Callus induction and micropropagation improved by colchicine and phytoregulators in Kappaphycus alvarezii (Rhodophyta, Solieriaceae)

Tissue culture techniques were applied for micropropagation of the red alga Kappaphycus alvarezii in order to select the best strain and experimental system for in vitro culture. Five strains were tested: brown (BR), green (GR) and red (RD) tetrasporophytes, brown female gametophyte (BFG), and a strain originating from tetraspore germination (""Edison de Paula"", EP). The effects of three culture media were tested on callus formation, regeneration from explants and from callus in the three tetrasporophytic and EP strains: seawater enriched with half-strength of von Stosch`s (VS 50) and Guillard & Ryther`s (F/2 50) solutions, plus synthetic ASP 12-NTA medium, with or without gelling agent. Explants of the EP strain were treated with glycerol and the phytoregulators indole-3-acetic acid (IAA); 2,4-diclorophenoxyacetic acid (2,4-D); and benzylaminopurine (BA), alone or in combination. The effects of colchicine (0.01%) during 24, 48, 72 hours and 14 days were analyzed in the BFG and EP strains. The EP strain showed the highest percentage of explants forming callus and regeneration from explants in VS 50, indicating its high potential for micropropagation in comparison to the other strains. Regeneration from callus was very rare. Treatments with glycerol and IAA:BA (5:1 mg L(-1)) stimulated the regeneration from explants. Significant differences were observed in the percentages of regeneration of EP strain explants treated with colchicine for 14 days. Our results indicate that IAA and BA stimulated the regeneration process, and that colchicine produced explants with high potential for regeneration, being useful for improving the micropropagation of K. alvarezii.

Laurdan Spectrum Decomposition as a Tool for the Analysis of Surface Bilayer Structure and Polarity: a Study with DMPG, Peptides and Cholesterol

The highly hydrophobic fluorophore Laurdan (6-dodecanoyl-2-(dimethylaminonaphthalene)) has been widely used as a fluorescent probe to monitor lipid membranes. Actually, it monitors the structure and polarity of the bilayer surface, where its fluorescent moiety is supposed to reside. The present paper discusses the high sensitivity of Laurdan fluorescence through the decomposition of its emission spectrum into two Gaussian bands, which correspond to emissions from two different excited states, one more solvent relaxed than the other. It will be shown that the analysis of the area fraction of each band is more sensitive to bilayer structural changes than the largely used parameter called Generalized Polarization, possibly because the latter does not completely separate the fluorescence emission from the two different excited states of Laurdan. Moreover, it will be shown that this decomposition should be done with the spectrum as a function of energy, and not wavelength. Due to the presence of the two emission bands in Laurdan spectrum, fluorescence anisotropy should be measured around 480 nm, to be able to monitor the fluorescence emission from one excited state only, the solvent relaxed state. Laurdan will be used to monitor the complex structure of the anionic phospholipid DMPG (dimyristoyl phosphatidylglycerol) at different ionic strengths, and the alterations caused on gel and fluid membranes due to the interaction of cationic peptides and cholesterol. Analyzing both the emission spectrum decomposition and anisotropy it was possible to distinguish between effects on the packing and on the hydration of the lipid membrane surface. It could be clearly detected that a more potent analog of the melanotropic hormone alpha-MSH (Ac-Ser(1)-Tyr(2)-Ser(3)-Met(4)-Glu(5)-His(6)-Phe(7)-Arg(8)-Trp(9)-Gly(10)-Lys(11)-Pro(12)-Val(13)-NH(2)) was more effective in rigidifying the bilayer surface of fluid membranes than the hormone, though the hormone significantly decreases the bilayer surface hydration.

Velocity-dependent KAM islands

We revisit the non-dissipative time-dependent annular billiard and we consider the chaotic dynamics in two planes of conjugate variables in order to describe the behavior of the growth, or saturation, of the mean velocity of an ensemble of particles. We observed that the changes in the 4-d phase space occur without changing any parameter. They occur depending on where the initial conditions start. The emerging KAM islands interfere in the behavior of the particle dynamics especially in the Fermi acceleration mechanism. We show that Fermi acceleration can be suppressed, without dissipation, even considering the non-dissipative energy context. (C) 2011 Published by Elsevier Ltd.

