The effectiveness of thidiazuron (TDZ) in promoting in vitro organogenesis in cultured embryo axes and cotyledons from two Spanish Mediterranean provenances of Pinus pinaster Aiton is reported. Application of 44.4 μM TDZ as a 6-d pulse on DCR medium and subsequent transference of maritime pine explants to the same medium without growth regulators, but supplemented with activated charcoal, produced between 17 and 19 axillary shoots per embryo axis with no differences between the two provenances. The same sequential treatment allowed adventitious shoot differentiation on 75% of the cotyledons, with a mean of 12.2 shoots regenerated per explant. Axillary and adventitious shoots were rooted ex vitro after an auxin pulse of indole-3-butyric acid and 1-naphthaleneacetic acid. The micropropagation protocols presented here should enable vegetative multiplication of selected families, and elite trees from these Spanish Mediterranean provenances of maritime pine.
Agriculture and modern biotechnology are increasingly becoming interdependent, and many new techniques have brought new opportunities for enhancing production and marketing. Germplasm storage is an alternative for the conservation of plant genetic diversity, contributing to the improvement and maintenance of propagation programs for species of interest. In this work, banana corms were collected as plant material from relatively young commercial plantations of three different cultivars: 'Williams', Valery (AAA genome; Cavendish subgroup), and 'Barraganete' (AAB genome; Plantain subgroup). Their shoot tips were introduced into in vitro conditions, and subcultured monthly to obtain the required number of shoots. The shoots were subsequently rooted and stimulated to invigoration in order to extract apical meristems (0.8-1.0 mm), which were prepared for cryopreservion in liquid nitrogen (−196 °C) following preconditioning in PVS2 vitrification solution. Thereafter, the explants were rapidly thawed and then recovered and regenerated using two different methods -by Panis (2009) and Korneva et al. (2009) -consisting of two different sets of recovery and subsequent regeneration media. Statistical analysis of the results showed that the banana cultivar 'Williams' demonstrated higher survival and regeneration rates after cryopreservation using the Korneva method, whereas in cultivars 'Valery' and 'Barraganete', there were no significant differences between the tested methods. The 'Barraganete' cultivar had the lowest survival and regeneration rates, regardless of the applied method.
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