Summary
Rice is one of the most important crops in the world with 35% of the total population (over two billion people) depending on it as their source of food. It is therefore essential to develop efficient methods for the transformation and regeneration of rice plants in order to delineate the exact regulatory sequences responsible for gene expression and to transfer beneficial genes into this plant. Here, for the first time, we present definitive evidence for the regeneration of a large number of transgenic rice plants after introduction of the bacterial β-glucuronidase gene into rice protoplasts. The presence of integrated copies of this gene was detected in the genome of transgenic plants by DNA hybridization analysis. Furthermore, under the control of regulatory regions from a maize alcohol dehydrogenase sequence, β-glucuronidase gene expression was detected in the roots of transgenic plants. This expression was stimulated up to six fold under anaerobic conditions.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abdullah R, Cocking EC, Thompson JA (1986) Efficient plant regeneration form rice protoplasts through somatic embryogenesis. Bio/Technology 4:1087–1090
Bevan MW, Chilton M-D (1982) T-DNA of the agrobacterium Ti and Ri plasmids. Annu Rev Genet 16:357–384
Coulibaly MY, Demarly Y (1986) Regeneration of plantlets from protoplasts of rice, Oryza saliva L. Z Pflanzenzücht 96:79–81
Fraley RT, Rogers SG, Horsch RB (1986) Genetics transformation in higher plants. CRC Crit Rev Plant Sci 4:1–46
Fujimura T, Sakurai M, Akagi H, Negishi T, Hirose A (1985) Regeneration of rice plants from protoplasts. Plant Tissue Culture Lett 2:74–75
Gheysen G, Dhaese P, Van Montagu M, Schell J (1985) Advances in plant gene research. In: Hohn B, Dennis ES (eds) Genetic flux in plants, vol. 2. Springer, Berlin Heidelberg New York, pp 11–47
Goldberg RB (1986) Regulation of plant gene expression. Phil Trans R Soc London Ser B 314:343–353
Goodman RM, Hauptli H, Crossway A, Knauf VC (1987) Gene transfer in crop improvement. Science 236:48–54
Jefferson RA, Burgess SM, Hirsh D (1986) β-glucuronidase from Escherichia coli as a gene-fusion marker. Proc Natl Acad Sci USA 83:8447–8451
Jefferson RA, Kavanaugh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907
Kao KN (1977) Chromosomal behavior in somatic hybrids of soybean-nicotiana glauca. Mol Gen Genet 150:225–230
Kao TH, Moon E, Wu R (1984) Cytochrome oxidase subunit II gene of rice has an insertion sequence within the intron. Nucleic Acids Res 12:7305–7315
Klein TM, Gradziel T, Fromm ME, Sanford JC (1988) Factors influencing delivery into Zea mays cells by high-velocity microprojectiles. Bio/Technology 6:559–563
Kuhlmeier C, Green PJ, Chua N-H (1987) Regulation of gene expression in higher plants. Annu Rev Plant Physiol 38:221–257
Kyozuka J, Hayashi Y, Shimamoto K (1987) High frequency plant regeneration from rice protoplasts by novel nurse culture methods. Mol Gen Genet 206:408–413
Moon E, Kao T-H, Wu R (1987) Rice Chloroplast DNA molecules are heterogeneous as revealed by DNA sequences of a cluster of genes. Nucleic Acids Res 15:611–630
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Negrutiu I, Shillito R, Potrykus I, Biasini G, Sala F (1987) Hybrid genes in the analysis of transformation conditions. Plant Mol Biol 8:363–373
Okimoto R, Sachs MM, Porter EK, Freeling M (1980) Patterns of polypeptide synthesis in various maize organs under anaerobiosis. Planta 150:89–94
Ou-Lee TM, Turgeon R, Wu R (1986) Expression of a foreign gene liked to either a plant virus or a Drosophila promoter after electroporation of protoplasts of rice, wheat and sorghum. Proc Natl Acad Sci USA 83:6815–6819
Schell JS (1987) Transgenic plants as tools to study the molecular organization of plant genes. Science 237:1176–1183
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Uchimiya H, Fushimi T, Hashimoto H, Harada H, Syono K, Sugawara Y (1986) Expression of a foreign gene in callus derived from DNA-treated protoplasts of rice (Oryza sativa L.) Mol Gen Genet 204:204–207
Yamada Y, Yang ZQ, Tang DT (1986) Plant regeneration from protoplast-derived callus of rice (Oryza sativa L.). Plant Cell Rep 5:85–88
Author information
Authors and Affiliations
Additional information
Communicated by P. Maliga
Rights and permissions
About this article
Cite this article
Zhang, W., Wu, R. Efficient regeneration of transgenic plants from rice protoplasts and correctly regulated expression of the foreign gene in the plants. Theoret. Appl. Genetics 76, 835–840 (1988). https://doi.org/10.1007/BF00273668
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00273668