Versatile microanalytical system with porous polypropylene capillary membrane for calibration gas generation and trace gaseous pollutants sampling applied to the analysis of formaldehyde, formic acid, acetic acid and ammonia in outdoor air

The analytical determination of atmospheric pollutants still presents challenges due to the low-level concentrations (frequently in the mu g m(-3) range) and their variations with sampling site and time In this work a capillary membrane diffusion scrubber (CMDS) was scaled down to match with capillary electrophoresis (CE) a quick separation technique that requires nothing more than some nanoliters of sample and when combined with capacitively coupled contactless conductometric detection (C(4)D) is particularly favorable for ionic species that do not absorb in the UV-vis region like the target analytes formaldehyde formic acid acetic acid and ammonium The CMDS was coaxially assembled inside a PTFE tube and fed with acceptor phase (deionized water for species with a high Henry s constant such as formaldehyde and carboxylic acids or acidic solution for ammonia sampling with equilibrium displacement to the non-volatile ammonium ion) at a low flow rate (8 3 nLs(-1)) while the sample was aspirated through the annular gap of the concentric tubes at 25 mLs(-1) A second unit in all similar to the CMDS was operated as a capillary membrane diffusion emitter (CMDE) generating a gas flow with know concentrations of ammonia for the evaluation of the CMDS The fluids of the system were driven with inexpensive aquarium air pumps and the collected samples were stored in vials cooled by a Peltier element Complete protocols were developed for the analysis in air of NH(3) CH(3)COOH HCOOH and with a derivatization setup CH(2)O by associating the CMDS collection with the determination by CE-C(4)D The ammonia concentrations obtained by electrophoresis were checked against the reference spectrophotometric method based on Berthelot s reaction Sensitivity enhancements of this reference method were achieved by using a modified Berthelot reaction solenoid micro-pumps for liquid propulsion and a long optical path cell based on a liquid core waveguide (LCW) All techniques and methods of this work are in line with the green analytical chemistry trends (C) 2010 Elsevier B V All rights reserved

Fully electrochemical hyphenated flow system for preconcentration, cleanup, stripping, capillary electrophoresis with stacking and contactless conductivity detection of trace heavy metals

Successful coupling of electrochemical preconcentration (EPC) to capillary electrophoresis (CE) with contactless conductivity detection (C(4)D) is reported for the first time. The EPC-CE interface comprises a dual glassy carbon electrode (GCE) block, a spacer and an upper block with flow inlet and outlet, pseudo-reference electrode and a fitting for the CE silica column, consisting of an orifice perpendicular to the surface of a glassy carbon electrode with a bushing inside to ensure a tight press fit. The end of the capillary in contact with the GCE is slant polished, thus defining a reproducible distance from the electrode surface to the column bore. First results with EPC-CE-C(4)D are very promising, as revealed by enrichment factors of two orders of magnitude for Tl, Cu, Pb and Cd ion peak area signals. Detection limits for 10 min deposition time fall around 20 nmol L(-1) with linear calibration curves over a wide range. Besides preconcentration, easy matrix exchange between accumulation and stripping/injection favors procedures like sample cleanup and optimization of pH, ionic strength and complexing power. This was demonstrated for highly saline samples by using a low conductivity buffer for stripping/injection to improve separation and promote field-enhanced sample stacking during electromigration along the capillary. (C) 2010 Elsevier B.V. All rights reserved.

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.

