Abstract
A highly efficient system for direct somatic embryogenesis is described. Leaf sections originating from young trifoliate leaves of Medicago falcata line 47/1–5 and Medicago sativa line No2/9R, directly produced embryos after cultivation in liquid B5IV induction medium. In comparison with indirect somatic embryogenesis the system omits the callus stage and thus allows shortening of the process of somatic embryogenesis in alfalfa by 35–40 days. It permits the avoidance of secondary changes occurring during the process of dedifferentiation. A modified B5/3H medium containing Polyethylene Glycol 6000 promoted embryo development from globular up to torpedo stage. It was clearly shown that 2.5% Polyethylene Glycol stimulated this process for both H. falcata 47/1–5 and M. sativa No 2/9R. Maturation of torpedo stage embryos was carried out on solidified or liquid abscisic acidcontaining medium. A 30μM abscisic acid concentration was optimal in allowing one embryo to yield one plant. Somatic embryo conversion to plants and plant regeneration was performed on Murashige and Skoog medium. Regenerated plants showed a normal morphology.
Similar content being viewed by others
Abbreviations
- ABA:
-
Abscisic acid
- B5:
-
Medium of Gamborg et al.(1968)
- COT:
-
Cotyledone stage embryos
- 2,4-D:
-
2,4-dichlorphenoxyacetic acid
- FW:
-
Fresh weight
- GA3:
-
Gibberellin A3
- MS:
-
Medium of Murashige and Skoog (1962)
- PEG:
-
Polyethylene Glycol
- POLY:
-
Polyembryos
References
Atanassov AI, Brown DC (1984) Plant Cell Tissue and Organ Culture 3:149–162
Borkird C (1987) Ph.D. Thesis, University of California, Berkeley
Brown DC, Atanassov AI (1985) Plant Cell Tissue and Organ Culture 4:111–122
Denchev PD (1987) Ph.D.Thesis, Institute of Genetics, Academy of Sciences, Moscow, USSR
Denchev PD, Atanassov AI (1988) In: Staszewski Z, Utrata A (eds) Unconventional methods in Lucerne breeding, Inst. Extention Service, Poland, pp 17–21
Dijak M, Brown DC (1987) Plant Cell Tissue and Organ Culture 9:121–130
Dos Santos AV, Putka DE, Cocking EC, Davey MR (1980) Z. Pflanzenphysiol. 99:261–270
Gamborg OL, Miller RA, Ojima K (1968) Exp. Cell Res. 50:151–158
Halperin W (1966) Am. J. Bot. 53:443–453
Kao KM, Michayluk MR (1980) Z. Pflanzenphysiol. 96:135–141
Kao KN, Michayluk MR (1981) In vitro 17:645–648
Little TM, Hills FJ (1978) In: Wiley J (ed) Agricultural Experimentation, Design and Analysis, John Wiley and Sons, Inc., New York, pp 57–77
Litz RE (1986) J. Am. Soc. Hort. Sci. 111:969–972
Lupotto E (1986) Annals of Botany 57:19–24
McCoy TJ, Bingham ET (1977) Plant Sci. Lett. 10:59–66
Murashige T, Skoog F (1962) Physiol.Plant. 15:473–497
Seitz MH, Bingham ET (1988) In vitro Cell. & Dev.Biol. 24:1047–1052
Senaratna T, McKersie BD, Bowley SR (1990) In Vitro Cell.Dev.Biol. 26:85–90
Strickland SG, Nichol JW, McCall CM, Stuart DA (1987) Plant Sci. 48:113–121
Stuart DA, Nelson J, McCall CM, Strickland SG, Walker KA (1985) In: Zaitlin M, Day P, Hallander A (eds) Biotechnol. in Plant Sci. Relevance to agriculture in the eighties Acad. Press, Orlando, pp 35–47
Stuart DA, Redenbaugh MK (1987) In: LeBaron HM, Mumma RO, Honeycutt RC, Dresing JH (eds) Biotechnology in Agricultural Chemistry, ASC Symp. Ser. 334, Ch 6, American Chemical Society, Washington, DC pp 87–96
Walker KA, Sato SJ (1981) Plant Cell Tissue and Organ Culture 1:109–121
Author information
Authors and Affiliations
Additional information
Communicated by N. Amrhein
Rights and permissions
About this article
Cite this article
Denchev, P., Velcheva, M. & Atanassov, A. A new approach to direct somatic embryogenesis in Medicago . Plant Cell Reports 10, 338–341 (1991). https://doi.org/10.1007/BF00193154
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00193154