Effect of ethyl methanesulfonate on growth and morphology of Doritis pulcherrima Lindl.

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Published: Jan 24, 2026
DOI: https://doi.org/10.63369/ijat.2026.22.1.529-546
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Plant tissue culture Orchid EMS Mutation and RAPD marker

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Supreeworakij, S.
School of Agricultural Technology, King Mongkut’s Institute of Technology Ladkrabang, Bangkok 10520, Thailand.
Saetiew, K.
School of Agricultural Technology, King Mongkut’s Institute of Technology Ladkrabang, Bangkok 10520, Thailand.
Montri, N.
School of Agricultural Technology, King Mongkut’s Institute of Technology Ladkrabang, Bangkok 10520, Thailand.

Abstract

Results showed that 0.5 mg/l BA and 1 mg/l NAA promoted the greatest seedling growth, with increased leaf number, leaf length, plant height, and root development. In the third experiment, protocorm-like bodies were treated with EMS at 0, 0.5, and 1.0% for 60 or 90 minutes. Survival declined with increasing concentration and time, with LD₅₀ values of 0.82% at 60 minutes and 0.54% at 90 minutes. Chlorophyll a, Chlorophyll b, and carotenoids were not significantly affected, but stomatal size changed. Stomatal width and length increased under 1.0% EMS, while density decreased from 220.17 to 149.47 stomata/mm². EMS treatment for 90 minutes showed an increased density relative to 60 min. RAPD analysis using 10 primers revealed high polymorphism indicating EMS induced genetic variation. Cluster analysis divided 22 samples into 9 groups at a similarity coefficient of 0.89, confirming genetic diversity despite no visible morphological changes.

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How to Cite
Supreeworakij, S., Saetiew, K., & Montri, N. (2026). Effect of ethyl methanesulfonate on growth and morphology of Doritis pulcherrima Lindl. International Journal of Agricultural Technology, 22(1), 529–546. https://doi.org/10.63369/ijat.2026.22.1.529-546
Issue
Vol. 22 No. 1 (2026): January
Section
Original Study
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This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

References

Ackerman, D. J., Phillips, D. R., Tremblay, L. R., Karremans, A., Reiter, N., Peter, I. C., Bogarín, D., Pérez-Escobar, A. O. and Liu, H. (2023). Beyond the various contrivances by which Orchids are pollinated: global patterns in orchid pollination biology. Botanical Journal of the Linnean Society, 202:295-324.

Arditti, J. (2008). Micropropagation of Orchid. In: Methods for Specific Genera, 2nd edn, Australia, Blackwell Publishing Ltd., pp.693.

Averyanov, L. V. (2009). Doritis pulcherrima var. apiculata (Orchidaceae). In: a New Variety from Southern Vietnam and Conditions of its Natural Habitat, Miami, USA, AOS Publishes, pp.9-16.

Cristension, E. A. (2001). Phalaenopsis: In a Monograph, Portland, Oregon, Timber Press, pp.330

Gnasekaran P., Xavier, R., Uma, R.S. and Sreeramanan, S. (2010). A study on the use of organic additives on the protocorm-like bodies (PLBs) growth of Phalaenopsis violacea orchid. Journal of Phytology, 2:29-33.

Jyothsna, B. S., S, V., Hallur, R. L. S., Srivastava, D. and Dey, S. (2024). Molecular, morphological, and biomolecular characterization of ethyl methanesulfonate-induced mutations in Aerides odoratum, an orchid. Journal of Applied Biology & Biotechnology, 12:136-143.

Kosir, P., Skof, S. and Luthar, Z. (2004). Direct shoot regeneration from nodes of Phalaenopsis orchids. Acta Agriculturae Slovenica, 83:233-242.

Lichtenthaler, H. K. (1987). Chlorophylls and carotenoids: Pigments of photosynthetic biomembranes. In Methods in enzymology on CD-ROM/Methods in enzymology., pp.350-382.

Martha, K. M., Dewanti, P., Restanto, D. P., Ratnasari, T. and Alfian, F. N. (2023). In Vitro Mutagenesis of Dendrobium gabriella suryajaya Using Ethyl Methane Sulfonate (EMS) and Plantlet Regeneration. Biosaintifika Journal of Biology & Biology Education, 15:87-96.

Mose, W., Daryono, B. S., Indrianto, A., Purwantoro, A. and Semiarti, E. (2020). Direct SomaticEmbryogenesis and Regeneration of anIndonesian orchid Phalaenopsis amabilis (L.) Blume under a Variety of Plant Growth Regulators, Light Regime, and Organic Substances. Jordan Journal of Biological Sciences, 13:509-518.

