Micropropagação de Croton antisyphiliticus Mart.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Cultura de embrião e indução de brotos in vitro para micropropagação do pinhão-manso

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

In vitro growth of Brassocattleya orchid hybrid in different concentrations of KNO3, NH4NO3 and benzylaminopurine

Entre as maiores aplicações da cultura de tecidos de plantas está a propagação massal de mudas de plantas ornamentais. O objetivo deste trabalho foi avaliar o cultivo in vitro de um híbrido de orquídea Brassocattleya em diferentes concentrações de NH4NO3, KNO3 e BAP. Foram utilizadas sementes do híbrido de orquídea Brassocattleya 'Pastoral' e as plantas foram cultivadas em meio MS com redução pela metade das fontes de P, Mg e Ca e adição de 25 g L-1 de sacarose, 100 mg L-1 de mio-inositol, 1,5 g L-1 de carvão ativo e 6,5 g L-1 de ágar-ágar, sendo o pH ajustado para 5,8. Como tratamentos foram usados quatro concentrações dos sais NH4NO3 e KNO3 (2x; 1x; ½ e ¼ do meio MS) e três concentrações de BAP (0,0; 0,5 e 1,0 mg L-1). Avaliou-se a multiplicação, o crescimento em altura, massa fresca e seca, além dos teores de açucares redutores na massa seca das mudas. Observou-se grande influência das doses de NH4NO3 e KNO3 sobre o crescimento em altura das mudas, massa fresca e seca e teores de açucares redutores em Bc. ('Pastoral' x Auto). A dose de ¼ da utilizada no meio MS promoveu aumento significativo do crescimento das plantas. Para multiplicação, houve melhor resultado com a dose de ½ dos sais NH4NO3 e KNO3 utilizados no meio MS e 1,0 mg L-1 de BAP.

Efeito de reguladores vegetais no desenvolvimento de mudas de Passiflora alata Curtis

O principal fator limitante à produção de mudas enxertadas de maracujazeiro é o elevado tempo para a sua formação. Assim, objetivou-se avaliar o efeito de reguladores vegetais no desenvolvimento de mudas de Passiflora alata Curtis, a serem empregadas como porta-enxerto, visando a reduzir o tempo para atingir o ponto de enxertia. O delineamento experimental empregado foi o inteiramente casualisado, em esquema fatorial 4x5 (4 reguladores vegetais x 5 concentrações), com 4 repetições de 25 plantas por parcela. Os tratamentos foram constituídos por 0,0 mg L-1 (testemunha); 25 mg L-1; 50 mg L-1; 75 mg L-1, e 100 mg L-1 de Benziladenina (BA), GA4+7 + Fenilmetil-aminopurina (GA4+7+CK), Ácido giberélico (GA3) e Cloreto de chlormequat (CCC). Avaliaram-se o comprimento e o diâmetro do caule, o número de folhas, o comprimento e a fitomassa seca de raíz, do caule, das folhas e total. Os dados foram submetidos à análise de variância e de regressão polinomial, sendo realizado desdobramento quando houve efeito significativo da interação.Os reguladores não promoveram respostas significativas para o comprimento e a massa seca de raiz, do caule, de folhas e total. Porém, GA4+7+Fenilmetil-aminopurina foi o regulador que incrementou o comprimento do caule, o diâmetro e o número de folhas, promovendo maior desenvolvimento às mudas e reduzindo o tempo para a formação do porta-enxerto, de três a oito meses, para dois meses (63 dias).

EFEITOS DE GIBERELINAS, CITOCININAS E do NITRATO DE POTÁSSIO NO CRESCIMENTO E DESENVOLVIMENTO do PORTA-ENXERTO DE LIMOEIRO `CRAVO'