Investigating the formation and the properties of monoalkyl carbonates in aqueous medium using capillary electrophoresis with capacitively coupled contactless conductivity detection

Although alkyl carbonic acids (ACAs) and their salts are referred to as instable species in aqueous medium, we demonstrate that a monoalkyl carbonate (MAC) can in fact be easily formed from bicarbonate and an alcohol even in the presence of a high amount of water. A CE system with two capacitively coupled contactless conductivity detectors (C(4)Ds) was used to obtain different parameters about these species and their reactions. Based on the mobilities obtained for a series of alcohols ranging from 1 to 5 carbons, the coefficients of diffusion and the hydrodynamic radii were calculated. When compared with the equivalent carboxylates, MACs have radii systematically smaller. Although the precise pK(a) values of the ACAs could not be obtained, because of the fast decomposition in acid medium, it was possible, for the first time, to show that they are below 4.0. This result suggests that the acidity of an ACA is quite similar to the first hydrogen of H(2)CO(3). Using a new approach to indirectly calibrate the C(4)D, the kinetic constants and the equilibrium constants of formation were also obtained. The results suggest that the increase in the chain length makes the MACs less stable and more inert.

Automated two-dimensional separation flow system with electrochemical preconcentration, stripping, capillary electrophoresis and contactless conductivity detection for trace metal ion analysis

This paper describes the automation of a fully electrochemical system for preconcentration, cleanup, separation and detection, comprising the hyphenation of a thin layer electrochemical flow cell with CE coupled with contactless conductivity detection (CE-C(4)D). Traces of heavy metal ions were extracted from the pulsed-flowing sample and accumulated on a glassy carbon working electrode by electroreduction for some minutes. Anodic stripping of the accumulated metals was synchronized with hydrodynamic injection into the capillary. The effect of the angle of the slant polished tip of the CE capillary and its orientation against the working electrode in the electrochemical preconcentration (EPC) flow cell and of the accumulation time were studied, aiming at maximum CE-C(4)D signal enhancement. After 6 min of EPC, enhancement factors close to 50 times were obtained for thallium, lead, cadmium and copper ions, and about 16 for zinc ions. Limits of detection below 25 nmol/L were estimated for all target analytes but zinc. A second separation dimension was added to the CE separation capabilities by staircase scanning of the potentiostatic deposition and/or stripping potentials of metal ions, as implemented with the EPC-CE-C(4)D flow system. A matrix exchange between the deposition and stripping steps, highly valuable for sample cleanup, can be straightforwardly programmed with the multi-pumping flow management system. The automated simultaneous determination of the traces of five accumulable heavy metals together with four non-accumulated alkaline and alkaline earth metals in a single run was demonstrated, to highlight the potentiality of the system.

Controle de espécies indesejáveis em pastagem nativa

O experimento foi desenvolvido em uma área de pastagem nativa representativa da transição entre a Serra do Sudeste e a Depressão Central do RS. Foram testados os efeitos de diferentes métodos de controle de plantas indesejáveis e adubação, sobre a produção de forragem, dinâmica da vegetação e freqüência das espécies presentes na pastagem. Na primeira etapa avaliou-se o efeito inicial (até 60 dias após aplicação) dos tratamentos: T1 – sem controle (SC), T2 – controle mecânico (roçada de primavera, CMP) e T3 – controle químico (Picloram + 2,4-D, na dosagem de 5 L/ha, CQT), os quais foram arranjados num delineamento em blocos ao acaso, com quatro repetições. Aos 60 dias pós-aplicação dos tratamentos, a eficiência de controle das espécies indesejáveis em termos de participação na matéria seca foi de 100% para o CQ e 44,5% para o CMP quando comparados ao SC. Quando a eficiência foi calculada em termos de freqüência de ocorrência, verificou-se que o tratamento CM não foi eficiente no controle do alecrim no estrato inferior e de caraguatá no estrato superior, os quais aumentaram sua freqüência. Na segunda etapa, incluiu-se o tratamento de roçada de outono (CMO) e adubação ou não de cada tratamento e a avaliação estendeu-se por um ano. O delineamento continuou em blocos, com quatro tratamentos (SC, CMP, CMO e CQT) e as parcelas subdivididas em adubado e não adubado. O CQT proporcionou controle total das espécies indesejáveis, mas deprimiu inicialmente as leguminosas (principalmente Desmodium incanum), cuja participação na massa de forragem foi substituída por gramíneas. O CMO revelou-se mais eficiente que o CMP no controle de espécies indesejáveis apesar deste ter sido aplicado duas vezes. A adubação não afetou a resposta aos métodos de controle em nenhuma das variáveis analisadas, mas propiciou maior massa total e de gramíneas no outono, primavera e verão. As espécies indesejáveis não alteraram sua freqüência em função da adubação (em média 8,2%), mas Trifolium polymorphum foi favorecido, independente dos tratamentos de controle.

