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.

The Response of Local White Guinea Yam Cultivars to the Vine Cutting Propagation Technique Using Locally Available Rooting Substrates

Good quality planting materials for yam cultivation is a major challenge and adapting the most preferred local varieties to high ratio propagation methods such as the vine cutting could help ameliorate this constraint. An experiment was conducted in the screen house at CSIR-Savanna Agricultural Research Institute to assess the response of ‘Labako’, ‘Nyamenti’ and ‘Kpamyo’ (check) to vine cutting under different substrates. The experiment was 3 x 6 factorial arranged in a completely randomized design with 3 replications. Single node cuttings were obtained from 3 months old plants and established in the various rooting substrates (carbonized rice husk; fermented rice bran; aged rice husk; top-soil ,cocopeat and carbonized rice husk + top-soil; 2:1).Highly significant differences (P<0.001) existed in main effects and interaction of the factors. All cultivars attained 100% survival in CRH (carbonized rice husk) and cocopeat. Kpamyo and Nyamenti had 89.6% regeneration in cocopeat, FRB (fermented rice bran) and CRH. Labako exhibited the maximum shoot growth (31 cm) in CRH whiles the minimum (10 cm) was observed in ARH (aged rice husk) for Kpamyo. Six nodes were produced by Nyamenti in CRH, whiles Kpamyo produced 3 nodes in the same substrate. Labako initiated 88% tuberization, higher than Kpamyo with 75%. Generally, the cultivars were successfully adapted to the vine cutting technique in CRH, FRB as well as cocopeat. CRH and FRB are locally available, hence less expensive to acquire and therefore recommended for cost-effective single node propagation of the popular local yam cultivars.

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.

Effects of Indole-3-Butyric Acid (IBA) and rooting media on rooting and survival of air layered wax apple (Syzygium samarangense) CV Jambu Madu / Efeitos do Ácido Indol-3-Butírico (AIB) e meio de enraizamento no enraizamento e sobrevivência de maçã de cera em camada de ar (Syzygium samarangense) CV Jambu Madu

The wax apple or jambu madu, is a non-climacteric tropical fruit from Myrtaceae family and widely cultivated in South East Asia. The limited availability of good quality seedlings of wax apple is the main problem to development of flourish it's market share in the current fruit industry. Therefore, in order to produce good quality planting materials, a study aimed at optimizing propagation and adventitious rooting technique and survivability of wax apple air layer was conducted. In this study, four different levels of Indole-3-Butyric Acid (IBA) concentration (0, 1000, 1500 and 2000 mg L-1) and three rooting media (sphagnum moss, vermicompost and garden soil) were applied after removal of bark (phloem) on the shoot to determine the effect on rooting and survivability of the wax apple air layer under field conditions. The results showed that the wax apple shoots treated with 2000 mg L-1 IBA produced the significantly higher number of roots, increased length of root, diameter of branch, length of branch, number of leaf and leaf area of air layers. In addition, the highest chlorophyll content and stomatal aperture were recorded in 2000 mg L-1 IBA treatment compared to other treatments including control. Vermicompost medium was better than garden soil and sphagnum moss in respect of rooting and survivability of air layers. The results showed that the combination of 2000 mg L-1 IBA and vermicompost as rooting media give the best combination to root initiation, root number, root length and survival rate (100%) of wax apple air layers. From this study, it can be concluded that 2000 mg L-1 IBA and vermicompost treatment enhance the root initiation, early establishment and survivability of wax apple air layered under field conditions.

A maçã de cera, ou jambu madu, é uma fruta tropical não climatérica da família Myrtaceae e amplamente cultivada no Sudeste Asiático. A disponibilidade limitada de mudas de macieira de boa qualidade é o principal problema para o desenvolvimento de sua participação de mercado na fruticultura atual. Portanto, com o objetivo de produzir materiais de plantio de boa qualidade, foi realizado um estudo visando otimizar a técnica de propagação e enraizamento adventício e a sobrevivência da camada aérea da cera de macieira. Neste estudo, quatro diferentes níveis de concentração de ácido indol-3-butírico (AIB) (0, 1000, 1500 e 2000 mg L-1) e três meios de enraizamento (musgo esfagno, vermicomposto e solo de jardim) foram aplicados após a remoção da casca (floema) na parte aérea para determinar o efeito no enraizamento e capacidade de sobrevivência da camada de ar da macieira em condições de campo. Os resultados mostraram que os brotos de macieira tratados com 2000 mg L-1 de AIB produziram significativamente maior número de raízes, maior comprimento de raiz, diâmetro de galho, comprimento de galho, número de folhas e área foliar das camadas aéreas. Além disso, o maior teor de clorofila e abertura estomática foram registrados no tratamento 2000 mg L-1 IBA em comparação com outros tratamentos, incluindo o controle. O meio de vermicomposto foi melhor do que o solo de jardim e o musgo esfagno em relação ao enraizamento e capacidade de sobrevivência das camadas aéreas. Os resultados mostraram que a combinação de 2000 mg L-1 de AIB e vermicomposto como meio de enraizamento proporciona a melhor combinação para iniciação radicular, número de raízes, comprimento radicular e taxa de sobrevivência (100%) das camadas aéreas de macieira. A partir deste estudo, pode-se concluir que 2.000 mg L-1 de AIB e tratamento com vermicomposto melhoram a iniciação radicular, o estabelecimento precoce e a capacidade de sobrevivência de macieiras em camadas de ar em condições de campo.

