Characterisation of Fruticose Lichen Genus Stereocaulon from Sabah Based on Morphology, Chemotyping, and Molecular Typing: Stereocaulon lichen of Sabah

Jing Kae LIM; Charles Santhanaraju VAIRAPPAN; Thor-Seng LIEW

doi:10.51200/jtbc.v22i.5673

Authors

Jing Kae LIM Universiti Malaysia Sabah
Charles Santhanaraju VAIRAPPAN
Thor-Seng LIEW

DOI:

https://doi.org/10.51200/jtbc.v22i.5673

Keywords:

Snow lichen, integrative taxonomy, alpine ecosystem, montane forest, thin-layer chromatography, high-performance thin-layer chromatography (HPTLC), mountain ecosystem, Borneo

Abstract

Lichens in the genus Stereocaulon are commonly found in high-elevation mountains in tropical regions. Taxonomy of Stereocaulon is always an ongoing topic discussed among lichenologists, especially for species-level identification due to polymorphism and chemical diversity of Stereocaulon. In this study, we examined Stereocaulon spp. from Mount Kinabalu, the highest mountain in Borneo (4095 m). A total of 42 Stereocaulon specimens were included in this study; 40 were newly collected between 1900 m and 3700 m a.s.l. along the summit trail of Mount Kinabalu, and two specimens from Mount Alab (1900 m) were previously published. We used integrative approaches to delimit the specimens into Operational Taxonomic Units (OTUs) based on macro-morphology, chemical profiles, and molecular phylogeny. Macro-morphological characters, including apothecia (reproductive organ), phyllocladia, pseudopodetia and cephalodia were examined, while chemical profiles were obtained from spot tests and High-performance thin-layer chromatography (HPTLC). A phylogenetic tree was constructed based on the ITS gene by using Bayesian and Maximum Likelihood analyses. A total of six Operational Taxonomic Units (OTUs) were identified and were provisionally named according to their diagnostic morphological and chemical characters, namely, RF (ramular-fluorescence), RN (ramular-nonfluorescence), GF (granular-fluorescence), GN (granular-nonfluorescence), RU1 (ramular-curved 1) and RU2 (ramular-curved 2). The monophyly of each of the OTUs is supported by phylogenetic analysis. Due to lack of reliable identification keys and reference genetic data for this genus, we could not determine the species identities for the OTUs revealed in this study. Nevertheless, the findings of this study provide a baseline for future studies on the taxonomy of Stereocaulon species in Mount Kinabalu, integrating morphology, chemistry, and genetics. The implementation of high-performance thin-layer chromatography (HPTLC) profiles show potential for distinguishing samples. However, further research with more Stereocaulon samples from different taxa and regions is necessary to verify the reliability of this method.

References

Athukorala SN, Doering J, Piercey-Normore MD (2016) Morphological and genetic polymorphism in two North American reindeer lichens: Cladonia arbuscula and C. rangiferina. Ceylon Journal of Science (Biological Sciences) 44(2): 55–65.

Badang D (1999) Tourism geology of Kinabalu Park, Sabah (in Malay). (M.Sc Thesis. Universiti Kebangsaan Malaysia (unpublished).

Del-Prado R, Cubas P, Lumbsch HT, Divakar PK, Blanco O, de Paz GA, Molina MC, Crespo A (2010) Genetic distances within and among species in monophyletic lineages of Parmeliaceae (Ascomycota) as a tool for taxon delimitation. Molecular Phylogenetics and Evolution 56(1): 125–133.

Del-Prado R, Divakar PK, Crespo A (2011) Using genetic distances in addition to ITS molecular phylogeny to identify potential species in the Parmotrema reticulatum complex: a case study. The Lichenologist 43(6): 569–583.

Din LB, Zakaria Z, Samsudin MW, Elix JA (2010) Chemical Profile of Compounds from Lichens of Bukit Larut, Peninsular Malaysia. Sains Malaysiana 39(6): 901–908.

Divakar PK, Leavitt SD, Molina MC, Del-Prado R, Lumbsch HT, Crespo A (2016) A DNA barcoding approach for identification of hidden diversity in Parmeliaceae (Ascomycota): Parmelia sensu stricto as a case study. Botanical Journal of the Linnean Society 180(1): 21–29.

Ekman S, Tønsberg T (2002) Most species of Lepraria and Leproloma form a monophyletic group closely related to Stereocaulon. Mycological Research 106(11): 1262–1276.

Fontaine KM, Stocker-Wörgötter E, Booth T, Piercey-Normore MD (2013) Genetic diversity of the lichen-forming alga, Diplosphaera chodatii, in North America and Europe. The Lichenologist 45(6): 799–813.

Fries TM (1858) Monographia Stereocaulorum et Pilophororum. Nova acta Regiae Societatis Scientiarum Upsaliensis 3(2): 307–380.

Gardes M, Bruns TD (1993) ITS primers with enhanced specificity for basidiomycetes – application to the identification of mycorrhizae and rusts. Molecular Ecology 2(2): 113–118.

Henssen A, Jahns HM, Santesson J (1974) Lichenes: eine Einführung in die Flechtenkunde. Thieme Georg Verlag. 468 pp.

