Embryogenic Callus Induction from Leaf Tip Explants and Protocorm-Like Body Formation and Shoot Proliferation of Dimorphorchis lowii: Borneon Endemic Orchid

Juddy E. Jainol, Jualang Azlan Gansau

Abstract


Dimorphorchis lowii is a rare epiphytic orchid endemic to Borneo and very popular due to its ornamental value. For the purpose of mass propagation, procedures for in vitro propagation through callus were developed. The objectives of this study were to determine the effects of plant growth regulators (PGRs) on callus induction from leaf tip explant and the effects of complex additives on PLBs formation and shoot development. The best combination of PGR was 3.0 mg L-1 Thidiazuron (TDZ) with 0.046 mg L-1 NAA supplemented in half-strength medium (½ MS) (Murashige & Skoog, 1962). The percentage of survival and callus formation obtained were 96.0 % ± 19.8 and 52.0 % ± 16.5 respectively. Maximum shoot proliferation from PLBs was observed in Knudson C (KC) medium enriched with 15 % (v/v) coconut water. In this treatment, 10.2 ± 6.2 shoots were produced from one callus explant. Shoots were rooted on KC medium containing 15 % (v/v) coconut water and transferred in a medium mixture containing sphagnum moss, charcoal, brick and coco peat with the ratio of 1:1:1:1. After 2 months being acclimatized in the glasshouse, 78 % of plantlets survived. Histology observations showed that embryogenic callus derived from leaf tip explants might originate from epidermis and mesophyll cells and was capable of developing into complete plantlets.

Keywords


callus; in vitro; orchid; protocorm like-bodies (PLBs)

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References


Arditti, J. (2008). Micropropagation of orchids (2nd ed., Vol. 1). Oxford, UK: Wiley-Blackwell.

Begum, A. A., Tamaki, M., Tahara, M., & Kako, S. (1994). Somatic embryogenesis in Cymbidium through In vitro culture of inner tissue of protocorm-like bodies. Journal of the Japanese Society for Horticultural Science, 63(2), 419-427. crossref

Bustam, S., Sinniah, U. R., Kadir, M. A., Zaman, F. Q., & Subramaniam, S. (2013). Selection of optimal stage for protocorm-like bodies and production of artificial seeds for direct regeneration on different media and short term storage of Dendrobium Shavin White. Plant Growth Regulation, 69(3), 215–224. crossref

Chen, J. T., & Chang, W. C. (2006). Direct somatic embryogenesis and plant regeneration from leaf explants of Phalaenopsis amabilis. Biologia Plantarum, 50(2), 169-173. crossref

Chugh, S., Guha, S., & Rao, I. U. (2009). Micropropagation of orchids: A review on the potential of different explants. Scientia Horticulturae, 122(4), 507–520. crossref

Cribb, P., & Bell, A. (2008). The Bornean endemic genus Dimorphorchis (Vandeae: Aeridinae). Malesian Orchid Journal, 2, 77-92.

Devi, H. S., Devi, S. I., & Singh, T. D. (2013). High frequency plant regeneration system of Aerides odorata Lour. through foliar and shoot tip culture. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 41(1), 169–176. Retrieved from https://www.researchgate.net/publication/236949935_High_Frequency_Plant_Regeneration_System_of_Aerides_odorata_Lour_Through_Foliar_and_Shoot_Tip_Culture PDF

Gantait, S., Bustam, S., & Sinniah, U. R. (2012). Alginate-encapsulation, short-term storage and plant regeneration from protocorm-like bodies of Aranda Wan Chark Kuan “Blue” Vanda coerulea Grifft. ex. Lindl. (Orchidaceae). Plant Growth Regulation, 68(2), 303–311. crossref

Gow, W. P., Chen, J. T., & Chang, W. C. (2009). Effects of genotype, light regime, explant position and orientation on direct somatic embryogenesis from leaf explants of Phalaenopsis orchids. Acta Physiologiae Plantarum, 31(2), 363–369. crossref

Jainol, E. J. (2016). Development of in vitro propagation of Dimorphorchis lowii (Orchidaceae) through semi-solid and liquid culture systems (Unpublished doctoral dissertation). University Malaysia Sabah, Kota Kinabalu, Sabah.

