說明
Permafrost, frozen ground cemented with ice, occupies about a quarter of the Earth’s hard surface and reaches up to a thousand meters depth. Due to constant negative temperatures, permafrost sediments represent a unique record of past epochs, whenever it comes to accumulated methane, oxygen isotope ratio, or stored mummies of animals. Permafrost is also a unique environment where cryptobiotic stages of different microorganisms are trapped and stored alive for up to hundreds of thousands of years. Several strains isolated from permafrost cores have been already described.
Here described is the collection of strains of clonal amoeboid protists of the Soil Cryology Lab, Pushchino, as well as permafrost samples from which the strains were isolated. The dataset presented contains different core characteristics and the taxonomy of isolated strains. Specimens of the collection are available from the Soil Cryology Lab upon request. The cores are stored frozen at −18 °C and may be used for further detailed studies and isolation attempts.
資料紀錄
此資源sampling event的資料已發佈為達爾文核心集檔案（DwC-A），其以一或多組資料表構成分享生物多樣性資料的標準格式。 核心資料表包含 19 筆紀錄。
亦存在 2 筆延伸集的資料表。延伸集中的紀錄補充核心集中紀錄的額外資訊。 每個延伸集資料表中資料筆數顯示如下。
此 IPT 存放資料以提供資料儲存庫服務。資料與資源的詮釋資料可由「下載」單元下載。「版本」表格列出此資源的其它公開版本，以便利追蹤其隨時間的變更。
版本
以下的表格只顯示可公開存取資源的已發布版本。
如何引用
研究者應依照以下指示引用此資源。:
Malavin S, Shmakova L (2020): Amoeboid protists isolated from ancient Siberian permafrost. v1.4. Institute of physicochemical and biological problems in soil science of the Russian Academy of Sciences. Dataset/Samplingevent. http://gbif.ru:8080/ipt/resource?r=ancientpermafrostamoeboidprotists&v=1.4
權利
研究者應尊重以下權利聲明。:
此資料的發布者及權利單位為 Institute of physicochemical and biological problems in soil science of the Russian Academy of Sciences。 This work is licensed under a Creative Commons Attribution Non Commercial (CC-BY-NC 4.0) License.
GBIF 註冊
此資源已向GBIF註冊，並指定以下之GBIF UUID: e11d99cb-4a96-4e9d-847e-d078cfd59f6c。 Institute of physicochemical and biological problems in soil science of the Russian Academy of Sciences 發佈此資源，並經由Participant Node Managers Committee同意向GBIF註冊成為資料發佈者。
關鍵字
Samplingevent; permafrost; protists; drilling and coring; carbon-14 analysis; DNA sequencing; microbiology
聯絡資訊
- 元數據提供者 ●
- 作者 ●
- 出處 ●
- 連絡人
http://orcid.org/0000-0003-0470-9378
- 作者
- 作者
- 研究主持人
地理涵蓋範圍
Permafrost samples for the strain isolation were obtained in North-Eastern Eurasia, from Yamal Peninsula to Chukotka.
| 界定座標範圍 | 緯度南界 經度西界 [65, 65], 緯度北界 經度東界 [75, 180] |
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時間涵蓋範圍
| 彙整期間 | 2000-ongoing |
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計畫資料
The project aims at expanding the view on the diversity of protists and their viruses that could be found in ancient (Pliocene to Holocene) Arctic permafrost.
