Endophytic association with Fusarium oxysporum enhances reintroduction survival of Aenhenrya rotundifolia (Blatt.) C.S. Kumar & F.N. Rasm.
DOI:
https://doi.org/10.25081/jp.2026.v18.9928Keywords:
Monotypic terrestrial orchid, Fusarium oxysporum, Endophytic fungi, Plant growth regulators, SymbiosisAbstract
Conservation efforts for the monotypic endangered jewel orchid, Aenhenrya rotundifolia requires a multifaceted approach. The present study outlines a dual strategy that combines an efficient in vitro propagation protocol with an investigation into the role of native endophytic fungi. The propagation phase of the study found that 1.0 mg/L meta-topolin was highly effective for inducing multiple shoots (74.3), while 0.5 mg/L Indole-3-Acetic Acid and 0.75 mg/L Naphthalene Acetic Acid were optimal for promoting shoot elongation (3.1 cm) and frequency of rooting (72.93%), respectively. Concurrently, four distinct endophytic fungi from the orchid’s rhizome were isolated and identified as Daldinia eschscholtzii, Fusarium oxysporum, Colletotrichum boninense, and Aspergillus sydowii. To assess their potential symbiotic benefits, in vitro raised plantlets were co-cultured with F. oxysporum, which has resulted in a significant increase in survival rates of about 91.58%.
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Abdalla, N., & Dobránszki, J. (2024). Meta-topolin as an effective benzyladenine derivative to improve the multiplication rate and quality of in vitro axillary shoots of húsvéti rozmaring apple scion. Plants, 13(11), 1568. https://doi.org/10.3390/plants13111568
Ábrahám, R., Baka, E., Al-Nussairawi, M., Táncsics, A., Farkas, M., Nagy, I., Kriszt, B., & Cserháti, M. (2025). Molecular insights into ochratoxin A biodegradation. Biologia Futura, 76, 315-328. https://doi.org/10.1007/s42977-025-00258-2
Ahamed Sherif, N., Senthil Kumar, T., & Rao, M. V. (2020). DNA barcoding and genetic fidelity assessment of micropropagated Aenhenrya rotundifolia (Blatt.) C.S. Kumar and F.N. Rasm.: a critically endangered jewel orchid. Physiology and Molecular Biology of Plants, 26, 2391-2405. https://doi.org/10.1007/s12298-020-00917-9
Avhad, A., Patil, N., & Zunjarrao, R. (2023). Isolation and identification of rhizospheric and endophytic fungi associated with Habenaria brachyphylla (Lindl.) Aitch.: An endemic and rare orchid of the Western Ghats of Maharashtra, India. Biological Forum, 15(5), 1535-1542.
Barnes, E. C., Jumpathong, J., Lumyong, S., Voigt, K., & Hertweck, C. (2016). Daldionin, an unprecedented binaphthyl derivative, and diverse polyketide congeners from a fungal orchid endophyte. Chemistry, 22(13), 4551-4555. https://doi.org/10.1002/chem.201504005
Benyon, F., & Summerell, B. A., & Burgess, W. (1996). Association of Fusarium species with root rot of Cymbidium orchids. Australasian Plant Pathology, 25, 226-228. https://doi.org/10.1071/AP96041
Bhattacharjee, A., & Chowdhery, H. J. (2012). Jewel orchids of India – An overview. In A. Bhattacharjee & H. J. Chowdhery (Eds.), Multidisciplinary Approaches of Angiosperm Systematics (Vol. 2, pp. 554-556). University of Kalyani.
