Skip to main navigation menu Skip to main content Skip to site footer

Vol. 19 No. 6 (2020)

Articles

THE INFLUENCE OF DISINFECTION METHODS AND LIQUID PHASE MEDIA ON Paphiopedilum insigne SEEDS GERMINATION AND MEDIA SUPPLEMENTS ON MORPHOLOGICAL FEATURES OF PROTOCORMS IN TISSUE CULTURE

DOI: https://doi.org/10.24326/asphc.2020.6.1
Submitted: November 19, 2020
Published: 2020-12-31

Abstract

Paphiopedilum is a very original orchid, which labellum resembles a slipper. It is cultivated as a cut flower or a pot plant. The aim of the presented work was to estimate the influence of a disinfection method and addition of a liquid media on germination of Paphiopedilum insigne seeds in vitro. The whole seed capsules were disinfected with the use of 0.5% of AgNO3 for 20 min, 0.1% of HgCl2 for 5 s, 1% NaOCl for 30 min or immersed in a 96% ethanol and burned in a direct flame. The disinfected seeds were placed on a 1/4 MS (Murashige and Skoog) solidified media on top of which a liquid phase containing 1/4 MS elements, GA3 in concentration of 400 mg·dm–3 or distilled sterile water were added. The media without the liquid phase was also tested. It was noted that all methods of disinfection used effectively reduced contaminations of Paphiopedilum insigne seeds. The highest germination rate was observed when capsules were direct flamed and the liquid phase was added on top of the solidified media. The influence of growth regulators (BA, TDZ, KIN and 2,4-D) and casein hydrolysate added to the media on the morphological features of the obtained plants was also tested. It was observed that the most regenerated protocorms of the best quality were obtained when the media was supplemented with 1 mg·dm–3 of BA + 2 mg·dm–3 of TDZ or 5 mg·dm–3 of KIN + 1 mg·dm–3 of BA.