Rescue and vegetative propagation of Eremanthus erythropappus (DC. ) Ma ish in natural stand / Resgate e propagação vegetativa de Eremanthus erythropappus (DC. ) MacLeish em povoamentos naturais

Abstract The native stands of 'candeia' (Eremanthus erythropappus) have been explored through management plans due to the economic potential of essential oil. The rescue of adult trees, as well as the application of silvicultural techniques that favor the restoration of the stand, can contribute to the genetic conservation of this species. This study's objective was to assess the efficiency of propagation techniques for the rescue of 26 matrices of 'candeia' in a natural managed stand and discussion about the rhizogenesis. In August 2017, trees were induced to regrowth by coppice, followed by exposure and scarification of roots. The emergence of shoots and morphology were evaluated according to the origin (i.e., stump or root). After that period, 19 matrices had their sprouts collected for the preparation of apical cuttings. Indole-3-butyric acid (IBA) was applied at the base of the cuttings. Cutting survival at greenhouse exit (GE), rooting at shade house exit (SHE), morphology and root anatomy were evaluated. In 189 days, the scarification of roots promoted 76.92% of budding. The percentage of sprouted matrices, number of shoots per matrice, length, diameter, and shoot length/diameter ratio increased over time. Only 12.2% of the cuttings survived in GE, and of these, 7.9% rooted in SHE. The cutting resulted in the formation of a clonal mini-garden of 'candeia', with seven of the 19 matrices submitted to propagation. The anatomical analyses showed that bud formation occurs from cell redifferentiation in the phloem parenchyma, and presence of crystals on the walls of the vessel elements of the secondary xylem. The shoots induction from scarification of roots could be used as a silvicultural practice for the reestablishment of the native fragments handle.

Resumo Os povoamentos nativos de candeia (Eremanthus erythropappus) vêm sendo explorados por planos de manejo devido ao potencial econômico do óleo essencial. O resgate de árvores adultas, bem como a aplicação de técnicas silviculturais que favoreçam o restabelecimento do povoamento podem contribuir para a conservação genética dessa espécie. O objetivo desse trabalho foi avaliar a eficiência de técnicas de propagação para o resgate de 26 matrizes de candeia em um povoamento natural manejado e discutir sobre a rizogênese. Em agosto de 2017, as árvores foram induzidas à rebrota por meio da decepa, seguida da exposição e escarificação das raízes. A emissão brotações e morfologia foram avaliadas de acordo com a origem (toco ou raiz). Após esse período, 19 matrizes tiveram as brotações recolhidas para o preparo de estacas apicais, que foram tratadas com ácido indolbutírico (AIB). A sobrevivência das estacas na saída da casa de vegetação (SCV), o enraizamento na saída da casa de sombra (SCS), a morfologia e a anatomia da raiz foram avaliados. Aos 189 dias, a escarificação das raízes resultou em 76,92% de emissão de brotos. O percentual de matrizes brotadas, número de brotos por matriz, comprimento, diâmetro e relação comprimento/diâmetro dos brotos aumentaram ao longo do período avaliado. Somente 12,2% das estacas sobreviveram na SCV e 7,9% enraizaram na SCS. A estaquia resultou na formação de um minijardim clonal de candeia com sete das dezenove matrizes submetidas à propagação. As análises anatômicas mostraram a diferenciação das células na região do parênquima floemático e a presença de cristais de inulina nas paredes dos elementos de vaso do xilema secundário. A indução de brotos radiculares pode ser usada como prática silvicultural visando o restabelecimento de fragmentos nativos manejados.

An efficient protocol for in vitro regeneration of peanut (Arachis hypogaea L ) cultivar L14

The present work aims to establish an efficient protocol for in vitro regeneration of peanut (Arachis hypogaea) cultivar L14. The study showed that de-embryonated cotyledon was a suitable explant for shoot multiplication on MS medium containing 4 mg/L BAP. The highest number of shoots per explant obtained after 4 weeks of culture was up to 6.8 shoots. Shoots in vitro were able to produce a large number of approximately 11 roots on MS medium supplemented with 0.5 mg/L NAA. These results will be very useful in establishing an in vitro regeneration protocol for peanut cultivar L14 during gene transfer in the next studies to improve their disease resistance.

