Effective Multiplication of Somatic Embryos Using Suspension Culture and Regeneration in Soybean

  • Kim, Young Jin (Honam Agricultural Research Institute, National Institute of Crop Science) ;
  • Park, Tae Il (Honam Agricultural Research Institute, National Institute of Crop Science) ;
  • Kim, Hyun Soon (Honam Agricultural Research Institute, National Institute of Crop Science) ;
  • Suh, Sug Kee (Honam Agricultural Research Institute, National Institute of Crop Science) ;
  • Kim, Hag Sin (Honam Agricultural Research Institute, National Institute of Crop Science) ;
  • Yun, Song Joong (Honam Agricultural Research Institute, National Institute of Crop Science)
  • Published : 2004.06.01

Abstract

The use of liquid-medium-based procedure relative to the solid media led to a 4.5-fold increase in the number of cotyledon-stage embryos. The most efficient system for multiplication and regeneration of somatic embryos was CP6 procedure with the media MSD40/MSD20/MSM6AC/FNL0S3S3GM. However, the rate of regeneration was lower. About 71% of the embryos with dicotyledon were continued to develop the roots after desiccation treatment and 92% of the germinated embryos produced shoots in 10 days. Of the four morphologically different types of embryos, dicotyledonous ones showed a high frequency of conversion, while only a few with fused and horn type cotyledon developed shoots. Mature somatic embryos were desiccated in empty petri dishes for 12-72 h. Embryo survival rate was the highest after 12 h of desiccation, but maximal germination was observed at 24 h. After desiccation, they were placed on MS medium without growth regulators for germination. Germinating embryos were transferred to small pots with vermiculite for plant regeneration. The etiolating the plants during the growth was resolved to add 1% activated charcoal on hormone-free MS medium.

Keywords

References

  1. Bailey MA, Boerma HR, and Parrott WA (1993) Genotype effects on proliferative embryogenesis and plant regenera-tion of soybean, In Vitro Cell Dev Biol 29: 102-108
  2. Buchhelm JA, Colburn SM, and Ranch JP (1989) Maturation of soybean somatic embryos and the transition to plantlet growth. Plant Physiol 89: 768-775
  3. Durham RE, Parrott WA (1992) Repetitive somatic embryogenesis from peanut cultures in liquid medium. Plant Cell Rep 11 12-125
  4. Finer JJ, Nagasawa A (1988) Development of an embryogenic suspension culture of soybean (Glycine max Merrill.). Plant Cell Tiss Org Cult 15: 125-136
  5. Fridborg G, Pedersen M, Landstrom L, Eriksson T, (1987) The effect of activated charcoal on tissue cultures: adsorption of metabolites inhibiting morphogenesis. Physio Plant 43:104-106
  6. Hadi MZ, McMullen MD, Finer JJ (1996) Transformation of 12 different plasmids into soybean via particle bombardment. Plant Cell Rep 15: 500-505
  7. GB (1989) Analysis of lipid composition and morphological characteristics in soybean regenerants. Plant Cell Rep 7: 701-703
  8. Komatsuda T, Ohyama K (1988) Genotypes of high competence for somatic embryogenesis and plant regeneration in soybean Glycine max. Theo Appl Genet 75: 695-700
  9. Li J, Grabau EA (1996) Comparison of somatic embryogenesis and embryo conversion in commercial soybean cultivars. Plant Cell Tiss Org Cult 44: 87-89
  10. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473-479
  11. Parrott WA, Dryden G, Vogt S, Hildebrand DF, Collins GB, Williams EG (1988) Optimization of somatic embryogenesis and embryo germination in soybean. In Vitro Cell Dev Biol 24: 817-820
  12. Ranch JP, Oglesby L, Zielinski AC (1986) Plant regeneration from tissue cultures of soybean by somatic embryogenesis. In: Vasil I.K. (ed.) Cell Culture and Somatic Cell Genetics of Plants pp. 97-110, Academic Press, New York
  13. Samoylov VM, Tucker DM, Parrott WA (1998) A liquid-medium-based protocol for rapid regeneration from embryogenic soybean cultures. Plant Cell Rep 18: 49-54
  14. Trick HN, Finer JJ (1998) Sonication-assisted Agrobacterium-mediated transformation of soybean [Glycine max (L.) Merrill] embryogenic suspension cultures. Plant Cell Rep 17: 482-488