Large scale propagation of an exotic edible bamboo, Phyllostachys pubescens Mazel ex H. De Lehale (Moso Bamboo) using seeds.

For ex vitro propagation, seeds of P.pubescens were treated with different concentrations of gibberellic acid (GA3) and germination of seeds was tested both in plastic pots as well as by direct sowing in the nursery beds. Maximum seed germination was achieved when treated with 200 mgL–1 (w/v) GA3. For in vitro propagation, an exposure of nodal explants from in vitro raised seedlings to 0.2 mgL–1 1–phenyl–3–(1,2,3–thiadiazol–5–yl) urea and 1 mgL–1 kinetin supplemented medium for 30 days and thereafter to hormone free Murashige and Skoog basal medium resulted in axillary shoot proliferation. For rooting, in vitro raised shoots were exposed to MS medium containing 2 mgL–1 indole-3-butyric acid for 15 days and then shifted to hormone free medium. On an average, 2.8 shoots were obtained in 75% of the cultures within 4 weeks. Such in vitro raised plants were successfully hardened and shifted to field conditions.

Crescimento in vitro e aclimatização de Cattleya loddigesii Lindley (Orchidaceae) com carvão ativado sob dois espectros luminosos / In vitro growth and acclimatization of Cattleya loddigesii Lindley (Orchidaceae) with actived charcoal in two light spectra

O objetivo deste estudo foi avaliar a adição de concentrações de carvão ativado em meio de cultura ½ MS (com metade da concentração dos macronutrientes) sob dois espectros luminosos para a obtenção de plântulas in vitro de Cattleya loddigesii. Plântulas com aproximadamente 90 dias foram subcultivadas em oito tratamentos, nos quais foi testada a adição ao meio de cultura ½ MS com carvão ativado (0; 0,5; 1,0 e 2,0g L-1) e combinados sob espectro de luz branca e luz vermelha. Após 180 dias da germinação, foram mensurados dados biométricos (raiz e parte aérea), massa fresca e teores de pigmentos fotossintéticos. Em plântulas aclimatizadas em casa de vegetação, foram avaliadas a taxa de sobrevivência após 120 dias. As concentrações de clorofila total, clorofila a e carotenoides foram maiores nos tratamentos sob luz branca, enquanto a luz vermelha influenciou significativamente maior clorofila b, plântulas com menos raízes e de menor comprimento e elevada mortalidade ex vitro. A adição de 2,0g L-1 de carvão ativado ao meio de cultura e o uso de luz branca proporcionaram maior eficiência de desenvolvimento tanto para as culturas in vitro quanto para a sobrevivência ex vitro das plantas.

The aim of this study was to evaluate the effect of different concentrations of activated charcoal in ½ MS (half concentration of macronutrients) culture medium under two light spectra on the in vitro growth of Cattleya loddigesii seedlings. Plantlets with approximately 90 days were subcultured under eight treatments, consisting of different active charcoal concentrations (0; 0.5; 1.0 and 2.0g L-1) in ½ MS medium combined with white and red light spectra. After 180 days of germination, biometric data, fresh weight, and the level of photosynthetic pigments were evaluated. Plantlets acclimatized in a greenhouse were evaluated for survival after 120 days. Total chlorophyll, chlorophyll a, and carotenoid concentrations were higher in treatments under white light, while red light promoted greater chlorophyll b, plantlets with fewer and shorter roots, and high ex vitro mortality. The addition of 2.0g L-1 of active charcoal to the culture medium and the use of white light provided greater development efficiency both on in vitro culture and ex vitro plant survival.

Enraizamento de microestacas de Lavandula angustifolia / Rooting of Lavandula angustifolia microcuttings

A eliminação da etapa de enraizamento in vitro na micropropagação de plantas é desejável do ponto de vista econômico, além de proporcionar a melhoria na qualidade do sistema radicial formado. Dois experimentos foram realizados com os objetivos de avaliar diferentes concentrações (0; 2,5; 5,0 e 10mM) de ácido indolbutírico (AIB) no enraizamento ex vitro de lavanda (L. angustifolia), cv. 'Provence Blue' e avaliar a capacidade de enraizamento ex vitro das cultivares 'Vera', 'Provence Blue', 'English' e 'Elegance Ice'. Após 30 dias, foi avaliado o número de microestacas enraizadas, comprimento das raízes principais, porcentagem de enraizamento e porcentagem de sobrevivência. A concentração de 5,0mM de AIB foi mais efetiva para o comprimento de raízes e porcentagem de enraizamento das microestacas de lavanda cv. 'Provence Blue', apesar de reduzir o número de raízes formadas. Entre as cultivares estudadas, a porcentagem de sobrevivência das plantas variou de 82 por cento a 100 por cento. As cultivares apresentaram diferenças no enraizamento ex vitro das microestacas, sendo as maiores médias de porcentagem de enraizamento registradas na 'Provence Blue' e 'Elegance Ice'. Conclui-se que a microestaquia pode ser uma técnica eficiente para a propagação de lavanda, pelo tratamento das microestacas com 5,0mM de AIB, por proporcionar alta porcentagem de enraizamento e sobrevivência das plantas.