Huang MR (2008) Noteworthy species of Stereocaulon from China. Mycosystem 27(1): 58–90.

Huang MR (2010) Altitudinal patterns of Stereocaulon (Lichenized Ascomycota) in China. Acta Oecologica. 36(2): 173–178.

Huyen VT, Tram NTT, Cuong NM, Dam NP, Françoise LLD, Joël B (2017) Phytochemical and cytotoxic investigations of the lichen Stereocaulon evolutum Graewe. Vietnam Journal of Chemistry 55(4): 429–432.

Hyde KD, Abdel-Wahab MA, Abdollahzadeh J, Abeywickrama PD, Absalan S, Afshari N, et al. (2023) Global consortium for the classification of fungi and fungus-like taxa. Mycosphere 14(1): 1960–2012.

Ismed F, Lohézic-Le Dévéhat F, Guiller A, Corlay N, Bakhtiar A, Boustie J (2018) Phytochemical review of the lichen genus Stereocaulon (Fam. Stereocaulaceae) and related pharmacological activities highlighted by a focus on nine species. Phytochemistry Reviews 17: 1165–1178.

Jørgensen PM (1998) Acantholichen pannarioides, a new basidiolichen from South America. Bryologist 101 (3): 444–447.

Kirk PM, Cannon PF, David JC, Stalpers JA (2001) Ainsworth and Bisby’s Dictionary of the Fungi, 9th edn. CABI Publishing, Wallingford. 655 pp.

Kitayama K (1991) Actual Vegetation of Mount Kinabalu Park, Sabah, Malaysia. Vegetation map, scale 1: 100 000. Project Paper, Protected Areas and Biodiversity. East-West Center, Honolulu.

Kumar S, Stecher G, Li M, Knyaz C, Tamura K (2018) MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution 35: 1547–1549.

Lamb IM (1951) On the morphology, phylogeny, and taxonomy of the lichen genus Stereocaulon. Canadian Journal of Botany 29(5): 522–584.

Lamb IM (1977) A conspectus of the lichen genus Stereocaulon (Schreb.) Hoffm. The Journal of the Hattori Botanical Laboratory 43: 191–355.

Lamb IM (1978) Keys to the species of the lichen genus Stereocaulon (Schreb.) Hoffm. The Journal of the Hattori Botanical Laboratory 44: 209–250.

Leavitt S, Fernández-Mendoza F, Pérez-Ortega S, Sohrabi M, Divakar P, Lumbsch T, Clair LS (2013) DNA barcode identification of lichen-forming fungal species in the Rhizoplaca melanophthalma species-complex (Lecanorales, Lecanoraceae), including five new species. MycoKeys 7: 1–22.

Lim J-K (2019) Diversity of Secondary Metabolites in Stereocaulon of Mount Alab. BSc thesis, University Malaysia Sabah. Conservation Biology Programme. Faculty of Science and Natural Resources Universiti Malaysia Sabah. 73 pp.

Lõhmus A, Motiejūnaitė J, Lõhmus P (2023) Regionally varying habitat relationships in lichens: The concept and evidence with an emphasis on North-Temperate ecosystems. Journal of Fungi 9(3): 341.

Lumbsch, HT (2002) Analysis of phenolic products inlichens for identification and taxonomy. In Protocolsin Lichenology. Culturing, Biochemistry, Ecophysiologyand Use in Biomonitoring (I. Kranner, R. Beckett &A. Varma, eds): 281–295.

McCune B, Vančurová L, Myllys L (2023) Stereocaulon tomentosoides, a new combination for a western North American endemic species with cyanobiont and chemotype polymorphisms. Plant and Fungal Systematics 68(2): 364–377.

Myllys L, Lohtander K, Tehler A (2001) β-tubulin, ITS and group I intron sequences challenge the species pair concept in Physcia aipolia and P. caesia. Mycologia 93(2): 335–343.

Orange A, James PW, White FJ (2001) Microchemical Methods for the Identification of Lichens. British Lichen Society. 101 pp.

Oset M (2014) The lichen genus Stereocaulon (Schreb.) Hoffm. in Poland–a taxonomic and ecological study. Monographiae Botanicae 104. 81 pp.

Park JS, Park CH, Park SY, Oh SO, Jayalal U, Hur JS (2018) Revision of the lichen genus Stereocaulon (Stereocaulaceae, Ascomycota) in South Korea. Mycobiology 46(2): 101–113.

Sipman HJM (1993) Lichens from Mount Kinabalu. Tropical Bryology 8: 281–314.

Tønsberg T (1977) The chemical strains in Stereocaulon rivulorum and their distribution. Norwegian Journal of Botany 24: 231–234.

Torres JM, Torres VDO, Rodrigues AS, Gianini AS, Micheletti AC, Honda NK, Spielmann AA, Lorenz AP (2023) Lineages of the lichen-forming fungus Stereocaulon alpinum and their photobionts in southern South America and maritime Antarctica. Polar Biology 46(9): 865–879.

White TJ, Bruns T, Lee SJWT, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR protocols: a guide to methods and applications 18(1): 315–322.