Effects of kinetin and nitrogen on growth rates, pigment and protein contents in wild and phycoerythrin-deficient strains of Hypnea musciformis (Rhodophyta)

Hypnea musciformis (Wulfen in Jacqu.) J.V. Lamour. is the main source for carrageenan production in Brazil and strains with selected characteristics could improve the production of raw material. The effects of kinetin on growth rates, morphology, protein content, and concentrations of pigments (chlorophyll a, phycoerythrin, phycocyanin, and allophycocyanin) were assessed in the wild strain (brown phenotype) and in the phycoerythrin-deficient strain (green phenotype) of H. musciformis. Concentrations of kinetin ranging from 0 to 50 mu M were tested in ASP 12-NTA synthetic medium with 10 mu M nitrate (N-limited) and 100 mu M nitrate (N-saturated). In N-limited condition, kinetin stimulated growth rates of the phycoerythrin-deficient strain and formation of lateral branches in both colour strains. Kinetin stimulated protein biosynthesis in both strains. However, differences between both nitrogen conditions were significant only in the phycoerythrin-deficient strain. In the wild strain, effects of kinetin on concentrations of phycobiliproteins were not significant in both nitrogen conditions, except for chlorophyll content. However, the phycoerythrin-deficient strain showed an opposite response, and kinetin stimulated the phycobiliprotein biosynthesis, with the highest concentrations of phycoerythrin in N-saturated medium, while the highest concentrations of allophycocyanin and phycocyanin were observed in N-limited medium. These results indicate that the effects of kinetin on growth, morphology, protein and phycobiliprotein contents are influenced by nitrogen availability, and the main nitrogen storage pools in phycoerythrin-deficient strain of H. musciformis submitted to N-limited conditions were phycocyanin and allophycocianin, the biosynthesis of which was enhanced by kinetin.

Efficacy of Cosmetic Formulations Containing Dispersion of Liposome with Magnesium Ascorbyl Phosphate, Alpha-Lipoic Acid and Kinetin

The present study aimed to evaluate the photoprotective effects of cosmetic formulations containing a dispersion of liposome with magnesium ascorbyl phosphate (MAP), alpha-lipoic acid (ALA) and kinetin, as well as their effects on the hydration and viscoelastic skin properties. The photoprotection was determined in vitro (antioxidant activity) and in vivo on UV-irradiated hairless mouse skin. The hydration effects were performed with the application of the formulations under study on the forearm of human volunteers and skin conditions were analyzed before and after a single application and daily applications during 4 weeks in terms of transepidermal water loss (TEWL), skin moisture and viscoelastic properties. The raw material under study possessed free-radical scavenging activity and the formulation with it protected hairless mouse skin barrier function against UV damage. After 4 weeks of application on human skin, the formulation under study enhanced stratum corneum skin moisture and also showed hydration effects in deeper layers of the skin. Thus, it can be concluded that the cosmetic formulation containing a dispersion of liposome with MAP, ALA and kinetin under study showed photoprotective effects in skin barrier function as well as pronounced hydration effects on human skin, which suggests that this dispersion has potential antiaging effects.

Polyamine patterns in haploid and diploid tobacco tissues and in vitro cultures

The aim of this work was to determine PAs levels in pith tissues and callus cultures from haploid and diploid tobacco plants, explanted from the apical and basal regions of the stem. These explants were cultured in an RM-64 medium supplied with IAA and kinetin, under light or in the dark, during successive subcultures. PAs levels followed a basipetal decrease in diploid and an increase in haploid, pith tissues. A similar pattern of total PAs (free + conjugated) was observed for the callus of diploid and haploid plants maintained in the light, and for the haploid callus in the dark, whereas the diploid callus in the dark showed a constant increase in total PAs levels until the end of culture. The PA increase in the diploid callus in the dark was related to free Put levels increase. The ploidy status of the plants could express different PA gradients together with the plant pith and in vitro callus cultures.

Multiple shoot formation in lentil (Lens culinaris) seeds

A protocol based on seed culture was developed for efficient in vitro propagation of lentil (Lens culinaris Medik). Benzyladenine (BA), thidiazuron (TDZ), and kinetin all induced multiple shoot formation. In terms of the number of long shoots (>2.0 cm) produced per seed, BA and TDZ at optimum concentrations (0.2-0.4 and 0.1 mg/litre, respectively) had similar efficiency, whereas kinetin produced less shoots. Murashige and Skoog (MS) salt composition was better than that of Gamborge (B5) for shoot induction. Increasing calcium (Ca) concentration was necessary to overcome shoot-tip necrosis. For shoot elongation, fresh medium of the same composition of shoot induction medium could be used for stumps from medium with low BA (

