DOI QR코드

DOI QR Code

Protoplast Fusion of Panax ginseng Callus and Aralia Continentalis Mesophyll

인삼 캘러스와 독활 엽육조직의 원형질체 융합

  • Park, Jong-Bum (Department of Biological Sciences, Silla University)
  • 박종범 (신라대학교 생물과학과)
  • Published : 2008.02.28

Abstract

Protoplasts of Panax ginseng C. A. Meyer and Aralia continentalis K. (Araliaceae) were isolated from callus cells and mesophyll cells, respectively. The maximum yield of protoplasts isolated from callus cells of P. ginseng were obtained by incubation for 3 hrs in the enzyme mixture of 0.5% macerozyme, 1.5% cellulase, and 0.5 M mannitol as an osmoticum. In the case of mesophyll cells of A. continentalis, the highest yield of protoplasts were obtained by incubation for 5 hrs in the enzyme mixture of 1% macerozyme, 2% cellulase, and 0.6 M mannitol. A polyethylene glycol (PEG) treatment induced an intergeneric fusion of the protoplasts. The fusion products, that is, heterokaryocytes were obtained by treatment of 50% PEG containing 0.05 M Ca salts.

Keywords

Panax ginseng;Aralia continentalis;Protoplast;Fusion;PEG;Heterokaryocyte

References

  1. Gresshoff P. M., 1980, In vitro culture of white clover, callus, suspension, protoplast culture, and plant regeneration, Bot. Gaz., 141, 157-164 https://doi.org/10.1086/337138
  2. Cocking E. C., 1960, A method for the isolation of plant protoplasts and vacuoles, Nature, 187, 962-963 https://doi.org/10.1038/187962a0
  3. Grambow H. J., Kao K. N., Miller R. A., Gamborg O. L., 1972, Cell division and plant development from protoplasts of carrot cell suspension culture, Planta, 103, 348-355 https://doi.org/10.1007/BF00386707
  4. Horine R. K., Ruesink A. W., 1972, Cell wall regeneration around protoplasts isolated from Convolvulus tissue culture, Plant Physiol., 50, 438-445 https://doi.org/10.1104/pp.50.4.438
  5. Shepard J. E., Totten R. E., 1975, Isolation and regeneration of tobacco mesophyll cell protoplasts under low osmotic conditions, Plant Physiol., 55, 689-694 https://doi.org/10.1104/pp.55.4.689
  6. Evans D. A., Flick C. E., 1980, Somatic hybrid plants between species of the genus Nicotiana, In vitro, 16, 217
  7. Gleba Y. Y., Monot V. P., Cherep N. N., Skarzynskaya M. V., 1982, Intertribal hybrid cell lines of Atropa belladonna (x) Nicotiana chinensis obtained by cloning individual protoplast fusion products, Theor. Appl. Genet., 62, 75-79
  8. Vardi A., Speigel-Roy P., Galun E., 1982, Plant regeneration from Citrus protoplasts: Variability in methodological requirements among cultivars and species, Theor. Appl. Genet., 62, 171-176 https://doi.org/10.1007/BF00293354
  9. Dorion N., Chupeau Y., Bourgin J. P., 1975, Isolation, culture and regeneration into plants of Ranunculus sceleratus L. leaf protoplasts, Plant Sci. Lett., 5, 325-331 https://doi.org/10.1016/0304-4211(75)90060-7
  10. Kohlenbach H. W., Bohnke E., 1975, Isolation and culture of mesophyll protoplasts of Hyoscyamus niger L. var. annuus Sims., Experientia, 34, 1281-1283
  11. Saxena P. K., Gill R., Rashid A., Maheshwari S. C., 1981, Isolation and culture of protoplasts of Capsicum annuum L., Protoplasma, 103, 357-361
  12. Binding H., Nehls R., Kock R., Finger J., Mordhorst G., 1981, Comparative studies on protoplast regeneration in herbaceous species of the Dicotyledoneae class, Z. Pflanzenphysiol., 101, 119-130 https://doi.org/10.1016/S0044-328X(81)80046-3
