Brotações adventícias de abacaxizeiro ornamental sob o efeito de benzilaminopurina, ácido naftalenoacético e períodos de subcultivo Adventitious shoot of ornamental pineapple under benzylaminopurine, naphthalene acetic acid and subculture period effect

O objetivo deste trabalho foi avaliar os efeitos da adição de benzilaminopurina (BAP), em combinação com o ácido naftalenoacético (ANA), e dos períodos de subcultivo, na formação de brotações in vitro de abacaxizeiro ornamental. O meio básico líquido consistiu de sais minerais MS e vitaminas. O delineamento experimental utilizado foi o inteiramente casualizado, em esquema fatorial 6x2x4, com seis concentrações de BAP (0, 0,125, 0,25, 0,5, 1 e 2 mg L-1) e duas de ANA (0 e 0,1 mg L-1), e quatro períodos de subcultivo (30, 60, 90 e 120 dias). Os explantes consistiram de brotações individuais de 1 cm de comprimento, oriundas da cultura in vitro. A formação de novas brotações foi observada em meio suplementado com BAP, em todos os períodos de subcultivo. O maior número médio de brotos por explante foi obtido em meio com a concentração de 1,2 mg L-1 de BAP e 120 dias de subcultivo.<br>The objective of this work was to evaluate the effect of benzylaminopurine (BAP) additions, in combination with naphthaleneacetic acid (NAA), and of subcultures periods on in vitro shoot formation of ornamental pineapple. The basal liquid medium consisted of MS salts and vitamins. The experiment was a completely randomized design, in a factorial arrangement 6x2x4, with six concentrations of BAP (0, 0.125, 0.25, 0.5, 1, e 2 mg L-1) and two of NAA (0 and 0.1 mg L-1), and four subculturing periods (30, 60, 90 e 120 days). The explants consisted of individualized shoots with 1 cm length from in vitro cultures. Shoot formation was observed in medium supplemented with BAP in all subculturing periods. The greatest average number of shoots per explant was obtained with BAP concentration of 1.2 mg L-1 and 120 days of subcultive

In vitro morphogenesis and cell suspension culture establishment in Piper solmsianum C. DC. (Piperaceae)

In vitro morphogenesis and cell suspension culture establishment in Piper solmsianum C. DC. (Piperaceae)). Piper solmsianum is a shrub from Southeast Brazil in which many biologically active compounds were identified. The aim of this work was to establish a cell suspension culture system for this species. With this in mind, petiole and leaf explants obtained from in vitro plantlets were cultured in the presence of different plant growth regulator combinations (IAA, NAA, 2,4-D and BA). Root and indirect shoot adventitious formation, detected by histological analysis, was observed. Besides the different combinations of plant growth regulators, light regime and the supplement of activated charcoal (1.5 mg.l(-1)) were tested for callus induction and growth. Cultures maintained in light, on a 0.2 mg.l(-1) 2,4-D and 2 mg.l(-1) BA supplemented medium, and in the absence of activated charcoal, showed the highest calli fresh matter increment. From a callus culture, cell suspension cultures were established and their growth and metabolite accumulation studied. The achieved results may be useful for further characterization of the activated secondary metabolites pathways in in vitro systems of P. solmsianum

Alterações morfológicas foliares em abacaxizeiros cv. IAC "Gomo-de-mel" micropropagados e aclimatizados em diferentes condições de luminosidade Morphological alterations in leave of micropropagated pineapple plants cv. IAC "Gomo-de-mel" acclimatizated in different conditions of luminosity

Plantas micropropagadas geralmente apresentam dificuldades de adaptação ao meio ex vitro, sendo muitas vezes submetidas a processos de rustificação para minimizar os impactos decorrentes da mudança de ambiente. Uma vez que a folha e seus anexos destacam-se como importantes indicativos das estratégias adaptativas das plantas a ambientes adversos, este trabalho teve por objetivo comparar a anatomia foliar de plantas de abacaxi cv. IAC "Gomo-de-mel" cultivadas in vitro com microplantas aclimatizadas em diferentes condições de luminosidade, sob telado com 50% de sombreamento e em pleno sol para verificar a necessidade do processo de rustificação para este cultivar. Avaliações por meio de microscopia de luz e eletrônica de varredura da epiderme foliar, demonstraram aumento na densidade de escamas em ambas as superfícies das folhas, em microplantas dos ambientes ex vitro, principalmente em relação às que foram expostas diretamente ao sol. Observou-se ainda, aumento no espessamento da cutícula, na sinuosidade das células epidérmicas, e na distribuição e quantidade de fibras no mesofilo evidenciando a interferência das condições de luminosidade nas características morfológicas das microplantas. Essas alterações não prejudicaram o desenvolvimento das microplantas, indicando que não são necessárias etapas de rustificação para a aclimatização deste cultivar.<br>Microprapagated plants usually show difficulties to adapt to ex vitro conditions, and many times are submitted to the rustication process to aim the reduction of all the impacts resulting from the environmental changes. Once the leaf and its annexes are important indicators of adaptability strategies of the plants to adverse environmental conditions, the objective of this work was to compare the leaf anatomy of pineapple cv. IAC "Gomo-de-mel" in vitro cultivated plants with microplants acclimatized in different conditions of luminosity, under mesh, with 50 % of shading and directly exposed to sunlight, to verify the needed of rustication process on this cultivar. Evaluations of the leaf epidermis using light and electronic scanning microscopy showed an increase on scale density in both leaves surfaces of the ex vitro microplants, mainly related to the ones directly exposed to sunlight. Subsequent observations showed an increase on cuticle thickness, on wavy contours of epidermal cells, and on the distribution and quantity of mesophyll fibers, evidencing the light conditions interference in morphological characteristics of these microplants. These alterations had not harmed microplant development, showing that are not need of rustication stages on the acclimatization process of this cultivar

The introduction of transgenes to control blackheart in pineapple (Ananas Comosus L.) cv. Smooth Cayenne by microprojectile bombardment

A transformation technique for the introduction of transgenes to control blackheart by particle bombardment has been developed for pineapple cv. Smooth Cayenne. Leaf callus cultures capable of high frequency organogenesis with a short regeneration time were used as explant material. Gus and gfp reporter genes were used to observe and determine transient and stable expression. The ppo gene, isolated from pineapple, was introduced to control blackheart. Co-transformation occurred with constructs containing the nptII gene conferring geneticin resistance. We have recovered 15 independent transgenic gus and gfp lines each from 8 separate experiments and 22 ppo lines from 11 experiments. Gus, gfp, ppo and nptII positive plants have been regenerated, which have been shown by Southern blot analysis to be stable transgenics containing multiple copies of the introduced genes. These results show that biolistic gene delivery in pineapple can be successfully achieved at an acceptable efficiency of 0.21-1.5% for genetic improvement of 'Smooth Cayenne', the industry standard throughout the world