Resumo Apuleia leiocarpa Vogel J. F. Macbr. é uma espécie arbórea nativa do Brasil que possui madeira de alta qualidade e, atualmente, se encontra ameaçada de extinção. Mudas seminais de grápia dificilmente são comercializadas, sendo técnicas de propagação vegetativa, a exemplo da micropropagação, uma alternativa a ser considerada para produção de mudas. No entanto, há uma falta de informação disponível sobre a introdução in vitro de segmentos nodais e pouca informação está disponível sobre o enraizamento adventício de grápia. Como tal, existem lacunas de conhecimento que precisam ser abordadas para definir um protocolo de micropropagação para essa espécie. Assim, o objetivo deste estudo foi avaliar diferentes soluções de desinfestação na introdução de segmentos nodais e de diferentes tipos e concentrações de auxinas no enraizamento in vitro de explantes de grápia. Para a introdução, foram testados diferentes tipos de solução de desinfestação e tempos de imersão. Para o enraizamento, foram avaliadas diferentes concentrações e tipo de auxina. O melhor tratamento de desinfestação foi a imersão em solução de hipoclorito de sódio (NaOCl) a 2,5% de cloro ativo por 5 minutos. Para o enraizamento, o tratamento com 1,0 mg L-1 de ácido naftalenoacético (ANA, C10H7CH2CO2H) resultou nas maiores taxas enraizamento adventício. Os segmentos nodais de grápia podem ser estabelecidos in vitro, consistindo em boa fonte de propágulos para a micropropagação, e o enraizamento pode ser favorecido pela aplicação de ANA.

Abstract Apuleia leiocarpa Vogel J. F. Macbr. is a tree species native to Brazil that has high wood quality, and is currently threatened with extinction. Seminal seedlings of grápia are rarely commercialized, and vegetative propagation techniques, such as micropropagation, are considered as alternative methods for the production of grápia trees. However, there is a lack of available information about the in vitro introduction of nodal segments, and little information is available regarding in vitro rooting of grápia trees. As such, there are knowledge gaps that need to be addressed in order to define a micropropagation protocol for this species. Thus, the objective of this study was to evaluate different disinfestation solutions in the introduction of nodal segments and of different types and concentrations of auxins in the in vitro rooting of grápia explants. For the introduction of nodal segments, different types of disinfestation solutions and immersion times were evaluated. For rooting, different concentrations and types of auxins were evaluated. The best disinfestation treatment was found to be immersion in sodium hypochlorite (NaOCl) solution with 2.5% active chlorine for 5 minutes. For rooting, treatment with 1.0 mg L-1 of naphthalene acetic acid (NAA, C10H7CH2CO2H) resulted in the highest rates of adventitious rooting. Grápia nodal segments can be established in vitro, and provide a good source of propagules for micropropagation, and rooting can be favored through the application of NAA.

Abstract: This study aimed to evaluate half-sibling progenies for developing breeding strategies and to select mate clones with adventitious root competence, required for vegetative propagation by mini-cuttings. For rooting, single-bud mini-cuttings of 2 cm length were treated with indolebutyric acid at 2000 mg L-1. At 60 days of cultivation, the mini-cuttings were evaluated for percentages of survival and rooting, number and mean length of the three largest roots, and number of rooted mini-cuttings per mini-stump. These data were analyzed considering the mixed model described for a completely randomized design, an environment, half-sibling progenies, and a single plant per plot, by using the SELEGEN-REML/BLUP software. The 50 genotypes with the highest number of rooted mini-cuttings were selected. The genetic gain was 48.41% for the number of rooted mini-cuttings per mini-stump. Selection for adventitious rooting based on the number of rooted mini-cuttings can be used in mate breeding programs for vegetative propagation by mini-cuttings.

