Fottea 2018, 18(1):72-85 | DOI: 10.5507/fot.2017.022

Myrmecia israeliensis as the primary symbiotic microalga in squamulose lichens growing in European and Canary Island terricolous communities

Patricia Moya1, Salvador Chiva1, Aránzazu Molins1, Iva Jadrná2, Pavel ©kaloud2, Ondøej Peksa3, Eva Barreno1
1 Universitat de València, Fac. CC. Biológicas, ICBIBE, Botánica. C/ Dr. Moliner 50, 46100-Burjassot, Valencia, Spain; *Corresponding author e-mail: patricia.moya@uv.es
2 Charles University in Prague, Faculty of Sciences, Department of Botany, Benátská 2, 128 01 Praha 2, Czech Republic
3 The West Bohemian Museum in Pilsen, Kopeckého sady 2, 30100-Plzeò, Czech Republic

Myrmecia israeliensis has been traditionally considered as a green coccoid free-living microalga. This microalga was previously suggested as the primary phycobiont in the lichens Placidium spp., Heteroplacidium spp., and Psora decipiens. However, due to the absence of ITS rDNA sequences (barcode information) published along with these investigations, the symbiotic nature of M. israeliensis might be confirmed by using the DNA barcoding and different microscopic examinations both in the symbiotic state and in culture. The aim of this study was to settle the presence of M. israeliensis as the primary microalga in squamulose lichens growing in terricolous communities (Psora spp., Placidium spp. and Claviscidium spp.) in 32 localities within European and Canary Island ecosystems by using both molecular and ultrastructural techniques. The lichen-forming fungi were identified using ITS rDNA as a barcode, and in the case of P. decipiens specimens, the mycobiont analyses showed an unexpected variability. Phycobiont phylogenetic analyses were made using both chloroplast (LSU rDNA) and nuclear (ITS rDNA) molecular markers. Our results proved that M. israeliensis is the primary symbiotic microalga in all the chosen and analyzed lichens. In addition, fluorescence microscopy, transmission electron microscopy and scanning electron techniques were used to characterize M. israeliensis. Finally, the presence of this microalga in lichen thalli was verified using different microscopic observations. A combination of different techniques, both molecular and microscopic, allowed for the accurate identification of this symbiotic microalga, beforehand mainly known as free living. Here, we suggest the combination of these techniques to prevent incorrect identification in microalgal lichen studies.

Keywords: Clavascidium spp., ITS rDNA (barcoding), LSU rDNA, Myrmecia israeliensis, phycobiont, Placidium spp., Psora spp., ultrastructure

Received: June 20, 2017; Accepted: October 10, 2017; Prepublished online: February 27, 2018; Published: March 31, 2018Show citation

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Moya, P., Chiva, S., Molins, A., Jadrná, I., ©kaloud, P., Peksa, O., & Barreno, E. (2018). Myrmecia israeliensis as the primary symbiotic microalga in squamulose lichens growing in European and Canary Island terricolous communities. Fottea, 18(1), 72-85. doi: 10.5507/fot.2017.022
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    References

    1. Ahmadjian, V. (1993): The lichen symbiosis. - John Wiley & Sons, New York.
    2. Akaike, H. (1974): A new look at the statistical model identification. - IEEE transactions on automatic control 19: 716-723. Go to original source...
    3. Akaike, H. (2011): Akaike's information criterion. - In: Heidelberg, S. B. (ed.): International Encyclopedia of Statistical Science. - pp. 25-25, Springer, Berlin. Go to original source...
    4. Arnold, A.E.; Miadlikowska, J.; Higgins, K.L.; Sarvate, S.D.; Gugger, P.; Way, A.; Hofstetter, V.; Kauff, F. & Lutzoni, F. (2009): A phylogenetic estimation of trophic transition networks for ascomycetous fungi: Are lichens cradles of symbiotrophic fungal diversification? - Syst. Biol. 58: 283-297. Go to original source...
    5. Belnap, J. & Lange, O.L. (2001): Structure and functioning of biological soil crusts: A synthesis. - In: Belnap, J. & Lange, O.L. (eds): Biological soil crusts: structure, function, and management. - pp. 471-479, Springer, Berlin. Go to original source...