Murashige, T. and Skoog, F. (1962). A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum, 15:473-497.

Park, S. Y., Murthy, H. N. and Paek, K. Y. (2003). Protocorm-like body induction and subsequent plant regeneration from root tip cultures of Doritaenopsis. Journal of Plant Sciences, 164:919-923.

Paul, S., Kumaria, S. and Tandon, P. (2012). An effective nutrient medium for asymbiotic seed germination and large-scale in vitro regeneration of Dendrobium hookerianum, a threatened orchid of northeast India. AoB Plants, 2012:032.

Rademacher, W. (2015). Plant growth regulators: backgrounds and uses in plant production. Journal of Plant Growth Regulation, 34:845-872.

Ritti, W., Chourykaew, B., Fangmuang, W., Chotimit, K. and Boonlua, S. (2022). Breeding Systems and Fruit Set of Habenaria lindleyana Steud. (Orchidaceae). PBRU Science Journal, 18:12-21.

Srivastava, D., MC, G. and Sarangi, S. (2018). In vitro mutagenesis and characterization of mutants through morphological and genetic analysis in orchid Aerides crispa Lindl. Indian Journal of Experimental Biology, 56:385-394.

Stewart, S. L. and Zettler, L. W. (2002). Symbiotic germination of three semi-aquatic rein orchids (Habenaria repens, H. quinqueseta, H. macroceratitis) from Florida. Aqutic Botany, 72:25-35.

Stewart, S. L. and Kane, M. E. (2006). Asymbiotic seed germination and in vitro seedling development of Habenaria macroceratitis (Orchidaceae), a rare Florida terrestrial orchid. Plant Cell Tissue Organ Culture, 86:147-158.

Tang, C., Luo, M., Zhang, S., Jia, G., Tang, S., Jia, Y., Zhi, H. and Diao, X. (2022). Variations in chlorophyll content, stomatal conductance, and photosynthesis in Setaria EMS mutants. Journal of Integrative Agriculture, 22:1618-1630.

Tao, J., Yu, L., Kong, F. and Zhao, D. (2011). Effects of plant growth regulators on in vitro propagation of Cymbidium faberi Rolfe. African Journal of Biotechnology, 10:15639-15646.

Thipwong, J., Kongton, K. and Samala, S. (2022). Micropropagation and somaclonal variation of Doritis pulcherrima (Lindl.). Plant Biotechnology Reports, 16:401-408.

Trigiano, R. N. and Gray, D. J. (2000). Plant Tissue Culture Concepts and Laboratory Exercises. New York, CRC Press LLC, pp.27-75.

Trigiano, R. N. and Gray, D. J. (2004). Plant Development and Biotechnology. In: Boca Raton. pp.376.

Twaij, B. M., Jazar, Z. H. and Hasan, M. N. (2020). Trends in the use of tissue culture, applications and future aspects. International Journal of Plant Biology, 11:8385.

Utami, E. S. W. and Hariyanto, S. (2019). In vitro seed germination and seedling development of a rare indonesian native orchid Phalaenopsis amboinensis J. J. Sm. Scientifica, 2019:1-6.

Vacin, E. and Went, F. (1949). Some pH changes in nutrient solution. Botanic Gardens Conservation News, 110:605-613.

Yoosumran, V., Saetiew, K., Ruamrungsri, S., Akarapisarn, A.and Teerarak, M. (2025). Induced mutation of curcuma hybrid cv. sweetmemory through tissue culture by ethyl methanesulphonate (EMS). International Journal of Agricultural Technology, 21(1):381-396

Zeng, S. J., Chen, Z. I., Wu, K.L., Zhang, J. X., Bai, C. K., Teixeira da, Silva, J. A. and Duan, J. (2011). Asymbiotic seed germination, induction of calli and protocorm-like-bodies, and in vitro seedling development of rare and endangered Nothodoritis zhejiangensis Chinese Orchid. Horticultural Science, 46:460-46.

Zeng, S. J. (2009). Nothodoritis zhejiangensis Z. H. Tsi—A rare and endangered epiphytic orchid in China. Flower. Horticultural Science, 46:460-465.

Zeng, S. J., Chen, Z. I., Wu, K. L., Zhang, J. X., Bai, C. K., Teixeira da Silva, J. A. and Duan, J. (2011). Asymbiotic seed germination, induction of calli and protocorm-like-bodies, and in vitro seedling development of rare and endangered Nothodoritis zhejiangensis Chinese Orchid. Horticultural Science, 46:460-465.