Abstract
Two finely-dispersed, homogeneous and regenerable cell suspension cultures were established from embryogenic callus derived from immature and mature embryos in barley. The quality and viability of suspension cells obtained were determined using differential-interference-contrast microscope and fluorescence microscope. Cell suspension cultures, maintained in modified liquid CC medium, showed a 10-fold increase in dry weight after two weeks with a doubling time of about 3 days. Addition of l-proline and casein hydrolysate in the medium had positive effect on the growth of cell cultures. Subculture interval significantly affected mitotic index. Both cell lines established were able to regenerate plants by somatic embryogenesis, but cell line Z-IM showed much higher regeneration capacity than cell line Z-M. Comparatively high frequencies of variations in chromosome number and structure were found in both lines, and a correlation between karyotype and morphogenic capacity was noticed.
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Abbreviations
- KT:
-
kinetin
- BAP:
-
benzylaminopurine
- FDA-PI:
-
fluorescein diacetate and propidium iodide
- 2,4-d :
-
2,4-dichlorophenoxyacetic acid
References
Dodds, JH & Roberts, LW (1985) Experiments in Plant Tissue Culture, second edition (pp 198–199). Cambridge University Press, Cambridge
Green, CE (1982) Somatic embryogenesis and plant regeneration from the friable callus of Zea mays L. In: A.Fujiwara (Ed) Plant Tissue Culture (pp 107–108) Maruzen, Tokyo
Huang, CN, Cornejo, MJ, Bush, DS & Jones, RL (1986) Estimating viability of plant protoplasts using double and single staining. Protoplasma 135: 80–87
Jähne, A, Lazzeri, PA, Jäger-Gussen, M & Lörz, H (1991a) Plant regeneration from embryogenic cell suspension derived from anther cultures of barley (Hordeum vulgare L.) Theor. Appl. Genet. 82: 74–80
Jähne, A, Lazzeri, PA & Lörz, H (1991b) Regeneration of fertile plants from protoplasts derived from embryogenic cell suspensions of barley (Hordeum vulgare L.). Plant Cell Rep. 10: 1–6
Kamo, KK & Houges, TK (1986) Establishment and characterization of long-term embryogenic maize callus and cell suspension cultures. Plant Sci. 45: 111–117
Kott, LS & Kasha, KJ (1984) Initiation and morphological development of somatic embryoids from barley cell cultures. Can. J. Bot. 62: 1245–1249
Lee, M & Phillips, RL (1988) The chromosomal basis of somaclonal variation. Ann. Rev. Plant Mol. Biol. 39: 413–437
Lu, CY & Vasil, IK (1981) Somatic embryogenesis and plant regeneration from freely-suspended cells and cell groups of Panicum maximum. Ann. Bot. 48: 543–547
Lührs, R & Lörz, H (1989) Initiation of morphogenic cell suspension and protoplast cultures of barley. Planta 157: 71–81
Muller, B, Schulze, J & Wegner, U (1989) Establishment of Barley cell suspension cultures of mesocotyl origin suitable for isolation of dividing protoplasts. Biochem. Physiol. Pflanzen. 185: 123–130
Polikarpochkina, RT, Gamburg, KI & Khavikin, EE (1979) Cell suspension culture of maize. Z. Pflanzenphysiol. 95: 57–67
Potrykus, I, Harms, CT & Lörz, H (1979) Callus formation from cell culture protoplasts of corn. Theor. Appl. Genet. 54: 209–214
Rengel, Z (1987) Embryogenic calles induction and plant regeneration from cultured Hordeum vulgare mature embryos. Plant Physiol. Biochem. 25(1): 43–48
Rhodes, CA, Pierce, DA, Mettler, IJ, Mascarenhas, D & Detmer, JJ (1988) Genetically transformed maize plants from protoplasts. Science 240: 204–207
Srinivasan, C & Vasil, IK (1986) Plant regeneration from protoplasts of Sugarcane. J. Plant Physiol. 126: 41–49
Terada, R, Kyozuka, J, Nishibayashi, S & Shimamoto, K (1987) Plantlet regeneration from somatic hybrids of rice (Oryza sativa L.) and barnyard grass (Echinochloa oryzieola, Vasing). Mol. Gen. Genet. 210: 39–43
Thomas, MR & Scott, KJ (1985) Plant regeneration by somatic embryogenesis from callus initiated from immature embryos and immature inflorescence of Hordeum vulgare. J. Plant Physiol. 121: 159–169
Vasil, V & Vasil, IK (1986) Plant regeneration from friable embryogenic callus and cell suspension cultures of Zea mays. J. Plant Physiol. 124: 399–408
Wang, HB, Li, XH, Sun, YR, Chen, J & Zhu, Z (1989) Protoplast culture of wheat-high frequency of cell cluster formation and plant regeneration. Chinese Science B. 1989: 828–834
Yamada, Y, Yang, ZQ & Tang, DT (1986) Plant regeneration from protoplast-derived callus of rice (Oryza sativa L.) Plant Cell Rep. 5: 85–88
Yan, QS, Zhang, XQ, Shi, JB & Li, JM (1991) Green plant regeneration protoplasts of barley (Hordeum vulgare L.) Chinese Science Bulletin 36: 923–935
Zhang, HM, Yang, H, Rech, EL, Golds, TJ, Davis, AS, Mulligan, GJ, Cocking, EC & Davey, MR (1988) Transgenic rice plants produced by electroporation-mediated plasmid up-take into protplasts. Plant Cell Rep. 7: 397–384
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Huang, C., Yan, H., Yan, Q. et al. Establishment and characterization of embryogenic cell suspension cultures from immature and mature embryos of barley (Hordeum vulgare L.). Plant Cell Tiss Organ Cult 32, 19–25 (1993). https://doi.org/10.1007/BF00040111
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DOI: https://doi.org/10.1007/BF00040111