Pülümür'deki (Tunceli) bir Hipersalin Kaynak Suyunun Prokaryotik Çeşitliliği

Year 2020, Volume: 25 Issue: 3, 127 - 134, 30.12.2020

Seval Çınar , Mehmet Burçin Mutlu

Abstract

Tuz göllerinin ve solar tuzlaların mikrobiyal çeşitliliği kapsamlı bir şekilde çalışılmıştır, ancak hipersalin kaynak sularının mikrobiyotaları hakkında sadece birkaç rapor vardır. Bu çalışmada, Pülümür'deki bir hipersalin kaynak suyunda yaşayan arkeal ve bakteriyal gruplara odaklandık. Kültür-bağımlı çalışmalara ek olarak, 16S rRNA genlerinin PCR ile amplifiye edilmiş fragmentlerinin denatüre edici gradiyent jel elektroforezi (DGGE) ve floresan in situ hibridizasyon (FISH) analizi gerçekleştirildi. DAPI boyası kullanılarak gerçekleştirilen toplam hücre sayımında kaynak suyunda ~105 hücre/mL saptandı. FISH analizi, bakteri hücrelerinin toplam mikrobiyal topluluğun ~%72'sini oluşturduğunu, arkeal hücrelerin ise topluluğun küçük bir bölümünü oluşturduğunu göstermiştir. İzolatlar, Rhodovibrio, Spiribacter, Salinivibrio, Halomonas, Idiomarina, Marinobacter, Natrinema ve Halohasta cinsleriyle filogenetik olarak ilişkiliydi. Spiribacter filotipleri ve Owenweeksia ile uzaktan ilişkili filotipler DGGE analizinden elde edilmiştir.

Keywords

Hipersalin çevreler, Halofilik mikroorganizmalar, DGGE, FISH

Prokaryotic Diversity of a Hypersaline Spring Water in Pülümür (Tunceli)

Abstract

The microbial diversity of the solar salterns and salt lakes has been extensively studied, but there are only a few reports on microbiota of hypersaline spring waters. In this study, we focused on the archaeal and bacterial groups inhabiting a hypersaline spring water in Pülümür. In addition to culture-dependent studies, denaturing gradient gel electrophoresis (DGGE) of PCR-amplified fragments of 16S rRNA genes and fluorescence in situ hybridization (FISH) analyzes were performed. The total cell count performed using 4',6-diamidino-2-phenylindole (DAPI) stain revealed ~105 cells mL-1 in spring water. The FISH analysis showed that bacterial cells accounted for ~72% of the total microbial community, while archaeal cells made up a small portion of the community. Isolates were phylogenetically related with the genera Rhodovibrio, Spiribacter, Salinivibrio, Halomonas, Idiomarina, Marinobacter, Natrinema and Halohasta. Phylotypes of Spiribacter and phylotypes distantly associated with Owenweeksia were obtained from DGGE analysis.

