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Master's Dissertation
DOI
https://doi.org/10.11606/D.11.2019.tde-20191218-175543
Document
Author
Full name
Mariza Monteiro
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 2000
Supervisor
Title in Portuguese
Regeneração de plantas a partir de protoplastos de alfafa (Medicago sativa L.) e otimização da técnica de fusão química entre M. sativa L. e M. arborea L.
Keywords in Portuguese
ALFAFA
PROTOPLASTOS
REGENERAÇÃO
VARIEDADES VEGETAIS
Abstract in Portuguese
Foram utilizados protocolos de cultura e regeneração de plantas a partir de protoplastos derivados de explantes foliares e cotiledonares das variedades de alfafa: Crioula, Alfa 200, Valley Plus, Semit 921 e Rangelander. Após 28 dias de cultivo, obteve-se microcalos que foram submetidos a dois tratamentos: (a) meio líquido K8 (60 dias) onde houve formação de estruturas diferenciadas nas variedades Rangelander e Semit 921, identificadas por análises anatômicas como sendo embriões somáticos e raízes, respectivamente; (b) meio sólido B5h (28 dias), subcultivo em meio SHb (21 dias) para a indução de embriogênese, e subcultivo em meio Boi2Y (28 dias) no qual observou-se diferenciação de embriões na superfície dos calos de M. sativa var. Rangelander. Este meio também favoreceu a maturação dos embriões somáticos que desenvolveram parte aérea e radicular. As plântulas foram aclimatadas em casa de vegetação onde se desenvolveram. Para o estabelecimento dos protocolos de fusão, foram isolados protoplastos de calos de M. sativa var. Rangelander e de folhas de M. arborea. O tratamento em solução de PEG (20 a 30%) por 15 min. favoreceu a formação de heterocários (4 a 8%) sem haver redução drástica da viabilidade celular.
Title in English
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.
Abstract in English
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.
 
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MonteiroMariza.pdf (7.54 Mbytes)
Publishing Date
2019-12-19
 
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