Vegetative propagation of an ornamental conifer: some techniques in podocarpo cutting / Propagação vegetativa de uma conífera ornamental: algumas técnicas no estaqueamento de podocarpo

ABSTRACT: The asexual propagation of conifers, such as Podocarpus macrophyllus (Thunb.) Sweet, faces difficulties. Among the factors that can improve the efficiency of asexual propagation are the types of cuttings used and lesions performed at the base of the cuttings, and the use of plant regulators, such as indolebutyric acid (IBA). This study determined the types of lesions to be performed at the cutting base, the type of cuttings used, the method to convey IBA at the cutting base, and the concentration of IBA required to obtain a proper rooting condition for the species. Two experiments were performed from March to July, 2018, in randomized blocks and with a factorial scheme, as follows: 2 x 4 (IBA application in solution and talc x IBA at concentrations of 0, 500, 1,000, and 1,500 mg L-1 IBA); and 2 x 3 (basal and apical cuttings x bevel, wedge, and horizontal lesions at the cutting base). In both experiments, the cuttings used were arranged on a masonry bed containing medium-textured sand. After 120 days of experimentation, the study concluded that the rooting percentage was favored by the use of basal cuttings with wedge lesions at the cutting base and treated with IBA in solution at a concentration of approximately 550 mg L-1.

RESUMO: A propagação assexuada de coníferas, como o Podocarpus macrophyllus, apresenta dificuldades. Entre os fatores que podem tornar mais eficientes a propagação assexuada tem-se o tipo da estaca utilizada, lesões na base da estaca e uso de fitoreguladores, como o ácido indolbutírico (AIB). Objetivou-se determinar o tipo de lesão a ser efetuado na base da estaca e o tipo de estaca bem como método de veiculação do AIB na base da estaca e a concentração do AIB visando obter melhor enraizamento para a espécie. Dois experimentos foram realizados, no período de março a julho de 2018, em blocos casualizados e esquema fatorial, sendo o primeiro 2 x 4 [aplicação AIB em solução e em talco x concentração de 0, 500, 1000 e 1500 mg L-1 de ácido indolbutírico (AIB)] e o segundo 2 x 3 [estaca basal e apical x lesões na base da estaca do tipo bisel, cunha e horizontal].Em ambos experimentos as estacas utilizadas foram dispostas em canteiro de alvenaria contendo areia de textura média. Após 120 dias de experimentação conclui-se que o uso de estacas basais com lesões na base da estaca em cunha e tratadas com AIB em solução na concentração de aproximadamente 550 mL L-1favorecem a porcentagem de enraizamento.

Clonal bamboo production based on in vitro culture / Produção de plantas clonais de bambu a partir do cultivo in vitro

Bamboo species are an alternative for the composition of forest plantations. However, their potential has not been explored due to the hard time in producing large-scale clonal plants. Thus, the aim this work was to evaluate the in vitro establishment, bud multiplication and ex vitro rooting of Bambusa vulgaris. The first experiment tested different systemic and contact fungicide solutions, based on exposure time, during the establishment phase. Established explants were subjected to evaluation of residual fungicide effect on subcultures during the multiplication and elongation phases. The second experiment evaluated the influence of activated carbon on ex vitro survival and on adventitious rooting. Explant immersion in liquid culture medium added with 1.0 mL of fungicide for 120 hours has favored the in vitro establishment and reduced fungal contamination. On the other hand, it favored the shoot emission of shoots per explant during the multiplication phase. Both rooting induction culture medium and mini-incubator system use were effective in enabling adventitious root formation. The presence of activated carbon in the rooting induction culture medium resulted in a higher clonal plant survival rate.

As espécies de bambus são uma alternativa para a composição de plantios florestais. Entretanto, esse potencial não tem sido explorado devido à dificuldade de produção de mudas clonais em larga escala. Assim, objetivo deste trabalho foi avaliar o estabelecimento in vitro, a multiplicação e o enraizamento ex vitro de Bambusa vulgaris. No primeiro experimento foram testadas diferentes soluções de fungicida sistêmico e de contato em relação ao tempo de exposição durante a fase de estabelecimento. Os explantes estabelecidos foram avaliados quanto ao efeito residual do fungicida durante subcultivos nas fases de multiplicação e alongamento. No segundo experimento, foi avaliada a influência do carvão ativado sobre a sobrevivência e enraizamento ex vitro. Durante o estabelecimento in vitro, a imersão de explantes em meio de cultura líquido contendo alíquota de 1,0 mL de fungicida durante 120 horas favoreceu o estabelecimento e reduziu a contaminação fúngica, enquanto na fase de multiplicação, houve o favorecimento da emissão de brotos por explante. O meio de cultura de indução ao enraizamento e uso de sistema de mini-estufim foram efetivos para a formação de raízes adventícias e a presença de carvão ativado resultou em uma maior sobrevivência das mudas clonais.

