Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Effects of carbohydrates and osmoticum on the somatic embryogenesis and cotyledon morphology of Codonopsis lanceolata L.

더덕의 체세포발생과 자엽형태에 미치는 탄수화물과 삼투제의 영향

  • Choi, Pil Son (Department of Oriental Pharmaceutical Development, Nambu University)
  • 최필선 (남부대학교 한방제약개발학과)
  • Received : 2020.05.14
  • Accepted : 2020.06.01
  • Published : 2020.06.30
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Abstract

Embryogenic calli from Codonopsis lanceolata L. were cultured in MS liquid media and supplemented with various concentrations of primary carbon sources to study the effects of carbohydrates and osmoticum on somatic embryogenesis and somatic embryo morphology. Sucrose, glucose, and a combination of 3% sucrose and various concentrations of sorbitol or mannitol as osmoticum were used as carbon supplements. The maximum number of somatic embryos per flask was greater in media exclusively supplemented with 3% sucrose (128.29) than exclusively glucose-supplemented media (47.67) and either supplement combination of 3% sucrose and osmoticum (95.67 with mannitol and 114.00 with sorbitol). The number of somatic embryos gradually decreased in media with increasing concentrations of combined osmoticum supplement. Decreases also occurred in the highest concentrations of sucrose- and glucose-supplemented media. The total frequency of somatic embryos with two cotyledons was slightly higher in medium with 3% + mannitol (24.09%) compared with exclusively sucrose (21.52%), glucose (21.22%), or 3% sucrose + sorbitol (22.13%). As concentrations of sucrose and glucose increased, the occurrence of two cotyledons and trumpet cotyledons gradually decreased and the occurrence of polycotyledon and globular stage embryos increased. Furthermore, as concentrations of 3% sucrose and osmoticum increased, the occurrence of trumpet cotyledon and globular stage embryos increased and the occurrence of polycotyledon gradually decreased. These results demonstrated that the somatic embryogenesis and occurrence of cotyledon morphology were influenced by the concentration of carbohydrates and combinations of 3% sucrose and osmoticum supplements.

체세포배 발생과 자엽 형태에 미치는 탄수화물과 삼투제의 영향을 알아보기 위하여 배발생캘러스를 탄소원으로 sucrose와 glucose를 그리고 삼투제로서 mannitol 또는 sorbitol을 3% sucrose와 조합하여 첨가한 MS액체배지에 배양하였다. 플라스크당 체세포배 최대 형성수(128.29)는 glucose단독 또는 삼투제와 3% sucrose를 조합처리한 배지보다 3% sucrose를 첨가한 배지에서 얻을 수 있었고, 그리고 탄수화물과 삼투제의 농도가 증가할수록 체세포배 형성수는 감소하였다. 2개의 자엽을 갖는 전체 체세포배 형성빈도(24.09)는 sucrose 단독(21.52), glucose단독(21.22) 그리고 sorbitol과 3% sucrose의 조합처리보다 mannitol과 3% sucrose조합처리배지에서 약간 높게 나타났다. 특히, sucrose와 glucose농도가 높을수록 2개의 자엽과 트럼펫 모양의 자엽빈도는 점차 감소하였고, 반면 다자엽과 생장이 지연된 구형기 체세포배 빈도는 증가하였다. 한편 삼투제와 3% sucrose의 조합처리 농도가 높을수록 트럼펫 모양과 생장이 지연된 구형기 체세포배 빈도는 뚜렷이 증가하였고, 반면 다자엽 체세포배의 빈도는 점차 감소하였다. 이와 같이 체세포배 발생과 체세포배의 자엽형태는 탄수화물과 삼투제의 처리농도에 의하여 영향을 받을 수 있음을 보여주었다.

