Anatomical analysis of peach palm (Bactris gasipaes) leaves cultivated in vitro, ex vitro and in vivo

(Anatomical analysis of peach palm (Bactris gasipaes) leaves cultivated in vitro, ex vitro and in vivo). The present work characterized and compared the anatomical structures of the leaves of Bactris gasipaes (Arecaceae) plants grown under different cultivation conditions (in vitro, ex vitro and in vivo) with the goal of identifying the origins of the difficulties encountered in acclimatizing micro-plants. The Quant program was used to determine leaf tissue thicknesses and areas, and histochemical tests were performed on leaf sections and analyzed using light microscopy. Stomatal and trichome densities were determined using the epidermal impression method and by scanning electronic microscopy. Our results indicated that there were no discernible alterations of the anatomical characteristics of the leaves of micro-plants cultivated under differing conditions and that the thickening of the mesophyll and the vascular fibers indicated adaptive responses to ex vitro conditions. As such, the observed difficulties in acclimatizing peach palm micro-plants to ex vitro conditions cannot be attributed to plant anatomical characteristics acquired during in vitro cultivation.

Difference in competence for in vitro proliferation and ex vitro growth of genetically identical mature and juvenile clones of apomictic Malus species

The apomictic system in Malus wits Used Is a model to examine rejuvenation by generating genetically identical tissue culture lines that had two entirely different developmental origins: either embryo-derived tissues (juvenile clones) or somatic tissue from the adult/mature tree (mature clones). These two lines were then subsequently used to examine in vitro difference between mature (M) and juvenile (J) tissues in potential for shoot, root proliferation and ex vitro (glasshouse) growth. The M clones of M. hupehensis and M. toringoides in vitro had significantly fewer total shoots and shoot more than 2 cm in length per proliferating explant than the J clones and also rooted less efficiently. Ex vitro (glasshouse) juvenile clones had shorter internodes, a greater number of leaves and more dry weight compared to their mature counterparts.

Auxin-producing bacteria isolated from the roots of Cattleya walkeriana, an endangered Brazilian orchid, and their role in acclimatization

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Germinação de sementes in vitro e desenvolvimento de plantas ex vitro de algodãozinho-do-campo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

MICROPROPAGAÇÃO do KIWI CV. HAYWARD

O presente trabalho teve como objetivo desenvolver um protocolo para a obtenção de mudas de kiwi (Actinidia chinensis Planch), cv. Hayward, por meio do cultivo in vitro de cotilédones. Utilizou-se o meio de MURASHIGE & SKOOG (1962) -- MS, suplementado com dois tipos de auxina (AIA e AIB) e uma citocinina (BAP). Foram verificados os efeitos de três doses de auxinas (0,125; 0,250 e 0,375 mg.L-1), combinadas com três doses de citocinina (0,5; 1,0 e 1,5 mg.L-1) na capacidade morfogênica dos explantes. Procedeu-se o estudo histológico dos órgãos das plântulas obtidas in vitro, e verificou-se, também, a capacidade de aclimatação das mudas ex vitro. A menor dose de AIB (0,125 mg.L-1), independentemente das doses de BAP, foi a mais eficaz na morfogênese dos explantes. Não foram verificadas alterações histológicas e anatômicas das plântulas obtidas in vitro. Aos três meses após o cultivo ex vitro dos explantes, verificou-se a sobrevivência de 88% das plantas transplantadas em condições de campo.

Isolamento, identificação e inoculação de bactérias produtoras de auxinas associadas às raízes de orquídeas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

In vitro conservation and low cost micropropagation of Cochlospermum regium (Mart. Ex. Scharank)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Concentrações de sacarose no desenvolvimento in vitro e na aclimatização de Cattleya loddigesii Lindley

This work aimed to study the influence of sucrose in the culture medium for in vitro growth and acclimatization of the epiphytic orchid Cattleya loddigesii. Five sucrose treatments (absence, 10, 20, 30 and 40g L-1) were used in a randomic experimental design. Mature seeds were sowed in 1/2 MS culture medium and after 90 days the plantlets (1.0 +/-0.2 cm) were inoculated between the treatments, whereby they were remained more 90 days. After 180 days of the beginning of the experiment the plantlets were removed from the flasks and evaluated the number of roots, shoot length, number of leafs, total dry weight and photosynthetic pigments. Survival percentage was evaluated after 75 days of acclimatization. The data of biometric variables were analyzed by Anovaand polynomial regression (p<0.05). Theothers data were submitted to the Anova and the means compared by the Tukey test (p<0.05). The sucrose concentration of 20g L-1 favored the in vitro growth in all evaluated parameters, showed higher production of chlorophyll a, total chlorophyll and carotenoids, in addition to increased survival under ex vitro condition. The sucrose concentration of 20g L-1 in the culture medium was the most efficient among the tested concentrations both for in vitro growth and ex vitro establishment of Cattleya loddigesii.

Paclobutrazol no crescimento e desenvolvimento in vitro e na aclimatização de Sophronitis cernua (Lindl.) Lindl. e Brassavola flagellaris Barb. Rodr. (Orchidaceae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Paclobutrazol no crescimento e desenvolvimento de Zygopetalum crinitum Lodd. e Cattleya schilleriana Rchb.f. (Orchidaceae) durante as fases in vitro e de aclimatização

Pós-graduação em Agronomia (Produção Vegetal) - FCAV

Desenvolvimento in vitro e criopreservação de sementes de orquídeas

Pós-graduação em Agronomia (Produção Vegetal) - FCAV

Simplificação do processo de multiplicação in vitro da oliveira "Olea europaea L."