  13. Power J. B., Frearson E. M., George D., Evans P. K., Berry S. F., Hayward C., Cocking E. C., 1976, Isolation, culture and regeneration of leaf protoplasts in the genus Petunia, Plant Sci. Lett., 7, 51-55 https://doi.org/10.1016/0304-4211(76)90045-6
  14. Mii M., Seeni S., Fowke L. C., King J., 1985, The isolation and cultivation of protoplasts from cell suspensions of a pantothenote-requiring auxotroph of Datura, Can. J. Bot., 63, 779-783 https://doi.org/10.1139/b85-099
  15. Melchers G., Sacristan M. D., Holder A. A., 1978, Somatic hybrid plants of potato and tomato regenerated from fused protoplasts, Carlsberg Res. Commun., 43, 203-218 https://doi.org/10.1007/BF02906548
  16. Flick C. E., Evans D. A., 1983, Isolation, Culture, and plant regeneration from protoplasts isolated from flower petals of ornamental Nicotiana species, Z. Pflanzenphysiol., 109, 379-382 https://doi.org/10.1016/S0044-328X(83)80120-2
  17. Liu F., Ryschka U., Marthe F., Kloche E., Schumann G., Zhao H., 2007, Culture and fusion of pollen protoplasts of Brassica oleracea L. var. italica with haploid mesophyll protoplasts of B. rapa L. ssp., Protoplasma, 231, 89-97 https://doi.org/10.1007/s00709-006-0228-5
  18. Durieu P., Ochatt S. J., 2000, Efficient intergeneric fusion of pea (Pisum sativum L.) and grass pea (Lathyrus sativus L.) protoplasts, J. Exp. Bot., 51, 1237-1242 https://doi.org/10.1093/jexbot/51.348.1237
  19. Szabados L., Gaggero C., 1985, Callus formation from protoplasts of a sugarbeet cell suspension culture, Plant Cell Reports, 4, 195-198 https://doi.org/10.1007/BF00269287
  20. Wang Y. P., Sonntag K., Rudloff E., 2003, Development of rapeseed with high erucic acid content by asymmetric somatic hybridization between Brassica napus and Crambe abyssinica, Theor. Appl. Genet., 106, 1147-1155 https://doi.org/10.1007/s00122-002-1176-x
  21. Gupta P. K., Durzan D. J., 1986, Isolation and cell regeneration of protoplasts from sugar pine, Plant Cell Reports, 5, 346-348 https://doi.org/10.1007/BF00268598
  22. Murashige T., Skoog F., 1962, A revised medium for rapid growth and bioassays with tobacco tissue cultures, Physiol. Plant., 15, 473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  23. Park J. B., 1998, An electron microscopic study on the cell wall regeneration of cultured Panax ginseng callus protoplast, Kor. J. Plant Tissue Culture, 25, 495-500
  24. Brisa M. C., Segura J., 1987, Isolation, culture and plant regeneration from mesophyll protoplasts of Digitalis obscura, Physiol. Planta., 69, 680-686 https://doi.org/10.1111/j.1399-3054.1987.tb01984.x
  25. Hou S. W., Jia J. F., 2004, Plant regeneration from protoplasts isolated from embryogenic calli of the forage legume Astragalus melilotoides Pall., Plant Cell Reports, 22, 741-746 https://doi.org/10.1007/s00299-004-0760-8
  26. Jiang S. H., Guan R. Z., Tang S. Y., Xin R. Y., Zhang H. S., Zhao L. Q., Pan Q. Y., 2007, Somatic hybrids between Rorippa indica (Linn.) Hiern and Brassica napus L. through protoplast-fusion system, Yi Chuan, 29, 745-750 https://doi.org/10.1360/yc-007-0745
  27. Chatterjee G., Sikder S. R., Sen S. K., 1985, Regeneration of plantlets from mesophyll protoplasts of Brassica juncea (L.) Czern., Plant Cell Reports, 4, 245-247 https://doi.org/10.1007/BF00269368
