Embryo Culture of Taxus wallichiana (Zucc.)

  • Datta Mukul Manjari (Centre of Advanced Study, Department of Botany, Calcutta University) ;
  • Jha Sumita (Centre of Advanced Study, Department of Botany, Calcutta University)
  • Published : 2004.12.01

Abstract

Zygotic embryos were excised from immature and mature seeds of the Himalayan yew, Taxus wallichiana. The embryos germinated precociously when kept in darkness for 5 weeks and developed into full seedlings within 10-12 weeks. The highest rate of embryo germination ($81\%$) was obtained in modified Lloyd & McCown' s woody plant medium containing macro and micronutrients at half strength supplemented with $1\%$ activated charcoal, which supported both the best embryonic growth ($43\%$) and seedling development ($32\%$). However, the supplementation of basal media with kinetin, thidiazuron, 6-benzyl aminopurine or $GA_3$ had no effect on the germination of the embryos. The embryos derived from immature seeds germinated but the frequency of embryonic growth was better in mature seeds. Stratification of seeds effected precocious germination of embryos. Seeds kept at $4^{\circ}C$ for 1 week germinated earlier and at a higher frequency irrespective of the stage of seed maturity, while the germination rate declined with prolonged cold treatment for 1 month at that same temperature. Analysis of taxanes in germinating seedlings revealed that root tissues contained high levels of taxol, 10-deacetyl-baccatin ill and baccatin ill as compared to shoots. Thus embryo culture technique appears to overcome the lengthy dormancy requirement of T. wallichiana seeds.

Keywords

References

  1. Anonymous (2002) Tree Conservation Information Service. UNEP. WCMC, 1-2
  2. Ahuja MR (1993) Micropropagation of Woody Plants. Kluwer Academic Publishers
  3. Behera MD, Srivastava S, Kushwaha SPS, Roy PS (2000) Stratification and mapping of Taxus baccata L. bearing forests in Talle Valley using remote sensing & GIS. Current Science 78: 1008-1013
  4. Chang SH, Yang JC (1996) Enhancement of plant formation from embryo cultures of Taxus mairei using suitable culture medium and PVP. Bot. Bull. Acad. Sin. 37: 35-40
  5. Chang SH, Ho CK, Tsai JY (1998) Micropropagation of Taxus mairei seedlings at different ages and recoverability of their plagiotropic shoots. Taiwan J. Forest Sci. 13: 29-39
  6. Chee PP (1994) In vitro culture of zygotic embryos of Taxus species. Hort.Sci. 29: 695-697
  7. Choi MS (2000) In vitro germination and propagation by embryo culture of Taxus cuspidata for the taxol production. J. Plant Biotech. 2: 29-33
  8. Flores HE, Sgrignoli PJ (1991) In vitro culture and precocious germination of Taxus embryos. In Vitro Cell. Dev. BioI. 27:139-142
  9. Flores T, Wagner LJ, Flores HE (1993) Embryo culture and taxane production in Taxus spp. In Vitro Cell. Dev. BioI. 29: 160-165
  10. Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soyabean root cells. Exp. Cell Res. 50: 151-158
  11. Gomez KA, Gomez AA (1984) Statistical Procedures for Agricultural Research, second ed. Wiley, New York
  12. Hezari M, Croteau R (1997).Planta Med. 63: 291-295
  13. Ho RH (1987) Embryo Culture. In: Bonga, J.M., Durzan, D.J. (eds.). Specific principles and Methods: Growth and Development. Cell and Tissue Culture in Forestry.vol. 2. pp.137-167. Martinus Nijhoff Publishers, Dordercht
  14. Hu CY, Wang L, Wu B (1992) In vitro culture of immature Taxus embryos. Hort. Sci. 27: 210
  15. Jha S, Sanyal D, Ghosh B, Jha TB (1998). Improved taxol yield in cell suspension culture of Taxus wallichiana (Himalayan Yew). Planta Medica. 64:270-272
  16. Li XK, HuangYQ, Deng lB, Su lM, Li JF (2000) The propagation and growth dynamics of seedlings of Taxus chinensis (Pilger) Rehd. J. Plant Res. Environ. 9: 48-50
  17. Li lL (2001) Embryo culture of Taxus chinensis (Pilger) Rehd. and Taxus brevifolia Nutt. in vitro. J. Plant Res. Environ. 10: 62-63
  18. Lloyd G, Mc Cown,B (1980) Commercially feasible micropropagation of mountain laurel, Klamia latifolia by use of shoot tip culture. Proc. Inter. Plant Prop. Soc. 30: 421-427
  19. Majada JP, Sierra MI, Sanchez T R (2000) One step more towards taxane production through enhanced Taxus propagation. Plant Cell Rep.19, 825-830
  20. Miller RW, Powell RG, Smith CR.Jr, Arnold E, Clardy, J (1981)
  21. Antileukemic alkaloids from Taxus wallichiana lucc. J. Org. Chem. 46: 1469
  22. Mitter H, Sharma A (1999) Propagation of T baccata Linn by stem cuttings. Ind. Forester. 125:159-162
  23. MurashigeT, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15 473-497
  24. Pilz D (1996a) Propagation of Pacific Yews from Seed, (Part I) American Conifer Society Bulletin. Winter Issue. 13: 13-18
  25. Pilz, D., 1996b. Propagation of Pacific Yews from Seed, (Part II) American Conifer Society Bulletin. Spring Issue. 13, 80-83
  26. Rajewski M, Lange S, Hattemer HH (2000) Problems of reproduction in the genetic conservation of rare tree species: The example of common yew (T baccata L). Forest Snow and Landscape Res. 75: 251-266
  27. Rikhari HC, Sharma S, Palni LMS, Sharma S, Nandi SK (1998) Himalayan Yew: stand structure, canopy damage, regeneration and conservation strategy. Environ. Conserv. 25: 334-341
  28. Saini RP (2001) Vegetative propagation in Silviculture (Hills) Division, Darjeeling (West Bengal). Ind. Forester. 127: 389-408
  29. Schenk RV, Hildebrandt AC (1972) Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures. Can.J.Bot. 50: 199-204
  30. Shi QW, Oritani T, Kiyota H, Zhao D (2000) Taxane diterpenoids from Taxus yunnanensis and Taxus cusoidste. Phytochem. 54: 829-834
  31. Shigemori H, Sakurai C A, Hosoyama H, Kobayashi A, Kajiyama S, Kobayashi J (1999) Taxezopidines J, K, and L, new taxoids from Taxus cuspidata inhibiting $Ca^2^+$-induced depolymerization of microtubules. Tetrahedron. 55: 2553-2558
  32. Zhang ZQ, Luo XT, Yang JX, Wu YW (1998) Seed development and seedling growth of Taxus chinensis (Piger) Rehd. J. Plant Res.Environ. 7: 12-15