Embryo rescue from interspecific crosses in apple rootstocks

The objetive of this work was to rescue immature embryos of apple rootstocks Malus prunifolia (Marubakaido) and Malus pumila (M9) after 40-60 days of pollination and to put them into MS culture media supplemented with agar (6 g L-1) and casein hydrolysate (500 mg L-1). Embryos originated from interspecific crosses and open pollination showed differences in the in vitro responses, depending on the female parent, the developmental stage of the embryo, and the culture medium composition. Embryos of the M. pumila rootstock, rescued within 40 days after pollination and put in culture medium supplemented with indolacetic acid (IAA), gibberellic acid (GA3), kinetin and maltose, resulted in a normal development of plantlets. However, embryos originating from hand-pollination, cultivated in medium supplemented with 14 µM IAA, 5 µM kinetin and 1.5 µM Ga3 (MS1), mainly those of M. prunifolia x M. pumila, showed a high percentage of rusted embryos (96.2%). Embryos from open pollination of M. prunifolia and M. pumila formed calluses. It was possible to identify the influence of the female parent by the enhanced development of M. pumila shoots derived from open or hand-pollination. The crossing of responsive species and the use of the technique of embryo culture provided a rapid and uniform germination and, consequently, the development of fully normal seedlings.

Leaf and root volatiles produced by tissue cultures of Alpinia zerumbet (pers.) Burtt & Smith under the influence of different plant growth regulators

Volatiles produced by plantlets of Alpinia zerumbet were obtained by means of simultaneous distillation-extraction (SDE). The effects of indole-3-acetic acid, kinetin, thidiazuron and 6-benzylaminopurine on leaf and root volatile composition obtained by tissue cultures were investigated. A higher content of b-pinene and a lower content of sabinene were observed in leaf volatile of plantlets cultured in control, IAA and IAA+ TDZ media, as compared with those of donor plants. In vitro conditions were favorable to increase caryophyllene content. Volatile compounds from the root were characterized mainly by camphene, fenchyl-acetate and bornyl acetate; which constitute about 60% of total volatile.

In vitro multiplication of Tabernaemontana fuchsiaefolia L. (Apocynaceae)

This study describes a simple and promising for in vitro multiplication of Tabernaemontana fuchsiaefolia, a species abundantly found in southern Brazil utilized for medicinal purposes and as a source of compounds that may be used to develop new synthetic drugs. Apical and hypocotyl explants were cultured in MS medium containing different concentrations of the cytokinins benzylaminopurine (BA) and 6-furfurylaminopurine (kinetin), supplemented with phloroglucinol (1, 3, 5-hydroxybenzene) to stimulate growth and shoot proliferation. Cytokinin added to the culture media positively influenced the micropropagation of T. fuchsiaefolia.and kinetin induced more shoots per explant than BA cytokinin. A favorable effect of phloroglucinol on apical and lateral buds from hypocotyls was also achieved in medium containing no kinetin or in all kinetin concentrations tested. Short pulses of auxin 3-indolebutyric acid (IBA) 5.0 mg/l resulted in satisfactory rooting in apical microcuttings. The addition of phloroglucinol to MS medium induced rhizogenesis in 29% of the nodal segments transferred to MS medium in the absence of IBA and in 50% of the nodal segments transferred to MS medium containing 0.5 mg/l IBA and in nodal segments previously submitted to short pulses of IBA.

MICROPROPAGATION AND ACCLIMATIZATION OF Aegiphila verticillata Vell.: AN ENDANGERED WOODY SPECIES

The objective of this work was to establish an efficient protocol for in vitro multiplication and rooting, as well as ex vitroacclimatization of Aegiphila verticillata, a woody species found in Brazilian rocky fields. Aseptic cultures were established by seeds and two multiplication analyses were performed. In the first, we employed 6-benzylaminopurine (BAP – 0, 2.5, 5 and 7.5 μM) + α-naphthalene acetic acid (NAA – 0, 0.2, 0.4 and 0.6 μM) and, in the second, were studied adenine sulfate, kinetin and thidiazuron (0, 5, 7.5, 10 and 12.5 μM). After 90 days, we assessed the quantitative and qualitative shoot propagation. There were more than 90% seed germination and low contamination (2%). In multiplication phase, the culture medium that promoted the best quantitative and qualitative culture development was supplemented with 7.5 μM BAP + 0.4 μM NAA. In the rooting assay, were used NAA, indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) (0, 0.1, 0.2, 0.3 or 0.4 μM). After 90 days, the root number and rooting quality were evaluated. In this analysis, differences were not found between the control and the other treatments. Rooted plantlets were acclimatized in styrofoam trays for 30 days, after which they were transferred to pots in the greenhouse. Only 3% of the plants subjected to initial acclimatization died and 70% of the plants transferred to the field conditions survived and showed normal development. The results founded in this work are the first involving in vitro propagation and ex vitroacclimatization of Aegiphila verticillata and provide a continuous supply of this medicinal native species, endangered due anthropogenic activities.