Abstract Leaves of mate is one of the main non-timber forest products marketed in South America, which makes establishment of new plantations of great interest. However, vegetative propagation of mate plantlets presents difficulties, which may be associated with the complexity of adventitious root formation. The aims of this study were to anatomically characterize the adventitious roots of mate-clone mini-cuttings and investigate the relationship of phenols and starch with adventitious rooting competence in mini-cuttings treated or not with indole-butyric acid (IBA). The mini-cuttings of four clones were collected at 0, 30, and 60 days of cultivation, fixed in a solution containing 1% glutaraldehyde and 4% formaldehyde, pre-infiltrated and infiltrated in (2-hydroxyethyl) methacrylate, and sectioned in a microtome. Ferric chloride and toluidine blue were used to detect phenolic compounds and lugol to identify starch. Adventitious roots formed in mini-cuttings treated with IBA presented disorganized xylem and phloem and poles irregularly but exhibited sclerenchyma vessel elements and tracheid cells indicating functionality. Differences in the rhizogenic ability of mate clones mini-cuttings were not due to the presence of anatomical barriers or the accumulation of phenolic compounds but be associated with the presence and distribution of starch grains in vegetative propagules.

Resumo O objetivo deste trabalho foi avaliar o efeito de épocas do ano e de clones na produtividade das minicepas e no enraizamento de miniestacas de erva-mate (Ilex paraguariensis A. St.-Hil.) tratadas ou não com ácido indolbutírico (AIB). Com esta finalidade, nas quatro épocas do ano realizou-se a coleta de brotos das minicepas de quatro clones de erva-mate, sendo que em cada coleta foi avaliada a porcentagem de sobrevivência das minicepas e o número de miniestacas obtidas por minicepa. Posteriormente, os brotos foram seccionados em miniestacas de gema única, as quais foram ou não tratadas com 2000 mg L-1 de AIB. Aos 30 e 60 dias foram avaliadas as porcentagens de sobrevivência, enraizamento e calogênese, número e comprimento das raízes. As minicepas dos quatro clones de erva-mate apresentaram alta sobrevivência, além de produção constante de miniestacas dos clones 06SM17, 06SM12 e 06SM15 durante as quatro épocas do ano. O tratamento com AIB não incrementa o enraizamento das miniestacas de erva-mate. O processo rizogênico das miniestacas varia conforme o genótipo e a sazonalidade. O clone 06SM15 apresenta a maior capacidade rizogênica nas quatro épocas do ano. As melhores porcentagens de enraizamento são verificadas nas épocas do outono e/ou inverno para os quatro genótipos avaliados.

Abstract The objective of this work was to evaluate the effect of season and clone in the productivity of the mini-stumps and the rooting of mini-cuttings of erva mate (Ilex paraguariensis A. St.-Hil.) treated or not with indolbutyric acid (IBA). Mini-cuttings were collected from mini-stumps of four clones during the four seasons of the year. In each collection, it was evaluated the percentage of mini-stumps survival and the number of mini-cuttings obtained per mini-stump. Single-budded mini-cuttings were treated or not with 2000 mg L-1 IBA before planting. At 30 and 60 days, the percentages of survival, rooting and callus formation in mini-cuttings, the number and length of roots were evaluated. The mini-stumps of the four erva mate clones showed high survival. The clones 06SM17, 06SM12 and 06SM15 had a uniform production of mini-cuttings per mini-stump during the four collection times. IBA treatment is not necessary for mini-cutting rooting of these clones. The mini-cutting rooting varied with the evaluated clone and collection time. The clone 06SM15 had the highest rhizogenesis capability in the four collection times. The highest rooting percentage was obtained during fall and winter seasons for the four clones.

RESUMO 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 acrescido 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.

ABSTRACT The aim of this work was to evaluate the in vitro productivity of grapia (Apuleia leiocarpa) micro-stumps cultivated in different media conditions to maximize its multiplication rate. Micro-stumps from aseptic seedlings were cultivated in WPM medium added with 0, 2.2, 4.4, 6.6 and 8.8 µM of 6-benzylaminopurine (BAP) and subcultured in WPM medium without cytokinin. Micro-stumps were also cultivated in WPM, MS or ½ MS base media with or without the addition of 1.5 g L-1 of activated charcoal and submitted to drastic pruning at 30, 60 and 90 days of cultivation. The in vitro cultivation of grapia micro-stumps in WPM or ½ MS media added with 8.8 µM of BAP and the subcultivation in media without cytokinin increase the number and length of shoots. The cultivation of grapia micro-stumps in media with activated charcoal increases the productivity of micro-cuttings per micro-stumps, but in the absence of activated charcoal induces callus formation and indirect organogenesis of shoots. The in vitro conservation of grapia microstumps maximizes the multiplication rate, being a feasible technique for the production of grapia plantlets.

