Alterações celulares em ápices caulinares e estabilidade genética de plantas de Eucaliptos submetidos à criopreservação / Cellular alterations in kaolinitic apexes and genetic stability of Eucalyptus plants submitted to cryopreservation
DOI:
https://doi.org/10.34117/bjdv6n12-042Keywords:
Criolesão, Crioprotetores, Citometria de Fluxo, Microscopia Eletrônica.Abstract
O eucalipto é amplamente explorado, principalmente para produção de papel e na construção civil , o que gera uma alta demanda por mudas de eucalipto, tendo-se a necessidade de buscar técnicas biotecnológicas para aumentar a produtividade e conservar o germoplasma de híbridos elite. Dentre essas técnicas a criopreservação apresenta alto potencial para conservação em longo prazo de genótipos de interesse. Os objetivos deste trabalho foram desenvolver um protocolo de criopreservação por meio da técnica droplet-vitrification para o híbrido comercial Eucalyptus grandis X Eucalyptus urophylla, identificar alterações celulares nos ápices caulinares cripreservados, e verificar estabilidade genética das plantas. Ápices caulinares foram pré-cultivados em meio MS com sacarose a 0,2 M e 0,5 M, por 24 horas. Posteriormente, os ápices foram dispostos em papel alumínio contendo uma gota de PVS2 à 0 ºC, antes da imersão em nitrogênio líquido (NL), em diferentes tempos (20, 40, 60, 80, 100 e 120 minutos). Os ápices foram reaquecidos por 10 minutos em solução de descarregamento a 25ºC e transferidos para o meio de regeneração MS suplementado com 0,14 µM de BAP. Após 60 dias, avaliaram-se a sobrevivência, brotações e a estabilidade genética das plantas por citometria de fluxo. Os ápices caulinares nos tempos: recém excisados, submetidos à criopreservação, e após uma semana de recuperação foram coletados para análise de histologia e Microscopia Eletrônica de Varredura. O tempo de 60 minutos de PVS2 seguido de imersão em nitrogênio líquido proporcionou as maiores taxas de sobrevivência (66%), e número de brotações (6 brotos/ápice). O conteúdo estimado de DNA nuclear das plantas de eucalipto tratadas com PVS2 e criopreservadas não apresentaram diferenças estatísticas, quando comparado com as plantas do controle. As observações histológicas e ultraestruturais revelaram que o protocolo de Droplet-vitrification estabelecido com 60 minutos de exposição ao PVS2 causou menores danos celulares dos ápices caulinares.
Alterações celulares em ápices caulinares e estabilidade genética de plantas de Eucaliptos submetidos à criopreservação /
Cellular alterations in kaolinitic apexes and genetic stability of Eucalyptus plants submitted to cryopreservation
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