Promoção de enraizamento de microestacas de um clone de Eucalyptus sp. por Trichoderma spp.

Este trabalho teve como objetivo o emprego de isolados antagonistas de fungos visando à promoção do enraizamento de microestacas de um clone de Eucalyptus sp. Utilizaram-se no teste de promoção de enraizamento de microestacas um isolado não-patogênico de Cylindrocladium spp. e mais três isolados antagonistas de Trichoderma spp. (E15, S2 e St), os quais apresentaram as melhores notas de antagonismo em teste in vitro, pelo método de confrontação direta contra isolado patogênico de Cylindrocladium spp., sendo inoculados no substrato de desenvolvimento das microestacas sob condições de estufa. Observou-se aumento de sobrevivência das microestacas na presença dos isolados de Trichoderma spp. e Cylindrocladium spp., em comparação com a testemunha, em ambiente naturalmente infestado por Botrytis cinerea. O tratamento com os isolados ST, E15 e S2 de Trichoderma spp. e Cyl de Cylindrocladium spp. aumentou a sobrevivência de microestacas de Eucalyptus sp. O isolado E15 promoveu o enraizamento de microestacas, apresentando aumento significativo na porcentagem de enraizamento (62,25%) em relação ao tratamento-testemunha (28,77%).

Drying and storage of cauliflower (Brassica oleracea var. botrytis) hydroprimed seeds

Literature has documented beneficial effects of seed priming on speed, synchronization and uniformity of germination. often leading to improved stand establishment. However. doubts still persist about the possible reversal effects, after drying and during storage of primed seeds that could overcome, partial or totally, the improved performance. The objectives of this research were to identify drying and storage procedures that would maintain the physiological performance achieved after seed priming, without negative effects on storability. First. hydroprimed cauliflower Seeds cv. Sharon and cv. Teresopolis Gigante, each represented by three seed lots were submitted to fast drying, slow drying, and treatments of pre-drying incubation (exposure to 35 degrees C, to a polyethylene glycol 6000 solution or a heat shock) followed by fast drying. In the second phase of this study, hydroprimed seed samples were submitted to fast drying (30-35 degrees C and 40-50% R.H.) and stored under laboratory conditions or in a chamber at 20 degrees C and 50% relative humidity for six months. Seed physiological potential was evaluated after 60-day intervals for germination (speed and percentage), Seedling emergence and saturated salt accelerated aging tests. All drying treatments efficiently preserved the favourable priming effects, except for the incubation at 35 degrees C for 96-144 hours. The beneficial priming effects followed by fast drying persisted for four months under controlled conditions (20 degrees C and 50% relative humidity).

Saprophytic microorganisms with potential for biological control of Botrytis cinerea on Geraldton waxflower flowers

Saprophytic bacteria, yeasts and filamentous fungi were isolated from Geraldton waxflower flowers and screened to identify potential antagonism towards Botrytis cinerea. Isolates from other sources (e.g. avocado) were also tested. Isolates were initially screened in vitro for inhibition of B. cinerea conidial germination, germ tube elongation and mycelial growth. The most antagonistic bacteria, yeasts and fungi were selected for further testing on detached waxflower flowers. Conidia of the pathogen were mixed with conidia or cells of the selected antagonists, co-inoculated onto waxflower flowers, and the flowers were sealed in glass jars and incubated at 20 degreesC. The number of days required for the pathogen to cause flower abscission was determined. The most antagonistic bacterial isolate, Pseudomonas sp. 677, significantly reduced conidial germination and retarded germ tube elongation of B. cinerea. None of the yeast or fungal isolates tested was found to significantly reduce conidial germination or retard germ tube elongation, but several significantly inhibited growth of B. cinerea. Fusarium sp., Epicoccum sp. and Trichoderma spp. were the most antagonistic of these isolates. Of the isolates tested on waxflower, Pseudomonas sp. 677 was highly antagonistic towards B. cinerea and delayed waxflower abscission by about 3 days. Trichoderma harzianum also significantly delayed flower abscission. However, as with most of the fungal antagonists used, inoculation of waxflower flowers with this isolate resulted in unsightly mycelial growth.

