美济礁沉积砂可培养细菌多样性分析

doi:10.11978/2023183

Abstract

Abstract:

The special environment of reefs in the South China Sea has created a rich and colorful microbial world, where a large number of microorganisms are yet to be excavated and utilized. Due to geographical conditions and other factors, there are few studies on them. In this paper, culturable microorganisms were selectively isolated from the sedimentary sands of the Meiji Reef by using oligotrophic media combined with high salt and high temperature conditions. And their diversity and distribution were also studied, a variety of functional media were used to explore the functions of these microorganisms. The 674 isolates belong to 5 phyla, 6 classes, 26 orders, 38 families, 62 genera, 146 species, respectively, and 42 potential new species with less than 98.65% similarity composes around 28.7% all of them. Among the cultured bacteria obtained, the Phyla of Actinobacteriota and Pseudomonadota are the most abundant, accounting for 61.6% of the total isolated strains. Functional analysis revealed that 2 species have the phospho-reducing activity, 36 species have the urease-producing activity and 57 species have the iron-bearing activity. The research results indicate that there are microbial abundance, much new species resources and high diversity of microorganism with special ecology functions in the reef environment. In study, these accumulates rich microbial species resources, laying the foundation for the deeply exploration of microbial resources utilization and the ecological restoration of the coral-sands ecosystem in the Meiji Reef.

Key words: coral sand on islands and reefs, oligotrophic medium, high temperature and salinity, culturable bacterial, functional diversity

CLC Number:

Q938.8

JIANG Rouyun, JIAN Lili, SHI Songbiao, TIAN Xinpeng. Diversity of culturable bacteria in the sedimentary sands of the Meiji Reef[J].Journal of Tropical Oceanography, 2024, 43(6): 170-180.

Figures/Tables 13

Tab. 1

Typic media and their components used in this study"

培养基	成分
R₂A	胰蛋白胨0.25g, 可溶性淀粉0.5g, 酵母浸粉0.5g, K₂HPO₄ 0.3g, MgSO₄ 0.1g, 丙酮酸钠0.3g, 蛋白胨0.25g, 葡萄糖0.5g, 酸水解酪蛋白0.5g, 稀释20倍, 琼脂15g, 水1L
2216E	酵母浸粉1.0g, 柠檬酸铁0.1g, NaCl 19.45g, MgCl₂ 5.98g, Na₂SO₄ 3.24g, CaCl₂ 1.8g, KCl 0.55g, Na₂CO₃ 0.16g, KBr 0.08g, SrCl₂ 0.034g, H₃BO₃ 0.022g, Na₂O·SiO₂ 0.004g, NaF 0.002 4g, NH₄NO₃ 0.001 6g, Na₂HPO₄ 0.008g, 蛋白胨5.0g, 稀释20倍, 琼脂15g, 水1L
SN	NaNO₃ 0.75g, K₂HPO₄ 0.015 9g, EDTA二钠0.005 6g, Na₂CO₃ 0.010 4g, VitaminB₁₂ 0.001g(过滤除菌), 1×10^-6微量盐(乙酸6.25g, 柠檬酸铁铵6.0g, Na₂MoO₄·2H₂O 0.39g, MnCl₂·4H₂O 1.4g, Co(NO₃)₂·6H₂O 0.025g, ZnSO₃·7H₂O 0.222g, 单独灭菌), 琼脂15g, 海水500mL, 水500mL
R	NaCl 25g, MgSO₄·7H₂O 9g, CaCl₂·2H₂O 0.14g, KCl 0.7g, Na₂HPO₄·12H₂O 0.25g, Na₂-EDTA 30mg, H₃BO₃ 34mg, FeSO₄·7H₂O 10mg, FeCl₃·6H₂O 1.452mg, MnCl₂·4H₂O 4.32mg, CoCl₂·6H₂O 0.12mg, Na₂MoO·2H₂O 0.63mg, RbCl 0.141 5mg, LiCl 0.61mg, NaBr 6.4mg KI 0.006 55mg, SrCl₂·6H₂O 3.04mg, V₂O₅ 0.001 785mg, ZnCl₂ 0.312mg, RPMI 500mg, EagleMedium 500mg, L-glutamine 15mg, NaHCO₃ 100mg, 海盐18g, 琼脂15g, 水1L
无机磷固体	MnSO₄·4H₂O 0.03g, (NH₄)₂SO₄ 0.5g, NaCl 0.3g, MgSO₄·7H₂O 0.3g, KCl 0.3g, FeSO₄·4H₂O 0.036g, 葡萄糖10g, 有机磷植酸钙2.0g [或无机磷Ca₃(PO₄)₂10g], 水1L, 琼脂15g, pH 7.0
亚历山大硅酸盐	蔗糖5g, Na₂HPO₄2g, FeCl₃0.005g, CaSO₄0.1g, MgSO₄·7H₂O 0.5g琼脂15g, 水1L, pH 7.0~7.5
DF	KH₂PO₄ 4g, MgSO₄·7H₂O 0.2g, 葡萄糖2g, 葡萄糖酸2g, 柠檬酸2g, Na₂HPO₄ 6g, H₃BO₃ 0.01mg, MgSO₄ 0.011 2mg, ZnSO₄ 0.124 6 mg, CuSO₄ 0.078 2mg, MoO₃ 0.01mg, 水1L, 琼脂15g, pH 7.0
ADF	KH₂PO₄ 4g, Na₂HPO₄ 6g, (NH₄)₂SO₄2g, MgSO₄·7H₂O 0.2g, 葡萄糖2g, 葡萄糖酸2g, 柠檬酸2g, H₃BO₃ 0.01mg, MgSO₄ 0.011 2 mg, ZnSO₄ 0.124 6 mg, MoO₃ 0.01mg, 1-氨基羰酰-1-环丙烷羧酸(ACC)5mmol/L, FeSO₄·7H₂O 0.1mg, CuSO₄ 0.078 2mg, 水1L, 琼脂15g, pH 7.0
CAS	CAS 0.605g, Na₂HPO₄·12H₂O 1.213 5g, HDTMA 0.0729g, NaH₂PO₄·2H₂O 0.295 25g, FeCl₃·6H₂O 0.000 3g, NHCl₄ 0.125g, KH₂PO₄ 0.037 5g, NaCl 0.625g, 琼脂9g, 水1L, pH 6.7~6.9
产脲酶	蛋白胨1g, NaCl l5g, KH₂PO₄ 2g, 葡萄糖1g, 酚红0.012g, 琼脂15g, 水1L, pH 7.0
三丁酸甘油酯	三丁酸甘油酯40mL,阿拉伯树胶4g, 琼脂15g, 水1L, pH 7.0

