Morpho-physiological and phytohormonal changes during the induction of adventitious root development stimulated by exogenous IBA application in Magnolia biondii Pamp

Abstract Magnolia biondii Pamp is an important ornamental tree species widely grown and used as a rootstock in the propagation of different Magnolia varieties. In the current studies, anatomical, physiological and endogenous hormones were studied to check the effect of IBA 750 mg/L on the adventitious rooting and to provide theoretical and technical support for the propagation of Magnolia biondii Pamp through stem cuttings. Two thousand stem cuttings were prepared and divided into two groups i.e., IBA treated cuttings and water control. For the evaluation of antioxidant enzyme activities, and endogenous hormones levels, samples were collected on the day of planting and each 5th day and further steps were carried out in the laboratory according to the protocols and proper precautions. For the anatomical observations, samples were collected on the 13th, 15th, and 17th day for IBA treated cuttings while 21st, 23rd, and 25th day for control. Collected samples were preserved in the FAA solution and further observations were carried out in the laboratory. Anatomical observations showed that it took 13 days for the differentiation of root primordia to the appearance of young adventitious roots in IBA treated cuttings, while it took 21 days to develop primordia in the control. Antioxidant enzyme activities involved in ROS were significantly higher in the IBA treated cuttings compared to control. POD showed a peak on the 13th day before the emergence of roots in IBA treated cuttings while it showed a peak on the 21st day in the control. PPO showed a peak on the 21st day in the IBA treated cuttings while it showed a peak on the 29th day in the control. SOD showed a peak on the 17th day in IBA treated cuttings, while it showed a peak on the 25th day in the control. Exogenous application of IBA enhanced the endogenous IAA and GA3 levels compared to CK, while it reduced the levels of ABA continuously at the time of rooting and then increased gradually. Inclusively, our study suggests that IBA 750 mg/L is efficient for the rooting of Magnolia biondii Pamp cuttings, as it enhanced the process of antioxidant enzyme activities, endogenous hormones levels and reduced the time of root formation which is evident from the anatomical observations.

Resumo Magnolia biondii Pamp é uma importante espécie de árvore ornamental muito cultivada e utilizada como porta-enxerto na propagação de diferentes variedades de Magnolia. Nos estudos atuais, hormônios anatômicos, fisiológicos e endógenos foram estudados para verificar o efeito do AIB na dose de 750 mg / L no enraizamento adventício e fornecer suporte teórico e técnico para a propagação de M. biondii Pamp por meio de estacas. Duas mil estacas foram preparadas e divididas em dois grupos, ou seja, tratadas com AIB e controle de água. Para a avaliação das atividades das enzimas antioxidantes e dos níveis de hormônios endógenos, as amostras foram coletadas no dia do plantio e a cada 5 dias, enquanto as demais etapas foram realizadas em laboratório de acordo com os protocolos e os devidos cuidados. Para as observações anatômicas, as amostras foram coletadas no 13º, 15º e 17º dias para estacas tratadas com AIB e no 21º, 23º e 25º dias para o controle. As amostras coletadas foram preservadas em solução FAA, e outras observações foram realizadas em laboratório. Observações anatômicas mostraram a necessidade de 13 dias para a diferenciação dos primórdios radiculares até o aparecimento de raízes adventícias jovens em estacas tratadas com AIB e de 21 dias para o desenvolvimento dos primórdios no controle. As atividades das enzimas antioxidantes envolvidas nas ROS foram significativamente maiores nas estacas tratadas com AIB em comparação com o controle. A POD apresentou pico no 13º dia antes da emergência das raízes nas estacas tratadas com AIB, enquanto no 21º dia apresentou pico no controle. A PPO teve pico no 21º dia nas estacas tratadas com AIB e no 29º dia no controle. A SOD apresentou pico no 17º dia nas estacas tratadas com AIB e no 25º dia no controle. A aplicação exógena de AIB aumentou os níveis endógenos de IAA e GA3 em relação ao controle, enquanto reduziu os níveis de ABA continuamente no momento do enraizamento e, em seguida, aumentou gradativamente. Inclusive, nosso estudo sugere que o AIB na dose de 750 mg / L é eficiente para o enraizamento de estacas de M. biondii Pamp, visto que potencializou o processo de atividades de enzimas antioxidantes e os níveis de hormônios endógenos, além de reduzir o tempo de formação de raízes, o que fica evidente nas observações anatômicas.

