Summary
Improvement of Hibiscus rosa-sinensis for increased frost tolerance has been attempted through somatic hybridization with the frost tolerant Lavatera thuringiaca. Cell suspensions from Hibiscus and Lavatera were transformed with A. tumefaciens harboring plasmids containing selectable genes coding for kanamycin and hygromycin resistance, respectively. We provided evidence that H. rosa-sinensis and L. thuringiaca were transformed by strong selection of transformed calluses in medium containing antibiotics, by GUS activity determination in protein extracts and by molecular confirmation of chromosomal integration and expression of the selectable genes. Protoplasts isolated from a kanamycinresistant Hibiscus callus and from a hygromycin-resistant Lavatera callus were fused and selected in medium containing both antibiotics. We determined unambiguously that the regenerated double-antibiotic resistant clones obtained are indeed somatic hybrids through analysis of acid phosphatase zymograms and nuclear DNA content. Plant regeneration through somatic embryogenesis was accomplished from both isolated protoplasts and transgenic calluses of L. thuringiaca. However, regeneration from the double-antibiotic resistant fusant calluses was unsuccessful. Analysis of the somatic hybrids at the callus level showed that chilling and freezing tolerance are governed by independent genetic components. The somatic hybrids displayed significant improvement for chilling tolerance at conditions lethal to H. rosa-sinensis, although frost tolerance was not expressed.
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References
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1987) Current Protocols in Molecular Biology. John Wiley & Sons, USA
Arumuganathan K, Earle ED (1991) Plant Mol Biol Reporter 9: 229–233
Chomczynski P, Sacchi N (1987) Anal Biochem 162: 156–159
Frearson EM, Power JB, Cocking EC (1973) Dev Biol 33: 130–137
Guy CL (1990) Annu Rev Plant Physiol Plant Mol Biol 41: 187–223
Hurt J, Hsu JL, Dougall WC, Visner GA, Burr IM, Nick HS (1992) Nucleic Acids Res 20: 2985–2990
Jefferson RA, Kavanagh TA, Bevan MW (1987) The EMBO J 6: 3901–3907
Karlsson MG, Heins RD, Gerberick JO, Hackmann ME (1991) Scientia Horticulturae 45: 323–331
Levitt J (1980) Responses of Plants to Environmental Stresses, Vol I: Chilling, Freezing, and High Temperature Stresses. Academic Press, New York
Menczel L, Nagy F, Kiss ZSR, Maliga P (1981) Theor Appl Genet 59: 191–195
Menczel L, Wolfe K (1984) Plant Cell Reports 3: 196–198
Palta JP, Simon G (1993) Breeding potential for improvement of freezing stress resistance: Genetic separation of freezing tolerance, freezing avoidance, and capacity to cold acclimate. In: Li PH, Christersson L (eds) Advances in Plant Cold Hardiness. CRC Press, Boca Raton, FL., pp299–310
Potrykus I, Shillito R (1986) Methods in Enzymol 118: 549–578
Stone JM, Palta JP, Bamberg JB, Weiss LS, Harbage JF (1993) Proc Natl Acad Sci USA 90: 7869–7873
Sung ZR, Okimoto R (1981) Proc Natl Acad Sci USA 78: 3683–3687
Sung ZR, Okimoto R (1983) Proc Natl Acad Sci USA 80: 2661–2665
Sutka J (1981) Theor Appl Genet 59: 145–152
Thomzik JE, Hain R (1988) Theor Appl Genet 76: 165–171
Towill LE, Mazur P (1975) Can J Bot 53: 1097–1102
Vazquez-Tello A, Hidaka M, Uozumi T (1995) Plant Cell Tissue Organ Culture 40: 169–177
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Communicated by A. Komamine
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Vazquez-Thello, A., Li Yang, J., Hidaka, M. et al. Inherited chilling tolerance in somatic hybrids of transgenic Hibiscus rosa-sinensis x transgenic Lavatera thuringiaca selected by double-antibiotic resistance. Plant Cell Reports 15, 506–511 (1996). https://doi.org/10.1007/BF00232983
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DOI: https://doi.org/10.1007/BF00232983