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As the energizing and therapeutic effects of infusions of mate (Ilex paraguariensis A.St.-Hil) leaves are increasingly recognized, rising global consumption is also increasing the demand for plantlets with high genetic and morphological qualities. Mini-cuttings are an alternative for clonal plantlets; however, more details are needed on factors that affect the success of this technique. Here, toward maximizing production, we evaluated adventitious rooting competence of mini-cuttings of different sizes and the morphophysiological quality of the resultant plantlets. Small (one bud; length up to 2.5 cm), medium (multiple buds; length 2.6–5.0 cm), and large (multiple buds; length 5.1–10.0 cm) mini-cuttings were planted in a mixture of commercial substrate, vermiculite, and coarse sand (1:1:1 v:v:v), grown in a humidity chamber, then evaluated after 45, 60, 75, and 90 days for survival, percentage of callus and shoot formation, number and length of shoots, percentage of rooted cuttings, and number and length of roots. Any rooted mini-cuttings were then cultivated in commercial substrate and subsurface soil (2:1 v:v) and evaluated after 30, 60, 90 and 120 days of cultivation for morphophysiology of the plantlets, percentage of survival, number of leaves, shoot height, stem diameter, and ratio of shoot height to stem diameter. After 120 days, the plantlets were evaluated for the ratio of shoot and root dry mass, total length, surface area, total volume, and mean diameter of roots and Dickson quality index. Adventitious rooting of the mini-cuttings and the morphophysiological quality of the plantlets were not influenced by the size of the vegetative propagule. Therefore, single-bud mini-cuttings have similar adventitious rooting competence, and result in plantlets with adequate morphophysiological quality, based upon leaf development, shoot height, stem diameter and length, and surface area, volume and diameter of roots. Single-bud mini-cuttings up to 2.5 cm long can thus be used to maximize production of mate plantlets by mini-cuttings.

The objective of this work was to define an early selection strategy to identify Cabralea canjerana (Meliaceae) clones with high multiplication rate. A mini-garden of 109 clones of canjerana seedlings was established in a completely randomized design, in an acclimatized greenhouse. From seedlings, the mini-stumps and mini-cuttings were obtained. Mini-cuttings were collected at five different times, and the number of mini-cuttings per mini-stump, rooting percentage, and number of rooted mini-cuttings were quantified. The number of rooted mini-cuttings per mini-stump was the only trait that showed high correlation with the others. Five groups of clones based on the number of rooted mini-cuttings per mini-stump were separated using k-means clustering, and the genetic gain from selection and Pearson correlation were estimated. The selection of the two best groups in each evaluation period resulted in high genetic gains from selection for all evaluated traits. Early selection for the number of rooted mini-cuttings discarded 65% of the evaluated clones, which increases experimental precision in evaluations of traits associated with plantlet growth and quality. Early selection for the number of rooted mini-cuttings per mini-stump at different times allows the identification of Cabralea canjerana clones with high multiplication rate. Resumo: O objetivo deste trabalho foi definir uma estratégia de seleção precoce para identificar clones de Cabralea canjerana (Meliaceae) com alta taxa de multiplicação. Um minijardim com 109 clones de plântulas de canjerana foi estabelecido com um delineamento inteiramente casualizado em casa de vegetação climatizada. Das plântulas, foram obtidas as minicepas e as miniestacas. As miniestacas foram coletadas em cinco épocas, e foram quantificados o número total de miniestacas por minicepa, a percentagem de enraizamento e o número de miniestacas enraizadas. O número de miniestacas enraizadas por minicepa foi o único caráter que apresentou alta correlação com os demais. Foram estabelecidos cinco grupos de clones com base no número de miniestacas enraizadas por minicepas, pelo agrupamento das k-médias, e estimados o ganho genético de seleção e a correlação de Pearson. A seleção dos dois melhores grupos em cada época resultou em altos ganhos de seleção para os caracteres avaliados. A seleção precoce para o número de miniestacas enraizadas descartou 65% dos clones avaliados, o que aumenta a precisão experimental em avaliações de caracteres associados ao crescimento e à qualidade das mudas. A seleção precoce do número de miniestacas enraizadas por minicepas, em diferentes épocas, permite identificar clones de Cabralea canjerana com alta taxa de multiplicação.