Abtract Apuleia leiocarpa is an endangered tree species with high quality timber. Propagation by seed is difficult due to irregular fruiting and water-impermeable tegument, so asexual propagation is a promising alternative. This study aimed to evaluate in vitro and ex vitro rooting of A. leiocarpa and acclimatization of micropropagated plantlets. The hypothesis was that type of explant, indolebutyric acid (IBA) concentration and substrate influence the induction of adventitious roots in apuleia. For in vitro rooting, nodal segments and micro-cuttings were grown in Wood Plant Medium (WPM) with 0, 4.9, 9.8, 14.7 and 19.6 μM IBA. Compositions of commercial substrate + vermiculite + coarse sand and commercial substrate + vermiculite, in equal proportions, were tested for acclimatization. For ex vitro rooting, nodal segments and micro-cuttings were treated with 0 and 4920 μM IBA for 10 s and grown in equal proportions of: commercial substrate + vermiculite + coarse sand, commercial substrate + vermiculite, and commercial substrate + coarse sand. Nodal segments presented greater rooting in vitro than micro-cuttings without need for IBA supplementation in the culture medium. Substrate composition did not affect survival or growth of in vitro rooted plantlets during acclimatization. The best ex vitro rooting was found in nodal segments and explants treated with 4920 μM IBA and grown in commercial substrate + vermiculite + coarse sand. Ex vitro rooting is a promising technique for producing apuleia plantlets; however, adjustments to environmental conditions in both ex vitro rooting and acclimatization are necessary in order to maximize survival of micropropagated A. leiocarpa plantlets.

Resumen Apuleia leiocarpa es una especie de árbol en peligro de extinción con madera de alta calidad. Su propagación por medio de semillas es difícil por que la aparición de frutos es irregular y por el tegumento impermeable al agua; así la propagación asexual es una alternativa prometedora. El objetivo de este estudio fue evaluar el enraizamiento in vitro y ex vitro de A. leiocarpa y la aclimatación de plántulas micropropagadas. La hipótesis fue que el tipo de explante, la concentración de ácido indolbutírico (IBA) y el sustrato influyen en la inducción de raíces adventicias en A. leiocarpa. Para el enraizamiento in vitro, se cultivaron segmentos nodales y microcortes en un Medio de Plantación de Árboles (Wood Plant Medium) (WPM) con 0, 4.9, 9.8, 14.7 y 19.6 μM de IBA. Para la aclimatización se probaron mezclas de sustrato comercial + vermiculita + arena gruesa y sustrato comercial + vermiculita, en proporciones iguales. Para el enraizamiento ex vitro, se trataron segmentos nodales y microcortes con 0 y 4920 μM de IBA durante 10 s y se cultivaron en proporciones iguales de: sustrato comercial + vermiculita + arena gruesa, sustrato comercial + vermiculita y sustrato comercial + arena gruesa. Los segmentos nodales presentaron mayor enraizamiento in vitro que los microcortes sin suplemento de IBA en el medio de cultivo. La composición del sustrato no afectó la supervivencia o el crecimiento de las plántulas enraizadas in vitro durante la aclimatación. El mejor enraizamiento ex vitro se encontró en segmentos nodales y explantes tratados con 4920 μM IBA y cultivados en sustrato comercial + vermiculita + arena gruesa. El enraizamiento ex vitro es una técnica prometedora para producir plántulas de apuleia; sin embargo, se necesitan ajustes a las condiciones ambientales, en el enraizamiento ex vitro y en la aclimatación para maximizar la supervivencia de las plántulas micropropagadas de A. leiocarpa.

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.

ABSTRACT: 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.