Micropropagación de Prosopis pallida (Humb &Bonpl. Ex Willd.) Kunth a partir de yemas apicales / Micropropagation of Prosopis pallida (Humb &Bonpl. Ex Willd.) Kunth from shoot tips

RESUMEN Prosopis pallida, conocido como algarrobo, es una especie emblemática de los bosques secos del norte del Perú. Es de gran importancia económica por su uso en la producción de leña y carbón, así como en la producción de algarrobina proveniente de sus frutos. Actualmente las actividades humanas han deforestado grandes poblaciones de algarrobo en el bosque seco, por lo que es muy importante una propagación masiva para planes de reforestación a gran escala en esos ecosistemas, con la finalidad de conservar la especie y también las características genéticas de individuos élite. El objetivo de la investigación fue establecer un protocolo para la propagación in vitro de algarrobo. Previo a la siembra in vitro, se evaluaron tres tratamientos pregerminativos para poder acelerar la germinación de las semillas con la finalidad de obtener mayor material de propagación. Luego se evaluó el efecto del medio de plantas leñosas con la adición de cuatro concentraciones (0; 0,5; 1,0 y 1,5 mg/L) de citoquininas (BAP y ZEA) sobre la propagación in vitro de Prosopis pallida, habiendo realizado tres ensayos debido a la poca efectividad de brotación de las yemas apicales, resultando mejor la ausencia de citoquininas en yemas apicales con sus dos cotiledones, usando tapas de algodón en los tubos de ensayo. En esta etapa se evaluó el número de nudos, altura de plántula y número de brotes. Para el enraizamiento se ensayó con tres concentraciones (0; 0,5 y 1,0 mg/L) de auxinas (NAA, IBA y IAA), y se evaluó el porcentaje de enraizamiento, longitud de la raíz y número de raíces; obteniéndose mejores resultados con 0,5 mg/L IBA. En la fase de aclimatación se evaluó el porcentaje de aclimatación en dos tipos de sustratos, obteniéndose mejores resultados con sustrato comercial de turba y vermiculita.

ABSTRACT Prosopis pallida, known as algarrobo, is an emblematic species of the dry forests of northern Peru. It is of great economic importance for its use in the production of firewood and coal, as well as in the production of carob from its fruits. Currently human activities have deforested large populations of algarrobo in the dry forest, so it is very important a massive propagation for large-scale reforestation plans in these ecosystems, in order to conserve the species and also the genetic characteristics of elite individuals. The objective of the research was to establish a protocol for the in vitro propagation of algarrobo. Before in vitro propagation, It was evaluated the effect of three seeds treatments to accelerate seeds germination in order to get more propagation material. Then, the effect of woody plants medium with the addition four concentrations (0, 0.5, 1.0 and 1.5 mg /L) of cytokinins (BAP and ZEA) on the in vitro propagation of Prosopis was evaluated and it has performed three trials for the ineffective budding of apical buds, resulting in a better absence of cytokinins in apical buds with their two cotyledons, and using cotton lids in the test tubes. In this stage the number of nodes, seedling height and number of shoots was evaluated. For rooting, it was tested with three concentrations (0, 0.5 and 1.0 mg /L) of auxins (NAA, IBA and IAA), and the percentage of rooting, root length and amount of roots was evaluated, obtaining better results with 0.5 mg / L IBA. In the acclimation phase, the acclimation percentage was evaluated using two substrates, and the best results were commercial substrate of peat and vermiculite.

Rapid in vitro adventitious rooting and proliferation by leaf and nodal cultures of Momordica cymbalaria Fenzl.

An effective approach for rapid in vitro rooting and proliferation of leaf and nodal cultures of Momordicacymbalaria has been developed. To the ability of induction of rhizogenesis, both leaf and nodal explants wereused in culture on Murashige and Skoog (MS) medium. The effects of auxins such as α-naphthaleneaceticacid (NAA), indole-3-butyric acid (IBA), and indole-3-acetic acid (IAA) at different concentrations have beenstudied. The maximum number of roots was produced from nodal explants containing 1.5 mg/L of NAA (9.3 ±0.61), 1.0 mg/L of IBA (6.5 ± 0.41), and 1.0 mg/L of IAA (3.5 ± 0.66), and in leaf explants containing 1.0 mg/Lof NAA (5.7 ± 0.56), 1.0 mg/L of IBA (6.9 ± 0.61), and 1.5 mg/L of IAA (5.0 ± 0.73) on the half-strength MSmedium. For the root induction, NAA is the very effective auxin in node explants of M. cymbalaria. Moreover,a large amount of quercetin bioactive compound is presented in the roots, which is used in anticancer drugs, andwe have described an effective method for the in vitro rhizogenesis of the M. cymbalaria.