Morpho-physiological and phytohormonal changes during the induction of adventitious root development stimulated by exogenous IBA application in Magnolia biondii Pamp / Alterações morfofisiológicas e fito-hormonais durante a indução do desenvolvimento radicular adventício estimulado pela aplicação exógena do AIB em Magnolia biondii Pamp

Magnolia biondii Pamp is an important ornamental tree species widely grown and used as a rootstock in the propagation of different Magnolia varieties. In the current studies, anatomical, physiological and endogenous hormones were studied to check the effect of IBA 750 mg/L on the adventitious rooting and to provide theoretical and technical support for the propagation of Magnolia biondii Pamp through stem cuttings. Two thousand stem cuttings were prepared and divided into two groups i.e., IBA treated cuttings and water control. For the evaluation of antioxidant enzyme activities, and endogenous hormones levels, samples were collected on the day of planting and each 5th day and further steps were carried out in the laboratory according to the protocols and proper precautions. For the anatomical observations, samples were collected on the 13th, 15th, and 17th day for IBA treated cuttings while 21st, 23rd, and 25th day for control. Collected samples were preserved in the FAA solution and further observations were carried out in the laboratory. Anatomical observations showed that it took 13 days for the differentiation of root primordia to the appearance of young adventitious roots in IBA treated cuttings, while it took 21 days to develop primordia in the control. Antioxidant enzyme activities involved in ROS were significantly higher in the IBA treated cuttings compared to control. POD showed a peak on the 13th day before the emergence of roots in IBA treated cuttings while it showed a peak on the 21st day in the control. PPO showed a peak on the 21st day in the IBA treated cuttings while it showed a peak on the 29th day in the control. SOD showed a peak on the 17th day in IBA treated cuttings, while it showed a peak on the 25th day in the control. Exogenous application of IBA enhanced the endogenous IAA and GA3 levels compared to CK, while it reduced the levels of ABA continuously at the time of rooting and then increased gradually. Inclusively, our study suggests that IBA 750 mg/L is efficient for the rooting of Magnolia biondii Pamp cuttings, as it enhanced the process of antioxidant enzyme activities, endogenous hormones levels and reduced the time of root formation which is evident from the anatomical observations.

Magnolia biondii Pamp é uma importante espécie de árvore ornamental muito cultivada e utilizada como porta-enxerto na propagação de diferentes variedades de Magnolia. Nos estudos atuais, hormônios anatômicos, fisiológicos e endógenos foram estudados para verificar o efeito do AIB na dose de 750 mg / L no enraizamento adventício e fornecer suporte teórico e técnico para a propagação de M. biondii Pamp por meio de estacas. Duas mil estacas foram preparadas e divididas em dois grupos, ou seja, tratadas com AIB e controle de água. Para a avaliação das atividades das enzimas antioxidantes e dos níveis de hormônios endógenos, as amostras foram coletadas no dia do plantio e a cada 5 dias, enquanto as demais etapas foram realizadas em laboratório de acordo com os protocolos e os devidos cuidados. Para as observações anatômicas, as amostras foram coletadas no 13º, 15º e 17º dias para estacas tratadas com AIB e no 21º, 23º e 25º dias para o controle. As amostras coletadas foram preservadas em solução FAA, e outras observações foram realizadas em laboratório. Observações anatômicas mostraram a necessidade de 13 dias para a diferenciação dos primórdios radiculares até o aparecimento de raízes adventícias jovens em estacas tratadas com AIB e de 21 dias para o desenvolvimento dos primórdios no controle. As atividades das enzimas antioxidantes envolvidas nas ROS foram significativamente maiores nas estacas tratadas com AIB em comparação com o controle. A POD apresentou pico no 13º dia antes da emergência das raízes nas estacas tratadas com AIB, enquanto no 21º dia apresentou pico no controle. A PPO teve pico no 21º dia nas estacas tratadas com AIB e no 29º dia no controle. A SOD apresentou pico no 17º dia nas estacas tratadas com AIB e no 25º dia no controle. A aplicação exógena de AIB aumentou os níveis endógenos de IAA e GA3 em relação ao controle, enquanto reduziu os níveis de ABA continuamente no momento do enraizamento e, em seguida, aumentou gradativamente. Inclusive, nosso estudo sugere que o AIB na dose de 750 mg / L é eficiente para o enraizamento de estacas de M. biondii Pamp, visto que potencializou o processo de atividades de enzimas antioxidantes e os níveis de hormônios endógenos, além de reduzir o tempo de formação de raízes, o que fica evidente nas observações anatômicas.

