南海热带岛礁生物土壤结皮中细菌的分离及其固砂特性初步研究

doi:10.11978/2022266

热带海洋学报 ›› 2023, Vol. 42 ›› Issue (6): 101-110.doi: 10.11978/2022266CSTR: 32234.14.2022266

南海热带岛礁生物土壤结皮中细菌的分离及其固砂特性初步研究

黄谕¹^,²(), 王琳¹, 麦志茂¹, 李洁¹, 张偲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.中国科学院大学, 北京 100049
3.南方海洋科学与工程广东省实验室(广州), 广东广州 511458

收稿日期:2022-12-29 修回日期:2023-02-27 出版日期:2023-11-10 发布日期:2023-03-06
作者简介:
黄谕(1997—), 女, 海南省文昌市人, 硕士研究生, 从事岛礁生物土壤结皮微生物生态学研究。email: huangyu20@mails.ucas.ac.cn
基金资助:
国家自然科学基金青年科学基金项目(42206155); 广东省自然科学基金面上项目(2022A1515011889); 广州市科技局基础与应用基础研究项目(202201010499)

Isolation and characterization of sand fixation ability of bacteria in biological soil crusts of the tropical islands, South China Sea

HUANG Yu¹^,²(), WANG Lin¹, MAI Zhimao¹, LI Jie¹, ZHANG Si¹^,³()

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

Received:2022-12-29 Revised:2023-02-27 Online:2023-11-10 Published:2023-03-06
Supported by:
National Natural Science Foundation of China(42206155); Natural Science Foundation of Guangdong Province, China(2022A1515011889); Guangzhou Basic and Applied Basic Research Foundation(202201010499)

摘要/Abstract

摘要：

南海热带岛礁生物土壤结皮中有大量可分泌胞外多糖的微生物, 这一特性对固砂具有重要的促进作用。本研究利用TSA、改良TSA、MA三种培养基对采自南海永暑礁及三亚鹿回头海岸带的生物土壤结皮进行可培养细菌的分离纯化。本研究共分离纯化70株细菌, 并进行基于16S rRNA基因序列的鉴定, 发现所分离的菌株属于3门5纲12目19科25属, 厚壁菌门(Firmicutes)为优势门, 芽孢杆菌属(Bacillus)为优势属。此外, 有22株菌与已知物种16S rRNA基因相似性低于98.65%, 为潜在的新物种。纯化菌株的胞外多糖采用乙醇沉淀法提取、苯酚硫酸法检测含量, 发现19株菌胞外多糖含量高于0.013mg·mL^-1。选取其中胞外多糖含量最高的9株菌进行土壤团聚能力的测定, 发现将菌株SCSIO 17111(Lysobacter sp.)菌液喷洒于珊瑚砂表面能够使土壤团聚体保持稳定。针对粒径小于0.2mm的细砂, 菌株SCSIO 17111固结的砂土在经过干筛处理后, 保留量可达92.7%。本研究可为岛礁珊瑚砂的固定、减少土壤流失提供高产胞外多糖菌株资源, 还可以为促进岛礁生物土壤结皮的形成提供新的研究思路和方法。

关键词: 南海岛礁, 可培养细菌多样性, 胞外多糖, 固砂

Abstract:

There are many microbial resources present in biological soil crusts of tropical reefs in the South China Sea, and the extracellular polysaccharides secreted by these microorganisms play an important role in promoting sand consolidation. In this study, three media, TSA, modified TSA and MA, were used to isolate and purify culturable bacteria from biological soil crusts in the Yongshu Reef and Luhuitou, Sanya, South China Sea. A total of 70 bacterial strains were isolated and purified in this study. And based on 16S rRNA gene identification, these isolated strains belonged to 3 phyla, 5 classes, 12 orders and 19 families, 25 genera. The dominant phylum was Firmicutes and the dominant genus was Bacillus. In addition, 22 strains had less than 98.65%16S rRNA gene similarities with known species, which were potential novel species. The extracellular polysaccharide content of purified strains was measured by ethanol precipitation method and phenol sulfate method, and the contents of 19 strains were more than 0.013 mg·mL^-1. Finally, 9 strains with the highest extracellular polysaccharide content were selected to determine the soil agglomeration ability. The results showed that strain SCSIO 17111 (Lysobacter) was the only strain which was able to maintain the stability of the biological soil crust clods by spraying. Our study provides high extracellular polysaccharide-producing strain resources for island reef management.