Direct in vitro regeneration of castor bean plants (Ricinus communis) using epicotyls / Regeneração direta in-vitro de plantas de mamona ( Ricinus communis ) usando epicótilos

A regeneration protocol for castor bean plant (Ricinus communis) was successfully developed using epicotyl sections obtained from in vitro seedlings. The maximum number of induced shoots (4.3 shoots/explant) and highest shoots frequency (75,56%) was obtained in WPM medium supplemented with TDZ (1 mg/L) and zeatin (0.5 mg/L), whereas the minimum number (0.8 shoots/explant) and lowest shoots frequency (37,78%) was obtained in medium containing TDZ (1 mg/L) and BAP (0.5 mg/L). The highest percentage of rooting (93.3%) was obtained in a medium containing IBA (1 mg/L). These plants were transplanted in a mesh house and achieved a high adaptability to acclimatization, reaching 77% survival. On the other hand, the maximum elongation (height) during this stage was 7.9 cm in plants supplemented with WPM nutrients, whereas it was only 4.38 cm in control plants

Foi desenvolvido com sucesso um protocolo de regeneração para a planta de Mamona (Ricinus communis) utilizando seções de epicótilos, obtidas a partir de mudas in vitro. O número máximo de brotações induzidas (4.3 brotos/explante), assim como a maior frequência de brotações (75,56%), foi obtido em meio WPM suplementado com TDZ (1 mg/L) e zeatina (0,5 mg/L). Enquanto que o número mínimo (0,8 brotos/explante), como a menor freqüência de rebentos (37,78%), foi obtido em meio contendo TDZ (1 mg/L) e BAP (0,5 mg/L). Adicionalmente, a maior percentagem de enraizamento (93,3%) foi obtida em um meio contendo IBA (1 mg/L). Depois da regeneração, as plantas foram transplantadas em casa de vegetação e conseguiram uma alta adaptabilidade e aclimatização, atingindo 77% de sobrevivência. Por outro lado, oalongamento máximo (altura) durante este estágio foi de 7,9 cm em plantas suplementadas com nutrientes de WPM, enquanto as plantas de controle presentaram apenas 4,38 cm

Rooting of black raspberry with plant growth regulator / Enraizamento de estacas caulinares da framboeseira-negra com o uso de fitormônios

ABSTRACT: The framboseira-negra has a potential for cultivation in Brazil, due to the fact that this fruit produces high quantities of fruits, which confer a high quality jelly. However, their propagation is an obstacle to the establishment of commercial plantations. Production and expansion of a fruit is directly linked to the propagation method and the quality of its development. Given the above, the aim of the present study was to evaluate the use of stem cuttings and IBA in asexual propagation of black raspberry. Two experiments being conducted in greenhouse conditions were performed. Stem cuttings were taken from didactic orchard, with matrix plant of four years, which were the experiments. The first experiment was carried out in factorial 3x2, with cuttings taken from three regions of the plant branch matrix (apical, median and basal) x two treatments with IBA (without IBA and 2000mg L-1 IBA), containing four repetitions and fifteen cuttings per repetition. The second experiment consisted of a 4x2, with four sizes of cuttings (5, 10, 15 and 20cm) x two treatments with IBA (without IBA and 2000mg L-1 IBA), with four replications and fifteen cuttings by repetition. After eighty days, the percentage of rooted cuttings (%), average number of roots and shoots for cuttings, the longest root length (cm) and fresh and dry biomass of shoots (g) were assessed. Best results in the formation of root system were observed in middle and apical stem cuttings. Stem Cuttings between 10 and 20cm obtained better results for rooting. There is no need to use IBA rooting of cuttings of black raspberry.

