A molecular approach to identification of protonemata helps assess biodiversity of extremely acidic freshwaters
Macroscopic fuzzy clumps of green filaments resembling filamentous algae were found on multiple sampling occasions in the water close to the shore of the extremely acidic (pH < 3) Hromnice Lake in Czechia. Microscopic investigation revealed that these filaments were moss protonemata. In order to identify the moss, we sequenced chloroplast (rbcL), mitochondrial (nad5), and nuclear (ITS2) molecular markers of these filaments. In addition, we sampled adult mosses growing on the wet substrate soaked with lakewater. The sequences of protonemata matched those of the adults, which were morphologically identified as Dicranella sp. Phylogenetic analysis of the rbcL gene showed a sister relationship with D. heteromalla, generally known for growing in acidic habitats, and other protonemata occurring in acidic rivers in Japan. The nad5-based phylogeny revealed that the studied protonemata belonged to the species D. cerviculata, and the same taxonomic affiliation was confirmed by the ITS2 rDNA sequence and its secondary structure. The extreme environment of Hromnice Lake prevents the further development of protonemata which, in turn, are capable of surviving acidic conditions in the prolonged protonemal stage. Due to their macroscopic similarity to filamentous algae, protonemata might be more common in extremely acidic waters than originally thought.
KeywordsAcidic pit lakes Protonema Dicranella Molecular phylogeny
This study was funded by the National Museum in Prague, grant no. P17/01IG-BA. We thank Jan Kučera (Department of Botany, University of South Bohemia, Czechia) for reading the draft of the manuscript.
- Bonfim Santos M, Fedosov VE, Hartman T, Ignatov M, Siebel H, Stech M (2017) Dicranella, a case study of misunderstood diversity of inconspicuous haplolepideous mosses. In: XXI Cryptogamic Botany Symposium; 20–24 June 2017, Aranjuez, Spain. Abstract SIST-08, p 100Google Scholar
- Geller W, Schultze M, Kleinmann B, Wolkersdorfer C (2013) Acidic pit lakes: the legacy of coal and metal surface mines. Springer, HeidelbergGoogle Scholar
- Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98Google Scholar
- Kučera J, Váňa J, Hradílek Z (2012) Bryophyte flora of the Czech Republic: updated checklist and Red List and a brief analysis. Preslia 84:813–850Google Scholar
- Lanfear R, Frandsen PB, Wright AM, Senfeld T, Calcott B (2017) PartitionFinder 2: new methods for selecting partitioned models of evolution for molecular and morphological phylogenetic analyses. Mol Biol Evol 34:772–773Google Scholar
- Marshall NL (1919) Mosses and lichens: a popular guide to the identification and study of our commoner mosses and lichens, their uses, and methods of preserving. Garden City, New YorkGoogle Scholar
- Rachna P, Vashistha BD (2017) Effects of heavy metals on protonemal growth and bud formation in the moss Hydrogonium arcuatum. Agric Sci Digest 37:117–121Google Scholar
- Rambaut A (2007) Figtree, a graphical viewer of phylogenetic trees. http://tree.bio.ed.ac.uk/software/figtree/
- White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis M, Gelfand D, Sninsky J, White T (eds) PCR protocols: a guide to methods and applications. Academic, Orlando, pp 315–322Google Scholar
- Whitton BA, Diaz BM (1981) Influence of environmental factors on photosynthetic species composition in highly acidic waters. Verh Internat Verein Limnol 21:1459–1465Google Scholar