Key words: South China Sea reefs, culturable bacterial diversity, extracellular polysaccharide, sand fixation

黄谕, 王琳, 麦志茂, 李洁, 张偲. 南海热带岛礁生物土壤结皮中细菌的分离及其固砂特性初步研究[J]. 热带海洋学报, 2023, 42(6): 101-110.

HUANG Yu, WANG Lin, MAI Zhimao, LI Jie, ZHANG Si. Isolation and characterization of sand fixation ability of bacteria in biological soil crusts of the tropical islands, South China Sea[J]. Journal of Tropical Oceanography, 2023, 42(6): 101-110.

图/表 9

图1

图2

图3

表1

表2

图4

表3

图5

表4

参考文献 35

[1]	艾雪, 2015. 沙漠结皮中耐盐碱细菌的分离及其固沙特性研究[D]. 兰州: 兰州交通大学.
	AI XUE, 2015. Isolation of Saline-alkali-tolerant bacterial strains from desert crust and its characteristics in sand fixation[D]. Lanzhou: Lanzhou Jiaotong University (in Chinese with English abstract).
[2]	艾雪, 王艺霖, 张威, 等, 2015. 柴达木沙漠结皮中耐盐碱细菌的分离及其固沙作用研究[J]. 干旱区资源与环境, 29(10): 145-151.
	AI XUE, WANG YILIN, ZHANG WEI, et al, 2015. Screening of halotolerant and alkalitolerant bacteria from the desert crust in the Qaidam and their effects of sand aggregation[J]. Journal of Arid Land Resources and Environment, 29(10): 145-151 (in Chinese with English abstract).
[3]	董群, 郑丽伊, 方积年, 1996. 改良的苯酚-硫酸法测定多糖和寡糖含量的研究[J]. 中国药学杂志, (9): 38-41.
	DONG QUN, ZHENG LIYI, FANG JINIAN, 2015. Modified phenol-sulfuric acid method for determination of the content of oligo- and polysaccharides[J]. Chinese Pharmaceutical Journal, (9): 38-41 (in Chinese with English abstract).
[4]	毛龙江, 张永战, 魏灵, 等, 2006. 海南岛三亚湾海滩研究[J]. 第四纪研究, 26(3): 477-484.
	MAO LONGJIANG, ZHANG YONGZHAN, WEI LING, et al, 2006. Study on beach characteristics in Sanya area of Hainan Island[J]. Quaternary Sciences, 26(3): 477-484 (in Chinese with English abstract).
[5]	南沙海域环境质量研究专题组, 1996. 南沙群岛及其邻近海域环境质量研究[M]. 北京: 海洋出版社: 1-111.
	Nansha sea area environmental quality research theme group, 1996. The research on environment quality in the Nansha Islands and adjacent sea area[M]. Beijing: China Ocean Press: 1-111 (in Chinese).
[6]	钱琨, 王新志, 陈剑文, 等, 2017. 南海岛礁吹填钙质砂渗透特性试验研究[J]. 岩土力学, 38(6): 1557-1564.
	QIAN KUN, WANG XINZHI, CHEN JIANWEN, et al, 2017. Experimental study on permeability of calcareous sand for islands in the South China Sea[J]. Rock and Soil Mechanics, 38(6): 1557-1564 (in Chinese with English abstract).
[7]	任玉宾, 2016. 南海钙质砂渗透特性试验研究[D]. 大连: 大连理工大学.
	REN YUBIN, 2016. Experimental study on the permeability characteristics of calcareous sand in South China Sea[D]. Dalian: Dalian University of Technology (in Chinese with English abstract).
[8]	王琳, 2020. 南海典型岛礁生物土壤结皮的微生物组及培育技术研究[D]. 广州: 中国科学院南海海洋研究所.
	WANG LIN, 2020. Study on microbiome and cultivation techniques of biological soil crusts on the typical reef islands of the South China Sea[D]. Guangzhou: South China Sea Institute of Oceanology, Chinese Academy of Sciences (in Chinese with English abstract).
[9]	王新志, 2008. 南沙群岛珊瑚礁工程地质特性及大型工程建设可行性研究[D]. 武汉: 中国科学院研究生院 (武汉岩土力学研究所).
	WANG XINZHI, 2008. Study on engineering geological properties of coral reefs and feasibility of large project construction on Nansha islands[D]. Wuhan: Institute of Rock and Soil Mechanics, Chinese Academy of Sciences (in Chinese with English abstract).
[10]	吴楠, 潘伯荣, 张元明, 2004. 土壤微生物在生物结皮形成中的作用及生态学意义[J]. 干旱区研究, (4): 444-450.
	WU NAN, PAN BORONG, ZHANG YUANMING, 2004. Effects and ecological significance of soil-inhabiting microorganisms in the formation of biological soil crusts[J]. Arid Zone Research, (4): 444-450 (in Chinese with English abstract).
[11]	谢作明, 2006. 荒漠藻类对紫外辐射的响应及其结皮形成的研究[D]. 武汉: 中国科学院研究生院(水生生物研究所).
	XIE ZUOMING, 2006. Studies on the responses of desert algae to ultraviolet radiation and the algal crust formation under field conditions[D]. Wuhan: Institute of Hydrobiology, Chinese Academy of Sciences (in Chinese with English abstract).
[12]	余劲聪, 2016. 海藻寡糖在农业领域的应用研究进展[J]. 南方农业学报, 47(6): 921-792.
	YU JINCONG, 2016. Research progress in application of seaweed oligosaccharides in agriculture[J]. Journal of Southern Agriculture, 47(6): 921-927 (in Chinese with English abstract).
[13]	张偲, 王琳, 李洁, 2018. 一株产胞外多糖可固沙的溶杆菌SCSIO 17111及其应用[P]. 广东: CN108795807A, 2018-11-13 (in Chinese).
[14]	张偲, 王琳, 李洁, 2021. 一种适用于南海珊瑚岛礁钙质砂土壤的藻菌结皮培育方法[P]. 广东: CN113287382A, 2021-08-24 (in Chinese).
[15]	张文平, 李昆太, 黄林, 等, 2017. 产胞外多糖菌株的筛选及其对土壤团聚体的影响[J]. 江西农业大学学报, 39(4): 772-729.
	ZHANG WENPING, LI KUNTAI, HUANG LIN, et al, 2017. Screening of exopolysaccharide-producing bacteria and their effects on aggregation in soil[J]. Acta Agriculturae Universitatis Jiangxiensis, 39(4): 772-729 (in Chinese with English abstract).
[16]	郑楠, 邵阳, 罗敏, 等, 2022. 土壤团聚体制备方法对其稳定性及固碳潜力评价的影响研究[J]. 中国环境科学, 42(6): 2821-2827.