RESUMO: A framboseira-negra possui potencial de cultivo no Brasil pelo fato dessa fruteira produzir altas quantidades de frutos que conferem uma geleia de alta qualidade. No entanto, a sua propagação é um entrave no estabelecimento de plantios comerciais. Diante do exposto, objetivou-se, com o presente trabalho, avaliar a utilização da estaquia caulinar e ácido indolbutírico na propagação assexuada de framboeseira-negra. Foram realizados dois experimentos, sendo conduzidos em condições de telado. Estacas caulinares foram retiradas de pomar didático de plantas matrizes de quatro anos, as quais constituíram os experimentos. O primeiro experimento foi elaborado em esquema fatorial 3x2, sendo as estacas retiradas de três regiões do ramo da planta matriz (apical, mediana e basal) x dois tratamentos com ácido indolbutírico (sem AIB e 2000mg L-1 de AIB), contendo quatro repetições e quinze estacas por repetição. O segundo experimento constituiu-se de esquema fatorial 4x2, sendo quatro tamanhos de estacas (5, 10, 15 e 20cm) x dois tratamentos com ácido indolbutírico (sem AIB e 2000mg L-1 de AIB), contendo quatro repetições e quinze estacas por repetição. Após oitenta dias, avaliaram-se a percentagem de estacas enraizadas (%); número médio de raízes e de brotações por estacas; comprimento da maior raiz (cm) e biomassa fresca e seca das brotações (g). Melhores resultados na formação do sistema radicular foram observados em estacas caulinares medianas e apicais. Estacas caulinares entre 10 e 20cm obtiveram melhores resultados para enraizamento. Não há necessidade de utilização de AIB no enraizamento de estacas caulinares de framboeseira-negra.

Vegetative propagation of Plukenetia polyadenia by cuttings: effects of leaf area and indole-3-butyric acid concentration / Propagação vegetativa de Plukenetia polyadenia por estacas: efeitos de área foliar e concentração de ácido indol-3-butírico

Abstract The seeds of Plukenetia polyadenia have high levels of unsaturated fatty acids and are used as medicine and food for native people in the Peruvian and Brazilian Amazon. The objective of this study was to develop a method for vegetative propagation of Plukenetia polyadenia by rooting of cuttings. The experiment was laid out in a randomized complete block design with 12 treatments and 3 replications of 8 cuttings, in a 3 × 4 factorial arrangement. The factors were: 3 levels of leaf area (25, 50 and 75%) and 3 indole-3-butyric acid - IBA concentrations (9.84, 19.68 and 29.52mM) and a control without IBA. Data were submitted to analysis of variance and means were compared by Tukey test at 5% probability. Our results show that the use of cuttings with 50% of leaf area and treatment with 29.52mM of IBA induced high percentages of rooting (93%) and the best root formation. Vegetative propagation of Plukenetia polyadenia by cuttings will be used as a tool to conserve and propagate germplasm in breeding programs.

Resumo As sementes de Plukenetia polyadenia têm altos níveis de ácidos graxos insaturados e são utilizadas como medicamentos e alimentos para as pessoas nativas da Amazônia Peruana e Brasileira. O objetivo do trabalho foi desenvolver um método de propagação vegetativa de Plukenetia polyadenia por meio do enraizamento de estacas em câmeras de sub-irrigação. Foi utilizado um delineamento de blocos ao acaso com 12 tratamentos e 3 repetições de 8 estacas, e esquema fatorial 3 × 4. Os fatores foram: 3 níveis de área foliar (25, 50 e 75%) e 3 doses de ácido indol-3-butírico - AIB (9,84; 19,68 e 29,52mM) e um controle sem AIB. Os dados foram submetidos à análise de variância e as médias foram comparadas pelo teste de Tukey a 5% de probabilidade. A maior taxa de enraizamento de estacas (93%) foi obtida com 29,52mM de AIB como indutor hormonal e estacas com área foliar de 50%. A propagação vegetativa de Plukenetia polyadenia por estacas será usada como ferramenta para conservar e propagar germoplasma em programas de melhoramento.

Caipira: cultura, resistência e enraizamento / Caipira: culture, resistance and rooting

RESUMO O presente artigo trata do surgimento da música caipira como uma expressão da cultura popular brasileira e como essa música, desde o momento em que foi gravada em disco e divulgada pelo rádio, ajudou no processo de reenraizamento dos migrantes caipiras na Grande São Paulo.

ABSTRACT This article deals with the emergence of Brazilian country folk (caipira) music as an expression of the country's popular culture and how this music, from the moment it was recorded and broadcast on radio, helped in the process of re-rooting the country's migrants in the city of São Paulo.

Protocorm proliferation and regeneration of Dendrobium officinale / 中草药

Objective To establish an efficient tissue culture and rapid propagation system, and study the protocorm proliferation and regeneration conditions using seeds as explants in Dendrobium officinale. Methods Seeds of D. officinale were used as explants, protocorm was induced on inducement medium. After proliferation, the protocorm were transferred to the regeneration medium. Then the regenerated shoots were transferred into rooting medium to induce rooting of plantlets, and developing complete plant. Results The basic culture medium for D. officinale growth was 1/2 MS; The best culture medium formula for inducing protocorms was 1/2 MS +1.0 mg/L 6-BA + 0.2 mg/L NAA + 50 g/L mashed potato; The optimal proliferation medium for protocorm was 1/2 MS + 1.0 mg/L 6-BA + 0.5 mg/L NAA, and the maximum multiplication factor could reach 23. The optimal regeneration medium was 1/2 MS + 2.0 mg/L 6-BA + 0.2 mg/L NAA, regeneration rate can reach 95%; The best culture medium for seedling rooting was 1/2 MS + 0.3 mg/L NAA + 50 g/L potato juice, and rooting rate reached 100%. Conclusion This research provides an effective way for keeping good varieties of features and rapid propagation of D. officinale, at the same time helps to solve some theoretical problems in factory production of D. officinale.