Davis, B. J., Growns, D., Stevens, J. C., & Scalzo, A. A. (2025). Protocorm-like body induction and proliferation as a conservation tool for the threatened Queen of Sheba orchid, Thelymitra variegata (Lindl.) F. Muell.. Plant Cell, Tissue and Organ Culture, 163, 19. https://doi.org/10.1007/s11240-025-03226-9
Devi, K. S., Devi, H. H., Singh, N. S., & Devi, H. S. (2025). Meta-topolin enhanced in vitro regeneration and genetic fidelity assessment of an endangered orchid, Pelatantheria insectifera using scot and ISSR. Vegetos. https://doi.org/10.1007/s42535-025-01401-z
Diengdoh, R. V., Kumaria, S., Tandon, P., & Das, M. C. (2017). Asymbiotic germination and seed storage of Paphiopedilum insigne, an endangered lady’s slipper orchid. South African Journal of Botany, 112, 215-224. https://doi.org/10.1016/j.sajb.2017.05.028
Ezeonuegbu, B. A., Abdullahi, M. D., Whong, C. M. Z., Sohunago, J. W., Kassem, H. S., Yaro, C. A., Hetta, H. F., Mostafa-Hedeab, G., Zouganelis, G. D., & Batiha, G. E. (2022). Characterization and phylogeny of fungi isolated from industrial wastewater using multiple genes. Scientific Reports, 12, 2094. https://doi.org/10.1038/s41598-022-05820-9
Jiang, J., Zhang, K., Cheng, S., Nie, Q., Zhou, S., Chen, Q., Zhou, J., Zhen, X., Li, X., Zhen, T., Xu, M., Hsiang, T., Sun, Z., & Zhou, Y. (2019). Fusarium oxysporum KB-3 from Bletilla striata: an orchid mycorrhizal fungus. Mycorrhiza, 29, 531-540. https://doi.org/10.1007/s00572-019-00904-3
Kaladharan, S., Mariappan, M., & Thiruppathi, S. K. (2024b). An effective in vitro propagation of Coelogyne mossiae Rolfe: An endangered endemic orchid of Western Ghats, India. South African Journal of Botany, 173, 217-224. https://doi.org/10.1016/j.sajb.2024.08.026
Kaladharan, S., Rengasamy, A., Chinnaiyan, R., Mariappan, M., & Thiruppathi, S. K. (2024a). In vitro asymbiotic seed germination and micropropagation of Dendrobium heyneanum Lindl. –an endemic orchid of Western Ghats, India. Plant Cell, Tissue and Organ Culture, 157, 31. https://doi.org/10.1007/s11240-024-02758-w
Koppada, U., Matam, P., & Parvatam, G. (2022). In vitro-derived plants grown from low nitrate medium produced quality tubers in Decalepis hamiltonii Wight & Arn. 3 Biotech, 12, 200. https://doi.org/10.1007/s13205-022-03256-6
Lal, A., Pant, M., Kumar, A., Palni, L. M. S., Singh, A., Siddiqui, M. R., & Husen, A. (2025). DNA barcoding of epiphytic orchids for species level identification. The Journal of Horticultural Science and Biotechnology, 100(1), 127-137. https://doi.org/10.1080/14620316.2024.2371591
Lee, Y.-I., Zahn, F. E., Xie, Q.-Y., Wu, S.-H., & Gebauer, G. (2025). Diverse mycorrhizal associations and nutrition in Didymoplexis orchids. Mycorrhiza, 35, 34. https://doi.org/10.1007/s00572-025-01208-5
Long, Y., Nong, Q., Xie, L., Zhang, W., Chen, Y., & Zhang, Y. (2022). Tiankengomelania guangxiense, gen. et sp. nov., a dark septate endophytic fungus, promotes the growth of the medicinal orchid Dendrobium officinale. Fungal Biology, 126(5), 333-341. https://doi.org/10.1016/j.funbio.2022.03.005
Ma, X., Nontachaiyapoom, S., Jayawardena, R. S., Hyde, K. D., Gentekaki, E., Zhou, S., Qian, Y., Wen, T., & Kang, J. (2018). Endophytic Colletotrichum species from Dendrobium spp. in China and Northern Thailand. MycoKeys, 43, 23-57. https://doi.org/10.3897/mycokeys.43.25081
Manokari, M., Priyadharshini, S., Jogam, P., Dey, A., & Shekhawat, M. S. (2021). Meta-topolin and liquid medium mediated enhanced micropropagation via ex vitro rooting in Vanilla planifolia Jacks. ex Andrews. Plant Cell, Tissue and Organ Culture, 146, 69-82. https://doi.org/10.1007/s11240-021-02044-z
Manuel, G., & Jeannette, N.-R. I. (2020). Preliminary evaluation of the potential of macromycetes as mediators in bio-hardening of orchid plantlets. Acta Scientific Agriculture, 4(1), 141-146.