Os efeitos de reguladores vegetais do grupo das giberelinas e citocininas, bem como do nitrato de potássio, foram estudados no crescimento e desenvolvimento do porta-enxerto de limoeiro `Cravo' (Citrus limonia Osbeck). As sementes foram extraídas de frutos maduros, lavadas, secas à sombra e armazenadas em câmara fria (4 - 5oC) durante sessenta dias, até o mês de maio/1993, quando foram semeadas nos canteiros de semeadura. As pulverizações com os fitorreguladores foram realizadas no viveiro e tiveram início com as plantas aos 8 meses de idade, contados a partir da semeadura, sendo efetuadas um total de 4 pulverizações, realizadas a intervalos quinzenais. Os tratamentos corresponderam a: 25, 50 e 75 ppm de GA3; 25, 50 e 75 ppm de GA4+GA7+fenilmetil-aminopurina; 20 ppm de fenilmetilpiranil-aminopurina; 0,2% de KNO3 e testemunha (sem pulverização). Os resultados obtidos permitiram concluir que as pulverizações com os reguladores vegetais e com o nitrato de potássio, realizadas no período de janeiro a março (verão), não tiveram efeito na diminuição do tempo de formação das plantas jovens de limoeiro `Cravo', tendo inclusive, o tratamento com KNO3 0,2%, exercido efeito depressivo no desenvolvimento das mesmas.

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

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

Desiccation tolerance in seeds of Annona emarginata (Schldtl.) H. Rainer and action of plant growth regulators on germination

Annona emarginata (Schldtl.) H. Rainer (araticum-de-terra-fria) is used as a rootstock for several species of Annonaceae. It is suggested that these seeds should be sown immediately after extraction and, therefore, they could be intolerant to desiccation. There are several mechanisms involved with desiccation tolerance. Soluble sugars, for example, can accumulate and act as osmoprotectants for the membrane system during desiccation. The aim of this study is to assess desiccation tolerance in A. emarginata seeds. In addition, we examined the soluble sugars involved in desiccation tolerance. Finally, we determined the effect of gibberellic acid (GA4+7) and N-(phenylmethyl)-aminopurine in promoting the germination of seeds with different water contents. The experiment consisted of a randomized 4×5 factorial design (desiccation levels × concentration of growth regulators). After drying, seeds containing 31 (control), 19, 12 and 5% water were incubated in different concentrations of GA4+7 N-(phenylmethyl)-aminopurine (0, 250, 500, 750 and 1000 mg L-1) for 60 hours. The experiment was conducted in a germination chamber with alternating temperature and photoperiod of 20oC for 18 hours of darkness and 30oC for 6 hours of light. We analyzed electrical conductivity, germination rate, mean germination time, germination speed, frequency and uniformity of germination, percentage of dormant seeds and soluble sugar profile in intact seeds through high-performance liquid chromatography (HPLC). The data were subjected to analysis of variance, and the means were compared using Tukey's test at a threshold of p<0.05. The results showed that seeds of A. emarginata appears to be desiccation tolerant and, also, that sucrose increases when seed water content is reduced to values as low as 12%, exogenous GA4+7+N-(phenylmethyl)-aminopurine improves its germinability.

Micropropagação e conservação in vitro de Croton antisyphiliticus Mart

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo agronômico, químico e biológico de Cochlospermum regium (mart. ex. Scharank): uma planta medicinal do Cerrado

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

Reguladores vegetais na emergência e no desenvolvimento de plantas de macadâmia (Macadamia integrifolia Maiden & Betche)

Pós-graduação em Ciências Biológicas (Botânica) - IBB

Protocolo para micropropagação de bananeira ‘Thap maeo’

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Aplicação de regulador vegetal e poda nas relações fisiológicas na produção do tomateiro

Pós-graduação em Agronomia (Horticultura) - FCA

Nitric Oxide Mediates the Hormonal Control of Crassulacean Acid Metabolism Expression in Young Pineapple Plants

