ABSTRACT Background: We have developed a micropropagation technique methodology to clonal micro-garden 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 mini-incubator 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.

Resumo 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. Observou-se 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.

Abstract Here, we deployed a genetic diversity analysis using ISSR markers in two Eucalyptus species, as a fundamental step toward breeding strategies. The selected species were Eucalyptus urophylla and Eucalyptus microcorys, both with economic potential in forestry. The studied individuals belong to a species and the provenance test that was installed in 1974 and until now remains exempt from silvicultural treatments. Nine universal ISSR primers were used in the analyses. Intra and interspecific variation through the polymorphism percentage, polymorphism information content (PIC) and Euclidean distances among individuals were computed. The Euclidean distance among the individuals was used to perform a principal coordinate analysis (PCoA), as well as the permutation analysis of multivariate dispersion (PermDisp), followed by the Tukey’s test. A high percentage of polymorphic bands were detected, with 57.14% for Eucalyptus microcorys and 80.95% for Eucalyptus urophylla. PIC values were higher than 0.5 for four primers (UBC827, UBC835, UBC841, and UBC842). The genetic variability was significantly higher within the Eucalyptus urophylla population than in Eucalyptus microcorys, which may be associated with the greater ability of Eucalyptus urophylla to naturally hybridize, therefore, enabling the introduction of new alleles to their populations.

ABSTRACT The relevance of Eucalyptus dunnii has been evidenced mainly for its wood quality and cold tolerance among cultivated subtropical eucalypts. However, rooting is a challenge for its propagation, particularly when adult material is involved. This study aimed to assess the mixotrophism on the in vitro elongation and adventitious rooting phases in Eucalyptus dunnii microcutting. The experimental material used was obtained from a ministumps of Eucalyptus dunnii clones. In order to evaluate gas exchange and sucrose supplementation on in vitro elongation and adventitious rooting, the experiment was prepared in a 3×4 factorial arrangements with three forms of sealing (rigid polypropylene caps with no membrane (0/M), with a membrane (1/M), with three membranes (3/M) and four sucrose concentrations (0, 10, 20 and 30 g.L-1). At 30 days in the elongation phase it was evaluated (length, number of shoots per explant, oxidation, bud vigor, pigment content, leaf area and anatomy) and rooting (length, root diameter and rooting). Results show that sucrose should be added in the culture medium for in vitro elongation and can be reduced to concentrations between 10 and 20 g.L-1. In vitro rooting requires the use of 30 g.L-1 of sucrose. The use of flasks with membranes that allow gas exchange is an effective alternative to promote the in vitro elongation and adventitious rooting of Eucalyptus dunnii microcutting.

ABSTRACT Khaya senegalensis, is a promising species for forest production, as it has excellent timber properties and adapts well to local edaphoclimatic conditions. To achieve better results in plant production and growth, some factors are important, such as nutrients, water and light. Light interferes directly with photosynthesis, plant growth and biomass. This study aims to investigate the morphological response and nutrient accumulation in seedlings of Khaya senegalensis under luminosity levels. An experiment was installed in the forest nursery during 150 days. The seedlings were distributed in a completely randomized design with five treatments in luminosity levels (full sun, 70%, 50%, 30% and natural shade), and 16 replicates. Height and diameter were measured in 30 days intervals. At the end of the experiment, mass weights (leaf, stem, root and total) were measured, and the root-shoot ratio and seed quality index proposed by Dickson were calculated. The accumulated macro and micronutrient contents were determined from the dry leaf material. The highest plants, diameters and total dry matter were found at levels of 70%, 50%, 30%, respectively. Plants exposed to natural shade grew less, in relation to plants with higher luminosity levels. As for nutrient accumulation, plants under natural shade accumulated higher levels of iron, while plants in the full sun accumulated more nitrogen contents. Plants with partial light conditions (70%, 50%, and 30%) were heavier in total biomass. Thus, the initial growth of Khaya senegalensis in low light conditions is compromised, whereas partial luminosities favor the growth and quality of seedlings.