In vitro symbiotic seed germination in Vanda wightii, an endemic orchid species of Western Ghats, India supported by Ceratobasidiaceae isolates
DOI:
https://doi.org/10.14719/pst.2367Keywords:
Endomycorrhiza, orchid mycorrhiza, notified orchids, symbiotic fungiAbstract
Symbiotic seed germination for conservation and cultivation of orchids holds colossal merit as
mycorrhizal fungus in its system improves their growth and adaptability. Symbiotic activity is
highly specific in some species, but in other cases the same fungus from one species is effective in a
series of related species. The present work describes inter-specific activity of three fungal isolates
from seedling root of Vanda thwaitesii to support seed germination and seedling growth of V.
wightii, a closely related species from India. Among the three isolates, two designated as Wyd2 and
Idk were identified as Ceratobasidium sp through sequencing of ITS1 and ITS4 regions. One
isolate designated as Wyd1 did not clad with any described genera, but remained as an out-group
under the family Ceratobasidiaceae. All the three isolates possessed binucleate hyphae producing
ellipsoidal, oval or barrel shaped monilioid cells and supported 80–95% seed germination,
transforming 70–85% of them into protocorms in 30 days duration. Symbiotic seedling
development starting with the promeristem formation, first leaf development and second leaf
initiation occurred in 95% of the protocorms in a time interval of 60 days compared to 90–120 days
through asymbiotic method. The fungal isolates from Vanda thwaitesii evaluated are proved
effective in V. wightii for its symbiotic seed germination and thus useful to mycorrhiza assisted
conservation.
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References
Arditti J, Ghani AKA. Numerical and physical properties of orchid seeds and their biological
implications. New Phytol. 2000;145:367–421. https://doi.org/10.1046/j.1469-8137.2000.00587.x
Rasmussen HN, Whigham DF. Pheonology of roots and mycorrhiza in five orchid species
differing in phototropic strategy. New Phytol. 2002;154:797–807.
https://doi.org/10.1046/j.1469-8137.2002.00422.x
Alghamdi AA. Influence of mycorrhizal fungi on seed germination and growth in terrestrial
and epiphytic orchids. Saudi J Biol Sci. 2019;26:495-502.
https://doi.org/10.1016%2Fj.sjbs.2017.10.021
Swarts ND, Dixon KW. Terrestrial orchid conservation in the age of extinction. Ann Bot.
;104:543–556. https://doi.org/10.1093/aob/mcp025
Zettler LW. Terrestrial Orchid Conservation by symbiotic seed germination: techniques and
preservatives. Selbyana. 1997;18:188–194
Otero JT, Ackerman JD, Bayman P. Differences in mycorrhizal preferences between two
tropical orchids. Mol Ecol. 2004;13:2393–2404. https://doi.org/10.1111/j.1365-
X.2004.02223.x
Stewart SL, Kane ME. Orchid conservation in the Americas- lessons learned in Florida.
Lankesteriana. 2007;7:382–387. https://doi.org/10.15517/lank.v7i1-2.19571
Steinfort U, Verdugo G, Besoain X, Cisternas MA. Mycorrhizal association and symbiotic
germination of the terrestrial orchid Bipinnula fimbriata (Poepp.) Johnst (Orchidaceae). Flora.
;205:811–817. https://doi.org/10.1016/j.flora.2010.01.005
Rasmussen HN. Recent developments in the study of orchid mycorrhiza. Plant Soil.
;244:149–163. https://doi.org/10.1023/A:1020246715436
Hollick PS. Mycorrhizal Specificity in Endemic Western Australian Terrestrial Orchids (Tribe
Diurideae): Implications for Conservation. [theses]. Murdoch (WA): Murdoch University;
Limansela B, Yogendra K, Jauti S. Biodiversity and status of Vanda Jones Ex R.Br. In:
Orchids of India Iii. New Delhi: Diya Publishing; 2002. p.22.
India: Ministry of Environment and Forests, The Gazette of India 2009; 648:4.
Decruse SW. Extended distribution of Vanda wightii Rchb.f., an endangered orchid of Western
Ghats revealed by ecological niche modelling. J. Orchid Soc. India. 2014; 28:15–21.
Decruse SW, Neethu RS, Pradeep NS. Seed germination and seedling growth promoted by a
Ceratobasidiaceae clone in Vanda thwaitesii Hook. f., an endangered orchid species endemic to
South Western Ghats, India and Sri Lanka. S Afr J Bot. 2018;16:222–229.
https://doi.org/10.1016/j.sajb.2018.04.002
Peterson RL, Uetake Y, Zelmer C. Fungal symbiosis with orchid protocorms. Symbiosis.
