新疆达坂城盐湖嗜盐细菌分离鉴定及活性分析

doi:10.6048/j.issn.1001-4330.2023.07.024

新疆农业科学 ›› 2023, Vol. 60 ›› Issue (7): 1766-1772.DOI: 10.6048/j.issn.1001-4330.2023.07.024

新疆达坂城盐湖嗜盐细菌分离鉴定及活性分析

李怡歆¹^,²(), 陈勇³, 刘晓禄², 艾尼江·尔斯满¹, 徐李娟¹^,⁴, 刘倩倩⁵, 包晓玮²(), 宋素琴¹()

1.新疆农业科学院微生物应用研究所/新疆特殊环境微生物重点实验室,乌鲁木齐 830091
2.新疆农业大学食品科学与药学学院,乌鲁木齐 830052
3.新疆农业科学院土壤肥料与农业节水研究所,乌鲁木齐 830091
4.新疆农业大学农学院,乌鲁木齐 830052
5.新疆生产建设兵团第十三师农业科学研究所,新疆额敏 839000

收稿日期:2022-10-07 出版日期:2023-07-20 发布日期:2023-07-11
通信作者: 包晓玮(1969-),女,浙江松阳人,副教授,博士,研究方向为食品营养与安全,(E-mail)408531623@qq.com；
宋素琴(1976-),女,山西长治人,副研究员,研究方向为微生物活性产物及作物病害,(E-mail)suqin_song@163.com
作者简介:李怡歆(1997-),女,新疆伊犁人,硕士研究生,研究方向为放线菌活性,(E-mail)2463514716@qq.com
基金资助:
国家自然科学基金项目(NSFC-新疆联合基金)“新疆盐湖放线菌来源的星孢菌素类化合物及其PKCθ抑制活性研究”(U1903101);新疆农业科学院科技创新重点培育专项“马铃薯疮痂病病原菌致病毒素thaxtomins产量差异分析及其对新疆农田杂草的除草性能研究”(xjkcpy-202203)

Isolation, identification and activity of halophilic bacteria from Dabancheng Salt Lake, Xinjiang

LI Yixin¹^,²(), CHEN Yong³, LIU Xiaolu², Ainijiang Ersiman¹, XU Lijuan¹^,⁴, LIU Qianqian⁵, BAO Xiaowei²(), SONG Suqin¹()

1. Xinjiang key Laboratory of Special Environmental Microorganisms / Research Institute of Applied Microbiology, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China
2. College of Food and Pharmaceutical Sciences, Xinjiang Agricultural University, Urumqi 830052,China
3. Institute of soil,Fertilizer and Agricultural Water Conservation,Xinjiang Academy of Agricultural Aciences,Urumqi 830091,China
4. College of Agronomy, Xinjiang Agricultural University, Urumqi 830052, China
5. Agricultural Science Research Institute of the 13th Division of XPPC,Emin Xinjiang 839000,China

Received:2022-10-07 Online:2023-07-20 Published:2023-07-11
Correspondence author: BAO Xiaowei(1969-),female,Zhejian somgyang county, associate professor, major in food nutrition and safety,(E-mail)408531623@qq.com；
SONG suqin(1976-),female,Shanxi changzhi county, associate researcher, major in microbial active products and crop diseases,(E-mail)suqin_song@163.com
Supported by:
National Natural Science Foundation of China (NSFC-Xinjiang Joint Foundation):Study on the inhibitory activity of staurosporine and PKCθ from Actinomycetes in Xinjiang Salt Lake(U1903101);Xinjiang Academy of Agricultural Sciences and Technology Innovation Key cultivation Special Project“Yield Difference Analysis of the Pathogenic toxin thaxtomins and its herbicidal performance on agricultural weeds in Xinjiang”(xjkcpy-202203)

摘要/Abstract

摘要：

【目的】研究新疆盐湖嗜盐微生物资源及发掘菌株发酵液的农用和医药活性。【方法】采集达坂城区达坂城盐湖湖边的淤泥样品进行菌株分离鉴定,采用4种不同分离培养基,使用平板稀释涂布法结合16S rRNA进行分离鉴定;并用平板对峙法对分离到的放线菌进行植物病原菌拮抗活性研究,用HepG2细胞增殖测定其抑制肿瘤活性。【结果】获得细菌18株,鉴定为14个属,其中芽孢杆菌属(Bacillus)、链霉菌属(Streptomyces)、拟诺卡氏菌属(Nocardiopsis)为优势种群。有3株放线菌具有拮抗植物病原菌活性,其中菌株Tsukamurella sinensis DBC26和Nocardiopsis crassaminis DBC31有拮抗棉花枯萎菌活性, Streptomyces longispororuberDBC5有拮抗水稻恶苗菌活性。菌株DBC5、DBC9和DBC10对HepG2细胞增殖的抑制率分别为96.4%、91.8%和87.8%。【结论】达坂城盐湖细菌在数量和种类上存在丰富的多样性,且稀有放线菌种类较为丰富,并且其发酵液具有拮抗植物病原菌活性和抑制肿瘤细胞增殖活性。

