Effect of ascorbic acid and glutathione on somatic embryogenesis induction in Picea pungens Engelmann
Abstract
Effect of exogenic antioxidants – ascorbic acid (AA) and reduced glutathione (GSH) – on initiation and proliferation processes of somatic embryos of blue spruce (Picea рungens) in vitro embryos culture in dependence of stage development, genotype of donor trees, medium composition was shown using a cytological analysis. Initiation of somatic embryogenesis was carried out on basic media ½ DSR and ½ LV supplemented with AA (0 and 300 mg/l) and/or GSH (0 and 300 mg/l). The highest frequency of callus formation (from 60 up to 100 %) was obtained when establishing in zygotic embryo culture at the cotyledon stage. Formation of embryonal-suspensor masses (ESM) was detected at the initiation stage with the help of cytological analysis. The highest frequency of ESM formation achieved 8,4 % on ½ DSR medium and 28 % on ½ LV medium. Active proliferation was observed in only one of the genotypes studied. Application of antioxidants stimulated the frequency both of callus formation and ESM, however, AA and GSH affected on development of somatic embryos in different ways. Application of AA at the initiation and proliferation stages promoted formation of somatic embryos; GSH, on the contrary, reduced their development. Formation of the great number of somatic embryos was achieved with optimal combination of antioxidants: culture incubation at the initiation stage with AA and addition of GSH in culture medium at proliferation stage.
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