Tab. 1

Fig. 1

Fig. 2

Fig. 3

Tab. 2

Tab. 3

Fig. 4

Fig. 5

Fig. 6

Tab. 4

Tab. 5

Fig. 7

Fig. 8

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样品	纲	种或亚种
MJ16	芽孢杆菌纲Bacilli	Cytobacillus pseudoceanisediminis
MJ16	放线菌纲Actinomycetes	Streptomyces radiopugnans
MJ17	芽孢杆菌纲Bacilli	Bacillus altitudinis、Bacillus tequilensis
MJ18	芽孢杆菌纲Bacilli	Bacillus altitudinis、Bacillus xiamenensis、Bacillus tequilensis
MJ19	芽孢杆菌纲Bacilli	Bacillus altitudinis、Bacillus xiamenensis、Bacillus tequilensis、Bacillus safensis subsp. safensis
MJ20	芽孢杆菌纲Bacilli	Fredinandcohnia onubensis、Paenibacillus cisolokensis
MJ20	放线菌纲Actinomycetes	Streptomyces radiopugnans

培养基	纲	种
5% 2216E	Actinomycetia	Streptomyces radiopugnans
	Alphaproteobacteria	Acuticoccus kandeliae、Metabacillus endolithicus
	Bacilli	Bacillus altitudinis、Bacillus tequilensis、Cytobacillus pseudoceanisediminis
5% R2A	Alphaproteobacteria	Acuticoccus kandeliae
5% R2A	Actinomycetia	Streptomyces radiopugnans
R	Actinomycetia	Streptomyces chumphonensis、Streptomyces genisteinicus、Streptomyces radiopugnans
	Alphaproteobacteria	Microvirga flocculans、Pararhizobium herbae
	Bacilli	Bacillus altitudinis、Bacillus aequororis、Bacillus tequilensis、Halobacillus dabanensis、Metabacillus halosaccharovorans、Virgibacillus kapii、Mesobacillus boroniphilus
SN	Actinomycetia	Streptomyces chumphonensis、Streptomyces ginkgonis、Streptomyces wuyuanensis
	Alphaproteobacteria	Celeribacter ethanolicus、Lysobacter korlensis、Glycocaulis abyssi、Phenylobacterium terrae
	Bacilli	Bacillus mesophilus
	Gammaproteobacteria	Luteimonas pelagia

Diversity of culturable bacteria in the sedimentary sands of the Meiji Reef

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条件	样品	门	纲	目	科	属	种	株
总条件	MJ16	3	4	14	20	30	54	178
	MJ17	5	6	15	24	35	57	157
	MJ18	3	4	6	7	9	14	56
	MJ19	3	4	5	7	12	18	50
	MJ20	4	5	16	19	29	53	233
寡营养	MJ16	3	4	11	16	26	36	103
	MJ17	5	6	14	20	27	40	100
	MJ18	3	4	6	7	8	11	33
	MJ19	3	3	4	6	10	12	27
	MJ20	4	5	14	17	26	41	159
高盐(150‰ NaCl)	MJ16	3	3	10	14	20	31	73
	MJ17	3	3	8	11	15	26	51
	MJ18	3	4	4	4	5	6	8
	MJ19	3	3	3	3	4	6	7
	MJ20	3	4	12	13	17	24	60
高温(50℃)	MJ16	2	2	2	2	2	2	2
	MJ17	1	1	1	1	1	2	6
	MJ18	1	1	1	1	1	3	15
	MJ19	1	1	1	1	1	3	16
	MJ20	2	2	2	2	3	3	14

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