Nguyen, D. Q., Li, H., Tran, T. T., Sivasithamparam, K., Jones, M. G. K., & Wylie, S. J. (2020). Four Tulasnella taxa associated with populations of the Australian evergreen terrestrial orchid Cryptostylis ovata. Fungal Biology, 124(1), 24-33. https://doi.org/10.1016/J.FUNBIO.2019.10.006
Palaniyappan, S., Aazhivaendhan, G., Silambarasan, R., Tinoammini, N., Rajasekar, C., & Senthil Kumar, T. (2025). Phytosociology and habitat suitability studies on Endemic jewel Orchid Aenhenrya rotundifolia in Southern Western Ghats, India. Vegetos. https://doi.org/10.1007/s42535-024-01129-2
Parthibhan, S., & Ramasubbu, R. (2020). Mycorrhizal and endophytic fungal association in Paphiopedilum druryi (Bedd.) Stein - A strict endemic and critically endangered orchid of the Western Ghats. Ecological Genetics and Genomics, 16, 100059. https://doi.org/10.1016/J.EGG.2020.100059
Parthibhan, S., Rao, M. V., & Senthil Kumar, T. (2017). Culturable fungal endophytes in shoots of Dendrobium aqueum Lindley – An imperiled orchid. Ecological Genetics and Genomics, 3-5, 18-24. https://doi.org/10.1016/J.EGG.2017.06.004
Ramírez-Mosqueda, M. A., Cadena-Zamudio, J. D., Bautista-Aguilar, J. R., Cruz-Cruz, C. A., Cruz-Gutiérrez, E. J., & Gutiérrez-Mora, A. (2025b). Effect of meta-topolin on the in vitro propagation of Vanilla planifolia Jacks. in RITA® systems. In Vitro Cellular & Developmental Biology - Plant. https://doi.org/10.1007/s11627-025-10610-9
Ramírez-Mosqueda, M. A., Cadena-Zamudio, J. D., Cruz-Gutiérrez, E. J., Sandoval-Cancino, G., Cortés-Paz, A. L., & Tafolla-Arellano, J. C. (2025a). Meta-topolin increases the number of shoots and rhizogenesis during in vitro propagation of a Laeliocattleya hybrid. Plant Cell, Tissue and Organ Culture, 163, 101. https://doi.org/10.1007/s11240-025-03299-6
Rineksane, I. A., Astuti, A., Samidjo, G. S., Nugrahani, P., & Sumarsih (2026). Characterization of novel rhizosphere bacteria isolated from Vanda tricolor Lindl. var. Suavis Orchids in Mount Merapi Indonesia. Emirates Journal of Food and Agriculture, 38, 1-13. https://doi.org/10.3897/ejfa.2026.159327
Sukamto, D. S., Avivi S., Nurcahyanti S. D., & Restanto, D. P. (2025). Symbiotic potential of endophytic fungi from native orchids with histological insights and growth responses in Dendrobium hybrid. Biodiversitas, 26(12), 6389-6398. https://doi.org/10.13057/biodiv/d261236
Sutthinon, P., Rungwattana, K., Suwanphakdee, C., Himaman, W., & Lueangjaroenkit, P. (2021). Endophytic fungi from root of three lady’s slipper orchids (Paphiopedilum Spp.) in Southern Thailand. Chiang Mai Journal of Science, 48(3), 853-866.
Tavares, A. R., Young, J. L. M., Ori, S. S., Kanashiro, S., Lima, G. P. P., Chu, E. P., & Suzuki, R. M. (2012). Orchid in vitro growth as affected by nitrogen levels in the culture medium. Horticultura Brasileira, 30(1), 119-124. https://doi.org/10.1590/S0102-05362012000100020
Thiyam, S., Uchoi, A., Singh, S. R., Sharma, G. R., Rama, T., Stina, K., & Khangjarakpam, G. (2025). Effect of plant growth regulators and growth additives on the in vitro micropropagation of Aerides odorata var. Alba from immature pod seeds. Indian Journal of Agricultural Research, 59(11), 1656-1661. https://doi.org/10.18805/IJARe.A-6326
Tinoammini, N., Aazhivaendhan, G., & Senthil Kumar, T. (2024). Invitro germination and optimization of basal media for protocorm-like bodies proliferation in Dienia ophrydis (J. Koenig) Seidenf. Rhizosphere, 29, 100854. https://doi.org/10.1016/J.RHISPH.2024.100854
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Copyright (c) 2026 Palaniyappan Subramanian, Shivakrishnan Kaladharan, Ahamed Sherif Naseruddin, Senthil Kumar Thiruppathi
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