Superação de dormência das sementese controle químico de plantas de Momordicacharantia L

This experiment were performed to evaluate methods of seeds scarification for overcoming dormancy and the efficacy of six herbicides registered for sugarcane to control adult plants of Mcharantia. The experimental design was completely randomized, with four replications in both trials. The first trial is a factorial 2x8 (two times of seeds collection (2006 and 2007) with eight methods of breaking dormancy (mechanical scarification; 100% and 50% concentrated sulfuric acid; 2% potassium nitrate for three and six hours; heat dried at 60 degrees C; hot water at 60 degrees C and an untreated control). In the second trial evaluated the control of M. charantia through six different herbicides: Imazapic, Metsulfuron-methyl, Metribuzin, 2,4-D, Amicarbazone, Paraquat and a control without herbicide application. Thus, it can be concluded that the seeds were immersed in concentrated sulfuric acid for three minutes and potassium nitrate for three hours gave a higher percentage and germination speed and the herbicide Metribuzin reached, albeit belatedly, a total control of M. charantia.

DESENVOLVIMENTO DE CALOS em EXPLANTES DE CUPUAÇUZEIRO (Theobroma grandiflorum Schum.) em FUNÇÃO da CONCENTRAÇÃO DE AUXINAS E do MEIO LÍQUIDO

Objetivou-se estudar o efeito da concentração de auxina e do meio líquido sobre o desenvolvimento de calos de cupuaçuzeiro. Segmentos de eixos embrionários e cotilédones, obtidos de frutos de cupuaçu dos tipos Mamorana e Redondo, foram cultivados em 4 meios de cultura diferentes: 1) meio MS (50%), suplementado com 2,4-D (1; 2; 4; 8 mg/L); 2) sais N6 (SIGMA) (4 g/L), acrescidos de 2,4-D (0; 2; 4 mg/L) e ANA (0; 3; 5 mg/L); 3) igual ao anterior, suplementado apenas com ANA (3 mg/L); e 4) meio MS, acrescido com ANA (1 mM). Calos com aspecto branco e brilhante foram observados em segmentos de eixos embrionários e cotilédones, cultivados nas menores concentrações de meio 1 (1 e 2 mg/L), enquanto nas maiores concentrações (4 e 8 mg/L) se observou a formação de calos e massa calosa branco-opaca, em eixos embrionários e em segmentos cotiledonares, estas estruturas tornaram-se escuras dentro de oito semanas. Usando o meio 2, um grande número de raízes foram formadas, enquanto o mesmo meio suplementado apenas com ANA (3; 5 mg/L) originou uma massa calosa. A combinação de ANA e 2,4-D, 3 e 2 mg/L, respectivamente, promoveu a formação de calos brancos e raízes. A transferência das culturas para meio líquido, sem regulador de crescimento, promoveu aumento de tamanho dos explantes e escurecimento dos mesmos. O cultivo desses explantes no meio 3 resultou no aparecimento de calos amarelos, com aspecto friável, que permaneceram com a mesma aparência no meio 4.

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.