Janarthanam, B., & Seshadri, S. (2008). Plantlet regeneration from leaf derived callus of Vanilla planifolia Andr. In Vitro Cellular and Developmental Biology - Plant, 44(2), 84–89. crossref

Jheng, F. Y., Do, Y. Y., Liauh, Y. W., Chung, J. P., & Huang, P. L. (2006). Enhancement of growth and regeneration efficiency from embryogenic callus cultures of Oncidium “Gower Ramsey” by adjusting carbohydrate sources. Plant Science, 170(6), 1133–1140. crossref

Jitsopakul, N., Thammasiri, K., & Ishikawa, K. (2013). Efficient adventitious shoot regeneration from shoot tip culture of Vanda coerulea, a Thai orchid. ScienceAsia, 39(5), 449–455. crossref

Kalimuthu, K., Senthilkumar, R., & Murugalatha, N. (2006). Regeneration and mass multiplication of Vanilla planifolia Andr. - A tropical orchid. Current Science, 91(10), 1401–1403. Retrieved from http://www.iisc.ernet.in/~currsci/nov252006/1401.pdf PDF

Khoddamzadeh, A. A., Sinniah, U. R., Kadir, M. A., Kadzimin, S. B., Mahmood, M., & Sreeramanan, S. (2011). In vitro induction and proliferation of protocorm-like bodies (PLBs) from leaf segments of Phalaenopsis bellina (Rchb.f.) Christenson. Plant Growth Regulation, 65, 381. crossref

Kishor, R., & Devi, H. S. (2009). Induction of multiple shoots in a monopodial orchid hybrid (Aerides vandarum Reichb.f × Vanda stangeana Reichb.f) using thidiazuron and analysis of their genetic stability. Plant Cell, Tissue and Organ Culture (PCTOC), 97(2), 121-129. crossref

Kuo, H. L., Chen, J. T., & Chang, W. C. (2005). Efficient plant regeneration through direct somatic embryogenesis from leaf explants of Phalaenopsis “Little Steve.” In Vitro Cellular & Developmental Biology - Plant, 41(4), 453–456. crossref

Martin, K. P., & Madassery, J. (2006). Rapid in vitro propagation of Dendrobium hybrids through direct shoot formation from foliar explants, and protocorm-like bodies. Scientia Horticulturae, 108(1), 95–99. crossref

Mathews, V. H., & Rao, P. S., (1980). In vitro multiplication of Vanda hybrids through tissue culture technique. Plant Science Letters, 17(3), 383-389. crossref

Mayer, J. L. S., Stancato, G. C., & Appezzato-Da-Glória, B. (2010). Direct regeneration of protocorm-like bodies (PLBs) from leaf apices of Oncidium flexuosum Sims (Orchidaceae). Plant Cell, Tissue and Organ Culture, 103(3), 411–416. crossref

Mei, T. A., Danial, M., Mahmood, M., & Subramaniam, S. (2012). Exquisite protocol of callus induction and protocorm-like bodies (PLBs) regeneration of Dendrobium sonia-28. Australian Journal of Crop Science, 6(5), 793–800. Retrieved from http://www.cropj.com/sabrumanian_6_5_2012_793_800.pdf

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

Murdad, R., Hwa, K. S., Seng, C. K., Latip, M. A., Aziz, Z. A., & Ripin, R. (2006). High frequency multiplication of Phalaenopsis gigantea using trimmed bases protocorms technique. Scientia Horticulturae, 111(1), 73–79. crossref

Pant, B., & Thapa, D. (2012). In vitro mass propagation of an epiphytic orchid, Dendrobium primulinum Lindl. through shoot tip culture. African Journal of Biotechnology, 11(42), 9970–9974. crossref