    6. Belnap, J. (2003): The world at your feet: Desert biological soil crusts. - Front. Ecol. Environ. 1: 181-189. Go to original source...
    7. Belnap, J. & Büdel, B. (2016): Biological soil crusts as soil stabilizers. - In: Weber, B.; Belnap, J. & Büdel, B. (eds): Biological Soil Crusts: An Organizing Principle in Drylands. - pp. 305-320, Springer, Berlin. Go to original source...
    8. Bischoff, H.W. & Bold, H.C. (1963): Phycological studies IV. Some soil algae from enchanted rock and related algal species. - 95 pp., Publications No. 6318, University of Texas.
    9. Bold, H.C. (1949): The morphology of Chlamydomonas chlamydogama, sp. nov. - Bulletin of the Torrey Botanical Club 76: 101-108. Go to original source...
    10. Bowker, M.A.; Mau, R.L.; Maestre, F.T.; Escolar, C. & Castillo-Monroy, A.P. (2011): Functional profiles reveal unique ecological roles of various biological soil crust organisms. - Funct. Ecol. 25: 787-795. Go to original source...
    11. Büdel, B.; Colesie, C.; Green, T.A.; Grube, M.; Suau, R.L.; Loewen-Schneider, K.; Maier, S.; Peer, T.; Pintado, A. & Raggio, J. (2014): Improved appreciation of the functioning and importance of biological soil crusts in Europe: The soil crust international project (SCIN). - Biodivers. Conserv. 23: 1639-1658. Go to original source...
    12. Casano, L.M.; del Campo, E.M.; García-Breijo, F.J.; Reig-Armiñana, J.; Gasulla, F.; Del Hoyo, A.; Guéra, A. & Barreno, E. (2011): Two Trebouxia algae with different physiological performances are ever present in lichen thalli of Ramalina farinacea. Coexistence versus competition? - Environ. Microbiol. 13: 806-818. Go to original source...
    13. Castresana, J. (2000): Selection of conserved blocks from multiple alignments for their use in phylogenetic analysis. - Mol. Biol. Evol. 17: 540-552. Go to original source...
    14. Catalá, S.; del Campo, E.M.; Barreno, E.; García-Breijo, F.J.; Reig-Armiñana, J. & Casano, L.M. (2015): Coordinated ultrastructural and phylogenomic analyses shed light on the hidden phycobiont diversity of Trebouxia microalgae in Ramalina fraxinea. - Mol. Phylogenet. Evol. 94: 765-777. Go to original source...
    15. Chantanachat, S. & Bold, H.C. (1962): Phycological studies II. Some algae from arid soils. - 74 pp., Publications No. 6218, University of Texas.
    16. CHIVA, S. (2012): Especificidad y variabilidad de los ficobiontes identificados en líquenes de comunidades gipsícolas. - 68 pp., Master Dissertation, Universitat de València, Spain.
    17. Chiva, S.; Moya, P.; Molins, A.; Reig-Armiñana, J.; García-Breijo, F.J. & Barreno, E. (2015): Coexistence and prevalence of symbiotic microalgae in Buellia zoharyi lichen: are substrata and/or biogeographic barriers involved? - In: Costa Vieira; Pinto Hespanhol; Mendonça Marques; Vasconcelos & Arenas (eds): XX Simpósio de Botânica Criptogâmica. - pp.60, CIIMAR- CIBIO/InBIO, Oporto, Portugal. ISBN: 978-989-97443-5-6.
    18. Darriba, D.; Taboada, G.L.; Doallo, R. & Posada, D. (2012): jModelTest 2: More models, new heuristics and parallel computing. - Nat. Methods 9: 772-772. Go to original source...
    19. Del Campo, E.; Casano, L.; Gasulla, F. & Barreno, E. (2010): Suitability of chloroplast LSU rDNA and its diverse group I introns for species recognition and phylogenetic analyses of lichen-forming Trebouxia algae. - Mol. Phylogenet. Evol. 54: 437-444. Go to original source...