Keywords

References

  1. Ammirato PV (1983) Embryogenesis. In: Evans DA, Sharp WR, Ammirato PV, Yamada Y (eds.), Handbook of plant cell culture, Vol 1, Macmillan, New York, pp82-123
  2. Ashburner GR, Thompson WK, Burch JM (1993) Effects of a-naphthalene acetic acid and sucrose levels on the development of cultured embryos of coconut. Plant Cell Tiss Org Cult 35: 157-163 https://doi.org/10.1007/BF00032965
  3. Brown C, Brooks FJ, Pearson D, Mathias RJ (1989) Control of embryogenesis and organogenesis in immature wheat embryo callus using increased medium osmolarity and abscisic acid. J Plant Physiol 133:727-733 https://doi.org/10.1016/S0176-1617(89)80080-X
  4. Buchheim JA, Colburn SM, Ranch JP (1989) Maturation of soybean somatic embryos and the transition to plantlet growth. Plant Physiol 89:768-775 https://doi.org/10.1104/pp.89.3.768
  5. Choi JH, Kwon SY, Choi PS (2011) Anomalous somatic embryos formation and plant regeneration from the cultures of immature embryos of Camellia japonica L. J Plant Biotechnol 38:258-262 https://doi.org/10.5010/JPB.2011.38.4.258
  6. Choi PS, Kwon SY (2013a) Histological characteristics of somatic embryos in melon (Cucumis melo L.). Kor J Plant Res 26:511-515 https://doi.org/10.7732/kjpr.2013.26.4.511
  7. Choi PS, Kwon SY (2013b) Procambium differentiation and shoot apical meristem development in somatic embryos of soybean (Glycine max L.). J Plant Biotechnol 40:55-58 https://doi.org/10.5010/JPB.2013.40.1.055
  8. Choi PS, Soh WY, Cho DY, Liu JR (1994) Somatic embryogenesis in immature zygotic embryo cultures of Korean soybean (Glycine max L.) cultivars and effect of 2,4-dichlorophenoxy acetic acid somatic embryo morphology. Korean J Plant Tiss Cult 21:7-13
  9. Choi PS, Soh WY, Cho DY, Liu JR (2005) Relationship of cotyledon number with procambium differentiation in somatic embryogenesis of Codonopsis lanceolata L. Kor J Plant Biotechnol 32:135-138 https://doi.org/10.5010/JPB.2005.32.2.135
  10. Finer JJ (1987) Direct somatic embryogenesis and plant regeneration from immature embryos of hybrid sunflower (Helianthus annus L.) on a high sucrose-containing medium. Plant Cell Rep 6: 372-374 https://doi.org/10.1007/BF00269564
  11. Isabelle GT, Mauro MC, Sossountzov L, Miginiac E, Deloire A (1993) Arrest of somatic embryo development in grapevine: histological characterization and the effect of ABA, BAP and zeatin in stimulating plantlet development. Plant Cell Tiss Org Cult 33: 91-103 https://doi.org/10.1007/BF01997603
  12. Kageyama C, Komatsuda T, Nadajima K (1990) Effects of sucrose concentration on morphology of somatic embryos from immature soybean cotyledons. Plant Tiss Cult Lett 7:108-110 https://doi.org/10.5511/plantbiotechnology1984.7.108
  13. Komatsuda T (1992) Research on somatic embryogenesis and plant regeneration in soybean. Natl Inst Agrobiol Resources (Japan), Ann Rep No. 7:1-78
  14. Lazzeri PA, Hildebrand DF, Collin GB (1987) Soybean somatic embryogenesis: effects of nutritional, physical and chemical factors. Plant Cell Tiss Org Cult 10:209-220 https://doi.org/10.1007/BF00037305
  15. Lazzeri PA, Hildebrand DF, Sunega J, William EG, Collin GB (1988) Soybean somatic embryogenesis : interaction between sucrose and auxin. Plant Cell Rep 7:517-520 https://doi.org/10.1007/BF00272745
  16. Lee KS, Soh WY (1993a) Somatic embryogenesis and structural aberrancy of embryos in tissue cultures of Aralia cordata Thumb. Korean J Plant Tiss Cult 20:77-84
  17. Lee KS, Soh WY (1993b) Effects of cytokinins on the number of cotyledons of somatic embryos from cultured cells of Aralia cordata Thumb. Korean J Plant Tiss Cult 20:171-176
  18. Liu CM, Xu ZH, Chua NH (1993) Auxin polar transport is essential for the establishment of bilateral symmetry during early plant embryogenesis. The Plant Cell 5:621-630 https://doi.org/10.2307/3869805
  19. Merkle SA, Parrott WA, Flinn BS (1995) Morphogenic aspects of somatic embryogenesis. In: Thorpe TA (ed.), In vitro embryogenesis in plants, Kluwer Academic Publishers, Dordrecht, pp155-205
  20. Min SR, Yang SG, Liu JR, Choi PS, Soh W-Y (1992) High frequency somatic embryogenesis and plant regeneration on tissue cultures of Codonopsis lanceolata. Plant Cell Rep 10:621-623 https://doi.org/10.1007/BF00232383
  21. 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
  22. Soh WY, Choi PS, Cho DY, Liu JR (2001) Plant regeneration of somatic embryos with anomalous cotyledons formed in cell cultures of Codonopsis lanceolata. Phytomorphology 51:327-336
  23. Strickland SG, Mccall CM, Nichol JW, Stuart DA (1986) Enhanced somatic embryogenesis in Medicago sativa by addition of maltose and its higher homologs to the culture medium. In: Somers DA, Gegenbach BG, Biesboer DD, Hackett WP, Green CE (eds.), VI Internal Conf Plant Tiss Cult (abstracts), Menneapolis, Univ Minnesota. pp188
  24. Stuart DA, Nelson J, Strickland SG, Nichoil JW (1985) Factors affecting developmental process in alfalfa cell cultures. In: Henke RR (ed.), Tissue culture in forestry and agriculture, Plenum Press, New York, pp59-73
  25. Swedlund B, Locy RD (1993) Sorbitol as the primary carbon source for the growth of embryogenic callus of maize. Plant Physiol 103: 1339-1346 https://doi.org/10.1104/pp.103.4.1339