关键词: 盐湖; 放线菌; 多样性

Abstract:

【Objective】 To study the resources of halophilic microorganisms in Salt Lake of Xinjiang and explore the agricultural and medical activities of the fermentation broth of the strain.【Methods】 In this study, the sludge samples from Dabancheng Salt Lake in Daban City were collected for isolation and identification of bacterial strains.Four different separation media were used for isolation and identification by plate dilution coating method combined with 16S rRNA.The antagonistic activity of the isolated actinomycetes was studied by flat-plate antagonistic method, and the anticancer activity of the isolated actinomycetes was determined by HepG2 cell proliferation.【Results】 Eighteen strains of bacteria were obtained and identified as 14 genera, among which Bacillus, Streptomyces and Nocardiopsis were the dominant species.Three actinomycetes showed antagonistic activity against plant pathogens, among which strains Tsukamurella sinensis DBC26 and Nocardiopsis crassaminis DBC31 showed antagonistic activity against fusarium oxysporum. Streptomyces longispororuber DBC5 showed antagonistic activity against rice vaccinia.The anti-tumor activity of DBC5, DBC9 and DBC10 strains against HepG2 cell proliferation was 96.4%, 91.8% and 87.8%, respectively.【Conclusion】 This study showed that the number and species of bacteria in Dabancheng Salt Lake are rich in diversity, and rare actinomycetes are rich in species, and their fermentation broth has the activity of antagonizing plant pathogens and inhibiting tumor cell proliferation.

Key words: Salt Lake; actinomycetes; diversity

中图分类号:

S188

李怡歆, 陈勇, 刘晓禄, 艾尼江·尔斯满, 徐李娟, 刘倩倩, 包晓玮, 宋素琴. 新疆达坂城盐湖嗜盐细菌分离鉴定及活性分析[J]. 新疆农业科学, 2023, 60(7): 1766-1772.

LI Yixin, CHEN Yong, LIU Xiaolu, Ainijiang Ersiman, XU Lijuan, LIU Qianqian, BAO Xiaowei, SONG Suqin. Isolation, identification and activity of halophilic bacteria from Dabancheng Salt Lake, Xinjiang[J]. Xinjiang Agricultural Sciences, 2023, 60(7): 1766-1772.

图/表 7

参考文献 14

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培养基 Culture medium	嗜盐细菌属 Halophilic bacteria	嗜盐细菌种 Halophilic bacterial species
M-2(海藻糖-脯氨酸)	类芽孢杆菌属(Paenibacillus)	Paenibacillus peoriae
MM琼脂	代谢杆菌(Metabacillus)	Metabacillus sediminilitoris
M-7(甘油-天门冬酰胺)	类诺卡氏菌属(Nocardioides) 动性球菌属(Planococcus) 冢村氏菌属(Tsukamurella) 短杆菌属(Brevibacterium) 假黄色单胞菌属(Pseudoxanthomonas) 链霉菌属(Streptomyces) 红球菌属(Rhodococcus) 小单胞菌属(Micromonospora) 芽孢杆菌属(Bacillus) 细杆菌属(Microbacterium paraoxydans) 微球菌属(Micrococcus) 拟诺卡氏菌属(Nocardiopsis)	Nocardioides furvisabuli、Nocardioides LTZG-s、Tsukamurella conjunctivitidis、Brevibacterium epidermidis、Pseudoxanthomonas japonensis、Streptomyces longispororuber、Rhodococcus qingshengii、Micromonospora oryzae、Bacillus paramycoides、Microbacterium paraoxydans、Micrococcus antarcticus、Tsukamurella_sinensis、Nocardiopsis dassonvillei
M-1(TWYE)	链霉菌属(Streptomyces)	Streptomyces pratensis

培养基 Culture medium	嗜盐细菌属 Halophilic bacteria	嗜盐细菌种 Halophilic bacterial species
M-2(海藻糖-脯氨酸)	类芽孢杆菌属(Paenibacillus)	Paenibacillus peoriae
MM琼脂	代谢杆菌(Metabacillus)	Metabacillus sediminilitoris
M-7(甘油-天门冬酰胺)	类诺卡氏菌属(Nocardioides) 动性球菌属(Planococcus) 冢村氏菌属(Tsukamurella) 短杆菌属(Brevibacterium) 假黄色单胞菌属(Pseudoxanthomonas) 链霉菌属(Streptomyces) 红球菌属(Rhodococcus) 小单胞菌属(Micromonospora) 芽孢杆菌属(Bacillus) 细杆菌属(Microbacterium paraoxydans) 微球菌属(Micrococcus) 拟诺卡氏菌属(Nocardiopsis)	Nocardioides furvisabuli、Nocardioides LTZG-s、Tsukamurella conjunctivitidis、Brevibacterium epidermidis、Pseudoxanthomonas japonensis、Streptomyces longispororuber、Rhodococcus qingshengii、Micromonospora oryzae、Bacillus paramycoides、Microbacterium paraoxydans、Micrococcus antarcticus、Tsukamurella_sinensis、Nocardiopsis dassonvillei
M-1(TWYE)	链霉菌属(Streptomyces)	Streptomyces pratensis