Factores que afectan el enraizamiento adventicio y brotación en estacas de Hyptis australis (Lamiaceae) / Factors affecting adventitious rooting and sprouting in Hyptis australis (Lamiaceae) cuttings

Resumen Hyptis australis es una especie de la familia Lamiaceae, endémica del Bosque Atlántico, cuyo estado de conservación es crítico y se desconoce aún sus beneficios potenciales para el ecosistema y la sociedad. Con fines de rescate y conservación (ex situ/in situ), se plantea como necesidad, generar un protocolo de propagación vegetativa para facilitar y aumentar la disponibilidad de plantas. Con este objetivo, se estudiaron factores como el tipo de sustrato, tipo de estaca y hormona. Se utilizaron plantas de un año de edad, obtenidas de semillas cosechadas de las plantas ubicadas en la región sur de la provincia de Misiones (Argentina). Estacas apicales y subapicales fueron utilizadas en dos ensayos, en el primero se estudió la factibilidad de inducir raíces adventicias en los sustratos corteza de pino, arena y perlita. En el segundo ensayo, se evaluó la inducción de rizogénesis en estacas tratadas durante 30 minutos con una solución de 100 mg.kg -1 de ácido naftalenacetico (ANA) o ácido indolbutirico (AIB) en corteza de pino. Los mejores resultados se obtuvieron en el sustrato arena y en las estacas terminales. El ANA fue la hormona que generó mayor porcentaje de estacas apicales (80.00±14.14 %) y subapicales (84.00±16.73 %) con raíces. En las estacas subapicales el porcentaje de brotación fue mayor en ambos experimentos. La probabilidad de que una estaca de H. australis desarrolle raíces adventicia depende del tipo de sustrato, pero la adicción de ANA mejora notablemente el porcentaje de enraizamiento.

Abstract Hyptis australis is a species of the Lamiaceae family, endemic to the Atlantic Forest, whose conservation status is critical and its potential benefits for the ecosystem and society are still unknown. For rescue and conservation purposes (ex situ/in situ), the need arises to generate a vegetative propagation protocol to facilitate and increase the availability of plants. With this objective, factors such as the type of substrate, type of stake and hormone were studied. One-year-old plants were used, obtained from seeds harvested from plants located in the southern region of the province of Misiones (Argentina). Apical and subapical cuttings were used in two trials, the first studied the feasibility of inducing adventitious roots in pine bark, sand and perlite substrates. In the second test, the induction of rhizogenesis was evaluated in cuttings treated for 30 minutes with a solution of 100 mg.kg-1 of naphthalenacetic acid (ANA) or indole butyric acid (IBA) in pine bark. The best results were obtained in the sand substrate and in the terminal stakes. ANA was the hormone that generated the highest percentage of apical cuttings (80.00±14.14%) and subapical (84.00±16.73%) with roots. In the subapical cuttings the sprouting percentage was higher in both experiments. The probability that an H. australis cuttings will develop adventitious roots depends on the type of substrate, but the addition of ANA notably improves the rooting percentage.