O objetivo deste trabalho foi avaliar a produtividade de microcepas de grápia (Apuleia leiocarpa) mantidas em diferentes condições de cultivo in vitro, visando maximizar a taxa de multiplicação. Microcepas oriundas da poda drástica de plantas assépticas foram cultivadas em meio de cultura WPM acrescida de 0; 2,2; 4,4; 6,6 e 8,8 µM de 6-benzilaminopurina (BAP) e subcultivadas em meio de cultura WPM sem citocinina. Microcepas de grápia também foram cultivadas em meio de cultura WPM, MS ou ½ MS, acrescido ou não de 1,5 g L-1 de carvão ativado e submetidas a três coletas de brotos aos 30, 60 e 90 dias de cultivo. A manutenção das microcepas em meio de cultura WPM ou ½ MS suplementado com 8,8 µM de BAP, seguido do subcultivo em meio de cultura sem citocinina aumenta o número e o crescimento de brotações. O cultivo de microcepas de grápia em meio de cultura com carvão ativado aumenta a produção de microestacas por microcepas, enquanto a ausência de carvão ativado favorece a formação de calos e brotos por organogênese indireta. A manutenção in vitro de microcepas de grápia maximiza a taxa de multiplicação, constituindo-se de uma técnica promissora para a produção de plantas de grápia.

The objective of this work was to evaluate different solvents and times in the ultrasound-assisted extraction of polyphenols, in order to quantify the total phenolic and flavonoid contents in and the antioxidant activity of mate (Ilex paraguariensis) leaves. The experiment was carried out in a completely randomized design, in a 8x3 (solvent x time of extraction) factorial arrangement. The solvent extractors were distilled water and aqueous solutions of ethanol, acetone, and methanol, acidified or not with hydrochloric acid, and the time of ultrasound-assisted extractions were 15, 30, and 45 min. All extractions were finished with a mechanical agitation of 30 min. Samples treated with acidified hydroalcoholic solvents resulted in higher values of total phenolics and flavonoids and antioxidant activity. The extraction using acidified hydroethanol solvent was not affected by the time in the ultrasound. The contents of total phenolics and flavonoids were highly correlated with each other and with the antioxidant activity, especially when it was determined by the ferric-reducing antioxidant power method. Acidified hydroethanol solvent in ultrasound for 15 min is the most suitable extraction condition for the quantification of the contents of total phenolics and flavonoids in and the antioxidant activity of mate leaves. Resumo: O objetivo deste trabalho foi avaliar diferentes solventes e tempos de extração de polifenóis com ultrassom, para quantificar os compostos fenólicos e flavonoides totais e a capacidade antioxidante em folhas de erva-mate (Ilex paraguariensis). O experimento foi realizado em delineamento inteiramente casualizado, em arranjo fatorial 8x3 (solvente x tempo de extração). Os solventes foram água destilada e as soluções aquosas de etanol, acetona e metanol, acidificados ou não com ácido clorídrico, e os tempos de extração em banho ultrassônico foram de 15, 30 e 45 min. As extrações foram concluídas com agitação mecânica de 30 min. As amostras tratadas com solventes hidroalcoólicos acidificados apresentaram os maiores valores de compostos fenólicos e flavonoides totais e capacidade antioxidante. A extração com hidroetanol acidificado não foi afetada pelo tempo em ultrassom. Houve correlação entre os teores de compostos fenólicos e flavonoides totais e destes com a capacidade antioxidante, especialmente quando esta foi determinada pelo poder antioxidante redutor do íon ferro. O solvente hidroetanólico acidificado em ultrassom por 15 min é a condição de extração mais apropriada para a quantificação dos teores de compostos fenólicos totais e flavonoides e da capacidade antioxidante em folhas de erva-mate.