    20. Doyle, J. & Doyle, J. (1987): Genomic plant DNA preparation from fresh tissue-CTAB method. - Phytochem. Bull. 19: 11-15.
    21. Friedl, T. (1995): Inferring taxonomic positions and testing genus level assignments in coccoid green lichen algae: A phylogenetic analysis of 18S ribosomal RNA sequences from Dictyochloropsis reticulata and from members of the genus Myrmecia (Chlorophyta, Trebouxiophyceae cl. nov.) 1. - J. Phycol. 31: 632-639. Go to original source...
    22. Friedl T. & Büdel B. (2008): Photobionts. - In: Nash, T. H. (eds): Lichen biology. - pp. 9-26, Cambridge University Press, Cambridge. Go to original source...
    23. Galun, M.; Ben-Shaul, Y. & Paran, N. (1971): The fungus-alga association in the Lecideaceae: An ultrastructural study. - New Phytol. 70: 483-485. Go to original source...
    24. Galun, M. (1988): CRC handbook of lichenology. Vols I-III. - CRC Press, Boca Raton, FL, USA.
    25. Gardes, M. & Bruns, T.D. (1993): ITS primers with enhanced specificity for basidiomycetes-application to the identification of mycorrhizae and rusts. - Mol. Ecol. 2: 113-118. Go to original source...
    26. Gasulla, F.; Guéra, A. & Barreno, E. (2010): A simple and rapid method for isolating lichen photobionts. - Symbiosis 51: 175-179. Go to original source...
    27. Geitler, L. (1963): Über haustorien bei flechten und über Myrmecia biatorellae in Psora globifera. - Plant Syst. Evol. 110: 270-280. Go to original source...
    28. Katoh, K.; Misawa, K.; Kuma, K. & Miyata, T. (2002): MAFFT: A novel method for rapid multiple sequence alignment based on fast fourier transform. - Nucleic Acids Res. 30: 3059-3066. Go to original source...
    29. Katoh, K. & Standley, D.M. (2013): MAFFT multiple sequence alignment software version 7: Improvements in performance and usability. - Mol. Biol. Evol. 30: 772-780. Go to original source...
    30. Leavitt, S.D.; Kraichak, E.; Nelsen, M.P.; Altermann, S.; Divakar, P.K.; Alors, D.; Esslinger, T.L.; Crespo, A. & Lumbsch, T. (2015): Fungal specificity and selectivity for algae play a major role in determining lichen partnerships across diverse ecogeographic regions in the lichen-forming family Parmeliaceae (Ascomycota). - Mol. Ecol. 24: 3779-3797. Go to original source...
    31. Maestre, F.T.; Bowker, M.A.; Cantón, Y.; Castillo-Monroy, A.P.; Cortina, J.; Escolar, C.; Escudero, A.; Lázaro, R. & Martínez, I. (2011): Ecology and functional roles of biological soil crusts in semi-arid ecosystems of Spain. - J. Arid Environ. 75: 1282-1291. Go to original source...
    32. Meeßen, J., & Ott, S. (2013). Recognition mechanisms during the pre-contact state of lichens: I. Mycobiont-photobiont interactions of the mycobiont of Fulgensia bracteata. - Symbiosis 59: 121-130. Go to original source...
    33. Melkonian, M. & Berns, B. (1983): Zoospore ultrastructure in the green alga Friedmannia israelensis: An absolute configuration analysis. - Protoplasma 114: 67-84. Go to original source...
    34. Miller, M. A.; Pfeiffer, W. & Schwartz, T. (2010): Creating the CIPRES Science Gateway for inference of large phylogenetic trees. - In: Gateway Computing Environments Workshop (GCE), 2010 November. Go to original source...
    35. Molins, A.; García-Breijo, F.J.; Reig-Armiñana, J.; Del Campo, E.; Casano, L. & Barreno, E. (2013): Coexistence of different intrathalline symbiotic algae and bacterial biofilms in the foliose Canarian lichen Parmotrema pseudotinctorum. - Vieraea 41: 349-370. Go to original source...
