河西走廊黑戈壁生态系统中可培养细菌分布特征及抗辐射活性

doi:10.7522/j.issn.1000-694X.2020.00014

中国沙漠

2020, Vol. 40

Issue (4): 52-62 DOI: 10.7522/j.issn.1000-694X.2020.00014

河西走廊黑戈壁生态系统中可培养细菌分布特征及抗辐射活性

张振清¹^,⁴(

), 张昺林³, 张威¹(

), 刘光琇¹, 陈拓³, 刘阳²^,⁵, 陈警伟⁶, 田茂¹^,⁴

^1.中国科学院西北生态环境资源研究院，中国科学院沙漠与沙漠化重点实验室，甘肃兰州 730000
^2.中国科学院西北生态环境资源研究院，甘肃省极端环境微生物资源与工程重点实验室，甘肃兰州 730000
^3.中国科学院西北生态环境资源研究院，冰冻圈科学国家重点实验室，甘肃兰州 730000
^4.中国科学院大学，北京 100049
^5.西北师范大学，甘肃兰州 730000
^6.兰州大学，甘肃兰州 730000

Distribution characteristics and anti-radiation activity of culturable bacteria in black gobi ecosystem of the Hexi Corridor

Zhenqing Zhang¹^,⁴(

), Binglin Zhang³, Wei Zhang¹(

), Guangxiu Liu¹, Tuo Chen³, Yang Liu²^,⁵, Jingwei Chen⁶, Mao Tian¹^,⁴

^1.Key Laboratory of Desert and Desertification /, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Lanzhou 730000, China
^2.Gansu Province Key Laboratory of Extreme Environmental Microbial Resources and Engineering /, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Lanzhou 730000, China
^3.State Key Laboratory of Cryospheric Sciences, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Lanzhou 730000, China
^4.University of Chinese Academy of Sciences, Beijing 100049, China
^5.Northwest Normal University, Lanzhou 730000, China
^6.Lanzhou University, Lanzhou 730000, China

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摘要：

黑戈壁区域具有干旱、强辐射等极端环境条件，鲜有人类踪迹，相关生物学研究报道较少，其中微生物研究未见报道。本研究首次针对黑戈壁生态系统中微生物分布特征开展研究，对河西走廊黑戈壁生态系统中不同生境土壤样品进行可培养细菌分离。结果表明：河西走廊黑戈壁生态系统中每克土壤可培养细菌数量（CFU）为2.3×10⁴~1.49×10⁶，不同生境土壤的可培养细菌具有明显差异，可培养细菌主要富集于石下生境，黑戈壁中砾石下为微生物提供了相对较适宜的生境；土壤总碳是影响黑戈壁土壤细菌数量的主要因素。结合16 S rRNA基因序列比对，分析共鉴定可培养细菌118株菌株，菌株主要归类于放线菌门（Actinobacteria）、厚壁菌门（Firmicutes）、变形菌门（Proteobacteria）和异常球菌-栖热门(Deinococcus-Thermus)4个类群，其中放线菌门和厚壁菌门是优势门；芽孢杆菌属（Bacillus）、链霉菌属（Streptomyces）是优势属，10株细菌菌株为潜在新种。从分离菌株中筛选出了多株抗辐射活性较高的菌株，其中7株活性显著高于阳性对照耐辐射奇球菌（Deinocccus radiodurans），为进一步筛选研究细菌抗辐射机制及抗辐射活性物质提供菌株资源。

关键词： 黑戈壁; 微生物; 可培养细菌; 抗辐射

Abstract:

The black gobi region has been occupied by extreme environmental conditions such as drought and strong radiation. Unfortunately, the area is almost negligible for human activities and biological research. Therefore, particular studies are required that cover microbial distribution characteristics of the black gobi ecosystem together valuable information. In the current study, the culturable method was used to analyse different types of soil in the Hexi Corridor. The results showed that the number of culturable bacteria in the study area was 2.3×10⁴-1.49×10⁶ CFU·g^-1soil, where the differences in culturable bacteria of different types of soil were obviously varied. The culturable bacteria were mainly enriched in the subsoil habitat, which indicated that the gravel in the Black Gobi provided an ideal colonization site for microorganisms. Furthermore, the statistical analysis showed that soil organic carbon was the main influencing factor on the bacteria number.About,118 different bacterial strains were obtained through 16S rRNA gene sequencing. The strains were mainly classified into Actinobacteria, Firmicutes, Proteobacteria, and Deinococcus-Thermus, respectively. The Actinobacteria and Firmicutes were the dominant phyla， and Bacillus and Streptomyces were the dominant genera. Ten bacterial strains were found to be the potential new species. Among the higher anti-radiation activity screening, seven strains had shown higher activity than positive control of Deinococcus radiodurans. This study expanded the source of desirable strains and their further screening of active substances resistant to radiation.