Rescue and vegetative propagation of Eremanthus erythropappus (DC.) MacLeish in natural stand

Abstract The native stands of candeia (Eremanthus erythropappus) have been explored through management plans due to the economic potential of essential oil. The rescue of adult trees, as well as the application of silvicultural techniques that favor the restoration of the stand, can contribute to the genetic conservation of this species. This studys objective was to assess the efficiency of propagation techniques for the rescue of 26 matrices of candeia in a natural managed stand and discussion about the rhizogenesis. In August 2017, trees were induced to regrowth by coppice, followed by exposure and scarification of roots. The emergence of shoots and morphology were evaluated according to the origin (i.e., stump or root). After that period, 19 matrices had their sprouts collected for the preparation of apical cuttings. Indole-3-butyric acid (IBA) was applied at the base of the cuttings. Cutting survival at greenhouse exit (GE), rooting at shade house exit (SHE), morphology and root anatomy were evaluated. In 189 days, the scarification of roots promoted 76.92% of budding. The percentage of sprouted matrices, number of shoots per matrice, length, diameter, and shoot length/diameter ratio increased over time. Only 12.2% of the cuttings survived in GE, and of these, 7.9% rooted in SHE. The cutting resulted in the formation of a clonal mini-garden of candeia, with seven of the 19 matrices submitted to propagation. The anatomical analyses showed that bud formation occurs from cell redifferentiation in the phloem parenchyma, and presence of crystals on the walls of the vessel elements of the secondary xylem. The shoots induction from scarification of roots could be used as a silvicultural practice for the reestablishment of the native fragments handle.

Resumo Os povoamentos nativos de candeia (Eremanthus erythropappus) vêm sendo explorados por planos de manejo devido ao potencial econômico do óleo essencial. O resgate de árvores adultas, bem como a aplicação de técnicas silviculturais que favoreçam o restabelecimento do povoamento podem contribuir para a conservação genética dessa espécie. O objetivo desse trabalho foi avaliar a eficiência de técnicas de propagação para o resgate de 26 matrizes de candeia em um povoamento natural manejado e discutir sobre a rizogênese. Em agosto de 2017, as árvores foram induzidas à rebrota por meio da decepa, seguida da exposição e escarificação das raízes. A emissão brotações e morfologia foram avaliadas de acordo com a origem (toco ou raiz). Após esse período, 19 matrizes tiveram as brotações recolhidas para o preparo de estacas apicais, que foram tratadas com ácido indolbutírico (AIB). A sobrevivência das estacas na saída da casa de vegetação (SCV), o enraizamento na saída da casa de sombra (SCS), a morfologia e a anatomia da raiz foram avaliados. Aos 189 dias, a escarificação das raízes resultou em 76,92% de emissão de brotos. O percentual de matrizes brotadas, número de brotos por matriz, comprimento, diâmetro e relação comprimento/diâmetro dos brotos aumentaram ao longo do período avaliado. Somente 12,2% das estacas sobreviveram na SCV e 7,9% enraizaram na SCS. A estaquia resultou na formação de um minijardim clonal de candeia com sete das dezenove matrizes submetidas à propagação. As análises anatômicas mostraram a diferenciação das células na região do parênquima floemático e a presença de cristais de inulina nas paredes dos elementos de vaso do xilema secundário. A indução de brotos radiculares pode ser usada como prática silvicultural visando o restabelecimento de fragmentos nativos manejados.

Establishment of an Embryonic Tip Regeneration System of Soybean

ABSTRACT In order to establish an optimal soybean embryonic tip regeneration system that can serve as soybean genetic transformation receptor, and be used for the study of genetic function verification, the influences of single factor on the adventitious bud of embryonic tip induction, elongation and rooting stage, are researched and compared.The single factors includes seeds soaking time, different kinds of hormones, different concentration of hormone and different concentration of sucrose. By one-way ANOVA and LSD ad hoc test , the results show that, for the embryonic tip adventitious bud induction stage, 12h is the optimal seeds soaking time, 2.0mg·L-1 is the optimal concentration of 6-Benzyl Aminopurine(6-BA), for the embryonic tip adventitious bud elongation stage, 0.2mg·L-1 indole-3-butyric acid (IBA) is optimal and 2.0mg·L-1Gibberellic acid (GA3) is optimal, and for the adventitious bud of embryonic tip rooting stage, 2.0mg·L-1 IBA is optimal,the average rooting rate is 93.34%. An Optimal embryonic tip regeneration system is established, and optimum mediums in different stages are found.