;25: 29–55
Johnson TR, Stewart SL, Dutra D, Kane ME, Richardson L. Asymbiotic and symbiotic seed
germination of Eulophia alta (Orchidaceae) – Priliminary evidence for the symbiotic culture
advantage. Plant Cell Tissue Organ Cult. 2007;90:313–323. https://doi.org/10.1007/s11240-
-9270-z
Rasmussen HN, Anderson TF, Johansen B. Temperature sensitivity of in vitro germination and
seedling development of Dactylorhiza majalis (Orchidaceae) with and without a mycorrhizal
fungus. Plant, J Syst Evol. 1990;13:171–177. https://doi.org/10.1111/j.1365-
1990.tb01289.x
Zhu GS, Yu ZN, Gui Y, Liu ZY. A novel technique for isolating orchid mycorrhizal fungi.
Fungal Divers. 2008; 33:123–137
Clements MA, Muir H, Cribb PJ. A preliminary report on the symbiotic germination of
European terrestrial Orchids. Kew Bull. 1986; 41:437–445. https://doi.org/10.2307/4102957
Zelmer CD, Cuthbertson L, Currah RS. Fungi associated with terrestrial orchid mycorrhizas,
seeds and protocorms. Mycoscince. 1996;37:439–448. https://doi.org/10.1007/BF02461001
Bandoni R. Safranin O as a rapid nuclear stain for fungi. Mycologia. 1979;71:873–874.
https://doi.org/10.1080/00275514.1979.12021088
Tamura K, Nei M. Estimation of the number of nucleotide substitutions in the control region of
mitochondrial DNA in humans and chimpanzees. Mol Biol Evol. 1993;10:512-526
Tamura K, Stecher G, Kumar S. MEGA 11: Molecular Evolutionary Genetics Analysis
Version 11. Mol Biol Evol. 2021;38:3022-3027. https://doi.org/10.1093/molbev/msab120
Mitra GC, Prasad RN, Roy Chowdhury A. Inorganic Salts and differentiation of protocorms in
seed cultures of an orchid and correlated changes in its free amino acid content. Indian J Exp
Biol. 1976;14:350–351
Stewart SL, Kane ME. Symbiotic seed germination of Habenaria macroceratitis
(Orchidaceae), a rare Florida terrestrial orchid. Plant Cell Tiss Organ Cult. 2006; 86:159–167.
https://doi.org/10.1007/s11240-006-9104-4
Aggarwal S, Nirmala C, Rastogi S, Adholeya A. In Vitro symbiotic seed germination and
molecular characterization of associated endophytic fungi in a commercially important and
endangered Indian orchid Vanda coerulea Griff. ex Lindl. Eur J Environ Sci. 2012;2:33–42.
https://doi.org/10.14712/23361964.2015.36
Wu P, Ding DH, Doris CNC. Mycorrhizal symbiosis enhances Phalaenopsis orchid's growth
and resistance to Erwinia chrysanthemi. Afr J Biotechnol. 2011;10:10095- 10100
Bhatti SK, Verma J, Sembi K, Promila P. Symbiotic seed germination of Aerides multiflora
Roxb. A study in vitro. J Orchid Soc India. 2017;31:85–91
Bhatti SK, Jagdeep V, Jaspreet KS, Ajay K. Mycobiont Mediated In vitro Seed Germination of
an Endangered ‘Fox-tail’ Orchid, Rhynchostylis retusa (L.) Blume. J Pure Appl Microbiol.
;10:663–670
Duran-Lopez ME, Caroca – Caceres R, Jahreis K, Narvaez – Vera M, Ansaloni R, Cazar ME.
The Mycorrhizal fungi Ceratobasidium sp. and Sebacina vermifera promote seed germination
and seedling development of the terrestrial orchid Epidendrum secundum Jacq. S Afr J Bot.
;125:54–61. https://doi.org/10.1016/j.sajb.2019.06.029
Garcia VG, Onco M A P, Susan VR. Review Biology and Systematics of the form genus
Rhizoctonia. Span J Agric Res. 2006;4:55–79. https://doi.org/10.5424/sjar/2006041-178
Hossain MM, Parveen R, Arvind G, Madhu S. Improved ex vitro survival of asymbiotically
raised seedlings of Cymbidium using mycorrhizal fungi isolated from distant orchid taxa. Sci
Hortic. 2013;159:109–116. https://doi.org/10.1016/j.scienta.2013.05.003
Khamchatra N, Dixon KW, Tantiwiwat S, Piapukiew J. Symbiotic seed germination of an
endangered epiphytic slipper orchid, Paphiopedilum villosum (Lindl.) Stein. from Thailand. S
Afr J Bot. 2016;04:76–81. https://doi.org/10.1016/j.sajb.2015.11.012
Athipunyakom P, Manoch L, Piluek C. Isolation and identification of mycorrhizal fungi from
eleven terrestrial orchids. Nat Sci. 2004;38:216–228
Shan XC, Liew ECY, Weatherhead MA, Hodgkiss IJ. Characterization and Taxonomic
Placement of Rhizoctonia-like Endophytes from Orchid Roots. Mycologia. 2002;94:230–239.
https://doi.org/10.1080/15572536.2003.11833228
Gónzalez D, Marianela R, Teun B, Joost S, Eiko EK, Andreia KN, Rytas V, Marc AC.