Efeito da densidade de população nos teores de ácido ascórbico e carboidratos solúveis de couve-flor (Brassica oleracea var. botrytis)

Os teores de carboidratos solúveis, glucose, frutose, sacarose e ácido ascórbico foram determinados em amostras de couve-flor (Brassica oleracea var. botrytis), obtidas de tratamentos apresentando diversas densidades de plantais. O efeito do espaçamento não foi significativo para os dviersos constituintes analisados, embora o tratamento com 20.833 plantas/ha apresentasse maior teor de ácido ascórbico. O peso seco das amostras variou de 8,24 a 10,13% e ácido ascórbico de 93,1 a 113,5 mg/100 g de peso fresco. Os teores de carboidratos solúveis, expressos em g/100 g de peso fresco foram: totais (3,17-4,54) sacarose (0,26-0,66), glucose (0,92-1,50) e frutose (0,91-145).

Aplicação de alguns herbicidas na cultura da couve-flor (Brassica oleracea var. botrytis)

Foi conduzido na Área Experimental do Setor de Horticultura, DAH-ESALQ, Piracicaba, um experimento sobre o uso de alguns herbicidas na cultura da couve-flor (Brassica oleracea var. botrytis). Os herbicidas foram Dacthal (DCPA), Tenoran (cloroxuron), Afalon (linuron) e Ramrod (propacloro). O Dacthal, Ramrod, e o Tenoran não foram fitotóxicos à couve-flor, enquanto que o Afalon foi muito fitotóxico. O herbicida que melhor resultado apresentou para a couve-flor foi o Ramrod.

Teor de ácido ascórbico de couve-flor (Brassica oleracea var. botrytis) conservada à temperatura ambiente, em geladeira e em congelador

Amostras de couve-flor (Brassica oleracea var. botrytis) foram armazenadas à temperatura ambiente, a 5°C (geladeira) e a - 20°C (congelador) e foram feitas as análises para ácido ascórbico. O teor de ácido ascórbico não variou durante o período de resfriamento e as amostras de couve-flor congelada mostrou uma redução de 50% durante 12 dias de armazenamento. Amostras colocadas à temperatura ambiente apresentaram um pequeno aumento até os 3 dias. Não foram observadas diferenças significativas entre os tipos de embalagens usadas (garrafa de vidro, sacos plásticos e folhas de alumínio) quanto ao teor de ácido ascórbico.

Effects of population density on cauliflower crop (Brassica oleracea L. var. botrytis)

An experiment was carried out to study the effects of the following population densities cauliflowers (plants per ha): 20,833 (0.60 m x 0.80 m), 25,641 (0.60 m x 0.65 m), ....37.037 (0.60 m x 0.45 m) , 55.555 (.0.60 m x 0.30 m), and 111,111 (0,60 m x 0,15 m) ; variety Snow ball. It was concluded that the effects of plant population density are greater on curd quality (weight and size) than on production per ha. The best plant population density to produce cauliflowers curd for Brazil market is from 20,000 to 25,000 plants/ha while for mini-curd is above 55,000 plants/ha.

Simultaneous Evaluation of Intact Glucosinolates and Phenolic Compounds by UPLC-DAD-MS/MS in Brassica oleracea L. var. botrytis

A method for the simultaneous determination of intact glucosinolates and main phenolic compounds (flavonoids and sinapic acid derivatives) in Brassica oleracea L. var. botrytis was proposed. A simplified sample extraction procedure and a UPLC separation were carried out to reduce the total time of analysis. Brassica oleracea samples were added with internal standards (glucotropaeolin and rutin), and extracted with boiling methanol. Crude extracts were evaporated under nitrogen, redissolved in mobile phase and analyzed by UPLC with double detection (ESI--MRM for glucosinolates and flavonoids, and DAD for main sinapic acid derivatives). The proposed method allowed a satisfactory quantification of main native sinapic acid derivatives, flavonoids and glucosinolates with a reduced time of analysis.