Key words: black gobi microorganism culturable bacteria antiradiation

收稿日期: 2019-11-23 出版日期: 2020-09-01

Q938

基金资助: 国家自然科学基金项目(31870479);中国科学院对外合作重点项目(131B62KYSB20160014);中国科学院“西部之光”人才培养计划项目

通讯作者: 张威 E-mail: 1298576321@qq.com;ziaoshen@163.com

作者简介: 张威（E-mail: ziaoshen@163.com）
张振清（1994—），女，山东人，硕士研究生，研究方向为微生物生态学。E-mail：1298576321@qq.com

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引用本文:

张振清, 张昺林, 张威, 刘光琇, 陈拓, 刘阳, 陈警伟, 田茂. 河西走廊黑戈壁生态系统中可培养细菌分布特征及抗辐射活性[J]. 中国沙漠, 2020, 40(4): 52-62.

Zhenqing Zhang, Binglin Zhang, Wei Zhang, Guangxiu Liu, Tuo Chen, Yang Liu, Jingwei Chen, Mao Tian. Distribution characteristics and anti-radiation activity of culturable bacteria in black gobi ecosystem of the Hexi Corridor. Journal of Desert Research, 2020, 40(4): 52-62.

链接本文:

http://www.desert.ac.cn/CN/10.7522/j.issn.1000-694X.2020.00014 或 http://www.desert.ac.cn/CN/Y2020/V40/I4/52