	ZHENG NAN, SHAO YANG, LUO MIN, et al, 2022. Effects of soil aggregate preparation methods on the stability and carbon sequestration potential evaluation[J]. China Environmental Science, 42(6): 2821-2827 (in Chinese with English abstract).
[17]	BARGER N N, HERRICK J E, VAN ZEE J, et al, 2006. Impacts of biological soil crust disturbance and composition on C and N loss from water erosion[J]. Biogeochemistry, 77(2): 247-263. doi: 10.1007/s10533-005-1424-7
[18]	BLAY E S, SCHWABEDISSEN S G, MAGNUSON T S, et al, 2017. Variation in Biological Soil Crust bacterial abundance and diversity as a function of climate in cold steppe ecosystems in the Intermountain West, USA[J]. Microbial Ecology, 74(3): 691-700. doi: 10.1007/s00248-017-0981-3 pmid: 28409197
[19]	CHEN LANZHOU, ROSSI F, DENG SONGQIANG, et al, 2014. Macromolecular and chemical features of the excreted extracellular polysaccharides in induced biological soil crusts of different ages[J]. Soil Biology & Biochemistry, 78: 1-9. doi: 10.1016/j.soilbio.2014.07.004
[20]	DA ROCHA U N, CADILLO-QUIROZ H, KARAOZ U, et al, 2015. Isolation of a significant fraction of non-phototroph diversity from a desert biological soil crust[J]. Frontiers in Microbiology, 6: 277. doi: 10.3389/fmicb.2015.00277 pmid: 25926821
[21]	DUBOIS M, GILLES K A, HAMILTON J K, et al, 1956. Colorimetric method for determination of sugars and related substances[J]. Analytical Chemistry, 28(3): 350-356. doi: 10.1021/ac60111a017
[22]	FLEMMING H C, WINGENDER J, 2010. The biofilm matrix[J]. Nature Reviews Microbiology, 8(9): 623-633. doi: 10.1038/nrmicro2415
[23]	GODINHO A L, BHOSLE S, 2009. Sand aggregation by exopolysaccharide-producing microbacterium arborescens-AGSB[J]. Current Microbiology, 58(6): 616-621. doi: 10.1007/s00284-009-9400-4
[24]	KIM M, CHHETRI G, SO Y, et al, 2022. Characteristics and biological activity of exopolysaccharide produced by Lysobacter sp. MMG2 isolated from the roots of Tagetes patula[J]. Microorganisms, 10(7): 1257. doi: 10.3390/microorganisms10071257
[25]	KIM M, OH H -S, PARK S -C, et al, 2014. Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequence similarity for species demarcation of prokaryotes[J]. International Journal of Systematic and Evolutionary Microbiology, 64(Pt 2): 346-351. doi: 10.1099/ijs.0.059774-0 pmid: 24505072
[26]	KUMAR S, STECHER G, LI M, et al, 2018. MEGA X: molecular evolutionary genetics analysis across computing platforms[J]. Molecular Biology and Evolution, 35(6): 1547-1549. doi: 10.1093/molbev/msy096 pmid: 29722887
[27]	KUSKE C R, YEAGER C M, JOHNSON S, et al, 2012. Response and resilience of soil biocrust bacterial communities to chronic physical disturbance in arid shrublands[J]. Isme Journal, 6(4): 886-897. doi: 10.1038/ismej.2011.153 pmid: 22113374
[28]	LETUNIC I, BORK P, 2019. Interactive Tree Of Life (iTOL) v4: Recent updates and new developments[J]. Nucleic Acids Research, 47(W1): W256-W259. doi: 10.1093/nar/gkz239
[29]	MENG DAIZONG, WU JUN, CHEN KELI, et al, 2019. Effects of extracellular polymeric substances and microbial community on the anti-scouribility of sewer sediment[J]. Science of the Total Environment, 687: 494-504. doi: 10.1016/j.scitotenv.2019.05.387
[30]	RIMADA P S, ABRAHAM A G, 2003. Comparative study of different methodologies to determine the exopolysaccharide produced by kefir grains in milk and whey[J]. Lait, 83(1): 79-87. doi: 10.1051/lait:2002051
[31]	SAITOU N, NEI M, 1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees[J]. Molecular Biology and Evolution, 4(4): 406-425. doi: 10.1093/oxfordjournals.molbev.a040454 pmid: 3447015
[32]	SANDHYA V, ALI S Z, 2015. The production of exopolysaccharide by Pseudomonas putida GAP-P45 under various abiotic stress conditions and its role in soil aggregation[J]. Microbiology, 84(4): 512-519. doi: 10.1134/S0026261715040153
[33]	SCHULZ K, MIKHAILYUK T, DRESSLER M, et al, 2016. Biological soil crusts from coastal dunes at the baltic sea: cyanobacterial and algal biodiversity and related soil properties[J]. Microbial Ecology, 71(1): 178-193. doi: 10.1007/s00248-015-0691-7 pmid: 26507846
[34]	YANG KAI, ZHAO YUNGE, GAO LIQIAN, 2022. Biocrust succession improves soil aggregate stability of subsurface after “Grain for Green” Project in the Hilly Loess Plateau, China[J]. Soil and Tillage Research, 217: 105290. doi: 10.1016/j.still.2021.105290
[35]	ZHANG BINGCHANG, KONG WEIDONG, WU NAN, et al, 2016. Bacterial diversity and community along the succession of biological soil crusts in the Gurbantunggut Desert, Northern China[J]. Journal of Basic Microbiology, 56(6): 670-679. doi: 10.1002/jobm.201500751 pmid: 26947139