Sexual and vegetative propagation of the medicinal Mexican species Phyllonoma laticuspis (Phyllonomaceae) / Propagación sexual y vegetativa de la especie medicinal mexicana Phyllonoma laticuspis (Phyllonomaceae)

Abstract:Phyllonoma laticuspis leaves are used in Carrizal de Bravo, Guerrero, Mexico, to heal skin lesions such as injuries and smallpox sequelae and to treat diabetes mellitus type 2, and organic extracts of these leaves have been reported to exert antibacterial effects. High demand of P. laticuspis as a medicinal plant has decreased its natural populations, and propagation of the species has not yet been reported. Therefore, the purpose of this study was to assess the vegetative propagation of the species through cutting and air layering, as well as its sexual propagation in a preserved population. For this, concentrations of 1 000, 4 000 and 6 000 ppm of a commercial root enhancer, with indole butyric acid (IBA), and a control treatment without IBA, were applied to the cuttings and air layers. Germination was evaluated under light and dark conditions using lots of freshly collected seeds and lots of seeds that had been stored for three months at 4 °C or 24 ± 2 °C. All experiments were performed in a completely randomized design. The cuttings did not develop roots in any concentration, whereas 100 % of the air layers rooted, displaying vigorous roots in the presence of 4 000 ppm IBA, after four month of treatment application. Regarding germination, more than 60 % of the freshly collected seeds germinated, whereas less than 20 % of the seeds stored at 4 °C, and close to 50 % of the seeds stored at 24 ± 2 °C germinated under light and dark conditions. No significant differences were observed between light and dark conditions, so they were categorized as indifferent photoblastic seeds. The observed moisture content of 13.5 % and germination behaviour as the response to cold storage, suggest that the resultant seed quality was intermediate. P. laticuspis propagation for short-term foliage production can be carried out in air layers, in populations with a high density of adult plants as a source of plant material and for the restoration of disturbed areas, in the same locality. On the other hand, large-scale seedling production, medium-term foliage production and preservation of species variability can be achieved using seeds. Rev. Biol. Trop. 65 (1): 9-19. Epub 2017 March 01.

ResumenLas hojas de Phyllonoma laticuspis se utilizan en Carrizal de Bravo, Gro. México, para curar lesiones de la piel como heridas, secuelas de viruela y afecciones de la diabetes mellitus tipo 2. Los extractos orgánicos de hojas, mostraron efectos antibacterianos. La gran demanda de P. laticuspis como planta medicinal, ha disminuido sus poblaciones naturales, y no existen reportes de la propagación de la especie. Por lo tanto, el propósito del presente trabajo fue evaluar la propagación vegetativa de la especie a través de estacas y acodos, y su propagación sexual en una población conservada. En la propagación por estacas y acodos se aplicaron concentraciones de 1 000, 4 000 y 6 000 ppm de un enraizador comercial con ácido indolbutírico (AIB), y un tratamiento control sin AIB. La germinación se evaluó en luz y oscuridad mediante el uso de lotes de semillas recién recolectadas y lotes de semillas almacenadas por tres meses a 4 °C y a 24 ± 2 ºC. Todos los experimentos se realizaron en un diseño completamente al azar. Las estacas no desarrollaron raíces en ninguna de las concentraciones, mientras que el 100 % de los acodos enraizaron, produciendo raíces vigorosas con la concentración de 4 000 ppm de AIB, a los cuatro meses de la aplicación. Con respecto a la germinación bajo condiciones de luz y oscuridad, las semillas recién recolectadas germinaron más del 60 %, mientras que las semillas almacenadas a 4 ºC menos del 20 % y las almacenadas a 24 ± 2 ºC cerca del 50 %. No hubieron diferencias significativas entre la germinación en luz y oscuridad, por lo que se consideraron como semillas fotoblásticas indiferentes. El contenido de humedad de 13.5 % y el comportamiento de la germinación en respuesta al almacenamiento en frío, sugieren la cualidad de semillas de tipo intermedias. La propagación de P. laticuspis para la producción de follaje a corto tiempo, puede hacerse por acodos en poblaciones con alta densidad de plantas adultas como fuente de material vegetal y para la restauración de áreas alteradas en la misma localidad. Mientras que la producción de follaje a mediano plazo y la conservación de la variabilidad de la especie, se puede lograr con la obtención masiva de plántulas a través de semillas.

