Plant regeneration from protoplasts of alfalfa (Medicago sativa L.) and optimization of the chemical of fusion technique between M. sativa L. and M. arborea L.
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| Author(s): |
Mariza Monteiro
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | Piracicaba. |
| Institution: | Universidade de São Paulo (USP). Escola Superior de Agricultura Luiz de Queiroz (ESALA/BC) |
| Defense date: | 2001-01-24 |
| Advisor: | Maria Lucia Carneiro Vieira |
| Abstract | |
Protocols of culture and plant regeneration from leaf and cotyledonary protoplasts were used for the following alfalfa genotypes: Crioula, Alfa 200, Valley Plus, Semit 921 and Rangelander. Microcally were obtained after 28 days of culture which were submitted to two treatments: (a) liquid medium K8 (60 days) where differentiated structures were produced and identified by anatomical analysis as somatic embryos and roots for the genotypes Rangelander and Semit 921, respectively; (b) solidified medium B5h (28 days), subculture in SHb (21 days) for embryogenesis induction, and subculture in Boi2Y (28 days) where embryo differentiation was observed on the surface of the M. sativa cv. Rangelander calli. In this medium somatic embryo development with shoot and root formation occurred. Plantlet acclimatization was carried out in the greenhouse conditions where plant development occurred. For establishing the fusion procedures, protoplasts were isolated from M. sativa cv. Rangelander calli and from M. arborea mesophylls. The treatment using 20-30% PEG solution for 15 min. favoured the heterokarion formation (4-8%) without reducing the cellular viability. (AU) | |