Korean Journal of Plant Resources (한국자원식물학회지)

The Plant Resources Society of Korea (한국자원식물학회)
권호 표지

Cryopreservation of in vitro-cultured Axillary Shoot Tips of Japanese Bead Tree (Melia azedarach) using Vitrification Technique

  • Yang Byeong-Hoon (Department of Forestry, Kyungpook National University) ;
  • Kim Hyun-Tae (Department of Forestry, Kyungpook National University) ;
  • Park Ju-Yong (Department of Forestry, Kyungpook National University) ;
  • Park Young-Goo (Department of Forestry, Kyungpook National University)
  • Published : 2006.06.01
Download PDF
⟨ Previous Next ⟩

Abstract

In vitro-grown axillary buds of Melia aredarach were successfully cryopreserved by vitrification. On the MS medium supplemented with BA 1 mg/L, multiple shoots were developed within $4{\sim}5$ weeks. Plantlets of Melia azedarach were cold-hardened at $10^{\circ}C$ for a 16-hr photo-period for 6 weeks. Excised axillary shoot-tips from hardened plantlets were precultured on a solidified Murashige & Skoog agar medium (MS) supplemented with 0.7 M sucrose for 1 day at $25^{\circ}C$. Axillary shoot-tip meristems wert dehydrated using a highly concentrated vitrification solution (PVS2) for 60 min at $0^{\circ}C$ prior to a direct plunge into liquid nitrogen (LN). The PVS2 vitrification solution consisted of 30% glycerol (w/v), 15% ethylene glycol (w/v), 15% DMSO (w/v) in MS medium containing 0.4M sucrose. After short-term warming in a water bath at $40^{\circ}C$, the meristems were transferred into 2 ml of MS medium containing 1.2M sucrose for 15 min and then planted on solidified MS culture medium. Successfully vitrified and warmed meristems resumed growth within 2 weeks and directly developed shoots without intermediary callus formation. The survival rate of cold-hardened plantlets for 3 and 4 weeks was 90%. We did not find any difference in PCR-band patterns between control and cryopreserved plants. This method appears to be a promising technique for cryopreserving axillary shoot-tips from in vitro-grown plantlets of Medicinal plants.

Keywords

References

  1. Ascher, K.R.S., H. Schmutterer, C.P.W., Zebitz and S.N.H., Naqvi. 1995. Melia spp. In: Schmutterer H (ed) The neem tree: source of unique products for interested pest management, medicine, industry and other purposes. VCH., Weinheim, pp. 605-633
  2. Benson, E.E. 1994. Cryopreservation. In: Dixon RA, Gonzales A (eds) Plant cell culture. A practical approach. Oxford University Press, London. pp. 147-167
  3. Brison M., M,T. De Boucaud and F. Dosba. 1995. Cryopreservation of in vitro grown shoot tips of two interspecific Prunus rootstock. Plant Sci. 105: 235-242 https://doi.org/10.1016/0168-9452(94)04045-1
  4. CaccavaleA., M. Lambardi and A. Fabbri. 1998. Cryopreservation of woody plants by axillary bud vitrification: a first approach with poplar. Acta. Hortic. 457: 79-83
  5. Erwin, T.L. 1983. Tropical Forest Canopies: The Last Biotic Frontier. Bull. ESA, 29(1): 14-19
  6. Fahy, G.M., D.R.MacFarlane, C.A.Angell and H.T.Meryman. 1984. Vitrification as an approach to cryopreservation. Cryobiology. 21: 407-427 https://doi.org/10.1016/0011-2240(84)90079-8
  7. Grout, B. 1995. Introduction to the in vitro preservation of plant cells, tissue and organs. In: Grout B (ed) Genetic preservation of plant cells in vitro. Springer, Berlin Heidelberg New York, pp. 1-20
  8. Kuo, C.C. and R.D. Lineberger. 1985. Survival of in vitro cultured tissue of 'Jonathan' apples exposed to -196 $^{\circ}C$. HortScience. 4: 764-767
  9. Lambardi, M., A. Fabbri and A. Caccabale. 2000. Cryopreservation ofwhite poplar (Populus alba L.) by vitrification of in vitro-grown shoot tips. Plant Cell Reports. 19: 213-218 https://doi.org/10.1007/s002990050001
  10. Mayr, E. 1969. Principles of systematic zoology. McGraw-Hill, Net York. pp. 428
  11. Niino, T. 1995. Cryopreservation of germplasm of mulberry (Morus species). In: Bajaj YPS (ad) Cryopreservation of plant germplasm I; Biotechnology in agriculture and forestry, vol. 32. Springer, Berlin Heidelberg New York, pp. 102-266
  12. Niino, T., A. Sakai, H. Yakuwa, and K. Nojiri. 1992. Cryopreservation of in vitro-grown shoot tips of apple and pear by vitrification. Plant Cell Tissue Organ Culture. 28: 261-266 https://doi.org/10.1007/BF00036122
  13. Niino, T., K. Tashiro, M. Suzuki, S. Ohuchi, J. Magoshi and T. Akihama. 1997. Cryopreservation of in vitro-grown shoot tips of cherry and sweet cherry by one-step vitrification. Sci. Hortic. 70: 155-163 https://doi.org/10.1016/S0304-4238(97)00062-9
  14. Reed, B. 1990. Survival of in vitro grown apical meristems of Pyrus following cryopreservation. HortScience. 1: 111-113
  15. Reed, B., J. Denoma, J. Luo, Y. J. Chang and L. Towill. 1998. Cryopreservation and long term storage of pear germplasm. In vitro cellular and developmental biology. 34(3): 256-260 https://doi.org/10.1007/BF02822718
  16. Reid, W.V. and P. Miller. 1989. Keeping options alive: The scientific basis for conserving biodiversity. World Res. Inst., Washington, D.C
  17. Sakai, A. 1993. JICA, GRP, Ref., No.6, 5-26
  18. Sakai, A., S. Kobayashi and I.Oiyama. 1990. Cryopreservation of nucellar cells of navel orange (Citrus sinensis Osb. var. brasiliensis Tanaka) by vitrification. Plant Cell Reports. 9: 30-33
  19. Schmidt, G.H., A.I. Ahmed and M. Breuer. 1997. Effect of Melia azedarach extract on larval development and reproduction parameters of Spodoptera littoralis (Boised.) and Agrotis ipsilon (Hufu) (Lep. Noctuidae). Anz Schaedlingskd Pflanzen-Umweltschutz. 70: 4-12 https://doi.org/10.1007/BF02009609
  20. Stushnoff, C. 1987. Cryopreservation of apple genetic resources. Can. J. Plant Sci. 67: 1151-1154 https://doi.org/10.4141/cjps87-154
  21. Towill L.E. 1990. Cryopreservation of isolated mint shoot tips by vitrification. Plant Cell Reports. 9: 178-180
  22. Wilson, E.O. 1988. The current state of biodiversity. In: Biodiversity. E.O.Wilson (ed). National Academy Press,Washington, D.C
  23. Yaga, S. 1978. On the termite resistance of Okinawa timbers. Sci. Bull. Coll. Agric. Univ. Ryukyus Okinawa. 25: 555-613