Genotypic, developmental, and environmental factors converge to determine the degree of Crassulacean acid metabolism (CAM) expression. To characterize the signaling events controlling CAM expression in young pineapple (Ananas comosus) plants, this photosynthetic pathway was modulated through manipulations in water availability. Rapid, intense, and completely reversible up-regulation in CAM expression was triggered by water deficit, as indicated by the rise in nocturnal malate accumulation and in the expression and activity of important CAM enzymes. During both up-and down-regulation of CAM, the degree of CAM expression was positively and negatively correlated with the endogenous levels of abscisic acid (ABA) and cytokinins, respectively. When exogenously applied, ABA stimulated and cytokinins repressed the expression of CAM. However, inhibition of water deficit-induced ABA accumulation did not block the up-regulation of CAM, suggesting that a parallel, non-ABA-dependent signaling route was also operating. Moreover, strong evidence revealed that nitric oxide (NO) may fulfill an important role during CAM signaling. Up-regulation of CAM was clearly observed in NO-treated plants, and a conspicuous temporal and spatial correlation was also evident between NO production and CAM expression. Removal of NO from the tissues either by adding NO scavenger or by inhibiting NO production significantly impaired ABA-induced up-regulation of CAM, indicating that NO likely acts as a key downstream component in the ABA-dependent signaling pathway. Finally, tungstate or glutamine inhibition of the NO-generating enzyme nitrate reductase completely blocked NO production during ABA-induced up-regulation of CAM, characterizing this enzyme as responsible for NO synthesis during CAM signaling in pineapple plants.

Genetic transformation of three sweet orange cultivars from explants of adult plants

The difficulty in adult tissue genetic transformation in woody species is still an obstacle to be overcome, including in most sweet orange cultivars of the Brazilian citrus industry. This work reports that, after in vitro culture adjustments, transgenic adventitious buds of 'Hamlin', 'Pra', and 'Valencia' sweet oranges (Citrus sinensis L. Osbeck) were recovered using adult material as explant source, in genetic transformation experiments via Agrobacterium tumefaciens. The transgenic buds were identified by the GUS histochemical analysis and confirmed by PCR analysis, which indicated the presence of an amplified fragment of 817 bp corresponding to the uidA gene sequence. The efficiencies of genetic transformation for 'Hamlin', 'Pra', and 'Valencia' sweet orange cultivars were 2.5, 1.4, and 3.7%, respectively. Media supplemented with auxins and cytokinins during co-culture, and media with high concentrations of cytokinins (3 mg L-1) during transgenic selection led to the transformation and, consequently, the regeneration of adequate number of adventitious buds for the three cultivars. The use of sonication during the explant disinfection was not effective to reduce endophytic contamination and reduced transformation efficiency.

Gene expression during early somatic embryogenesis in Brazilian pine (Araucaria angustifolia (Bert) O. Ktze)

Brazilian pine (Araucaria angustifolia (Bert) O. Ktze) is the only native conifer species with economic importance in Brazil. Recently, due to intensive exploitation Brazilian pine was included in the official list of endangered Brazilian plants, under the "vulnerable" category. Biotechnology tools like somatic embryogenesis (SE) are potentially useful for mass clonal propagation and ex situ conservation strategies of commercial and endangered plant species. In spite of that, numerous obstacles still hamper the full application of SE technology for a wider range of species, including Brazilian pine. To enhance somatic embryogenesis in Brazilian pine and to gain a better understanding of the molecular events associated with somatic embryo development, we analyzed the steady-state transcript levels of genes known to regulate somatic embryogenesis using semiquantitative reverse transcription polymerase chain reaction (sqRT-PCR). These genes included Argonaute (AaAGO), Cup-shaped cotyledon1 (AaCUC), wushel-related WOX (AaWOX), a S-locus lectin protein kinase (AaLecK), Scarecrow- like (AaSCR), Vicilin 7S (AaVIC), Leafy Cotyledon 1 (AaLEC), and a Reversible glycosylated polypeptide (AaRGP). Expression patterns of these selected genes were investigated in embryogenic cultures undergoing different stages of embryogenesis, and all the way to maturation. Up-regulation of AaAGO, AaCUC, AaWOX, AaLecK, and AaVIC was observed during transition of somatic embryos from stage I to stage II. During the maintenance phase of somatic embryogenesis, expression of AaAGO and AaSCR, but not AaRPG and AaLEC genes was influenced by presence/ absence of plant growth regulators, both auxins and cytokinins. The results presented here provide new insights on the molecular mechanisms responsible for somatic embryo formation, and how selected genes may be used as molecular markers for Brazilian pine embryogenesis.