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  • Author or Editor: Elisabet Clavería x
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Micropropagation of Pistacia vera L. `Mateur' was improved with the addition of methyl jasmonate (MeJA) to the multiplication and rooting media. Shoot tip cultures established from grafted trees were maintained on a modified Murashige and Skoog medium containing 5μM BA and 0.05μM IBA. Addition of 1μM MeJA improved the multiplication rate but inhibited shoot growth when present at higher concentrations. Rooting experiments comparing the effects of IAA, NAA, or IBA at 0, 1, 3.2, 10, or 31.6 μM demonstrated a significant effect of temperature on auxin root induction for shoots maintained at 25 or 28°C. At 25°C NAA was better than IAA or IBA, whereas no differences among auxins were observed at 28°C. Addition of MeJA (0, 0.3, 1, 3.2, or 10 μM) to the best rooting media significantly improved the rooting percentage and root number. Greater than 80% rooting was obtained when 1 μM MeJA was added to both the root induction medium, containing 31.6 μM NAA, and the auxin-free medium. In addition, transfer to soil and acclimation was easier for plantlets rooted in MeJA-containing medium.

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Anthers from more than 17000 flowers of 19 bell pepper Capsicum annuum L. hybrids (provided by `Semillas Fitó S.A.') were cultured in a double layer modified H medium (Nitsch and Nitsch, 1969) supplemented with 0.5 % activated charcoal and 0.26 % Gelrite in the solid phase. Significant differences between genotypes were observed on embryogenesis (472.3 to 9.7 embryos / 100 flowers) and number of plants rescued (4.0 to 0.3 plants / 100 flowers). Trying out maltose, malt extract, and sucrose. as carbohydrates, at 20, 40, 60 or 80 g/l, gave significantly better results for maltose (20 or 40 g/l). In addition, maintaining the anther cultures in an atmosphere enriched with 600 ppm CO2 was beneficial for embryo number, embryo development and number of rescued plants. Isocitrate dehydrogenase zymograms from leaf extracts indicate the microspore origin of the acclimated plants. Flow citometry of nuclei was used to determined an early diploidization of 70 % of the acclimated plants.

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Callus and shoot organogenesis were obtained from anthers of Dianthus caryophyllus L. `Manon', `Amapola', `Elsy', and `IB212', harboring mid-uninucleated microspores. Significant differences between genotypes were observed on number of responsive anthers (10.4% to 72.1%) and rescued plants per responsive anthers (1.2% to 4.8%). A modified H medium (Nitsch and Nitsch, 1969) with 20 g/L maltose and 0.25% Gelrite, supplemented with 10 μM 2,4-D and 1 μM TDZ, was most appropriate for callus induction. Plants were regenerated after subsequent subculture to the same medium, but amended with 0.1 μM TDZ. Zymogram types for aminopeptidase (AAP) in polyacrilamide gel electrophoresis proved that all 40 plants regenerated from `Amapola', `Elsy', or `IB212' where heterozygous, and consequently not originated from the microspores but from somatic tissue. Alternatively, in situ-induced parthenogenesis through pollination with gamma-irradiated pollen and in vitro embryo rescue was tested. A total of 92 embryos, including normal and no cotyledonary embryos, were rescued from 38 fruits harvested out of 70 crosses between `Scania' and `Amapola'. Embryos were rescued 21 to 28 days after pollination by culture in a modified E20A (Sauton and Vaulx, 1987) medium. Phosphogluco isomerase (PGI) and Shikimic dehydrogenase (SDH) zymograms in starch gel electrophoresis, and AAP in polyacrilamide gel electrophoresis, indicated the parthenogenic origin of three of the regenerated plants. Flow cytometry of nuclei proved the early diploidization, during in-vitro micropropagation, of the parthenogenic carnation haploid plantlets.

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