Two experiments were carried out aiming to evaluate the ex vitro rooting of L. angustifolia cv. 'Provence Blue' treated with different concentrations (0, 2.5, 5.0 and 10mM) of indolebutyric acid (IBA) with talc as a vehicle to evaluated the ex vitro rooting of 'Vera', 'Provence Blue', 'English' and 'Elegance Ice' lavender cultivars. The experiments were carried out in a greenhouse using three concentrations of AIB plus control. After the 30th day, it was evaluated: surviving microcuttings percentage, percentage of rooted microcuttings, roots number, roots length. It was concluded that the microcutting can be an efficient technique for lavender plantlets production, by using micropropagated plants as explants donor. The concentration of 5.0mM IBA was more effective on root length and rooting rate of microcuttings lavender cv. 'Provence Blue', despite reducing the number of roots. Among the cultivars the percentage of plant survival ranged from 82 percent to 100 percent. The lavender cultivars evaluated showed differences in ex vitro microcuttings rooting. The cultivars 'Provence Blue' and 'Elegance Ice' achieved higher percentage of rooting. The cultivars had no influence on the percentage of microcuttings survival. It was concluded that the microcuttings can be an efficient technique for the propagation of lavender since microcuttings treated with 5.0mM IBA have higher survival and rooting percentage of rooting and survival.

Niveles de sacarosa en el enraizamiento in vitro y aclimatización ex vitro de plántulas del portainjerto de vid R110 (vitis rupestris' vitis berlandieri) / Sucrose levels in the in vitro rooting and ex vitro acclimatization of plantlets of the R110 (vitis rupestris x vitis berlandieri) grapevine rootstock / Níveis de sacarose no enraizamento in vitro e aclimatização ex vitro de plantas do porta-enxerto de videira R110 (vitis rupestris x vitis berlandieri)

La concentración de sacarosa y el intercambio gaseoso del contenedor son factores que influyen en las características de las plántulas micropropagadas. Brotes apicales del portainjerto de vid R110 (Vitis rupestris×Vitis berlandieri) fueron cultivados in vitro en medio con 0; 43,8; 87,6; 131,4 y 175,2mM de sacarosa, en contenedores con o sin intercambio gaseoso complementario. Después de 13 días de cultivo las plántulas fueron trasplantadas y colocadas en invernadero, donde permanecieron por nueve semanas. El porcentaje de enraizamiento, número y tamaño de raíces, tamaño de brotes, peso de materia fresca y contenido de agua de los explantes in vitro, disminuyeron al incrementar la concentración de sacarosa, en los contenedores con intercambio gaseoso. En la etapa de aclimatización, la concentración de sacarosa y tipo de contenedor influyeron en el número y tamaño de raíces. El tamaño de brotes, número de hojas y el contenido de agua fueron menores en las plántulas que crecieron con 175,2mM de sacarosa, en contenedores sin intercambio gaseoso. La supervivencia fue de 100% después de nueve semanas en plantas que se desarrollaron en medio con 43,8mM de sacarosa en contenedores sin intercambio; así como en 43,8 y 87,6mM en contenedores con intercambio gaseoso.

Sucrose concentration and container gas exchange are factors that influence the characteristics of micropropagated and acclimatized plantlets. Apical buds of the rootstock of the R110 (Vitis rupestris´Vitis berlandieri) grapevine were cultivated in media containing 0, 43.8, 87.6, 131.4, and 175.2mM sucrose, in containers with or without complementary gas exchange. After 13 days of growth, the plantlets were transplanted and placed in a greenhouse, where they remained for nine weeks. Percentage of rooting, number and length of the roots, bud size, weight of fresh material and water content in vitro, all decreased with the increase in the concentration of sucrose, the change being more pronounced in containers with gas exchange. In the acclimatization stage, sucrose concentration and type of container did not influence the number and length of the roots. However, bud size, number of leaves, and water content were lower in the plantlets growing with 175.2mM sucrose in containers with gas exchange. The survival rate was 100% after nine weeks with 43.8mM of sucrose in containers without exchange, and with 43.8 and 87.6mM in containers with gaseous exchange.