图1 河西走廊黑戈壁采样点位置示意图

图2 河西走廊黑戈壁土壤理化性质对同一采样点，不同小写字母表示差异显著（P<0.05），相同小写字母表示差异不显著

图3 河西走廊黑戈壁土壤可培养细菌数量分布特征对同一采样点，不同小写字母表示差异显著（P<0.05），相同小写字母表示差异不显著

门	属	编号	相似菌株	登录号	相似度/%	属	编号	相似菌株	登录号	相似度/%
Actinobacteria	Agrococcus	ST2LB-2	A. citreus ZBGKL11	KJ734880	99.71	Streptomyces	ST2SX-8	S. africanus E3SQ	MH472998	98.80
		ST1-1-2-2	A. jenensis DW414	KR856324	97.32		ST1-1-2-5	S. cacaoi Ru87	KY818662	97.87
		ST3-6	A. jenensis Y25	MK721042	99.34		ST3-3-2-5	S. capoamus JCM 4734	NR_040856	99.56
		ST3-4	A. jenensis Y14	MK721034	99.26		ST3-3-2-6-1	S. chryseus HBUM174847	EU841613	97.51
		ST3-3	A. terreus BT116	MH934923	99.85		ST2-S1	S. coeruleoaurantiacus K7	KR023963	99.85
	Amycolatopsis	ST3-18	A. nigrescens CSC17Ta-90	NR_043880	97.93		ST1-1-1-13	S. coeruleofuscus MR-18	KY753217	99.85
	Arsenicicoccus	ST1-1-1-17-1	A. bolidensis CCUG 47306	NR_025598	99.93		ST1SX-8	S. glaucescens NRRL B-2706	NR_115773	99.93
	Arthrobacter	ST3-16	A. agilis IHBB 11164	KR085842	99.01		ST1SX-14	S. gobitricini LMG 19910	AJ781335	100.00
		ST3-15	A. agilis II/11	KM036066	99.71		ST3-2-1-10-2	S. lavendulocolor NBRC 12881	NR_112317	99.93
		ST1-1-2-7	A. crystallopoietes MR-15	KY753214	99.93		ST3B1	S. litmocidini HBUM175011	FJ486428	98.65
	Auraticoccus	ST3-10	A. monumenti AL12	KU258209	98.17		ST1-1-2-4	S. lunaelactis 244-HNR7	MF077012	99.11
		ST3-2	A. monumenti MON 2.2	LT629688	99.55		ST3-24-1-1	S. lunaelactis MM109	CP026304	99.12
		ST3-21	A. sp. R-68201	KY386505	95.24		ST1-1-2-3	S. lunaelactis MMun143	MG980181	99.19
		ST3-22	A. monumenti AL12	KU258209	94.31		ST3-2-1-1-2-1	S. misionensis 12-4	KJ571074	99.49
	Blastococcus	ST1-1-1-16	B. aggregatus 1P10AnA	EU977831	99.63		ST3-2-1-8-1	S. misionensis cfcc3147	FJ792563	99.63
		ST3-5-2	B. capsensis R9	MK696404	99.63		ST1-1-2-10-1	S. palmae CMU-AB199	LC413945	98.02
		ST3-24-1-3	B. capsensis RQ2	MK696394	99.09		ST1-1-2-10	S. palmae CMU-AB204	NR_152026	97.47
		ST2B4	B. endophyticus YIM 68236	NR_108608	99.32		ST2SB-6	S. piloviolofuscus 174468	EU593715	97.66
		ST3-12-1	B. saxobsidens DD2	FO117623	99.78		ST3-2-2-2	S. pulveraceus MR-27	KY753226	96.65
		ST3-24	B. saxobsidens BC444	NR_117019	99.33		ST3-3-2-6	S. sp. Z594b	MN371360	97.72
		ST2SX-1	B. saxobsidens BC448	NR_025482	99.64		ST2SX-5-2	S. rimosus PSK5-20B	MN421096	96.81
	Crossiella	ST3SX-11	C. equi NRRL B-24104	NR_025088	99.18		ST1-3-2-2	S. sioyaensis W24	KP718602	99.12
	Janibacter	ST1-1-2-11	J. terrae CS12	NR_036868	99.85		ST3-2-1-9-1	S. spinoverrucosus 173372	EU570683	98.89
	Kineococcus	ST2SS-1	K. radiotolerans SRS30216	NR_074542	99.00		ST3-12	S. spinoverrucosus 174464	EU593714	99.05
	Kocuria	ST1-1-1-24-2	K. gwangalliensis SJ2	NR_116266	99.93		ST2SX7	S. spinoverrucosus NBRC 14228	NR_041159	97.81
		ST3-24-1-2	K. indica SJU27	MN511772	99.86		ST3SX-4	S. spinoverrucosus Ng2-6	MK519101	98.62
		ST2LB-1	K. rosea 5	KF923415	99.78		ST1-1-2-9	S. sp. NO8	KC200022	100.00
	Lechevalieria	ST1SX-6-2	L. atacamensis 41-HR6	MF077035	99.46		ST2-5	S. xinghaiensis S15077	MG563223	99.78
		ST2SX-6	L. atacamensis C61	NR_116354	95.81	Saccharothrix	ST2B-10-4	S. lopnurensis YIM LPA2h	NR_145947	98.62
		ST1SX-10	L. xinjiangensis R24	NR_044009	99.26	Saccharothrix	ST3-3-1	S. yanglingensis Hhs.015	NR_117283	99.55
	Microbacterium	ST2LB-3	M. kitamiense kitami C2	NR_112042	99.71	Modestobacter	ST3-14	M. caceresii KNN 45-2b	NR_137398	99.56