Avaliação da sensibilidade de diversas espécies de plantas daninhas aquáticas ao carfentrazone-ethyl, em ambiente controlado

Plantas aquáticas, especialmente macrófitas, tornam-se sério problema em hidrelétricas, afetando a múltipla utilização dos corpos d'água, incluindo produção de peixes e atividades de pesca, perdas d'água por evapotranspiração, esportes aquáticos, canoagem, irrigação e produção de energia nas usinas hidrelétricas. Com o objetivo de analisar o potencial de uso do carfentrazone-ethyl no controle das principais plantas daninhas aquáticas no Brasil, foi instalado um experimento em vasos com água. Utilizaram-se os seguintes tratamentos herbicidas (g i.a. ha-1): carfentrazone-ethyl a 15, 30 e 60; glyphosate a 4.536; 2,4-D a 4.690; imazapyr a 1.250; e uma testemunha sem herbicida. Esses tratamentos foram testados nas seguintes espécies: Eichhornia crassipes, Salvinia auriculata, Pistia stratiotes, Myriophyllum aquaticum, Brachiaria arrecta, Hydrocotyle umbellata, Typha sp. e Echinochloa polystachya. As avaliações foram efetuadas aos 7, 14, 21 e 28 dias após os tratamentos. Os resultados mostraram que o carfentrazone-ethyl foi eficiente no controle de E. crassipes (maior dose) e P. stratiotes (duas maiores doses), com efeito supressivo sobre S. auriculata. Foi observado que nas outras plantas daninhas estudadas não houve eficiência de controle.

Efeito de concentrações de herbicidas sobre aspectos biológicos de Fusarium sp. (isolado FCAV#940)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Segurança das condições de aplicação de herbicidas com aerobarco em plantas daninhas aquáticas no lago da Hidrelétrica de Jupiá

Os objetivos deste trabalho foram quantificar as exposições dérmicas (EDs) e respiratórias (ERs) proporcionadas ao piloto e ao seu ajudante nas aplicações de herbicidas para o controle de plantas daninhas aquáticas com aerobarco; classificar essas condições de trabalho em seguras ou inseguras; e calcular a necessidade de controle das exposições (NCE) e o tempo de trabalho seguro (TTS). O aerobarco utilizado tinha casco de alumínio (4,85 x 2,42 m) e acionamento por hélice acoplada a motor a gasolina de 350 HP. O equipamento de pulverização era composto por bomba de diafragma com fluxo máximo de 49,69 L min-1, pressão máxima de 25 kg cm-2, acionada por motor a gasolina de 4 HP, e tanque de calda de 189 L. A barra de pulverização de alumínio era composta de duas seções laterais de 3 m, posicionadas na linha entre o encosto do banco do piloto e o início da estrutura protetora da hélice. Cada seção da barra tinha seis bicos com pontas de jato plano com indução de ar AI 100 03, espaçados de 0,5 m, e uma ponta OC 20 fixada em cada extremidade. O conjunto de pontas pulverizava faixas de 6 m de largura e aplicava o volume de calda de 200 L ha-1. O sistema tinha gerenciador de fluxo, controlado por central eletrônica acoplada a DGPS (com precisão submétrica), para corrigir automaticamente a vazão em função de alterações na velocidade real da embarcação. As EDs e ERs aos herbicidas foram calculadas com os dados substitutos das exposições às caldas, avaliadas com os traçadores cobre e manganês adicionados às caldas. As exposições foram extrapoladas para uma jornada de trabalho de seis horas. A segurança das condições de trabalho foi determinada com o cálculo da margem de segurança (MS), utilizando-se a fórmula MS = (NOEL x 70)/(QAE x 10), em que QAE = quantidade absorvível da exposição. As condições de trabalho foram classificadas em seguras, se MS>1, ou inseguras, se MS<1. As exposições proporcionadas pelas condições de trabalho foram de 10,65 mL de calda por dia para o piloto e de 16,80 mL por dia para o ajudante, que fica sentado em uma cadeira a 2,0 m à frente do piloto e da barra de pulverização. Classificaram-se como seguras as aplicações dos herbicidas glyphosate (Rodeo, 6 L ha-1), 2,4D (DMA 806 BR, 8 L ha-1) e fluridone (Sonar AQ, 0,4 L ha-1), para o piloto e o seu ajudante. Classificou-se como insegura a aplicação do herbicida diquat (Reward, 4,0 L ha-1) para as duas condições de trabalho, cujas necessidades de controle das exposições calculadas foram de 65% para o piloto e de 78% para o ajudante do piloto.