A concentração de sacarose e o intercâmbio gasoso do contenedor são fatores que influencíam nas características das plantas micropropagadas e aclimatizadas. Devido ao anterior, se cultivaram in vitro brotes apicais do porta-enxerto de videira R110 (Vitis rupestris×Vitis berlandieri), en medio de cultivo com 0, 43,8; 87,6; 131,4 e 175,2mM de sacarose, em contenedores com ou sem intercâmbio gasoso complementar. Depois de 13 dias de cultivo, as plantas foram transplantadas e colocadas em estufa onde permaneceram por nove semanas. In vitro, a porcentajem de enraizamento, número e tamanho de raízes, tamanho de brotos, peso de materia fresca e conteúdo de água diminuiram com o incremento da concentração de sacarose, de manera más acentuada nos contenedores com intercâmbio gasoso. Na etapa da aclimatização, a concentração de sacarose e o tipo de contenedor não influenciaram no número e tamanho das raízes. Entretanto, o tamanho de brotos, número de folhas e o conteúdo de água foram menores nas plantas que cresceram com 175,2mM de sacarose em contenedores sem intercâmbio gasoso. A sobrevivência foi de 100%, depois de nove semanas, com 43,8mM de sacarose em contenedores sem intercâmbio e com 43,8 ou 87,6mM, em contenedores com intercâmbio gasoso.

Evaluación del efecto del medio MS y la temperatura en el desarrollo de propágulos de Marchantia polymorpha L (Marchantiaceae) bajo condiciones in vitro y ex vitro / Evaluating the effect of MS medium and temperature in developing Marchantia polymorpha L (Marchantiaceae) propagules in in vitro and ex vitro conditions

En este trabajo de investigación se propagaron, bajo condiciones in vitro, propágulos de Marchantia polymorpha evaluando su desarrollo vegetativo mediante observaciones y registros semanales a cada cultivo por tratamiento en función del tiempo de desarrollo. En este contexto, se evaluó cualitativamente la forma y las características del talo, las cuales se valoraron mediante observaciones y registros semanales. Se analizaron los resultados cuantitativos mediante un análisis de varianza con diseño al azar y arreglo factorial 4 x 2, mediante la prueba estadística de Fisher. Las condiciones apropiadas de cultivo para propágulos de M. polymorpha se establecieron en una concentración de 25% de sales minerales Murashige y Skoog (1962), incubados a una temperatura de 25 ± 1 ºC. Luego de 13 semanas de desarrollo bajo condiciones in vitro, se adaptaron a condiciones naturales mediante un cambio de sustrato, y controlando la temperatura y humedad en el lugar de desarrollo. En esta etapa se evaluó la sobrevivencia de plantas durante 10 semanas; posteriormente al cambio de condiciones, y como característica cualitativa, se tuvo en cuenta el vigor del talo. Este protocolo de propagación de M. polymorpha, pionero en Colombia, es un modelo que permite conservar y cultivar de manera masiva diferentes especies de briófitos, especialmente aquellos que se encuentran en vías de extinción en nuestro país.

Marchantia polymorpha propagules were propagated in this research in in vitro conditions to assess their growth in terms of development time. Development rate and contamination regarding each treatment were also evaluated by weekly observations and crop records for each experimental unit per treatment. Talus shape and characteristics were also qualitatively evaluated by weekly observations and records. Fisher’s statistical test was used for analysing quantitative results by variance analysis with random design and 4 x 2 factorial arrangement. Appropriate conditions for cultivating M. polymorpha propagules were established at 25% Murashige and Skoog mineral salt concentration (1962), incubated at 25ºC ± 1ºC. After 13 weeks development in in vitro conditions, they were adapted to natural conditions by changing development site substrate and controlling temperature and humidity. Plant survival was evaluated for 10 weeks during this stage. Conditions were then changed and (as qualitative characteristic) talus vigour was taken into account. In was also determined that the ex vitro cultivation level should allow for gradual adjustment to new humidity, temperature and substrate conditions, taking special care that these conditions were not so altered as to become insurmountable. This pioneering M. polymorpha propagation protocol in Colombia is a model for the future maintenance and mass development of different bryophyte species, especially those which are endangered in our country.