A oliveira tem sido multiplicada ao longo dos tempos por métodos convencionais de propagação vegetativa como a enxertia e a estacaria lenhosa e semilenhosa. No entanto, estes métodos revelam-se lentos ou ineficientes para determinadas cultivares. No caso da cv. ‘Galega Vulgar’, ainda com grande expressão no olival português, e de difícil enraizamento por estacaria semilenhosa, tem sido usada a micropropagação de modo a contornar essas limitações e assim obter um elevado número de plantas em curto período de tempo. O custo final de produção por este processo ainda é elevado, podendo comprometer a sua aplicação a nível comercial. Grande parte dos custos estão relacionados com a fase de enraizamento in vitro que carece de ambiente estéril e condições de assépsia para a sua execução. Com vista a uma redução de custos associados a esta fase de produção, pretendeu-se com este trabalho testar a viabilidade do enraizamento ex vitro, na ausência de condições de assépsia. Este método poderá permitir uma significativa redução da mão-de-obra, ao mesmo tempo que facilitará a aclimatização das plantas e a obtenção de um sistema radicular de melhor qualidade. Compararam-se as taxas de enraizamento in vitro (controlo), com as obtidas ex vitro. Foram utilizados explantes provenientes de dois clones da cv. ‘Galega Vulgar’, (cl. 1441 e cl. 2022) cultivados e mantidos in vitro há vários anos no Laboratório de Melhoramento e Biotecnologia da Universidade de Évora. Para além do clone foi avaliada a influência do tipo de estaca (basal e apical), da hormona de enraizamento (AIB e ANA), da sua concentração (540 e 3000 ppm) e ainda de dois substratos, Preformas Jiffy® e pastilhas de fibra coco. Os melhores resultados foram obtidos com o clone 1441 em pastilhas de fibra de coco prensada, com o uso de estacas basais. Quanto à auxina, não se observaram diferenças significativas entre a utilização de ANA na concentração de 540 ppm e AIB na concentração de 3000 ppm. A aclimatização das plantas foi conseguida com taxas elevadas de sucesso, independentemente do tratamento utilizado. Conclui-se que a aplicação do método de enraizamento ex vitro simplifica procedimentos e mantém taxas de enraizamento elevadas, conduzindo assim a uma efetiva redução de tempo e custos associados; Simplifying procedures for in vitro propagation of olive “Olea europaea L.” Abstract: The olive tree has been multiplied throughout the ages by conventional methods of vegetative propagation such as grafting and wood or softwood cuttings. These propagation methods are somehow inefficient for certain cultivars. For the CV. ‘Galega Vulgar‘, still with great expression in Portuguese olive orchards, propagation has been attempted by in vitro culture in order to circumvent these limitations and so obtain a large number of plants in short time period. The final production fees associated to this process are still high which may compromise its application to a commercial level. Most of this process fees are related to the in vitro rooting phase which lacks sterile and aseptic conditions for its implementation. Aiming to reduce the costs associated with this production phase, this work tested the feasibility of the ex vitro rooting in the absence of aseptic conditions, which can allow a significant reduction of the manpower involved and an easier plant acclimatization due to its transplant with a balled-root system. In vitro rooting rates (control) were compared with those obtained with the ex vitro experiments. Explants from two clones of the cv. ‘Galega Vulgar‘ (cl. 1441 and cl. 2022), grown and maintained in vitro for several years in the Laboratory of Biotechnology and Plant Breeding of the University of Évora, were used in the trials. In addition to the clone, the effect of the cutting type (basal and apical), the rooting hormone (AIB and ANA), their concentration (540 and 3000 ppm) and two substrates, Preformas Jiffy ® and pressed coco fiber pellets, were also evaluated. The best results were obtained with the clone 1441, when rooted in pressed coco fiber pellets, using basal cuttings. Under this conditions no significant differences were observed between the use of ANA at 540 ppm or AIB in the 3000 ppm. Acclimatization of plants was achieved with high rates of success, regardless of the treatment used. It can be concluded that the application of the ex vitro rooting method allows to maintain high rooting rates, contributing for an effective reduction of time and fees of the rooting process.

In vitro propagation of Corymbia torelliana x C. citriodora (Myrtaceae) via cytokinin-free node culture

Hybrids between Corymbia torelliana (F.Muell.) K.D.Hill & L.A.S.Johnson and C. citriodora subsp. variegata (F.Muell.) A.R.Bean & M.W.McDonald are used extensively to establish forestry plantations in subtropical Australia. Methods were developed for in vitro seed germination, shoot multiplication and plantlet formation that could be used to establish in vitro and ex vitro clone banks of juvenile Corymbia hybrids. Effects of sodium hypochlorite concentration and exposure time on seed contamination and germination, and effects of cytokinin and auxin concentrations on shoot multiplication and subsequent rooting, were assessed. A two-step surface sterilisation procedure, involving 70% ethanol followed by 1% sodium hypochlorite, provided almost no contamination and at least 88% germination. A novel method of cytokinin-free node culture proved most effective for in vitro propagation. Lateral bud break of primary shoots was difficult to induce by using cytokinin, but primary shoots rooted prolifically, elongated rapidly and produced multiple nodes in the absence of exogenous cytokinin. Further multiplication was obtained either by elongating lateral shoots of nodal explants in cytokinin-free medium or by inducing organogenic callus and axillary shoot proliferation with 2.2 µm benzyladenine. Plantlets were produced using an in vitro soil-less method that provided extensive rooting in sterile propagation mixture. These methods provide a means for simultaneous laboratory storage and field-testing of clones before selection and multiplication of desired genotypes.