The competence of adult plants of mate (Ilex paraguariensis St. Hil.) to undergo adventitious rooting is essential to their vegetative rescue by cuttings. This study aimed to evaluate the adventitious rooting competence of adult mate plants, and then rescue these plants by making cuttings from epicormic shoots induced in different trunk positions. The percent survival and rooting of cuttings from epicormic shoots induced by girdling in seven adult plants were evaluated. The two stock plants whose cuttings had the highest rooting competence were further pruned and girdled to evaluate the effect of the position of epicormic shoots on their adventitious rooting competence. The potential success of rescuing of the same adult plants was also studied with cuttings of epicormic shoots induced by coppicing. The shoots were sectioned in single-bud cuttings, either treated with 3,000 mg L-1 indolebutyric acid or untreated, and then cultivated in a humid chamber for 60 days, and then the percent survival and rooting of cuttings was evaluated. In conclusion, we found that it is possible to rescue adult mate plants by cuttings from epicormic shoots induced by pruning, girdling, or coppicing. The adventitious rooting competence of cuttings differs among adult mate plants, and depends on the position of the epicormic shoots on the trunk from which epicormic shoots are collected.

The aim of this research was to evaluate the feasibility of rescuing adult plants by cuttings from post-fire epicormic shoots. We evaluated the native tree species erva mate ( Ilex paraguariensis St. Hil.) and canjerana ( Cabralea canjerana Vell. Mart.), which present difficult seedling production and are economically and ecologically important. The plant material used for the preparation of cuttings consisted of shoots from bud differentiation of tissue from the stem collar of the trees. One-bud cuttings of erva mate were treated with 4000 or 8000mg L-1 of indolebutiric acid (IBA) and the checks were not treated with auxins. The cuttings were grown in three different substrates: carbonized rice husks, vermiculite and coarse sand. The experiment was a factorial with a completely random design and five replicates of five cuttings. One-bud cuttings from the basal, intermediate and apical parts of canjerna shoots were treated with 0 or 6000mg L-1 of IBA and planted in commercial substrate and coarse sand (3:2 v/v). The experiment was a factorial with a completely random design and ten replicates of three cuttings. Cuttings from post-fire epicormic shoots can be used for rescuing erva mate and canjerana adult plants. Erva mate cuttings treated with 8000mgL-1 IBA have increased rooting when grown in coarse sand. Canjerana cuttings from the basal part of epicormic shoots have greater survival and rooting than those from intermediate and apical parts. RESUMO: O objetivo deste trabalho foi avaliar a possibilidade de resgatar árvores adultas por estaquia de brotos epicórmicos de rebrota pós-fogo. Foram estudadas a erva-mate ( Ilex paraguariensis St. Hil.) e a canjerana ( Cabralea canjerana Vell. Mart.), que são espécies nativas de difícil propagação por sementes e de relevante importância econômica e ecológica. O material vegetativo utilizado para o preparo das estacas foi constituído de brotos, provenientes da rebrota da diferenciação dos tecidos de gemas do colo de árvores matrizes com, aproximadamente, 20 anos de idade. Para a erva-mate, estacas de gema única foram tratadas com 4000 e 8000mg L-1 de ácido indolbutírico (IBA), e as testemunhas que não receberam tratamento auxínico. As estacas foram cultivadas em três diferentes substratos: casca de arroz carbonizada, vermiculita e areia de granulometria grossa. O experimento foi um fatorial no delineamento inteiramente casualizado, com cinco repetições de cinco estacas. Para canjerana, estacas basais, medianas e apicais de gema única, foram ou não tratadas com 6000mg L-1 de IBA e cultivadas em substrato comercial e areia de granulometria grossa (3:2 v/v). O experimento foi um fatorial no delineamento inteiramente casualizado, com dez repetições de três estacas. A estaquia de rebrota pós-fogo pode ser utilizada para o resgate de árvores adultas de erva-mate e canjerana. Estacas de erva-mate tratadas com 8000mg L-1 de IBA enraízam melhor em areia grossa. Estacas de canjerana, obtidas da porção basal das brotações, apresentam maior sobrevivência e enraizamento do que as medianas e apicais.