    36. Molins, A.; Moya, P.; García-Breijo, F.J.; Reig-Armiñana, J. & Barreno, E. (2017): A multi-tool approach to assess microalgal diversity in lichens: isolation, Sanger sequencing, HTS and ultrastructural correlations. - The Lichenologist. doi:10.1017/S0024282917000664. Go to original source...
    37. Moya, P.; ©kaloud, P.; Chiva, S.; García-Breijo, F.J.; Reig-Arminana, J.; Vanèurová, L. & Barreno, E. (2015): Molecular phylogeny and ultrastructure of the lichen microalga Asterochloris mediterranea sp. nov. from Mediterranean and Canary Island ecosystems. - Int. J. Syst. Evol. Microbiol. 65: 1838-1854. Go to original source...
    38. Moya, P.; Chiva, S.; Molins, A.; Reig-Armiñana, J.; García-Breijo, F.J. & Barreno, E. (2016): Improved propagation method, rapid molecular identification and ultrastructural characterization as a multidisciplinary approach for Trebouxia species delimitation. - In: Meeting of the TREBOUXIA-WORKING group, September 2016 (http://www.symbioticgreenalgae.com/).
    39. Muggia, L.; Vanèurová, L.; ©kaloud, P.; Peksa, O.; Wedin, M. & Grube, M. (2013): The symbiotic playground of lichen thalli--a highly flexible photobiont association in rock-inhabiting lichens. - FEMS Microbiol. Ecol. 85: 313-323. Go to original source...
    40. Muggia, L.; Pérez-Ortega, S.; Kopun, T.; Zellnig, G. & Grube, M. (2014): Photobiont selectivity leads to ecological tolerance and evolutionary divergence in a polymorphic complex of lichenized fungi. - Ann. Bot. 114: 463-475. Go to original source...
    41. Muggia, L.; Leavitt, S. & Barreno, E. (2016): Report of the meeting of the trebouxia-working group. - International lichenological newsletter 49:35-37
    42. Osyczka, P. & Rola, K. (2013): Phenotypic plasticity of primary thallus in selected Cladonia species (lichenized Ascomycota: Cladoniaceae). - Biologia 68: 365-372. Go to original source...
    43. Peksa, O. & ©kaloud, P. (2008): Changes in chloroplast structure in lichenized algae. - Symbiosis (Rehovot) 46: 153-160.
    44. Pointing, S.B. & Belnap, J. (2012): Microbial colonization and controls in dryland systems. - Nat. Rev. Microbiol. 10: 551-562. Go to original source...
    45. Prieto, M.; Martínez, I.; Aragon, G.; Gueidan, C. & Lutzoni, F. (2012): Molecular phylogeny of Heteroplacidium, Placidium, and related Catapyrenioid genera (Verrucariaceae, lichen-forming ascomycota). - Am. J. Bot. 99: 23-35. Go to original source...
    46. Printz, H. (1921): Subaerial algae from South Africa. - Kong. Norsk. Vidensk. Selsk. Skrift. 1: 3-41.
    47. Puymaly, A. (1924): Recherches sur les algues vertes aériennes. - 274 pp., Thés. Fac. Sci. Univ. of Paris, Ser. A. Nº 991, Bordeaux.
    48. Rambaut, A. (2014): FigTree 1.4.2 software. Institute of Evolutionary Biology, Univ. Edinburgh.
    49. Ronquist, F.; Teslenko, M.; van der Mark, P.; Ayres, D.L.; Darling, A.; Hohna, S.; Larget, B.; Liu, L.; Suchard, M.A. & Huelsenbeck, J.P. (2012): MrBayes 3.2: Efficient bayesian phylogenetic inference and model choice across a large model space. - Syst. Biol. 61: 539-542. Go to original source...
    50. Rosentreter, R. (2007): Biotic soil crust lichens of the Columbia Basin. - Faculty Authored Books. 137. Northwest Lichenologists. Corvallis. http://scholarworks.boisestate.edu/fac_books/137
    51. Ruprecht, U.; Brunauer, G. & Türk, R. (2014): High photobiont diversity in the common European soil crust lichen Psora decipiens. - Biodivers. Conserv. 23: 1771-1785. Go to original source...
