定, 分析物种多样性, 并构建系统发育树。以模式生物秀丽隐杆线虫为模型考察菌株发酵提取物延缓衰老的作用。实验结果表明, 从4种样品中分离鉴定出22株细菌, 分别属于11目13科14属。16S rRNA基因序列相似性低于98.65%的细菌有2株, 疑似为潜在新种。GXIMD014 (Vibrio galatheae)和GXIMD112 (Kocuria kristinae)的500μg·mL^-1细菌粗提物具有延缓线虫衰老的活性, 能不同程度的延长线虫寿命, 提高线虫产卵量、运动能力、热激损伤能力、抗氧化损伤能力、体内酶活性以及改善虫体表皮形态。Khai岛和Pathiu岛珊瑚礁沉积物细菌具有较高的物种多样性, 蕴藏着丰富的微生物资源, 且部分细菌菌株具有延缓衰老的生物活性。

关键词: 珊瑚礁, 海洋细菌, 物种多样性, 延缓衰老活性

Abstract:

Two coral reef sediment samples were collected from the coral reef growth areas of Khai Island and Pathiu Island After the samples were pretreated, six kinds of separation medium were adopted to obtain the active Marine bacterial strains. Through PCR amplification and 16s rRNA gene sequencing, species diversity was analyzed and phylogenetic trees were constructed. The effect of fermentation extract of Caenorhabditis elegans strain on senescence was investigated. The results showed that 22 strains of bacteria were isolated and identified from four samples, belonging to 11 orders, 13 families and 14 genera. There were two strains of bacteria with 16S rRNA gene sequence similarity less than 98.65 %, which were suspected to be potential new species. The crude extracts of 500 μg·mL ^-1bacteria from GXIMD014 (Vibrio galatheae) and GXIMD112 (Kocuria kristinae) have the activity of delaying the aging of Caenorhabditis elegans, and can prolong the life of Caenorhabditis elegans to different degrees, improve the oviposition, motility, heat-induced injury, ability to resist oxidative damage, enzyme activity in vivo, and improve the epidermal morphology of Caenorhabditis elegans. Coral reef sediment bacteria of Khai Island and Pathiu Island have high species diversity and rich microbial and bacterial resources, and some bacterial strains have anti-aging biological activity.

Key words: coral reef, marine bacteria, species diversity, anti-aging activity

中图分类号:

P735.51

韩敏敏, 李蜜, 刘昕明, 刘永宏, 龙超, 钟振国, 易湘茜, 高程海. Khai岛和Pathiu岛珊瑚礁沉积物细菌多样性及细菌粗提物延缓秀丽隐杆线虫衰老活性研究*[J]. 热带海洋学报, 2020, 39(5): 19-29.

HAN Minmin, LI Mi, LIU Xinming, LIU Yonghong, LONG Chao, ZHONG Zhenguo, YI Xiangxi, GAO Chenghai. Studies on bacterial diversity in coral reef sediments in Khai Island and Pathiu Island and bacterial crude extract retards aging activity of Caenorhabditis elegans*[J]. Journal of Tropical Oceanography, 2020, 39(5): 19-29.

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类群	菌株编号	相似性最高模式菌株	16S rRNA基因序列相似性/%	GenBank登录号
Kocuria sp.	GXIMD112	K. Kristinae (BCSM01000028)	99.74	MT111730
Vibrio sp.	GXIMD014	V. Galatheae (JXXV01000023)	99.35	MT111742
Vibrio sp.	GXIMD099	V. Sagamiensis (AB428909)	98.75
Bacillus sp.	GXIMD115	B. Megaterium (JJMH01000057)	99.74
	GXIMD039	B. Aryabhattai (EF114313)	99.48
	GXIMD105	B. Altitudinis (ASJC01000029)	98.97
Pseudomonas sp.	GXIMD091	P. Pachastrellae (MUBC01000081)	98.97
Shewanel sp.	GXIMD013	S.Waksmanii (AY170366)	99.59
Staphylococcus sp.	GXIMD124	S. Gallinarum (D83366)	99.23
	GXIMD017	S. Nepalensis (UHDS01000001)	99.09
	GXIMD114	S. Haemolyticus (LILF01000056)	100.00
	GXIMD133	S. hominis subsp. Novobiosepticus (AB233326)	99.35
Achromobacter sp.	GXIMD088	A. marplatensis (EU150134)	98.83
Achromobacter sp.	GXIMD134	A. Ruhlandii (AB010840)	99.61
Leeuwenhoekiella sp.	GXIMD126	L. Palythoae (jgi.1059018)	98.83
Marinomonas sp.	GXIMD067	M. Vaga (X67025)	99.09
Gordonia sp.	GXIMD145-1	G. Bronchialis (CP001802)	99.23
Rhizobium sp.	GXIMD055	R. Roseltifomans (EU781656)	99.36
Erythrobacter sp.	GXIMD154	E. Pelagi (HQ203045)	99.87
Alteromonas sp.	GXIMD063	A. Macleodii (CP003841)	99.61
Alteromonas sp.	GXIMD003	A. Marina (AF529060)	97.93
Aestuariibacter sp.	GXIMD144	A. aggregatus (FJ847832)	96.68

菌株编号	平均寿命/d	最长寿命/d	菌株编号	平均寿命/d	最长寿命/d
1% DMSO	16.68±1.057	18.83	GXIMD039	19.75±1.060*	21.89
GXIMD055	14.50±0.844	16.21	GXIMD114	15.30±0.834	16.99
GXIMD099	15.50±0.938	17.40	GXIMD115	14.25±0.839	15.95
GXIMD154	16.56±1.126	18.84	GXIMD091	17.10±1.261	19.65
GXIMD144	14.30±0.818	15.96	GXIMD088	15.65±0.826	17.32
GXIMD112	20.75±1.417**	23.62	GXIMD145-1	14.27±0.891	16.08
GXIMD013	16.20±0.891	18.00	GXIMD014	21.75±1.119**	22.01
GXIMD124	16.45±1.041	18.56	GXIMD003	15.05±0.950	16.97
GXIMD017	17.22±1.250	19.78	GXIMD134	16.10±1.107	18.34
GXIMD126	15.20±0.925	17.07	GXIMD063	13.25±0.867	15.00
GXIMD067	13.15±0.671	14.51	GXIMD133	14.40±0.817	16.05
GXIMD105	14.22±1.010	16.26

属	种	菌株粗提物	菌株粗提物编号	浓度/(μg·mL^-1)	菌株粗提物编号
Vibrio sp.	Vibrio galatheae	GXIMD014细菌粗提物	S1	250	S1-1
				500	S1-2
				1000	S1-3
Kocuria sp.	Kocuria kristinae	GXIMD112细菌粗提物	S2	250	S2-1
				500	S2-2
				1000	S2-3
Bacillus sp.	Bacillus aryabhattai	GXIMD039细菌粗提物	S3	250	S3-1
				500	S3-2
				1000	S3-3

Khai岛和Pathiu岛珊瑚礁沉积物细菌多样性及细菌粗提物延缓秀丽隐杆线虫衰老活性研究*

Studies on bacterial diversity in coral reef sediments in Khai Island and Pathiu Island and bacterial crude extract retards aging activity of *Caenorhabditis elegans**

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