CHARACTERISATION OF ARABIDOPSIS THALIANA PERMEABLE CUTICLE MUTANTS SHOWING A STRONG RESISTANCE TO BOTRYTIS CINEREA

La cuticule des plantes, composée de cutine, un polyester lipidique complexe et de cires cuticulaires, couvre l'épiderme de la plupart des parties aériennes des plantes. Elle est constituée d'une barrière hydrophobique primaire qui minimise les pertes en eau et en soluté et protège l'organisme de différents stress environnementaux tels que les rayons UV, la dessiccation et l'infection par des pathogènes. Elle est aussi impliquée dans la délimitation des organes durant le développement. La cutine est un polyester qui, dans la plupart des espèces végétales, est principalement composé d'acides gras ω-hydroxylés composé de 16 à 18 carbones. Cependant, la cutine des feuilles d'Arabidopsis a une composition différente et est principalement constituée d'acides dicarboxyliques à 16-18 carbones. Les cires sont présentes dans le polyester de la cutine ou le recouvrent. Chez Arabidopsis, un nombre de mutants, tel que 1er, bdg, hth, att1, wbc11, et des plantes transgéniques avec différents changement dans la structure de la cuticule dans les feuilles et la tige, ont récemment été décrits et servent d'outils pour étudier la relation entre la structure et la fonction de la cuticule.7 mutants d'Arabidopsis ont été isolés par une méthode de coloration qui permet de détecter une augmentation dans la perméabilité cuticulaire. Ces mutants ont été appelés pec pour permeable cuticle.Pour la première partie de mon projet, j'ai principalement travaillé avec pec9/bre1 (permeable cuticle 9/botrytis resistance 1). PEC9/BRE1 a été identifié comme étant LACS2 (LONG CHAIN ACYL-CoA SYNTHETASE 2). Dans ce mutant, la cuticule n'est pas visible sous microscopie électronique et la quantité en acides gras omega- hydroxylés et en leurs dérivés est fortement réduite. Ces altérations conduisent à une plus grande perméabilité de la cuticule qui est mise en évidence par une plus grande sensibilité à la sécheresse et aux xénobiotiques et une coloration plus rapide par bleu de toluidine. Le mutant Iacs2 démontre aussi une grande capacité de résistance à l'infection du champignon nécrotrophique B. cinerea. Cette résistance est due à l'extrusion sur les feuilles d'un composé antifongique durant l'infection. Ce travail a été publié dans EMBO journal (Bessire et al., 2007, EMBO Journal).Mon second projet était principalement concentré sur pec1, un autre mutant isolé par le premier crible. La caractérisation de pec1 a révélé des phénotypes similaires à ceux de Iacs2, mais à chaque fois dans des proportions moindres : sensibilité accrue à la sécheresse et aux herbicides, plus grande perméabilité au bleu de toluidine et au calcofluor white, altération de la structure cuticulaire et résistance à B. cinerea à travers la même activité antifongique. PEC1 a été identifié comme étant AtPDR4. Ce gène code pour un transporteur ABC de la famille PDR ("Pleiotropic Drugs Resistance") qui sont des transporteurs ayants un large spectre de substrats. Le mutant se différencie de Iacs2, en cela que la composition en acides gras de la cuticule n'est pas autant altérée. C'est principalement le dihydroxypalmitate des fleurs dont la quantité est réduite. L'expression du gène marqué avec une GFP sous le contrôle du promoteur endogène a permis de localiser le transporteur au niveau de la membrane plasmique des cellules de l'épiderme, de manière polaire. En effet, la protéine est principalement dirigée vers l'extérieure de la plante, là où se trouve la cuticule, suggérant une implication d'AtPDR4 dans le transport de composants de la cuticule. Ce travail est actuellement soumis à Plant Cell.Une étude phylogénétique a aussi montré qu'AtPDR4 était très proche d'OsPDR6 du riz. Le mutant du riz a d'ailleurs montré des phénotypes de nanisme et de perméabilité similaire au mutant chez Arabidopsis.AbstractThe cuticle, consisting principally of cutin and cuticular waxes, is a hydrophobic layer of lipidic nature, which covers all aerial parts of plants and protects them from different abiotic and biotic stresses. Recently, the research in this area has given us a better understanding of the structure and the formation of the cuticle. The Arabidopsis mutants permeable cuticle 1 (peel) and botrytis resistance 1 (brel) were identified in two screens to identify permeable cuticles. The screens used the fluorescent dye calcofluor to measure permeability and also resistance to the fungal pathogen Botrytis. These mutants have highly permeable cuticle characteristics such as higher water loss, intake of chemicals through the cuticle, higher resistance to Botrytis cinerea infection, and organ fusion.BRE1 was cloned and found to be LACS2, a gene previously identified which is important in the formation and biosynthetic pathway of the cuticle. In brel, the amount of the major component of cutin in Arabidopsis leaves and stems, dicarboxylic acids, is five times lower than in the wild type. Moreover, the permeability of the cuticle allows the release of antifungal compounds at the leaf surface that inhibits the growth of two necrotrophic fungi: Botrytis cinerea and Sclerotinia sclerotiorum.PEC1 was identified as AtPDR4, a gene that codes for a plasma membrane transporter of the Pleiotropic Drug Resistance family, a sub-family of the ABC- transporters. AtPDR4 is strongly expressed in the epidermis of expanding tissues. In the epidermis it is located in a polar manner on the external plasma membrane, facing the cuticle. Analysis of the monomer composition of the cutin reveals that in this mutant the amount of hydroxy-acids and dihydroxy-palmitate is 2-3 times lower in flowers, in which organ these cutin monomers are the major components. Thus AtPDR4 is thought to function as a putative cutin monomer transporter.