| 計畫名稱 | Biodiversity and evolution of heterotrophic protists and giant viruses from Holocene and Pleistocene Arctic permafrost |
|---|---|
| 辨識碼 | RFBR-CNRS-a-17-54-150003 |
| 經費來源 | Russian Foundation for Basic Research (RFBR), grant 17-54-150003 |
| 研究區域描述 | Northern Siberia, from Yamal Peninsula to Chukotka |
| 研究設計描述 | Protists and viruses are aseptically isolated from permafrost samples, borehole cores and outcrop samples, taken using specially developed technique (Gilichinskiy, D.A., Khlebnikova, G.M., Zvyagintsev, D.G., Fedorov-Davydov, D.G., Kudryavtseva, N.N., 1989. Microbiology of sedimentary materials in the permafrost zone. International Geology Review 31, 847–858. https://doi.org/10.1080/00206818909465938. Shi, T., Reeves, R.H., Gilichinsky, D.A., Friedmann, E.I., 1997. Characterization of viable bacteria from Siberian permafrost by 16S rDNA sequencing. Microbial Ecology 33, 169–179. https://doi.org/10.1007/s002489900019). Isolated strains are studied using general microbiological techniques (cloning, axenization, DNA isolation and characterization, microscopy observations, physiological essays, taxonomic identification and description). |
參與計畫的人員:
- 處理者
取樣方法
The drilling was performed using a mobile drilling rig (core-drilling machine) UKB-12/25 (V.V. Vorovsky Machine-Building Plant, Moscow, Russia) operated without flushing and blowing. Flushing and blowing were shown to cause contamination of the cores by modern soil microorganisms. Cores were removed each 30—70 cm of the drilling. The core diameters were 115 to 75 mm, depending on the well depth (the deeper the well the smaller). A removed core was enveloped in a one-centimeter-thick coat of half-melted cuttings. This coat was removed with a knife, showing a completely frozen inner part. After a short lithological and glaciological description of the sediments, the core was passed to a clean field lab organized in a tent. Operations in the lab were conducted behind a gas-fired burner using disposable materials and gloves, following general microbiological practice. In the lab, the core was shaved with a sterile scalpel so that an approximately five mm outermost layer was removed. This remaining core was four to six cm in diameter, depending on the initial value. Immediately after shaving, the core was put into a sterile plastic bag and placed in a portable freezer, a cave dug into an ice wedge, or an empty borehole used as a freezer. In total, the “outdoor” stage of the process lasted five to ten minutes, depending on the current well depth. The “indoor” lab stage generally took around five minutes. All collected cores were kept at negative temperatures during the whole period of transportation to the stationary lab. Outcrops are natural exposures of ancient permafrost sediments formed at sea and river banks. The advantage of sampling from the outcrop wall is the possibility of visual inspection and description of the whole sediment layer. Samples from outcrops were taken from the frozen surface of the outcrop wall after the removal of melted material. In the wall, a hole of about 40 cm deep was made with either a hand-held drill, a chisel, or a knife. A cylinder sample of about 5 cm in diameter was carefully carved or drilled out from the bottom of the hole, treated with 95% ethanol, and immediately placed into a sterile plastic bag. As with the cores, all collected outcrop samples were kept frozen during the whole period of transportation. Buried terminal nesting chambers of the ground squirrel (Citellus) burrows are unique paleontological objects of Pleistocene Ice Complex sediments. They usually contain animal supplies made of seeds of surrounding grasses. Usually frozen in the living state, they are very well preserved. From the tissue of a Silene stenophylla seed, a viable flowering plant was grown (Yashina et al. 2012). Chambers also contain a diverse community of protists and fungi. Chambers were cut from the outcrop wall in one or several pieces ten or more cm in each dimension, put immediately in sterile plastic bags, and kept frozen until processing in the lab.