菌株编号	最近缘物种	16S rRNA基因相似性/%
SCSIO 17100	Microbacterium arborescens	98.55
SCSIO 17105	Microbacterium arborescens	98.55
SCSIO 17035	Microbacterium saccharophilum	98.48
SCSIO 17034	Bacillus zhanjiangensis	98.36
SCSIO 17007	Microbacterium pumilum	98.33
SCSIO 17026	Bacillus zhanjiangensis	98.29
SCSIO 17060	Devosia lucknowensis	98.28
SCSIO 17011	Microbacterium saccharophilum	98.26
SCSIO 17054	Bacillus oryzaecorticis	98.24
SCSIO 17069	Bacillus oryzaecorticis	98.23
SCSIO 17076	Microbacterium invictum	98.18
SCSIO 17016	Bacillus zhanjiangensis	98.15
SCSIO 17014	Skermanella aerolata	97.98
SCSIO 17004	Aquipuribacter nitratireducens	97.47
SCSIO 17099	Bacillus cohnii	97.21
SCSIO 17017	Bacillus salitolerans	97.02
SCSIO 17048	Nocardioides zeicaulis	96.94
SCSIO 17036	Mesorhizobium huakuii	96.93
SCSIO 17031	Bacillus herbersteinensis	96.85
SCSIO 17027	Devosia lucknowensis	96.66
SCSIO 17010	Bacillus abyssalis	96.56
SCSIO 17040	Kineosporia rhamnosa	95.90