A permeable cuticle in Arabidopsis leads to a strong resistance to Botrytis cinerea.

The plant cuticle composed of cutin, a lipid-derived polyester, and cuticular waxes covers the aerial portions of plants and constitutes a hydrophobic extracellular matrix layer that protects plants against environmental stresses. The botrytis-resistant 1 (bre1) mutant of Arabidopsis reveals that a permeable cuticle does not facilitate the entry of fungal pathogens in general, but surprisingly causes an arrest of invasion by Botrytis. BRE1 was identified to be long-chain acyl-CoA synthetase2 (LACS2) that has previously been shown to be involved in cuticle development and was here found to be essential for cutin biosynthesis. bre1/lacs2 has a five-fold reduction in dicarboxylic acids, the typical monomers of Arabidopsis cutin. Comparison of bre1/lacs2 with the mutants lacerata and hothead revealed that an increased permeability of the cuticle facilitates perception of putative elicitors in potato dextrose broth, leading to the presence of antifungal compound(s) at the surface of Arabidopsis plants that confer resistance to Botrytis and Sclerotinia. Arabidopsis plants with a permeable cuticle have thus an altered perception of their environment and change their physiology accordingly.

Enhanced Botrytis cinerea resistance of Arabidopsis plants grown in compost may be explained by increased expression of defense-related genes, as revealed by microarray analysis