Propagation studies in Smilax fluminensis Steud. (Smilacaceae) / Estudos de propagação em Smilax fluminensis Steud. (Smilacaceae)

Smilax L. species, popularly known in Brazil as salsaparrilha, have been used in folk medicine as tonic against rheumatism and as anti-syphilitic since the sixteenth century; however, until today, their roots have been explored in an extractive way. The aim of this study was to analyze the propagation of Smilax fluminensis aiming to meet the demand and to propose procedures for its sustainable management. It was analyzed the percentage of seed germination, rooting of aerial and subterranean stem cuttings and plant resprouting capacity after removal of the shoots. The monitoring lasted one year. Seeds were subjected to temperatures: 25°C, 30°C, 20-30°C and 20-35°C under a daily photoperiod of eight hours and in the absence of light. For rooting the stem cuttings, aerial and subterranean stems about 20cm long with two nodal regions were subjected to treatment with distilled water (control) and indolbutyric acid (IBA) at 100ppm. The best germination percentages obtained for S. fluminensis were 80 percent at 20-30°C under light and 85 percent at 30°C in the dark. Only subterranean stem cuttings showed significant difference concerning fresh and dry matter of roots with higher values in treatments with hormone compared to the control.

As espécies do gênero Smilax L., conhecidas popularmente como salsaparrilha, são empregadas na medicina popular como fortificante contra o reumatismo e antissifilítico desde o século XVI, porém até hoje as suas raízes são exploradas de maneira extrativista. O objetivo deste estudo foi analisar a propagação de Smilax fluminensis visando a auxiliar no atendimento da demanda e apontar propostas para o seu manejo sustentável. Foram analisados a porcentagem de germinação de sementes, o enraizamento de estacas de ramos aéreos e subterrâneos e o acompanhamento ao longo de um ano da capacidade de rebrotamento das plantas após a retirada dos ramos aéreos. As sementes foram submetidas às temperaturas: 25, 30, 20-30 e 20-35°C, sob fotoperíodo diário de oito horas e na ausência de luz. Para o enraizamento de estacas, foram utilizados ramos aéreos e subterrâneos com aproximadamente 20cm com duas regiões nodais submetidas ao tratamento com água destilada (controle) e com ácido indolbutírico (AIB) a 100ppm. As melhores porcentagens de germinação obtidas para S. fluminensis, 80 por cento em 20-30°C no claro e 85 por cento a 30°C no escuro. Apenas as estacas de ramos subterrâneos enraizaram, houve diferença significativa em relação à massa fresca e seca de raízes com valores maiores nos tratamentos com hormônio em relação ao controle.