An efficient regeneration and rapid micropropagation protocol for Almond using dormant axillary buds as explants.

An efficient in vitro protocol was standardized for Almond (Prunus dulcis) propagation using dormant axillary buds as explants. Explants were cultured on Murashige and Skoog (MS) and woody plant medium (WPM) supplemented with different concentration/combination(s) of phytohormones. MS basal medium showed lowest shoot induction and took longest duration for shoot initiation. Multiple shoots were induced in MS medium supplemented with the combination of BAP (0.5 mgL-1). Cultures showed poor response for rooting in all combinations of plant growth regulators (PGRs) and took 90 days for initiation. Rooting was higher in half strength of MS than in full-strength. The highest root induction (33.33%) was recorded in half MS medium supplemented with 0.1 mgL-1 IBA (indole-3-butyric acid) followed by full strength of MS medium (20%) supplemented with IBA (0.1 mgL-1). α-Naphthalene acetic acid (NAA) was less effective for rooting than IBA. The highest root induction (25%) was found in half strength of MS medium supplemented with 0.1 mgL-1 NAA followed by full strength of MS medium (20%). The protocol developed would be of use in mass propagation of almond and also support in vitro conservation.

Propagación vegetativa por estacas de dasyphyllum diacanthoidesmediante recursos endogenos. una aproximación agroecológica / Cutting vegetative propagation of dasyphyllum diacanthoides, with endogenous resources: an agroecological approach

La evidencia empírica obtenida por los agricultores del sector de Huechelepún (comuna de Melipeuco, Región de La Araucanía, Chile) indica que la especie Dasyphyllum diacanthoides, además de presentar propiedades medicinales, es una alternativa forrajera invernal. Esta propiedad junto con su condición de especie endémica de los bosques de Chile obliga a compatibilizar el uso y la conservación de esta especie. El objetivo de este estudio fue propagar D. diacanthoides mediante esquejes semilignificados y determinar el efecto de la presencia de espinas en los esquejes y la exposición Norte o Sur de las plantas madres en el enraizamiento. El estudio se realizó utilizando recursos endógenos, por lo que las variables evaluadas se determinaron sobre la base del conocimiento de los agricultores. El diseño experimental incluyó cuatro tratamientos: T1: estacas de plantas con exposición sur y espinas, T2: estacas de plantas con exposición sur sin espinas, T3: estacas de plantas con exposición norte sin espinas y T4: estacas de plantas con exposición norte con espinas. Cada tratamiento constó de 25 réplicas y se utilizó un diseño completo aleatorio para la ubicación de las estacas en el invernadero. Las estacas de plantas en laderas Norte tuvieron porcentajes mayores de sobrevivencia, enraizamiento y formación de callos que las de plantas en laderas S. La prueba U de Mann- Whitney evidenció diferencias significativas entre estacas con y sin espinas con exposición norte sólo para la longitud de raíces (p ≤ 0,05). La propagación vegetativa de D. diacanthoides mediante esquejes semilignificados es factible. Los mejores resultados se obtienen con estacas sin espinas de plantas madres con exposición Norte.

The empirical evidence obtained by farmers of Huechelepún area (commune of Melipeuco, La Araucania, Chile) indicates that the specie Dasyphyllum diacanthoides, besides presenting medicinal properties, is a winter forage alternative. This property along with its status as endemic forests of Chile requires reconciling the use and conservation of this species. The aim of this study was to propagate D. diacanthoides by semi lignified cuttings and determine the effect of the presence of thorns in the cuttings and North or South exposure mother plants in rooting. The study was performed using endogenous resources, so the evaluated variables were determined on the basis of knowledge of farmers. The experimental design included four treatments: T1: cuttings of plants with southern exposure and thorns, T2: Stakes southern exposure plants without thorns, T3: cuttings of plants with northern exposure spineless and T4: cuttings of plants with northern exposure with thorns. Each treatment consisted of 25 replicates and a complete randomized design for location of the cuttings in the greenhouse was used. Cuttings from plants in North slopes had higher percentages of survival, rooting and callus formation the plant on hillsides South. Mann-Whitney U test showed significant differences between stakes with and without spines north only exposure length roots (p ≤ 0.05). The vegetative propagation of cuttings D. diacanthoides by semi lignified is feasible. The best results are obtained with stakes thornless plants mothers with Northern exposure.