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In vitro symbiotic seed germination in Vanda wightii, an endemic orchid species of Western Ghats, India supported by Ceratobasidiaceae isolates

Authors

DOI:

https://doi.org/10.14719/pst.2367

Keywords:

Endomycorrhiza, orchid mycorrhiza, notified orchids, symbiotic fungi

Abstract

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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Author Biographies

Lekshmi Suresh, KSCSTE-JNTBGRI

Biotechnology and Bioinformatics Division

Reshma Raveendran, KSCSTE-JNTBGRI

Biotechnology and Bioinformatics Division

William Decruse, KSCSTE-JNTBGRI

Biotechnology and Bioinformatics Division

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

Published

28-06-2023

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How to Cite

1.
Lekshmi Suresh, Raveendran R, Decruse W. In vitro symbiotic seed germination in Vanda wightii, an endemic orchid species of Western Ghats, India supported by Ceratobasidiaceae isolates. Plant Sci. Today [Internet]. 2023 Jun. 28 [cited 2024 Dec. 21];. Available from: https://horizonepublishing.com/journals/index.php/PST/article/view/2367

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