	Microbacterium	ST3-6	M. oxydans 4-46-1-1-1	MK425667	99.85	Modestobacter	ST3-1-1-13	M. marinus BC501	FO203431	99.93
	Pseudonocardia	ST1SX-2	P. hierapolitana PM2084	NR_126236	99.26
Deinococcus-Thermus	Deinococcus	ST1SX-4	D. sp. 4B4	EU029131	97.07
Deinococcus-Thermus	Deinococcus	ST1-1-1-9	D. sp. 4B6	EU029132	99.83
Firmicutes	Bacillus	ST1-1-1-24-1	B. amyloliquefaciens HX2016004	MN176577	100.00	Bacillus	ST1SX-3	B. atrophaeus XAAS.xj4	MN187264	99.86
		ST1-1-1-17	B. amyloliquefaciens KB-82	KM269197	100.00		ST2SX-3	B. atrophaeus Y27	MK721044	100.00
		ST2SS3-1	B. atrophaeus HAB_5	MK310269	99.93		ST1-1-1-9	B. cereus LH8	KC248215	99.93
		ST1-1SX-3	B. atrophaeus MER_TA_30	KT719435	99.93		ST2-19-2	B. cereus st2	MF102134	99.93
		ST1-1-1-17-2	B. licheniformis D69	KU922147	99.86		ST1-1-2-6	B. foraminis CD5	MK216757	99.64
		ST2-19-1	B. mojavensis h	MG839268	100.00		ST3-1-1-3-1	B. halotolerans FJAT-45391	KY849471	100.00
		ST1SX8-1	B. paramycoides OOF5	MH542275	99.86		ST2-26	B. subtilis GuanMX	MN473282	100.00
		ST1-1-2-13	B. pumilus D51	JX293284	99.93		ST3-2	B. subtilis PSBnR5	MH257752	100.00
		ST1SX-5	B. safensis MDL5	MN493773.1	100.00		ST1-1-1-20	B. subtilis Sk01A	MH210872	99.79
		ST3-27	B. safensis YZ1709B01	MK748241.1	99.79		ST3-5	B. tequilensis 6MS1	MK713704	97.32
		ST2-29	B. simplex ZLynn1000-56	KY316470	99.57		ST1-1-2-1	B. vallismortis 70.LE.1	MN149347	99.93
		ST3SS4-1	B. subtilis 2/30	MN435586	100.00		ST2B8	B. vallismortis Y	MG839261	98.57
		ST3-2-2-12	B. velezensis N8	KX588164	100.00
	Paenibacillus	ST2-28	P. harenae NJY-3	MF101120	95.90	Enterococcus	ST3-2-1-9-3	E. sp. 79w3	AB675140	94.50
	Paenibacillus	ST3-24-2	P. polymyxa RCP6	GU369972	99.63	Staphylococcus	ST2SS2-1	S. saprophyticus subsp. saprophyticus zzx27	KJ009395	99.51
Proteobacteria	Azospirillum	ST3SX-5	A. palatum ww10	EU747318	94.87	Massilia	ST3-20	M. varians 66-LR14-2	MF077216	99.48
	Azospirillum	ST2SX5-1	A. sp. NCCP-699	LC193946	96.99	Methylorubrum	ST2SX-5-1	M. pseudosasae IMB16-188	MG190781	100.00
	Belnapia	ST1SX-20	B. moabensis CP2C	NR_042371	99.20	Microvirga	ST3-2-1-6	M. aerilata KBL26	MG576179	98.19
	Belnapia	ST3-2-1-11	B. rosea CPCC 100156	NR_109297	99.85		ST3-2-1	M. aerilata NBRC 106137	NR_114298	100.00
	Brevundimonas	ST2-2	B. diminuta 264AG7	KF836539	99.78		ST3-13	M. ossetica V5/5K	KX576554	98.36
		ST2-10	B. diminuta HMS9	MK696984	99.92		ST3-2-1-10	M. sp. R491-7	KX444133	98.89
		ST3SS-7	B. naejangsanensis 5S3	KM374767	99.69	Pseudomonas	ST2SS3-2	P. fluorescens psf14	MN256400	99.58
		ST3-27-2	B. vesicularis CX-89	MH368406	99.70		ST3-2-3-12-1-1	P. putida YP2	KP313537	100.00
	Candidimonas	ST1-3-2-5	C. bauzanensis BZ59	NR_108569	98.23		ST1-1-2-2-1	P. stutzeri SYJ1-8	KR262851	99.57
	Enterobacter	ST1SX-6-1	E. hormaechei SCEH020042 chromosome	CP028538	99.71	Roseomonas	ST3-1-1-14	R. oryzae JC288	NR_137403	98.34
	Herbaspirillum	ST3-2-1-12	H. sp. 1NM-18	JQ608328	97.76	Stenotrophomonas	ST3-3-1-2	S. rhizophila EGE-B-6	KP050794	99.71

表1 河西走廊黑戈壁土壤可培养细菌菌株

图4 河西走廊黑戈壁土壤菌株相对丰度气泡图SB:砾石上表面土;SX:砾石石下土;B:砾石间隙土;S:深层土

表2 河西走廊黑戈壁可培养细菌多样性指数

图5 河西走廊黑戈壁可培养细菌和土壤理化因子之间的典型对应分析（CCA）SB:砾石上表面土;SX:砾石石下土;B:砾石间隙土;S:深层土

图6 河西走廊黑戈壁部分可培养菌株抗辐射存活率（耐辐射奇球菌（Deinocccus radiodurans）作为阳性对照，大肠杆菌（Escherichia coli）作为阴性对照）不同小写字母表示差异显著（P<0.05），相同小写字母表示差异不显著

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