菌株编号	产量/(mg·mL^-1)
SCSIO 17048	6.4533
SCSIO 17023	6.4067
SCSIO 17046	6.2300
SCSIO 17003	6.1800
SCSIO 17036	6.0767
SCSIO 17037	6.0467
SCSIO 17054	6.0200
SCSIO 17111	5.9833
SCSIO 17100	5.9733
SCSIO 17096	5.9633
SCSIO 17049	5.8067
SCSIO 17060	5.6267
SCSIO 17061	5.6233
SCSIO 17019	5.5200
SCSIO 17051	5.4067
SCSIO 17076	5.4033
SCSIO 17099	5.1933
SCSIO 17097 SCSIO 17010	5.0767 5.0267

菌株编号	产量/(mg·mL^-1)	菌株来源	菌株属
SCSIO 17111	0.0754	永暑礁结皮底层砂	Lysobacter
SCSIO 17003	0.0539	永暑礁结皮层	Bacillus
SCSIO 17096	0.0403	三亚结皮层	Agromyces
SCSIO 17051	0.0307	三亚结皮层	Bacillus
SCSIO 17100	0.0305	三亚结皮层	Microbacterium
SCSIO 17049	0.0252	三亚结皮层	Bacillus
SCSIO 17010	0.0213	永暑礁结皮层	Bacillus
SCSIO 17061	0.0209	三亚结皮层	Pseudoxanthomonas
SCSIO 17048	0.0186	三亚结皮层	Nocardioides
SCSIO 17060	0.0185	三亚结皮层	Devosia
SCSIO 17097	0.0182	三亚结皮层	Brachybacterium
SCSIO 17023	0.0180	永暑礁结皮底层砂	Bacillus
SCSIO 17046	0.0170	三亚结皮层	Halobacillus
SCSIO 17076	0.0168	永暑礁结皮层	Microbacterium
SCSIO 17099	0.0165	永暑礁结皮底层砂	Bacillus
SCSIO 17036	0.0161	三亚结皮层	Mesorhizobium
SCSIO 17037	0.0158	三亚结皮层	Ornithinimicrobium
SCSIO 17054	0.0151	三亚结皮层	Bacillus
SCSIO 17019	0.0137	永暑礁结皮底层砂	Pseudomonas

菌株编号	土壤团聚体厚度/mm	干筛结果/g (粒径>1.25mm)	菌株来源	菌株属
SCSIO 17111	6.00±0.13	13.56	永暑礁结皮底层砂	Lysobacter
SCSIO 17023	3.89±0.14	0	永暑礁结皮底层砂	Bacillus
SCSIO 17096	4.44±0.14	0	三亚结皮层	Agromyces
SCSIO 17049	3.67±0.07	0	三亚结皮层	Bacillus
SCSIO 17051	3.78±0.10	0	三亚结皮层	Bacillus
SCSIO 17003	3.33±0.13	0	永暑礁结皮层	Bacillus
SCSIO 17061	3.78±0.07	0	三亚结皮层	Pseudoxanthomonas
SCSIO 17100	2.00±0.19	0	三亚结皮层	Microbacterium
SCSIO 17010	2.11±0.20	0	永暑礁结皮层	Bacillus

南海热带岛礁生物土壤结皮中细菌的分离及其固砂特性初步研究

Isolation and characterization of sand fixation ability of bacteria in biological soil crusts of the tropical islands, South China Sea

RichHTML

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

图/表 9

参考文献 35

相关文章 1

Metrics

本文评价

推荐阅读 0