Resprouting from roots in four Brazilian tree species

Previous studies pointed out that species richness and high density values within the Leguminosae in Brazilian forest fragments affected by fire could be due, at least partially, to the high incidence of root sprouting in this family. However, there are few studies of the factors that induce root sprouting in woody plants after disturbance. We investigated the bud formation on root cuttings, and considered a man-made disturbance that isolates the root from the shoot apical dominance of three Leguminosae (Bauhinia forficata Link., Centrolobium tomentosum Guill. ex Benth, and Inga laurina (Sw.) Willd) and one Rutaceae (Esenbeckia febrifuga (St. Hil.) Juss. ex Mart.). All these species resprout frequently after fire. We also attempted to induce bud formation on root systems by removing the main trunk, girdling or sectioning the shallow lateral roots from forest tree species Esenbeckia febrifuga and Hymenaea courbaril L. We identified the origin of shoot primordia and their early development by fixing the samples in Karnovsky solution, dehydrating in ethyl alcohol series and embedding in plastic resin. Serial sections were cut on a rotary microtome and stained with toluidine blue O. Permanent slides were mounted in synthetic resin. We observed different modes of bud origin on root cuttings: close to the vascular cambium (C. tomentosum), from the callus (B. forficata and E. febrifuga) and from the phloematic parenchyma proliferation (I. laurina). Fragments of B. forficata root bark were also capable of forming reparative buds from healing phellogen formed in callus in the bark’s inner side. In the attempt of bud induction on root systems, Hymenaea courbaril did not respond to any of the induction tests, probably because of plant age. However, Esenbeckia febrifuga roots formed suckers when the main trunk was removed or their roots were sectioned and isolated from the original plant. We experimentally demonstrated the ability of four tree species to resprout from roots after disturbance. Our results suggest that the release of apical dominance enables root resprouting in the studied species. Rev. Biol. Trop. 57 (3): 789-800. Epub 2009 September 30.

Estudios anteriores señalaron que la alta riqueza de especies de Leguminosae en fragmentos de bosques afectadas por el fuego podría deberse, al menos parcialmente, a la alta incidencia de brotes de raíz en esta familia. Sin embargo, existen pocos estudios. Investigamos la formación de brotes en tres Leguminosae (Bauhinia forficata Link., Centrolobium tomentosum Guill. ex Benth, e Inga laurina (Sw.) Willd) y una Rutaceae (Esenbeckia febrifuga (St. Hil.) Juss. ex Mart.). Todas estas especies suelen rebrotar después de los incendios. También hicimos cortes experimentales en árboles forestales: Esenbeckia febrifuga e Hymenaea courbaril L. Observamos tres orígenes: cerca del cambium vascular (C. tomentosum), desde el callo (B. forficata y E. febrifuga) y de parénquima (I. laurina). Hymenaea courbaril no respondió a ninguna de las pruebas de inducción, probablemente debido a la edad de la planta. Sin embargo, las raíces de Esenbeckia febrifuga forman retoños tras retirar el tronco o separar sus raíces. Hemos demostrado experimentalmente la capacidad de las cuatro especies de árbol de rebrotar a partir de sus raíces tras una perturbación grave.

Diferentes substratos e ambientes para enraizamento de mini-ixora (Ixora coccinea 'Compacta') / Different substrates and environments for mini-ixora (Ixora coccinea 'Compacta') rooting

A mini-ixora (Ixora coccinea 'Compacta') é uma espécie muito apreciada para uso em paisagismo, especialmente em jardins tropicais. A propagação é feita por estaquia, no entanto, a porcentagem de enraizamento é bastante baixa. Sendo assim, objetivou-se com este trabalho avaliar substratos e ambientes para o enraizamento da mini-ixora. Foram utilizadas estacas apicais com comprimento de 10 cm, as quais foram cultivadas em dois tipos de substratos: areia e o produto comercial Plantmax®. Utilizaram-se também três ambientes de enraizamento: câmara úmida, estufa e enraizador tradicional (telado com 50 por cento de sombreamento). O delineamento experimental utilizado foi o de parcelas subdivididas, sendo três ambientes nas parcelas e dois substratos nas subparcelas. Utilizaram-se 5 repetições e 10 estacas por parcela. Analisando-se todos os resultados 100 dias após a implantação do experimento, observou-se que a areia proporcionou maior porcentagem de enraizamento (68 por cento) quando comparada ao Plantmax (48 por cento). A câmara úmida também foi o ambiente que proporcionou melhores condições para a propagação da mini-ixora com maior porcentagem de enraizamento, 98 por cento, em relação a estufa (66 por cento) e ao enraizador tradicional (10 por cento). A areia e a câmara úmida também favoreceram a formação de raízes de melhor qualidade. Concluiu-se que o melhor substrato para propagação da mini-ixora foi areia devendo as estacas serem dispostas em câmara úmida.