Keywords

Hypersaline environments, Halophilic microorganisms, DGGE, FISH

References

• Antón, J., Llobet‐Brossa, E., Rodríguez‐Valera, F., & Amann, R. (1999). Fluorescence in situ hybridization analysis of the prokaryotic community inhabiting crystallizer ponds. Environmental Microbiology, 1, 517-523. doi: 10.1046/j.1462-2920.1999.00065.x
• Birbir, M., Calli, B., Mertoglu, B., Bardavid, R. E., Oren, A., Ogmen, M. N., & Ogan, A. (2007). Extremely halophilic Archaea from Tuz Lake, Turkey, and the adjacent Kaldırım and Kayacık salterns. World Journal of Microbiology and Biotechnology, 23, 309-316. doi: 10.1007/s11274-006-9223-4
• Çınar, S., & Mutlu, M. B. (2016). Comparative analysis of prokaryotic diversity in solar salterns in eastern Anatolia (Turkey). Extremophiles, 20, 589-601. doi: 10.1007/s00792-016-0845-7
• Çınar, S., & Mutlu, M. B. (2018). Sivas’ın solar tuzlalarındaki mikrobiyal toplulukların kültür bağımlı karakterizasyonu. Anadolu Üniversitesi Bilim ve Teknoloji Dergisi C-Yaşam Bilimleri ve Biyoteknoloji, 7, 1-11. doi: 10.18036/aubtdc.305855
• DeLong, E. F. (1992). Archaea in coastal marine environments. Proceedings of the National Academy of Sciences, 89, 5685-5689. doi: 10.1073/pnas.89.12.5685
• Dyall-Smith, M. (2009). The Halohandbook- Protocols for Haloarchaeal Genetics, Version 7.3. https://haloarchaea.com/halohandbook/ (date of access: 01.06.2020)
• Edgar, R. C. (2004). MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research, 32, 1792-1797. doi: 10.1093/nar/gkh340
• Kılıç, Ö., & Kılıç, A. M. (2005). Recovery of salt co-products during the salt production from brine. Desalination, 186, 11-19. doi: 10.1016/j.desal.2005.05.014
• Lane, D. J., Pace, B., Olsen, G. J., Stahl, D. A., Sogin, M. L., & Pace, N. R. (1985). Rapid determination of 16S ribosomal RNA sequences for phylogenetic analyses. Proceedings of the National Academy of Sciences, 82, 6955-6959. doi: 10.1073/pnas.82.20.6955
• León, M. J., Aldeguer-Riquelme, B., Antón, J., Sánchez-Porro, C., & Ventosa, A. (2017). Spiribacter aquaticus sp. nov., a novel member of the genus Spiribacter isolated from a saltern. International Journal of Systematic and Evolutionary Microbiology, 67, 2947-2952. doi: 10.1099/ijsem.0.002053
• Maturrano, L., Santos, F., Rosselló-Mora, R., & Antón, J. (2006). Microbial diversity in Maras salterns, a hypersaline environment in the Peruvian Andes. Applied and Environmental Microbiology, 72, 3887-3895. doi: 10.1128/AEM.02214-05
• Mou, Y. Z., Qiu, X. X., Zhao, M. L., Cui, H. L., Oh, D., & Dyall-Smith, M. L. (2012). Halohasta litorea gen. nov. sp. nov., and Halohasta litchfieldiae sp. nov., isolated from the Daliang aquaculture farm, China and from Deep Lake, Antarctica, respectively. Extremophiles, 16, 895-901. doi: 10.1007/s00792-012-0485-5
• Mutlu, M. B., Martínez-García, M., Santos, F., Peña, A., Guven, K., & Antón, J. (2008). Prokaryotic diversity in Tuz Lake, a hypersaline environment in Inland Turkey. Federation of European Microbiological Societies (FEMS) Microbiology Ecology, 65, 474-483. doi:10.1111/j.1574-6941.2008.00510.x
• Mutlu, M. B., & Güven, K. (2015). Bacterial diversity in Çamaltı saltern, Turkey. Polish Journal of Microbiology, 64, 37-45.
• Muyzer, G., De Waal, E. C., & Uitterlinden, A. G. (1993). Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Applied and Environmental Microbiology, 59, 695-700.
• Oren, A. (2008). Microbial life at high salt concentrations: phylogenetic and metabolic diversity. Saline Systems, 4, 2. doi:10.1186/1746-1448-4-2
• Oren, A. (2010). Diversity of halophiles. In Horikoshi, K., Antranikian, G., Bull, A. T., Robb, F. T., & Stetter, K. O. (Eds.), Extremophiles Handbook (pp. 310-325). Springer Science & Business Media.
• Özcan, B., Özcengiz, G., Colleri, A., & Cokmus, C. (2007). Diversity of halophilic Archaea from six hypersaline environments in Turkey. Journal of Microbiology and Biotechnology, 17, 985-992.
• Özdemir, K., Öğün, E., Ertaş, M., & Özok, N. (2018). Kozluk (Batman) Tuzla Gözü Tuzlasından İzole Edilen Ektraselüler İnulinaz Enzimi Üreten Salinivibrio T1 İzolatının Karakterizasyonu. Yüzüncü Yıl Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 23, 60-68.
• Güner, İ, Bekdemi̇r, Ü, Ertürk, M, & Şi̇mşek, O. (2000). Tuzluca kaya tuzlası. Doğu Coğrafya Dergisi, 6, 323-347.
• Stamatakis, A. (2014). RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics, 30, 1312-1313. doi: 10.1093/bioinformatics/btu033
• Ventosa, A., Márquez, M. C., Sánchez-Porro, C., & Rafael, R. (2012). Taxonomy of halophilic archaea and bacteria. In Vreeland, R. H. (Ed.), Advances in Understanding the Biology of Halophilic Microorganisms (pp. 59-80). Springer, Dordrecht.
• Yaşa, İ., Kahraman, Ö., Tekin, E., & Koçyiğit, A. (2008). Çamaltı tuzlasından ekstrem halofilik archaea izolasyonu ve moleküler karakterizasyonu. Ege University (EU) Journal of Fisheries & Aquatic Sciences, 25, 117-121.
• Yıldız, E., Özcan, B., & Calışkan, M. (2012). Isolation, characterization and phylogenetic analysis of halophilic Archaea from a salt mine in central Anatolia (Turkey). Polish Journal of Microbiology, 61, 111-117.