Apuleia leiocarpa (Vogel) J.F. MacBride is a hardwood species native to South America, which is at serious risk of extinction. Therefore, it is of prime importance to examine the genetic diversity of this species, information required for developing conservation, sustainable management, and breeding strategies. Although scarcely used in recent years, random amplified polymorphic DNA markers are useful resources for the analysis of genetic diversity and structure of tree species. This study represents the first genetic analysis based on DNA markers in A. leiocarpa that aimed to investigate the levels of polymorphism and to select markers for the precise characterization of its genetic structure. We adapted the original DNA extraction protocol based on cetyltrimethyl ammonium bromide, and describe a simple procedure that can be used to obtain high-quality samples from leaf tissues of this tree. Eighteen primers were selected, revealing 92 bands, from which 75 were polymorphic and 61 were sufficient to represent the overall genetic structure of the population without compromising the precision of the analysis. Some fragments were conserved among individuals, which can be sequenced and used to analyze nucleotide diversity parameters through a wider set of A. leiocarpa individuals and populations. The individuals were separated into 11 distinct groups with variable levels of genetic diversity, which is important for selecting desirable genotypes and for the development of a conservation and sustainable management program. Our results are of prime importance for further investigations concerning the genetic characterization of this important, but vulnerable species.

This study aimed to evaluate the rooting capability of apical and nodal mini-cuttings of ipe-roxo ( Handroanthus heptaphyllus ) in different substrates. Shoots produced from mini-stumps were fractioned into mini-cuttings 2-cm long apical or nodal mini-cuttings and treated with 1000 mg L-1 of indolebutyric acid (IBA). Four substrate combinations were evaluated: commercial substrate composed of pine bark and vermiculite in proportions of 1:1, 2:1 and 1:2 (v /v) and commercial substrate, vermiculite and sand in the proportion of 1:1:1 (v/v). The factorial experiment utilized a completely random design, with five replicates of four mini-cuttings. Rooting, shooting and survival percentage, number and length of shoots and roots were evaluated at 60 and 90 days of growth. Commercial substrate and vermiculite are a good combination for rooting mini-cuttings of ipe-roxo. Apical and nodal mini-cuttings exhibit similar rooting behavior and both are feasible for production of ipe- roxo plantlets. RESUMO: O objetivo deste estudo foi avaliar o enraizamento de miniestacas apicais e nodais de ipê-roxo ( Handroanthus heptaphyllus ) em diferentes substratos. Brotos coletados de minicepas foram seccionados em miniestacas de 2cm de comprimento, classificadas em apical ou nodal e tratadas com 1000mg L-1 de ácido indolbutírico (AIB). Foram também testadas as composições de substrato comercial à base de casca de pinus e vermiculita nas proporções de 1:1; 2:1 e 1:2 (v/v) e de substrato comercial, vermiculita e areia grossa (1:1:1 v/v). O experimento foi um fatorial no delineamento inteiramente casualizado, com cinco repetições de quatro miniestacas. A porcentagem de enraizamento, de brotação e de sobrevivência, o número e comprimento de brotos e raízes foram avaliados aos 60 e 90 dias. Miniestacas de ipê-roxo cultivadas em substrato comercial e vermiculita apresentam maior capacidade de enraizamento. Miniestacas apicais e nodais apresentam similar comportamento de enraizamento, podendo ambas serem utilizadas para a produção de mudas de ipê-roxo.