Abstract
Protoplasts were isolated from callus derived from a single homozygous seed of Oryza sativa L. var. Norin 8. Thirty protoclones were randomly selected and these showed variation in regeneration frequency ranging from 0–87% with an average of 52%. The potential for regeneration of each protoclone as reflected in the regeneration frequency was analyzed five times over a period of 250 days and showed that the protoclones can be classified into three types, namely: protoclones with high regeneration frequency; protoclones with low regeneration frequency, both of which maintained their respective levels of regeneration potential; and protoclones with gradually decreasing regeneration frequency. Secondary protoclones established from protoplasts isolated from some of these protoclones and regenerated 2–3 times for a period of 120 days also showed further reduction in regeneration frequency. The polypeptide composition analyzed by two dimensional gel electrophoresis suggests the presence of specific polypeptides related to regeneration potential. Analysis of ploidy level based on plant morphology and pollen size suggests the predominance of tetraploids among the regenerated plants.
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References
Abdullah R, Thompson JA, Khush GS, Kaushik RP & Cocking EC (1989) Protoclonal variation in the seed progeny of plants regenerated from rice protoplasts. Plant Sci. 65: 887–890
Abdullah R, Cocking EC & Thompson A (1986) Efficient plant regeneration from rice protoplasts through somatic embryogenesis. Bio/Technology 4: 1087–1090
Abe T & Futsuhara Y (1989) Selection of higher regenerable callus and change in isozyme pattern in rice (Oryza sativa L.). Theor. Appl. Genet. 78: 648–652
Anwar SY & Reddy GM (1980) Pollen grain size and fertility in certain polyploids of Oryza sativa L. Indian J. Exp. Biol. 18: 767–768
Brown PTH, Kyozuka J, Sukekiyo Y, Kimura Y, Shimamoto K & Lorz H (1990) Molecular changes in protoplast-derived rice plants. Mol. Gen. Genet. 223: 324–328
Chen CC & Chen CM (1980) Changes in chromosome number in microspore callus of rice during successive subcultures. Can. J. Genet. Cytol. 22: 607–614
Duncan DR, Williams ME, Zehr BE & Widholm JM (1985) The production of callus capable of plant regeneration from immature embryos of numerous Zea mays genotypes. Planta 165: 322–332
Fujimura T, Sakurai M, Akagi H, Negishi T & Hirose A (1985) Regeneration of rice plants from protoplasts. Plant Tiss. Cult. Lett. 2: 74–75
Gamborg OL, Miller RA & Ojima K (1968) Nutrient requirement of suspension cultures of soybean root cells. Exp. Cell Res. 50: 151–158
Hahne G & Hoffmann F (1984) Dimethyl sulfoxide can initiate cell divisions of arrested callus protoplasts by promoting cortical microtubule assembly. Proc. Natl. Acad. Sci. 81: 5449–5453
Hanzel JJ, Miller JP, Brinkman MA & Fendos E (1985) Genotype and media effects on callus formation and regeneration in barley. Crop Sci. 25: 27–31
Harms CT & Potrykus I (1978) Fractionation of plant protoplast types by iso-osmotic density gradient centrifugation. Theor. Appl. Genet. 53: 57–63
Kanda M, Kikuchi S, Takaiwa F & Oono K (1988) Regeneration of variant plants from rice (Oryza sativa L.) protoplasts derived from long term cultures. Jpn. J. Genet. 63: 127–136
Kyozuka J, Otto E & Shimamoto K (1988) Plant regeneration from protoplasts of indica rice: genotypic differences in culture response. Theor. Appl. Genet. 76: 887–890
Murashige T & Skoog F (1962) A revised medium for rapid growth and bioasays with tobacco tissue cultures. Physiol. Plant. 15: 473–497
Nishibayashi S, Hayashi Y, Kyozuka J & Shimamoto K (1989) Chromosome variations in protoplast derived calli and in plants regenerated from the calli of cultivated rice (Oryza sativa L). Jpn. J. Genet. 65: 355–361
O'Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250: 4007–4021
Ohira K, Ojima K & Fujiwara A (1973) Studies on the nutrition of rice cell culture I.A. simple, defined medium for rapid growth in suspension culture. Plant Cell Physiol. 14: 1113–1121
Oono K (1975) Production of haploid plants of rice (Oryza sativa L.) by anther culture and their use for breeding. Bull. Natl. Inst. Agric. Sci. D 26: 139–222
Oono K, Okuno K & Kawai T (1984) High frequency of somaclonal mutations in callus culture of rice. Gamma Field Symposia No. 23, Institute of Radiation Breeding (pp 71–95)
Prat D (1983) Genetic variability induced in Nicotiana sylvestris by protoplast culture. Theor. Appl. Genet. 64: 223–230
Thompson JA, Abdullah R & Cocking EC (1986) Protoplast culture of rice (Oryza sativa L.) using media solidified with agarose. Plant Sci. 47: 123–133
Toriyama K, Arimoto Y, Uchimiya H & Hinata K (1988) Transgenic rice plants after direct gene transfer into protoplasts. Bio/Technology 6: 1072–1074
Vasil IK (1987) Developing cell and tissue culture systems for the improvement of cereal and grass crops. J. Plant. Physiol. 128: 193–218
Zhang W & Wu R (1988) Efficient regeneration of transgenic plants from rice protoplasts and correctly regulated expression of the foreign gene in the plants. Theor. Appl. Genet. 76: 835–840
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Kawata, M., Harada, S., Antonio, B. et al. Protoclonal variation of plant regeneration in rice. Plant Cell Tiss Organ Cult 28, 1–10 (1992). https://doi.org/10.1007/BF00039909
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DOI: https://doi.org/10.1007/BF00039909