Park, S., Yeung, E., Chakrabarty, D., & Paek, K. (2002). An efficient direct induction of protocorm-like bodies from leaf subepidermal cells of Doritaenopsis hybrid using thin-section culture. Plant Cell Reports, 21(1), 46–51. crossref

Prando, M. A. S., Chiavazza, P., Faggio, A., & Contessa, C. (2014). Effect of coconut water and growth regulator supplements on in vitro propagation of Corylus avellana L. Scientia Horticulturae, 171, 91–94. crossref

Roy, A. R., Sajeev, S., Pattanayak, A., & Deka, B. C. (2012). TDZ induced micropropagation in Cymbidium giganteum Wall. Ex Lindl. and assessment of genetic variation in the regenerated plants. Plant Growth Regulation, 68(3), 435–445. crossref

Salam, P., Salam, J. S., & Mohanty, C. R. (2013). Effect of benzylamino purine and naphthalene acetic acid on callus and protocorm formation of Dendrobium cv. Banyat Pink. Journal of Cell & Tissue Research, 13(3), 3977-3981. Retrieved from http://www.tcrjournals.com/uploads/6980929._Salam.pdf

Shimura, H., & Koda, Y. (2004). Micropropagation of Cypripedium macranthos var. rebunense through protocorm-like bodies derived from mature seeds. Plant Cell, Tissue and Organ Culture, 78(3), 273–276. crossref

Tan, B. C., Chin, C. F., & Alderson, P. (2011). Optimisation of plantlet regeneration from leaf and nodal derived callus of Vanilla planifolia Andrews. Plant Cell, Tissue and Organ Culture (PCTOC), 105(3), 457–463. crossref

Tan, T. C., Cheng, L. H., Bhat, R., Rusul, G., & Easa, A. M. (2014). Composition, physicochemical properties and thermal inactivation kinetics of polyphenol oxidase and peroxidase from coconut (Cocos nucifera) water obtained from immature, mature and overly-mature coconut. Food Chemistry, 142, 121–128. crossref

Tariq, U., Ali, M., & Abbasi, B. H. (2014). Morphogenic and biochemical variations under different spectral lights in callus cultures of Artemisia absinthium L. Journal of Photochemistry and Photobiology B: Biology, 130, 264–271.

Tee, C., Wong, C., Lam, X., & Maziah, M. (2010). A preliminary study of protocorm-like-bodies (PLBs) induction using leaf ex-plants of Vanda and Dendrobium orchids. Asia Pacific Journal of Molecular Biology and Biotechnology, 18(1), 189–191. Retrieved from http://www.msmbb.org.my/apjmbb/html181/181as.pdf

Teixeira da Silva, J. A., & Tanaka, M. (2006). Multiple regeneration pathways via thin cell layers in hybrid Cymbidium (Orchidaceae). Journal of Plant Growth Regulation, 25(3), 203–210.

Vacin, E. F., & Went, F. W. (1949). Some pH changes in nutrient solutions. Botanical Gazette, 110(4), 605–613. Retrieved from https://www.jstor.org/stable/pdf/2472666.pdf

van Staden, J., Zažímalová, E., & George, E. F. (2008). Plant growth regulators II: Cytokinins, their analogues and antagonists. In E. F. George, M. A. Hall, & Geert-Jan de Klerk (Eds.), Plant propagation by tissue culture (3rd ed.) (205-226). Netherlands: Springer.

Wu, K., Zeng, S., Teixeira da Silva, J. A., Chen, Z., Zhang, J., Yang, Y., & Duan, J. (2012). Efficient regeneration of Renanthera Tom Thumb “Qilin” from leaf explants. Scientia Horticulturae, 135, 194–201.

Yam, T. W., & Arditti, J. (2009). History of orchid propagation: A mirror of the history of biotechnology. Plant Biotechnology Report, 3, 1–56.

Yong, J. W. H., Ge, L., Ng, Y. F., & Tan, S. N. (2009). The chemical composition and biological properties of coconut (Cocos nucifera L.) water. Molecules (Basel, Switzerland), 14(12), 5144–5164.




DOI: http://doi.org/10.17503/agrivita.v39i1.895

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