Alterações anatômicas de bananeiras micropropagadas em resposta a aclimatização ex vitro / Anatomic modifications of micropropagated banana plants in response to ex vitro acclimatization

Pesquisas acerca das modificações estruturais e fisiológicas inerentes ao processo de micropropagação são fundamentais para compreender os efeitos desta técnica, desenvolver protocolos mais eficientes e, sobretudo, reduzir perdas ex vitro. O objetivo neste trabalho foi avaliar e quantificar as modificações na anatomia foliar de bananeiras provenientes de micropropagação, durante a fase de aclimatização em casa de vegetação. Para tanto, brotações axilares de bananeira cv. Japira, provenientes da multiplicação in vitro, foram enraizadas em meio MS, acrescido de ANA (1mg L-1) e ágar (6g L-1), e mantidas à temperatura de 25°C±2°C e 16 horas de irradiância a 35µmol m-2 s-1, por 35 dias. Posteriormente, as plantas foram submetidas a diferentes períodos de aclimatização (zero, 21, 42, 63, 84 e 120 dias) e avaliadas quanto à anatomia, por meio de seções transversais e paradérmicas foliares. O delineamento experimental foi inteiramente casualizado. Verificou-se que as maiores alterações anatômicas ocorrem após 42 dias do transplantio ex vitro, com acentuado espessamento dos parênquimas clorofilianos e limbo foliar, bem como diferenciação da maioria dos tecidos. Quanto aos estômatos, estes estão distribuídos em ambas às faces da epiderme, com maior número na face abaxial e em folhas oriundas de primórdios foliares formados in vitro.

Researches about structural and physiological modifications in different stages of the micropropagation are fundamental to understand the effects of this technology to improve protocols and to reduce losses in the acclimatization. The objective of this study was to assess and to quantify the variations in the foliar anatomy of micropropagated banana plants during the ex vitro acclimatization in greenhouse. Thus, axillary buds from in vitro multiplication of Japira cultivar, were rooted in MS medium, added of NAA (1mg L-1) and agar (6g L-1), and kept at room temperature (25°C ±2°C) under 16 hours photoperiod and irradiation of 35µmol m-2 s-1, for 35 days. Subsequently, the plants were submitted to different acclimatization periods (zero, 21, 42, 63, 84 e 120 days) being the leaf anatomy of the plants evaluated by transversal and paradermal sections. A completely randomized design was used. The largest anatomical alterations it were verified after 42 days of the transplantation to ex vitro conditions, with pronounced thickness of chlorophyllian parenchyma and leaf blade, as well, as the differentiation of the majority of foliar tissues. The stomata were distributed on both sides of the leaves, with higher number on the undersurface and on leaves formed from in vitro foliar primordia.

Ex vitro and in vitro leaf anatomy of Tabebuia roseo alba (Ridl.) Sand. Bignoniaceae - DOI: 10.4025/actascibiolsci.v31i3.1937 / Anatomia foliar de ipê-branco (Tabebuia roseo Alba (Ridl.) Sand.) Bignoniaceae, proveniente do cultivo ex vitro e in vitro - DOI: 10.4025/actascibiolsci.v31i3.1937

Considering the importance of knowledge of anatomical structures in the protocol definition for the micropropagation of the ipê-branco, this study compared the in vitro and ex vitro internal structure of leaves. For anatomical evaluations, the first leaf in vitro with 30 days of growth and adult plants were used. The anatomical study of leaves was based on microscope examination from cross-sectional and paradermic sections from the leaf blade. The leaf structures from plants ex vitro show uniseriate skin, and mesophyll with dorsiventral organization. They are hipostomatics and trichome is present in all faces. Leaves ex vitro was thicker that culture in vitro in limb, central nervure, epidermis and palisade and spongious parenchyma. In leaves in vitro, cuticle and sclerenchyma are absent. Leaves of ex vitro presented minor numbers of stomata and greater number of trichromes when compared with culture in vitro. Stomata of in vitro are larger than stomata ex vitro.

Este estudo teve como objetivo comparar a estrutura interna de folhas de ipê-branco cultivadas in vitro e ex vitro. Para as avaliações anatômicas foram utilizadas folhas do primeiro nó de ramos, da base para o ápice, com 30 dias de cultivo in vitro e de plantas de campo. O estudo anatômico foi feito por meio das secções transversais e paradérmicas das folhas. As folhas apresentam epiderme uniestratificada e mesofilo dorsiventral. São hipostomáticas e apresentam tricomas em todas as faces. Folhas de plantas cultivadas ex vitro, quando comparadas com in vitro, tiveram espessura do limbo foliar, nervura central da epiderme adaxial e abaxial e os parênquimas paliçádico e esponjoso maiores. Em folhas oriundas do cultivo in vitro, a cutícula e o esclerênquima são ausentes. Folhas de plantas cultivadas ex vitro apresentaram menor número de estômatos e maior número de tricomas, quando comparadas com o cultivo in vitro. Os estômatos de folhas cultivadas in vitro são maiores que os de folhas ex vitro