Composts are the products obtained after the aerobic degradation of different types of organic matter waste and can be used as substrates or substrate/soil amendments for plant cultivation. There is a small but increasing number of reports that suggest that foliar diseases may be reduced when using compost, rather than standard substrates, as growing medium. The purpose of this study was to examine the gene expression alteration produced by the compost to gain knowledge of the mechanisms involved in compost-induced systemic resistance. A compost from olive marc and olive tree leaves was able to induce resistance against Botrytis cinerea in Arabidopsis, unlike the standard substrate, perlite. Microarray analyses revealed that 178 genes were differently expressed, with a fold change cut-off of 1, of which 155 were up-regulated and 23 were down-regulated in compost-grown, as against perlite-grown plants. A functional enrichment study of up-regulated genes revealed that 38 Gene Ontology terms were significantly enriched. Response to stress, biotic stimulus, other organism, bacterium, fungus, chemical and abiotic stimulus, SA and ABA stimulus, oxidative stress, water, temperature and cold were significantly enriched, as were immune and defense responses, systemic acquired resistance, secondary metabolic process and oxireductase activity. Interestingly, PR1 expression, which was equally enhanced by growing the plants in compost and by B. cinerea inoculation, was further boosted in compost-grown pathogen-inoculated plants. Compost triggered a plant response that shares similarities with both systemic acquired resistance and ABA-dependent/independent abiotic stress responses.

Potencial para o biocontrole de Botrytis cinerea por leveduras em sistema integrado de cultivo de lírio

O objetivo deste trabalho foi comparar uma população de leveduras do filoplano de lírio de um sistema convencional de cultivo com outra de sistema integrado, além de avaliar o antagonismo da levedura Sporidiobolus pararoseus a Botrytis cinerea. A comunidade de leveduras foi estimada pelo método da queda dos balistoporos nos terços inferior, médio e superior das plantas, bem como nas faces adaxial e abaxial das folhas. Nos discos de folhas de plantas do sistema integrado, foi alta a presença da população de leveduras, tendo sido maior na face adaxial das folhas do terço médio das plantas. Para avaliação do antagonismo, foram aplicadas aos discos de folhas as concentrações de 10(5), 10(6) e 10(7) células mL-1 da levedura, em três períodos de inoculação: 24 horas antes; simultânea a; e 24 horas após a inoculação de 10(4) esporos mL-1 de B. cinerea. A incidência, a percentagem de área de discos colonizados e a esporulação de B. cinerea foram avaliadas por meio de diagrama de notas, a partir do quarto dia após a inoculação. Todas as concentrações da levedura reduziram a esporulação de B. cinerea nos discos de folha, em comparação à testemunha, e a concentração de 10(7) esporos mL-1 foi a mais eficiente.

Vaporização de ácido acético para o controle pós-colheita de botrytis cinerea em uva 'Itália'

Visando a avaliar o efeito do vapor de ácido acético (AA) como medida alternativa no controle pós-colheita do mofo-cinzento (Botrytis cinerea) em uva 'Itália', foram conduzidos dois ensaios in vivo onde se avaliou o efeito direto e indireto do AA através do tratamento dos cachos antes e após a inoculação do patógeno, sendo: 1) cachos de uva foram inoculados e, após 4 h, vaporizados com AA (0,0; 0,25; 0,5; 1,0 ou 2,0 mL 100 L-1 vol. de câmara) em tambores herméticos (200 L), a 25±1 ºC / 70-80 % UR, por 30 min; 2) cachos foram, ou não, vaporizados com AA (1 mL 100 L-1 vol. de câmara) e, após 24; 48; 72 ou 96 h, inoculados com B. cinerea. Para inoculação, em cada cacho, 10 por tratamento, foram feridas 10 bagas, fazendo-se um furo por baga de ±2 mm de profundidade, procedendo-se, em seguida, aspersão de suspensão de esporos (±105 conídios mL-1). Após os tratamentos, os cachos foram mantidos a 25±1 ºC / 80-90 % UR e avaliados diariamente quanto à incidência e severidade da podridão. O efeito in vitro do vapor de AA no controle do patógeno foi avaliado com o intuito de verificar se tal agente estaria exercendo efeito direto sobre o crescimento micelial e a germinação de conídios de B. cinerea. Nos ensaios in vivo, o vapor de AA controlou a podridão de B. cinerea em uva 'Itália', nos cachos inoculados antes ou após o tratamento com AA, sendo as uvas inoculadas 48 h após o tratamento, as que apresentaram menor índice de doença. In vitro, o AA exerce efeito direto sobre o crescimento micelial e a germinação de conídios de B. cinerea.