Indução de calos embriogênicos em explantes de cupuaçuzeiro

Objetivou-se a indução de calos embriogênicos em cupuaçuzeiro, em função do tipo de explante e meio de cultura. Foram testados como explantes, segmentos cotiledonares e eixos embrionários divididos em três partes: região da plúmula, radícula e hipocótilo. Os explantes foram cultivados em 2 diferentes meios de cultura: 1) MS suplementado com 2,4-D (1 mg L-1) e Cinetina (0,25 mg L-1); 2) MS acrescido de ANA (5 mg L-1) e Cinetina (0,25 mg L-1). Constatou-se que a região do hipocótilo foi a parte mais responsiva do eixo embrionário, formando calos com aspecto branco e friável. As auxinas testadas nos meios não estimularam o processo embriogênico em calos de cupuaçuzeiro.

Micropropagation of fig (Ficus carica L.) 'Roxo de Valinhos' plants

An in vitro protocol for Ficus carica cv. 'Roxo de Valinhos' was optimized. Nodal explants containing two buds were excised from field-grown mature plants, and transferred to different proliferation media consisting of combinations of distinct concentrations of activated charcoal with benzyladenine (BA), kinetin with gibberellic acid (GA(3)), and WPM (woody plant medium) with kinetin. The regular strength of WPM in combination with 0.5 mg l(-1) kinetin was the best condition for shoot proliferation of Ficus carica 'Roxo de Valinhos' plants. The addition of activated charcoal in the medium completely inhibited shoot proliferation. The inclusion of BA in the medium induced excessive callus formation as well as small and vitrified shoots, while GA(3) induced excessive elongation associated with vitrification, chlorosis, and tip-burned shoots.

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)

Uso de regulador de crescimento em cultivares de feijão de inverno

A aplicação de reguladores vegetais, com a finalidade de aumentar a produtividade das culturas, tem merecido, nos últimos anos, grande atenção de pesquisadores. No entanto, alguns resultados têm se mostrado contraditórios. em face disto, este estudo teve como objetivo verificar os efeitos da aplicação de regulador vegetal sobre os componentes da produção e a produtividade de grãos de duas cultivares de feijão de inverno, em condições de Cerrado. O experimento foi realizado no outono-inverno de 2007, na Fazenda Experimental da Unesp, Campus de Ilha Solteira, localizada no município de Selvíria (MS). O delineamento experimental foi o de blocos casualizados, no esquema em faixas, com quatro repetições. Os tratamentos constituíram-se da combinação de cinco doses do regulador vegetal (0 L ha-1 0,5 L ha-1 1,0 L ha-1 1,5 L ha-1 e 2,0 L ha-1, que é composto por três hormônios vegetais (cinetina, ácido giberélico e ácido indolbutírico), em duas épocas de aplicação: no estádio vegetativo (V4) e reprodutivo (R5). Características vegetativas como altura das plantas de feijão, altura de inserção da primeira vagem, número de grãos por vagem e massa de 100 grãos não foram afetadas pela aplicação do produto. No entanto, sua aplicação no estádio reprodutivo (R5) aumentou o número de grãos por planta e a produtividade de grãos das cultivares de feijoeiro Carioca Precoce e IAC Apuã. Para este aumento, a melhor dose foi 2 L ha-1do regulador vegetal.

Emergência e desenvolvimento de plântulas de maracujazeiro azedo oriundas de sementes tratadas com bioestimulante

O objetivo do trabalho foi estudar os efeitos de bioestimulante na emergência e no desenvolvimento de plântulas de Passiflora edulis Sims.f. flavicarpa Deg. O experimento foi conduzido sob cultivo protegido, com temperatura controlada (25ºC), no Departamento de Botânica, Instituto de Biociências, UNESP, Câmpus de Botucatu-SP. As sementes receberam os tratamentos com as concentrações 0 (testemunha); 4; 8; 12; 16 e 20 ml de bioestimulante/kg de semente e foram semeadas em bandejas de isopor contendo substrato comercial. O bioestimulante empregado é constituído por 0,005% de ácido índolbutírico (auxina), 0,009% de cinetina (citocinina) e 0,005% de ácido giberélico (giberelina). O delineamento experimental foi inteiramente casualizado, com seis tratamentos e cinco repetições de 24 sementes. As avaliações de porcentagem de emergência de plântulas foram realizadas semanalmente, bem como o comprimento de caule e raiz, diâmetro do caule, número de folhas, área foliar e massa seca de raiz, caule e folha, aos 35 dias após a semeadura. Os dados foram submetidos à análise de variância e regressão polinomial, ao nível de 5% de probabilidade. As concentrações de 12 e 16 ml de bioestimulante/kg de semente aplicado às sementes promoveram as maiores porcentagens de emergência e desenvolvimento de plântulas.