References

  1. Alvarez-Pardo, V.M., Ferreira, A.G., Nunes, V.F. (2006). Seed disinfestation methods for in vitro cultivation of epiphyte orchids from Southern Brazil. Hortic. Bras., 24(2), 217–220. DOI: 10.1590/S0102-05362006000200019
  2. Asghar, S., Ahmad, T., Hafiz, I.A., Yaseen, M. (2011). In vitro propagation of orchid (Dendrobium nobile) var. Emma white [!]. Afr. J. Biotechnol., 10(16), 3097–3103. DOI: 10.5897/AJB10.401
  3. Bae, K.H., Kim, C.H., Sun, B.Y., Choi, Y.E. (2010). Structural changes of seed coats and stimulation of in vitro germination of fully mature seeds of Cypripedium macranthos Swartz (Orchidaceae) by NaOCl pretreatment. Propag. Ornam. Plants, 10(2), 107–113.
  4. Bliss, C.I. (1938). The transformation of percentages for use in the analysis of variance. Ohio J. Sci., 38(1), 9–12.
  5. Borkowska, B. (1997). Zastosowanie pożywek dwufazowych w pędowych kulturach in vitro. Rozmnażanie roślin in vitro. Materiały z konferencji, Skierniewice, 19 marca 1997, 5–11.
  6. Chmiel, H., Jędrzejuk, A. (2015). Przerywanie spoczynku, przyspieszanie i pędzenie roślin ozdobnych. In: Biologia roślin ozdobnych wybrane zagadnienia, Starck, Z., Rabiza-Świder, J. (eds.). Wyd. SGGW, Warszawa, 187–201.
  7. 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. S. Afr. J. Bot., 112, 215–224. DOI: 10.1016/j.sajb.2017.05.028
  8. Fakouri Ghaziani, M.V., Baker, A., Negahdar, N., Kaviani, B. (2014). Micropropagation of Orchis catasetum – a rare and endangered orchid. Sci. Papers. Ser. B, Horticulture, 58, 317–322.
  9. Guo, Y.Y., Luo, Y.B., Liu, Z.J., Wang, X.Q. (2015). Reticulate evolution and sea‐level fluctuations together drove species diversification of slipper orchids (Paphiopedilum) in South – East Asia. Molecular Ecol., 24(11), 2838–2855. DOI: 10.1111/mec.13189
  10. Jevšnik, T., Luthar, Z. (2015). Successful disinfection protocol for orchid seeds and influence of gelling agent on germination and growth. Acta Agric. Slov., 105(1), 95–102.
  11. Katsalirou, E., Gerakis, A., Haldas, X., Deconninck, G. (2017). Optimal disinfection times for seeds of Mediterranean orchids propagated on nutrient media. Eur. J. Env. Sci., 7(2), 119–124. DOI: 10.14712/23361964.2017.10
  12. Kaur, S., Bhutani, K.K. (2016). Asymbiotic seed germination and multiplication of an endengered orchid – Paphiopedilum venustum (Wall. ex Sims.). Acta Soc. Bot. Pol., 85, 1­–11. DOI: 10.5586/asbp.3494
  13. Khatun, H., Khatun, M.M., Biswas, M.S., Kabir, M.R., Al-Amin, M. (2010). In vitro growth and development of Dendrobium hybrid orchid. Bangladesh J. Agril Res., 35(3), 507–514. DOI: 10.3329/bjar.v35i3.6457
  14. Kulpa, D., Katroń, J. (2012). Seed germination and plant development of Bletilla striata in vitro. Folia Pomer. Univ. Technol. Stetin., 293(21), 51­–60.
  15. Kumar, A., Nandi, S.K., Bag, N., Palni, L.M.S. (2002). Tissue culture studies in two important orchid taxa: Rhynchostylis retusa (L.) Bl. and Cymbidium elegans Lindl. Gyanodaya Prakashan, Nainital, India, 113–124.
  16. Kutas, E., Ogorodnik, L. (2011). The influence of sterilizing compounds on the yield of viable explants of Rhododendron L. (Ericaceae). Int. J. Biodivers. Conserv., 3(1), 24–26.
  17. Murashige, T., Skoog, F. (1962). A revised medium for rapid growth and bioassays with Tobacco tissue cultures. Physiol. Plant., 15, 473–479. DOI: 10.1111/j.1399-3054.1962.tb08052.x
  18. Nisayan, E., Thepsithar, C., Thongpukdee, A. (2010). Multiple shoot induction from seed derived shoots of Paphiopedilum. I Int. Orchid Symp., 878, 199–204.
  19. Ng, C.Y., Saleh, N.M. (2011). In vitro propagation of Paphiopedilum orchid through formation of protocorm-like bodies. Plant Cell Tiss. Org. Cult., 105(2), 193–202. DOI: 10.1007/s11240-010-9851-0
  20. Oszkinis, K. (2004). Storczyki. PWRiL, Warszawa.
  21. Pedroza-Manrique, J., Fernandez-Lizarazo, Ch., Suarez-Silva, A. (2005). Evaluation of the effect of three growth regulators in the germination of Comparettia falcata seeds under in vitro conditions. In Vitro Cell. Dev. Biol. Plant, 41, 838–843. DOI: 10.1079/IVP2005698
  22. Pindel, A., Pindel, Z. (2004). Initiation of in vitro cultures of chosen endangered European species of orchids. Folia Hortic., 16(2), 111–117.
  23. Ponert, J., Vosolsobě, S., Kmecová, K., Lipavská, H. (2011). European orchid cultivation – from seed to mature plant. Eur. J. Environ. Sci., 1(2), 95–107. DOI: 10.14712/23361964.2015.52
  24. Sheelavanthmath, S.S., Murthy, H.N., Hema, B.P., Hahn, E.J., Paek, K.Y. (2005). High frequency of protocorm like bodies (PLBs) induction and plant regeneration from protocorm and leaf sections of Aerides crispum. Sci. Hortic., 106(3), 395–401. DOI: 10.1016/j.scienta.2005.04.012
  25. Sibin, N.T., Gangaprasad, A. (2012). Embryo culture of Rhynchostylis retusa, an exquisite orchid of Western Ghats. Int. J. Biol. Pharm. Allied Sci., 1(7), 973–981.
  26. Soonthornkalump, S., Nakkanong, K., Meesawat, U. (2019). In vitro cloning via direct somatic embryogenesis and genetic stability assessment of Paphiopedilum niveum (Rchb. f.) Stein: the endangered Venus’s slipper orchid. In Vitro Cell Dev. Biol. Plant, 55(3), 265–276. DOI: 10.1007/s11627-019-09981-7
  27. Talukder, S.K., Nasiruddin, K.M., Yasmin, S., Hassan, L., Begum, R. (2003). Shoot proliferation of Dendrobium Orchid with BAP and NAA. J. Biol. Sci., 3(11), 1058–1062. DOI: 10.3923/jbs.2003.1058.1062.
  28. Thompson, D.I., Edwards, T.J., Van Staden, J. (2007). A novel dual-phase culture medium promotes germination and seedling establishment from immature embryos in South African Disa (Orchidaceae) species. Plant Growth Regul., 53(3), 163–171. DOI: 10.1007/s10725-007-9215-x
  29. Vejsedova, H. (2006). Factors affecting seed germination and seedling growth of terrestial orchids cultured in vitro. Acta Biol. Cracov. Ser. Bot., 48(1), 109–113.
  30. Zaniecka, J., Łojkowska, E. (2004). Estabilishment of in vitro culture collection of endangered European orchids. Biul. Ogrod. Bot. Muz. Zbior., 13, 69–73.
  31. Zeng, S., Wu, K., Teixeira da Silva, J., Zhang, J., Chen, Z., Xia, N., Duan, J. (2012). Asymbiotic seed germination, seedling development and reintroduction of Paphiopedilum wardii Sumerh., an endangered terrestrial orchid. Sci. Hortic., 138, 198–209. DOI: 10.1016/j.scienta.2012.02.026
  32. Zeng, S., Huang, W., Wu, K., Zhang, J., Teixeria da Silva, J., Duan, J. (2016). In vitro propagation of Paphiopedilum orchids. Cr. Rev. Biotechnol., 36, 521–534. DOI: 10.3109/07388551.2014.993585

Downloads

Download data is not yet available.

Most read articles by the same author(s)

1 2 > >> 

Similar Articles

1 2 3 4 5 6 7 8 9 10 > >> 

You may also start an advanced similarity search for this article.