Abstract
Single-cell PCR and gene sequencing were conducted to evaluate species and genotypic compositions of Cryptomonas in the eutrophic Lake Hira, Japan. We determined the sequences of nuclear internal transcribed spacer 2 region from single Cryptomonas cells with a high success rate (83.3–97.9 %), excluding one case (56.3 %). A total of 325 sequences were obtained over eight sampling days from May 28, 2012, to October 3, 2012, and phylogenetic analysis indicated that all sequences were divided into six groups. Four groups were clustered together with known sequences of C. curvata, C. marssonii, C. pyrenoidifera or C. tetrapyrenoidosa, although the sequences of the other two groups did not show high similarity to known Cryptomonas species. Cryptomonas curvata dominated during the study period (45–98 %), and unidentified Cryptomonas species (group 2) became dominant at high water temperatures. The genotypic composition of C. curvata also varied temporarily, suggesting that the genotypic composition of Cryptomonas was susceptible to environmental changes. These results indicated that single-cell PCR can be used to analyze the species composition and ecology of Cryptomonas.
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Acknowledgments
Financial support of the present study was partially provided under the Environment Research and Technology Development Fund (D-0905) of the Ministry of the Environment Japan and Japan Science and Technology Strategic International Research Cooperative Program project ‘Fate of dissolved organic matter in lakes with special reference to loading and pollution’. S. D. Thottathil was supported by a scholarship from the Japanese Ministry of Education, Culture, Sports, Science and Technology.
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Nishino, H., Hodoki, Y., Thottathil, S.D. et al. Identification of species and genotypic compositions of Cryptomonas (Cryptophyceae) populations in the eutrophic Lake Hira, Japan, using single-cell PCR. Aquat Ecol 49, 263–272 (2015). https://doi.org/10.1007/s10452-015-9520-9
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DOI: https://doi.org/10.1007/s10452-015-9520-9