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• 方法与技术 • 上一篇    

水样保存方法对青藏高原湖泊可培养微生物的影响

郭春燕1,2,岳琳艳1,2,赵康1,2,邢鹏3,孔维栋1,2*   

  1. 1中国科学院青藏高原研究所高寒生态学与生物多样性重点实验室, 北京 100101;2中国科学院大学资源与环境学院, 北京 100190;3中国科学院南京地理与湖泊研究所湖泊生物与生态研究室, 南京 210008)
  • 出版日期:2018-08-10 发布日期:2018-08-10

Effect of preservation methods on culturable microorganisms from Tibetan Plateau lake waters.

GUO Chun-yan1,2, YUE Lin-yan1,2, ZHAO Kang1,2, XING Peng3, KONG Wei-dong1,2*#br#   

  1. (1Key Laboratory of Alpine Ecology and Biodiversity, Institute of Tibetan Plateau Research, Chinese Academy of Sciences, Beijing 100101, China; 2College of Resources and Environment, University of Chinese Academy of Sciences, Beijing 100190, China; 3Lake Biological and Ecological Research, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing 210008, China).
  • Online:2018-08-10 Published:2018-08-10

摘要: 青藏高原湖水中生存着大量适应极端环境的微生物,如耐冷及嗜冷菌。合适的湖水样品保存方法是野外取样的关键和开展深入研究的前提。本研究对比了冷冻和4 ℃冷藏两种保存方法对湖水中可培养微生物种类的影响。首先从不同保存方法样品中分离得到纯菌,然后对分离菌的16S rRNA测序进行菌种鉴定。结果表明,自冷冻保存湖水中分离得到7个不同的16S rRNA基因OUT(operational taxonomic unit),冷藏保存水样共分离得到14个不同的16S rRNA基因OTU。冷冻保存水样分离得到的可培养微生物包括芽孢杆菌、简单芽孢杆菌、微小杆菌、不动杆菌、苏云金芽孢杆菌;冷藏保存水样分离得到的微生物包括γ变形菌、金黄杆菌、芽孢杆菌、短小芽孢杆菌、寡养单胞菌、假炭疽杆菌、缺陷假单胞菌、细杆菌。研究表明,冷藏保存方法比冷冻保存方法可分离得到更多种类可培养微生物,这两种保存方法均分离得到了芽孢杆菌。

关键词: 景观连接度, 生态廊道, 白头叶猴, 最小费用距离

Abstract: A high diversity of psychrophilic and psychrotolerant bacteria have been identified in Tibetan Plateau lakes. Isolating pure strains of these bacteria can provide us essential knowledge on the mechanism of coldadaptation. However, it has always been a technical obstruct on how to transport these precious samples from the field to laboratories without reducing bacterial viability. Here, the influence of water sample preservation methods on the bacterial viability was investigated. We first isolated pure bacterial strains from water samples cryopreserved at -80  ℃ and fridgestored at 4  ℃, and classified these bacteria based on their 16S rRNA gene sequences. Seven unique 16S operational taxonomic units (OTUs) were recovered from the water samples stored at -80  ℃, which were identified asBacillus sp. D-U164(2010), Bacillus simplex, Exiguobacterium sp. Mkj28, Bacillus muralis, Acinetobactersp. L-HG-2BW, Exiguobacterium sp. AC-CS-C2, and Bacillus thuringiensis, respectively. In contrast, 14 OTUs were recovered from the water samples stored at 4  ℃, and were classified as Proteobacterium M71 D72, Chryseobacterium sp. 12 4K, Bacillus sp.HJ14, Bacillus pumilus, Stenotrophomonas sp. LL75,Bacillus muralis, Bacillus cereus, Brevundimonas bullata, Stenotrophomonas sp. Cza24, Microbacteriumsp. VKM Ac2016, Stenotrophomonassp. Ala15, Bacillussp. T4(2013), respectively. Our results indicated that higher diversity of culturable bacteria was recovered from fridgestored water samples than that of cryopreserved. Bacillussp. can be isolated from water samples cryopreserved at -80  ℃ and fridgestored at 4  ℃.

Key words: landscape connectivity, Presbytis leucocephalus., ecological corridor, least-cost path