The mini-ixora (Ixora coccinea 'Compacta') is a species highly appreciated for use in tropical gardens. Its propagation is made by cutting, however, its rooting percentage is very low so, the aim of this work was to evaluate substrates and environments for the mini-ixora rooting. Apical cuttings with 10 cm of length were used in two substrates: sand and Plantmax®. Three rooting environments were used: wet chamber, greenhouse and conventional rooting (shade cloth with 50 percent of shade). The experiment was carried out by using split plots with three different environments in the main plot and two substrates in the subplots. One also used five replications and ten cuttings per plot. Analyzing the results one hundred days after the experiment implantation one noticed that the sand had proportioned higher percentage of rooting (68 percent) compared to Plantmax (48 percent). The wet chamber also was the environment that provided better conditions for the propagation of the mini-ixora, with the highest percentage of rooting, 98 percent, compared to the greenhouse, 66 percent and to the conventional rooting, 10 percent. Moreover, sand and wet chamber also provided a better root quality. One concluded that the best substrate for mini-ixora propagation was sand and the cuttings should be placed in the wet chamber.

Efeito do substrato e da época de coleta dos ramos no enraizamento de estacas de ameixeira (Prunus salicina Lindl.) / Effect of substrate and time of cutting collection on rooting of cuttings of plum (Prunus salicina Lindl.)

O trabalho foi realizado em casa de vegetação do Departamento de Fitotecnia da Faculdade de Agronomia Eliseu Maciel, Universidade Federal de Pelotas, com o objetivo de verificar o efeito de diversos substratos no enraizamento de estacas de ameixeira em diferentes épocas de coleta dos ramos. Foram utilizadas estacas medianas da cultivar Frontier, com comprimento de 15cm e um par de folhas, coletadas nos meses de novembro de 1993, janeiro e março de 1994, obtidas em um pomar comercial com 8 anos de idade. Todas as estacas utilizadas foram tratadas com ácido indolbutirico na concentração de 3000ppm, introduziu-se em torno delem da base das estacas em ácido indolbutirico, na forma de pó e colocou-se em tubetes, acondicionados em bandejas de isopor, contendo diferentes substratos: areia média lavada; vermiculila de grânulos médios; cinza de casca de arroz e serragem de eucalipto, utilizados isolados e em misturas (1: 1v/v). O período de permanência das estacas na casa de vegetação foi de 60 dias. Observou-se maiores índices de enraizamento na coleta dos ramos realizadas em janeiro e março, e que o substrato areia + serragem proporcionou os maiores percentuais de estacas enraizadas com 68,22% e 65,99% nos meses de janeiro e março.

This work was carried out in a greenhouse with intermittent mist out at the plant Science Department, Eliseu Maciel Agronomy College of Federal University of Pelotas (Brazil), aiming to evaluate the effect of different substratos and time of cutting on rooting of pium cuttings. Mild cuttings with 15cm length andapair of ieaves of Frontier cultivar were coilected from a commercial orchard eight years old in November 1993, January and March 1994. All cuttings were treated with indolbutiric acid (IBA) at 3000ppm concentration introducing about 1.0cm of their basal end in IBA powder and placed in tubes on isopor trays containing different substratos: washed mild sand: vermiculite of medium grains; rice husk ash and saw dust, utilized as isolated or as mixtures (1:1 v/v). Cuttings were kept on substratos for 60 days. The results demonstrated that the higher rooted cuttings percentage were obtained for cuttings collected in January and in March. The mixture of sand and saw dust gave the higher number of cuttings rooting with 68.22% and 65.99% for January and March.