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Somatic Embryogenesis in Maize and Comparison of Genetic Variability Induced by Gamma Radiation and Tissue Culture Techniques
Authors:F J Novak    S Daskalov    H Brunner    M Nesticky    R Afza    M Dolezelova    S Lucretti    A Herichova  T Hermelin
Institution:Dr. F. J. Novak;, H. Brunner, R. Afza, T. Hermelin, Plant Breeding Unit, IAEA Laboratories Seibersdorf, P.O. Box 100, A-1400 Vienna (Austria) Dr. M. Nesticky;, A. Herichova, Maize Research Institute, Trnava Dr. M. Dolezelova;, Institute of Experimental Botany, Olomouc (Czechoslovakia) Dr. S. Daskalov;, Institute of Genetics, Sofia (Bulgaria) Dr. S. Lucretti;, ENEA, Casaccia (Italy)
Abstract:Sixteen inbred lines and one hybrid of manse were tested for their capability of somatic embryogenesis, and fully developed plants could be regenerated, from ten inbred, lines. The highest frequency of plant regeneration was expressed in the inbred line CHI 31, and when this line was crossed with a recalcitrant, non-regenerating line, the F1 and BC hybrids were regenerable. The results of reciprocal crosses demonstrated that dominant nuclear genes and cytoplasmic factors are primarily responsible for the heritable determination of embryogenic callus proliferation and in vitro regeneration of maize plants. Somaclonal and radiation-induced variability was studied in maize to assess their nature and potential contribution to plant breeding., The inbred line CHI 31 possessing a high in vitro capacity of somatic embryo formation was used as experiments.] material. CHI 31 plants were selfed and twelve-day old zygotic embryos irradiated with 60Co gamma radiation in situ. Mature caryopses were harvested and assigned as M1 material. In another series, immature zygotic embryos (size 1.2—1.5 mm) were cultured in vitro on N-6 medium supplemented with 2,4-D (2.5 μM), and somatic embryos regenerated into plants; these were transplanted into soil and self-pollinated. Regenerants from non-irradiated cultures were grown as R1 generation, while regenerants from irradiated explants were considered as M1R1 generation. The genetic variability was evaluated in the M2, R2 and M2R2 generations, respectively, and compared with a non-treated seed control. Irradiation induced a variety of chlorophyll and morphological variants segregating in the M; generation; however, the frequency of deviant types obtained in the R: generation (somaclonal variation) was significantly exceeding the one derived from the M2 populations. The combination of expert irradiation and in vitro regeneration was most effective for the manifestation of chlorophyll and morphological o if types in the M2R2 generation, and increased drastically the frequency of early flowering variants. Differences in the segregation patterns of mutant phenotypes amonsister somaclones in the R3 and M3R3 generations indicate a different genetic basis, of plants originating from the same explant. This phenomenon suggests a mutational sectoring of the callus during culture. Radiation induced and somaclonal variation exerted a similar spectrum of chlorophyll and morphological deviants.
Keywords:Zea mays                        in vitro plant regeneration  somaclonal variation  induced mutations
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