O objetivo deste trabalho foi avaliar a produtividade de minicepas de louro-pardo (Cordia trichotoma) de origem seminal e assexuada, bem como o efeito do ácido naftaleno acético (ANA) no enraizamento das miniestacas. Mudas de origem seminal e do enraizamento de estacas radiculares foram podadas para formar o minijardim clonal. As minicepas foram fertirrigadas com três concentrações de solução nutritiva e avaliadas quanto à sobrevivência e produtividade de miniestacas. As miniestacas foram ou não tratadas com 1000mg L-1 ANA e plantadas em substrato para o enraizamento. As miniestacas foram avaliadas para as porcentagens de sobrevivência e enraizamento e para o número e comprimento total de raízes, aos 30 dias de cultivo. Não houve diferença quanto à sobrevivência e o número de miniestacas obtidas de minicepas de origem seminal ou assexuada. Miniestacas oriundas de minicepas de origem assexuada apresentaram maior capacidade de enraizamento e sobrevivência. A aplicação de 1000mg L-1 de ANA não favorece o enraizamento das miniestacas de louro-pardo.

This study aimed to evaluate the production of cordia (Cordia trichotoma) ministumps of seminal and asexual origin, as well as the effect of the naphthaleneacetic acid (NAA) on the rooting of mini-cutitings. Seedlings and plantlets from rooted cuttings were pruned to form a clonal mini-hedge. The ministumps were fertirrigated with three concentrations of nutrient solution and they were evaluated for survival and production of mini-cuttings. The mini-cuttings were or not treated with 1000mg L-1 of NAA and planted in substrate for rooting. The mini-cuttings were evaluated for the percentages of survival and rooting, and number and total length of roots after 30 days of cultivation. There was no difference in survival and number of mini-cuttings obtained from seminal or asexual ministumps. Mini-cuttings from ministumps of asexual origin showed higher rooting capability. The use of 1000mg L-1 NAA did not increase the rooting percentage of cordia mini-cuttings.

O objetivo deste trabalho foi avaliar o estabelecimento e crescimento in vitro de plantas de grápia (Apuleia leiocarpa (Vogel) J. F. Macbr.) em diferentes condições de cultivo. Sementes de grápia tratadas com ácido sulfúrico (H2SO4) por 20min foram imersas em etanol a 70% por 30s e em hipoclorito de sódio (0; 2,5 ou 5,0% de cloro ativo) por 5, 10 ou 15min. Aos 30 dias de cultivo, as sementes foram avaliadas quanto às porcentagens de desinfestação e germinação. As plantas assépticas foram transferidas para os meios de cultura WPM, MS e ½MS e avaliadas quanto ao comprimento (cm) da parte aérea e total das raízes e ao número de segmentos nodais e folhas aos 15 dias. Sementes de grápia também foram semeadas em meio de cultura WPM, suplementado com 4, 5 ou 6g L-1 de agar combinado com 10, 20 ou 30g L-1 de sacarose, e mantidos sob duas condições de luminosidade: luz durante todo o período de estabelecimento e escuro durante sete dias após a semeadura. Foram avaliados a porcentagem de germinação das sementes, o comprimento (cm) da parte aérea e total das raízes e o número de segmentos nodais e folhas aos 15 dias. Concluiu-se que os tratamentos com ácido sulfúrico e etanol foram suficientes para o estabelecimento in vitro de plantas assépticas de grápia. Plantas assépticas podem ser cultivadas em meio de cultura WPM ou MS, suplementados com 10g L-1 de sacarose e 4g L-1 de agar.

The aim of this study was to evaluate the in vitro establishment and growth of apuleia (Apuleia leiocarpa (Vogel) J. F. Macbr.) seedlings under different culture conditions. Apuleia seeds were treated with sulfuric acid (H2SO4) for 20min and disinfected by the immersion in 70% of ethanol for 30s and sodium hypochlorite with 0, 2.5 and 5.0% of active chlorine for 5, 10 and 15min. The percentage of disinfection and germination were evaluated at 30 days. Aseptic seedlings were transferred to WPM, MS and ½ MS culture medium and evaluated for shoot and root (cm) growth and number of nodal segments and leaves at 15 days. Apuleia seeds were also sown in WPM medium supplemented with 4, 5 or 6g L-1 of agar combined with 10, 20 or 30g L-1 of sucrose. The cultures were kept under two luminescence conditions: light throughout the establishment period and dark during the first seven days. The percentage of germination, shoot and root (cm) growth and number of nodal segments and leaves were evaluated at 15 days. In conclusion, the sulfuric acid and ethanol treatments were enough for the in vitro production of apuleia aseptic seedlings. The aseptic seedlings can grow in both WPM and MS medium, supplemented with 10g L-1 of sucrose and 4g L-1 of agar.