序列号 NO.	菌株编号 NO.	最近相似菌株名称 The name of most similarity bacteria	相似度 Similarity	收藏号 Accession NO.
1	DBC1	Bacillus mobilis	99.51%	MACF01000036
2	DBC2	Pseudoxanthomonas japonensis	99.14%	AB008507
3	DBC3	Micromonospora oryzae	99.35%	AB981052
4	DBC4	Rhodococcus qingshengii	99.64%	LRRJ01000016
5	DBC5	Streptomyces longispororuber	99.71%	AB184440
6	DBC7	Brevibacterium epidermidis	99.60%	BCSJ01000023
7	DBC9	Tsukamurella conjunctivitidis	99.71%	MK583684
8	DBC25	LTZG_s	99.10%	LTZG01000007
9	DBC23	Nocardioides alpinus	97.43%	GU784866
10	DBC10	Streptomyces pratensis	99.85%	JQ806215
11	DBC12	Metabacillus sediminilitoris	98.20%	MN067806
12	DBC13	MRTF_s	98.90%	MRTF01000019
13	DBC18	Bacillus paramycoides	95.93%	MAOI01000012
14	DBC22	Microbacterium paraoxydans	99.22%	BCRH01000180
15	DBC24	Micrococcus antarcticus	98.93%	AJ005932
16	DBC26	Tsukamurella sinensis	99.64%	KP888571
17	DBC30	Nocardiopsis dassonvillei	100%	ABUI01000017
18	DBC31	Nocardiopsis crassaminis	99.82%	LR606207

序列号 NO.	菌株编号 NO.	最近相似菌株名称 The name of most similarity bacteria	相似度 Similarity	收藏号 Accession NO.
1	DBC1	Bacillus mobilis	99.51%	MACF01000036
2	DBC2	Pseudoxanthomonas japonensis	99.14%	AB008507
3	DBC3	Micromonospora oryzae	99.35%	AB981052
4	DBC4	Rhodococcus qingshengii	99.64%	LRRJ01000016
5	DBC5	Streptomyces longispororuber	99.71%	AB184440
6	DBC7	Brevibacterium epidermidis	99.60%	BCSJ01000023
7	DBC9	Tsukamurella conjunctivitidis	99.71%	MK583684
8	DBC25	LTZG_s	99.10%	LTZG01000007
9	DBC23	Nocardioides alpinus	97.43%	GU784866
10	DBC10	Streptomyces pratensis	99.85%	JQ806215
11	DBC12	Metabacillus sediminilitoris	98.20%	MN067806
12	DBC13	MRTF_s	98.90%	MRTF01000019
13	DBC18	Bacillus paramycoides	95.93%	MAOI01000012
14	DBC22	Microbacterium paraoxydans	99.22%	BCRH01000180
15	DBC24	Micrococcus antarcticus	98.93%	AJ005932
16	DBC26	Tsukamurella sinensis	99.64%	KP888571
17	DBC30	Nocardiopsis dassonvillei	100%	ABUI01000017
18	DBC31	Nocardiopsis crassaminis	99.82%	LR606207

植物病原菌Plant pathogen
嗜盐放线菌 Halophilic actinomycetes	南瓜枯萎病菌 F.oxysporum	水稻恶苗病菌 F.moliniforme	棉花枯萎病菌 F.oxysporum	核桃腐烂病菌 Cytcospora sp.	苹果树腐烂病菌 Valsa mali var.mali
DBC5	—	+	—	—	—
DBC23	—	—	—	—	—
DBC10	—	—	—	—	—
DBC26	—	—	+	—	—
DBC30	—	—	—	—	—
DBC31	—	—	+	—	—

新疆达坂城盐湖嗜盐细菌分离鉴定及活性分析

Isolation, identification and activity of halophilic bacteria from Dabancheng Salt Lake, Xinjiang

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菌株编号 Strain No.	抑制率 Inhibition rate
DBC5	96.4%
DBC10	87.8%
DBC9	91.8%
DBC3	NA
DBC26	83.2%
DBC24	16.7%

菌株编号 Strain No.	抑制率 Inhibition rate
DBC5	96.4%
DBC10	87.8%
DBC9	91.8%
DBC3	NA
DBC26	83.2%
DBC24	16.7%