    52. Schaper, T. & Ott, S. (2003): Photobiont selectivity and interspecific interactions in lichen communities. I. culture experiments with the mycobiont Fulgensia bracteata. - Plant Biol. 5: 441-450. Go to original source...
    53. Schmidle, W. (1901): Ueber drei algen genera. - Ber. Dtsch. Bot. Ges. 19: 10-24. Go to original source...
    54. Schoch, C.L.; Seifert, K.A.; Huhndorf, S.; Robert, V.; Spouge, J.L.; Levesque, C.A.; Chen, W.; Fungal Barcoding Consortium & Fungal Barcoding Consortium Author List. (2012): Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for fungi. - Proc. Natl. Acad. Sci. U.S.A. 109: 6241-6246. Go to original source...
    55. ©kaloud, P., Friedl, T., Hallmann, C., Beck, A. & Dal Grande, F. (2016): Taxonomic revision and species delimitation of coccoid green algae currently assigned to the genus Dictyochloropsis (Trebouxiophyceae, Chlorophyta). -J. Phycol. 52: 599-617. Go to original source...
    56. Stamatakis, A.; Hoover, P. & Rougemont, J. (2008): A rapid bootstrap algorithm for the RAxML web servers. - Syst. Biol. 57: 758-771. Go to original source...
    57. Stamatakis, A. (2014): RAxML version 8: A tool for phylogenetic analysis and post-analysis of large phylogenies. - Bioinformatics 30: 1312-1313. Go to original source...
    58. Thüs, H.; Muggia, L.; Pérez-Ortega, S.; Favero-Longo, S.E.; Joneson, S.; O'Brien, H.; Nelsen, M.P.; Duque-Thüs, R.; Grube, M. & Friedl, T. (2011): Revisiting photobiont diversity in the lichen family Verrucariaceae (Ascomycota). - Eur. J. Phycol. 46: 399-415. Go to original source...
    59. Timdal, E. (1984): The delimitation of Psora (Lecideaceae) and related genera, with notes on some species. - Nord. J. Bot. 4: 525-540. Go to original source...
    60. Timdal, E. (1986): A revision of Psora (Lecideaceae) in North America. - Bryologist 253-275. Go to original source...
    61. Tschermak-Woess, E. & Plessl, A. (1948): Über zweierlei typen der sukzedanen teilung und ein auffallendes teilungsverhalten des chromatophors bei einer neuen protococcale, Myrmecia pyriformis. - Österr. Bot. Z. 95: 194-207. Go to original source...
    62. Tschermak-Woess, E. (1980): Asterochloris phycobiontica, gen. et spec. nov., der phycobiont der Flechte Varicellaria carneonivea. - Plant Syst. Evol. 135: 279-294. Go to original source...
    63. Tschermak-Woess, E. (1988): The algal partner. In: Galun M (eds): CRC handbook of lichenology, vol. 1. - pp. 39-94, CRC Press, Boca Raton, FL, USA. Go to original source...
    64. U'Ren, J.M.; Riddle, J.M.; Monacell, J.T.; Carbone, I.; Miadlikowska, J. & Arnold, A.E. (2014): Tissue storage and primer selection influence pyrosequencing-based inferences of diversity and community composition of endolichenic and endophytic fungi. - Mol. Ecol. Res. 14: 1032-1048. Go to original source...
    65. Voytsekhovich, A. & Beck, A. (2015): Lichen photobionts of the rocky outcrops of Karadag massif (Crimean Peninsula). - Symbiosis 68: 9-24. Go to original source...
    66. Weber, B.; Belnap, J. & Büdel, B. (2016): Biological Soil Crusts: An Organizing Principle in Drylands. - 549 pp., Springer-Verlag, Berlin. Go to original source...
    67. White, T.J.; Bruns, T.; Lee, S. & Taylor, J. (1990): Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. - PCR protocols: a guide to methods and applications 18: 315-322. Go to original source...
    68. Williams, L.; Colesie, C.; Ullmann, A.; Westberg, M.; Wedin, M. & Büdel, B. (2017): Lichen acclimation to changing environments: Photobiont switching vs. climate-specific uniqueness in Psora decipiens. - Ecology and Evolution 7: 2560-2574. Go to original source...

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