| 研究範圍 | The samples were collected in the field and stored constantly frozen during all periods of transportation and processing. In the lab, a part of each sample was used to isolate protist strains. The remaining part has never been melted and is stored at −18 °C. The isolation was done in sterile conditions. Obtained samples were cloned and maintained as bacterized or axenic cultures on plastic or agar with liquid overlay. |
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| 品質控管 | During the development of the permafrost microbiological sampling technique, several tests for contamination of the core interior have been established at different phases of sampling and storage. Gilichinskiy et al. (2010) and Shi et al. (1997) used Serratia marcescens cultures (Bacteria) producing easy-to-notice red colonies. A drilling barrel was covered with a culture suspension 2 h prior to drilling. In a parallel experiment, frozen samples were seeded with the same suspension already in the lab and left intact at negative temperature for several hours to several months. In both experiments, the distribution of S. marcescens cells in a core has been investigated during the sample processing. In all tests, bacteria have been found exclusively in the outer layer and never inside the core. Later, Juck et al. (2005) used fluorescent latex beads (microspheres) 0.5 μm in diameter and a transformed Pseudomonas strain expressing the green fluorescent protein (GFP). Both beads and transformed bacteria were applied to the drilling equipment before drilling, similar way as described above for S. marcescens suspension. Fluorescence microscopy showed that neither beads nor bacteria penetrate into a sample to a depth exceeding one cm from the surface. Additionally, PCR revealed no amplification of the GFP gene from the inner part of the cores. Based on the negative results obtained for bacteria and fluorescent beads, i.e. particles around two μm in diameter or less, we consider it proven that resting protist cysts, which are at least five times bigger, can’t move inside the frozen ground and thus could not penetrate the sediments considerably later than they were deposited. Same way, the contamination of the inner part of the samples during sampling and laboratory processing is not possible either. |
方法步驟描述:
- Field sampling (see above)
- Isolation of protists in the lab For the isolation of protists, 1—5 grams of the sediment from the inner part of the frozen sample were placed in 90 mm Petri dishes filled with autoclaved mineral Prescott and James (PJ) medium (Prescott and James 1955). Negative controls (same procedures without sample inoculation) were performed simultaneously. The isolation was performed in a laminar flow hood using disposal or sterilized equipment. After 7—10 days of incubation, samples were examined using a Nikon TMF100 inverted microscope. Detected cells were transferred to the fresh medium by a glass capillary. Strains were cloned and further cultured in 60 mm Petri dishes using 0.01% Cerophyl infusion made on PJ medium (Smirnov and Brown 2004).
收藏資料
| 蒐藏名稱 | Коллекция образцов вечной мерзлоты лаборатории криологии почв ИФХиБПП РАН |
|---|---|
| 蒐藏編號 | SCL |
| 蒐藏名稱 | Коллекция амебоидных протистов лаборатории криологии почв ИФХиБПП РАН |
|---|---|
| 蒐藏編號 | SCL-Prot |
| 標本保存方法 | No treatment, Microscopic preparation, Deep frozen |
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引用文獻
- Malavin, S., Shmakova, L., 2020. Isolates from ancient permafrost help to elucidate species boundaries in Acanthamoeba castellanii complex (Amoebozoa: Discosea). European Journal of Protistology 73, 125671. https://doi.org/10.1016/j.ejop.2020.125671
- Shatilovich, A.V., Shmakova, L.A., Mylnikov, A.P., Gilichinsky, D.A., 2009. Ancient Protozoa isolated from permafrost, in: Margesin, R. (Ed.), Permafrost Soils. Springer Berlin Heidelberg, Berlin, Heidelberg, pp. 97–115. https://doi.org/10.1007/978-3-540-69371-0_8
- Shmakova, L., Bondarenko, N., Smirnov, A., 2016. Viable species of Flamella (Amoebozoa: Variosea) isolated from ancient Arctic permafrost sediments. Protist 167, 13–30. https://doi.org/10.1016/j.protis.2015.11.001
- Shmakova, L.A., Karpov, S.A., Malavin, S.A., Smirnov, A.V., 2018. Morphology, biology and phylogeny of Phalansterium arcticum sp. n. (Amoebozoa, Variosea), isolated from ancient Arctic permafrost. European Journal of Protistology 63, 117–129. https://doi.org/10.1016/j.ejop.2018.02.002
- Shmakova, L.A., Rivkina, E.M., 2015. Viable eukaryotes of the phylum Amoebozoa from the Arctic permafrost. Paleontological Journal 49, 572–577. https://doi.org/10.1134/S003103011506012X
- Демидов, Н.Э., Баранская, А.В., Дурденко, Е.В., Занина, О.Г., Караевская, Е.С., Пушина, З.В., Ривкина, Е.М., Спирина, Е., Спенсер, Р., 2016. Биогеохимия мерзлых толщ арктического побережья полуострова Гыдан. Проблемы Арктики и Антарктики 3, 34–49.
額外的詮釋資料
Empty cells represent "no data" N/A: "not applicable"
| 替代的識別碼 | e11d99cb-4a96-4e9d-847e-d078cfd59f6c |
|---|---|
| http://gbif.ru:8080/ipt/resource?r=ancientpermafrostamoeboidprotists |