Phylogenetic relationships of Rhizoctonia fungi within the Cantharellales. Fungal Biol. 2016;
(4):603–619. https://doi:10.1016/j.funbio.2016.01.012
Hietela AM, Korhonen K, Sen R. An Unknown Mechanism Promotes Somatic Incompatibility
in Ceratobasidium bicorne. Mycologia. 2003;95(2):239–250. https://doi.org/10.2307/3762035
Johnson JS, Daniel JS, Bo-Young H, Lauren MP, Patrick D, Lei C, Shana RL, Blake MH,
Hanako OA, Mark G, Erica S, George MW. Evaluation of 16S rRNA gene sequencing for
species and strain-level microbiome analysis. Nat Commun. 2019;10:5029.
https://doi.org/10.1038/s41467-019-13036
Curtis JT. The relation of specificity of orchid mycorrhizal fungi to the problem of symbiosis.
Am J Bot. 1939;26: 390–399. https://doi.org/10.1002/j.1537-2197.1939.tb09292.x
Hadley G. Non–specificity of symbiotic infection in orchid mycorrhiza. New Phytol. 1970;
:1015–1025. https://doi.org/10.1111/j.1469-8137.1970.tb02481.x
Otero JT, Ackerman JD, Bayman P. Diversity and host specificity of endophytic Rhizoctonia–
like fungi from tropical orchids. Am J Bot. 2002; 89:1852–1858.
https://doi.org/10.3732/ajb.89.11.1852
Suarez JP, Weib M, Abele A, Garnica S, Oberwinkler F. Diverse tulasnelloid fungi form
mycorrhizas with epiphytic orchids in an Andean cloud forest. Mycol Res. 2006;110:1257–70.
https://doi.org/10.1016/j.mycres.2006.08.004
Liu H, Yibo Luo Y, Liu H. Studies of Mycorrhizal Fungi of Chinese Orchids and Their Role in
Orchid Conservation in China—A Review, Bot Rev. 2010;6:241–262.
https://doi.org/10.1007/s12229-010-9045-9
Julou T, Burghardt B, Gebauer G, Berveiller D, Damesin C, Selosse AM. Mixotrophy in
orchids: Insights from a comparative study of green individuals and non-photosynthetic
individuals of Cephalanthera damasonium. New Phytol. 2005;166:639–53.
https://doi.org/10.1111/j.1469-8137.2005.01364.x
Dearnaley JDW. Further advances in orchid mycorrhizal research. Mycorrhiza. 2007;17:
–486. https://doi.org/10.1007/s00572-007-0138-1
Batty AL, Dixon KW, Bundrett M, Sivasithamparam K. Orchid conservation and mycorrhizal
associations. In: Sivasithamparam K, Dixon KW, Barret RL, editors, Microorganisms in plant
conservation and biodiversity. Kluwer Academic. 2002;205–235.
Curtis JT. Non-specificity of orchid mycorrhizal fungi. Proceedings of the Society for Exp Biol
Med. 1937;36:43–44. https://doi.org/10.3181/00379727-36-9109P
Arditti J, Ernst R, Yam TW, Glabe C. The contribution to orchid mycorrhizal fungi to seed
germination: A speculative review. Lindleyana. 1990;5:249–255.
Ramsay MM, Dixon KW. Propagation science, recovery and translocation of terrestrial
orchids. Orchid conserv. 2003;259-288. In Dixon KW, Kell SP, Barrett RL, Cribb PJ, editors,
Orchid Conservation. Kota Kinabalu Sabah: Natural History Publications. 2003;259–288.
Bonnardeaux Y, Brundrett M, Batty A, Dixon K, Kock J, Sivasithamparam K. Diversity of
mycorrhizal fungi of terrestrial orchid: compatibility webs, brief encounters, lasting
relationships and alien invasion. Mycol Res. 2007;111:51–61.
https://doi.org/10.1016/j.mycres.2006.11.006
Nontachaiyapoom S, Sasirat S, Monoch L. Symbiotic seed germination of Grammatophyllum
speciosum Blume and Dendrobium draconis Rchb.f., native orchids of Thailand. Sci Hortic.
;130:303–308. https://doi.org/10.1016/j.scienta.2011.06.040
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