  28. Kamlesh R. P., Shekhawat N. S., Berlyn G. P., Thorpe T. A., 1984, Isolation and culture of protoplasts from cotyledons of Pinus coulteri Don., Plant Cell Tissue Organ Culture, 3, 85-90 https://doi.org/10.1007/BF00033729
  29. Ruesink A. W., Thimann K. V., 1965, Protoplasts from the Avena coleoptile, Proc. Nat. Acad. Sci. U.S.A., 54, 56-64
  30. Barakat M. N., Cocking E. C., 1985, An assessment of the cultural capabilities of protoplast of some wild species of Linum, Plant Cell Reports, 4, 164-167 https://doi.org/10.1007/BF00571308
  31. Du L., Bao M., 2005, Plant regeneration from protoplasts isolated from embryogenic suspension cultured cells of Cinnamomum camphora L., Plant Cell Reports, 24, 462-467 https://doi.org/10.1007/s00299-005-0969-1
  32. Keller W. A., Melchers G., 1973, The effect of high pH and calcium on tobacco leaf protoplast fusion, Z. Naturforsch., 280, 737-741
  33. Kao K. N., Michayluk M. R., 1974, A method for high-frequency intergeneric fusion of plant protoplasts, Planta, 115, 355-367 https://doi.org/10.1007/BF00388618
  34. Uchimiya H., Murashige T., 1974, Evaluation of parameters in the isolation of viable protoplasts from cultured tobacco cells, Plant Physiol., 54, 936-944. https://doi.org/10.1104/pp.54.6.936
  35. Schenk R. U., Hildebrandt. A. C., 1969, Production of protoplasts from plant cells in liquid culture using purified commercial cellulase, Crop Sci., 9, 629-631 https://doi.org/10.2135/cropsci1969.0011183X000900050036x
  36. Takebe I., Nagata T., 1984, Isolation and culture of protoplasts: Tobacco. In Cell culture and somatic cell genetics of plants, Vasil, I. K. (ed.), Academic Press Inc. London, 328-339pp
  37. Nelson R. S., Creissen G. P., Bright S. W. J., 1983, Plant regeneration from protoplasts of Solanum brevidens, Plant Sci. Lett., 30, 355-362 https://doi.org/10.1016/0304-4211(83)90175-X
  38. Gosch G., Bajaj Y. P. S., Reinert J., 1975, Isolation, culture and induction of embryogenesis in protoplasts from cell suspensions of Atropa belladonna, Protoplasma, 86, 405-410 https://doi.org/10.1007/BF01287489
  39. Bui-Dang-Ha D., Mackenzie I. A., 1973, The division of protoplasts from Asparagus officinalis L. and their growth and differentiation, Protoplasma, 78, 215- 221 https://doi.org/10.1007/BF01275692
  40. Robertson D., Earle E. D., 1986, Plant regeneration from leaf protoplasts of Brassica oleracea var. italica cv. Green Comet broccoli, Plant Cell Reports, 5, 61-64 https://doi.org/10.1007/BF00269720
  41. Nishimaki T., Nozue M., 1985, Isolation and culture of protoplasts from high anthocyanin-producing callus of sweet potato, Plant Cell Reports, 4, 248-251 https://doi.org/10.1007/BF00269369
  42. Hu Q., Hansen N., Laursen J., Dixelius C., Andersen B., 2002, Intergeneric hybrids between Brassica napus and Orychophragmus violaceus containing traits of agronomic importance for oilseed rape breeding, Theor. Appl. Genet., 105, 834-840 https://doi.org/10.1007/s00122-002-1017-y
  43. Vardi A., Hutchison D. J., Galun E., 1986, A protoplast-to-tree system in Microcitrus based on protoplasts derived from a sustained embryogenic callus, Plant Cell Reports, 5, 412-414 https://doi.org/10.1007/BF00269629
  44. Yamagishi H., Landgren M., Forsberg J., Glimelius K., 2002, Production of asymmetric hybrids between Arabidopsis thaliana and Brassica napus utilizing an efficient protoplast culture system, Theor. Appl. Genet., 104, 959-964 https://doi.org/10.1007/s00122-002-0881-9