Background:We have developed a micropropagation technique methodology to clonal microgarden formation of Bambusa vulgaris selected adult plant. Collection site (i.e., stock plant cultivated in different environments) and seasonality of shoot collection (i.e., months of the year) effects on in vitro culture were evaluated. Explants that showed normal development were ex vitro rooted in miniincubator system, and three culture media (WPM, MS and deionized water + agar) supplemented with plant growth regulators (IBA, NAA and BAP) and antibiotic (streptomycin sulphate and culture medium free -control) were evaluated. Micropropagated plants were acclimatized in a shadow house and transferred to a semi-hydroponic system for establishment of a clonal micro-garden. The efficacy of the cloning protocol was determined with genetic fidelity analysis by ISSR molecular markers.Results: Considering all inoculations, 21.9% of nodal segments were in vitro established in nine shoot collections. The rooting percentage was 36.6%, and no interactions were observed between the use of culture medium and antibiotic. Culture medium free antibiotic resulted in 80.0% of survival and 50.0% of adventitious rooting. Micropropagated plants presented adequate growth and adaptation to ex vitro conditions in the clonal micro-garden. Molecular analyses by ISSR markers indicated the absence of genetic variations, and histological analyses revealed normal adventitious root formation originating from meristematic cells. Conclusion:The formation of a clonal micro-garden was demonstrated, proving the feasibility of the tested technique. Our results contribute to the development of a clonal propagation protocol for B. vulgaris selected adult plants.
Visando fornecer informações que sirvam de base para estudos de melhoramento genético de Eucalyptus foi realizada a análise da diversidade genética usando marcadores ISSR. As espécies estudadas foram Eucalyptus urophylla e Eucalyptus microcorys, ambas com potencial econômico florestal. Os indivíduos estudados pertencem a um teste de espécies e procedências instalado no ano de 1974 e permanecem isentos de tratos silviculturais. Para as análises foram utilizados nove primers ISSR universais. A partir dos resultados avaliou-se a existência de variação intra e interespecífica por meio da porcentagem de polimorfismo, conteúdo de informação polimórfica (PIC) e distância Euclidiana entre indivíduos. A fim de analisar a distância Euclidiana entre os indivíduos foram feitas a análise de coordenadas principais (PCoA) e análise permutacional de dispersão multivariada (PermDisp) seguida pelo teste de Tukey. Observouse elevada porcentagem de polimorfismo (57,14% para Eucalyptus microcorys e 80,95% para Eucalyptus urophylla). Considerando todos os grupos avaliados, os valores de PIC foram superiores a 0,5 para quatro primers (UBC827, UBC835, UBC841). A variabilidade interna foi significativamente maior na população de Eucalyptus urophylla em relação à Eucalyptus microcorys, o que pode estar associado à maior capacidade do Eucalyptus urophylla em hibridizar naturalmente.
Cordia trichotoma, popularly known in Brazil as 'louro-pardo', has numerous environmental and timber-related applications. This species is naturally propagated by seeds; however, this form of propagation results in restricted seedling production due to high genetic variability associated with a low degree of forest improvement. Thus, the cloning of selected adult trees is an alternative for producing high-quality plants. The aims of this work were to evaluate the emission of epicormic shoots from pruned branches and to define a protocol for the in vitro establishment of C. trichotoma selected adult trees. Propagules used in these experiments were derived from the selection of four C. trichotoma adult trees. The branches were placed in a greenhouse to induce epicormic shoots. Number of shoots according to the climatic season (autumn and spring) and shoot collection time were evaluated. Three sodium hypochlorite concentrations were evaluated for five minutes (C 1 = 0.58 % active chlorine, C 2 = 1.12 % active chlorine, and C 3 = 2.12 % active chlorine) for in vitro establishment. Results indicated that the epicormic shoot technique could be considered viable for shoot emission of C. trichotoma, regardless of the climatic season (autumn or spring). Sodium hypochlorite solution applied for 5 minutes (1.12 % of active chlorine) was effective for the in vitro establishment of nodal segments from epicormic shoots, resulting in 61.3 % of established explants. These results are promising for the cloning of adult trees during the mature stage of development in the field without the need to cut the selected tree.
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