Cultivars of auricula (Primulaϫpubescens Jacq.) are descendants of a natural cross between P. auricula and P. hirsuta, which is a large family of plants comprising many thousands of hybrids. They have become popular as pot flowers in not only England but also other countries. All cultivars of auricula are hybrids and vegetative propagated by taking offsets from donor plants. Establishment of system for plant regeneration from cultured cells and tissues is necessary for micropropagating new cultivars and conserving the valuable old cultivars in danger of extinction.There have been a few reports on plant regeneration from cultured cells and tissues of Primula species. Coumans et al. (1979) reported that young floral buds of P. obconica regenerated adventitious shoots when cultured on media with BA and NAA. Shimada et al. (1997) reported that regeneration of adventitious shoots and somatic embryos from leaf explants of P. cuneifolia var. hakusanensis was stimulated by TDZ or zeatin. Yamamoto et al. (1999) obtained regenerated plants from young expanding leaves of P. sieboldii on media with BA and NAA. Mizuhiro et al. (2001a) succeeded in plant regeneration from cell-suspension-derived protoplasts of P. malacoides and P. obconica, and produced somatic hybrids between them (Mizuhiro et al. 2001b). Plant regeneration from leaf calli of P. vulgaris and P. elatior was stimulated by a high level of TDZ Schwenkel 2002, Schween andSchwenkel 2003). In the present study, we investigated the effect of cytokinins on adventitious shoot regeneration from leaf explants of auricula cultivars.Four auricula cultivars, 'Borders Mixed', 'Border Stripes', 'Alpines Mixed' and 'Field House Mixed', were used in the present study. Seeds of each cultivar were kept in a refrigerator (4°C) until use. Seeds were surfacesterilized with a NaOCl solution containing 3% (v/v) active chlorine for 15 min followed by rinsing three times with sterilized distilled water, and then sown on plant growth regulator-free LS (Linsmaier and Skoog 1965) medium supplemented with 30 g l Ϫ1 sucrose and 2.5 g l Ϫ1 of gellan gum. After three-months of culture, seedlings grew into plantlets with 4 to 5 expanded leaves. Leaves were harvested from these plantlets, cut into 15ϫ15 mm pieces and placed on LS media supplemented with 30 g l Ϫ1 sucrose, 2.5 g l Ϫ1 gellan gum and various concentrations (0-2 mg l Ϫ1) and combinations of cytokinins (BA, Zeatin or TDZ) and NAA. For each medium, 30 leaf explants were cultured. Plant growth regulators were added to the medium and the medium was adjusted to pH 5.9 prior to autoclaving. All cultures were incubated at 20°C under a 14-h photoperiod with fluorescent lighting (about 3,000 lux). Abstract Adventitious shoots were regenerated from leaf explants of 4 cultivars of auricula (Primulaϫpubescens Jacq.) on media supplemented with various cytokinins combined with NAA. One to four adventitious shoots per leaf explant were regenerated. The highest frequency of adventitious shoot formation was 46.6% for 'Borders Mixed' on the medium c...
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