Fungal succession on leaves of Alchornea triplinervia (Spreng.) Muell. Arg. submerged in a stream of an Atlantic Rainforest in the state of São Paulo, Brazil

Leaves of Alchornea triplinervia (Spreng.) Muell. Arg. were submerged in a stream in an Atlantic Rainforest in São Paulo state, Brazil, from July/1988 to June/1989 and from July/1989 to May/1990. Fungi were isolated by the leaf disks washing technique followed by plating on culture media and also by using baiting techniques (using substrates with chitin, keratin and cellulose), what resulted on 565 fungal registers corresponding to 81 taxa. The most common species found during this study of the fungal succession were Trichoderma viride Pers. ex S.F. Gray and Fusarium oxysporum Schlecht emend. Snyd. & Hans. (23 registers), Penicillium hirsutum Dierckx (21 registers), Fusarium solani (Mart.) Appel & Wollenw. emend. Snyd. & Hans. (17), followed by 14 registers of: Cylindrocladium scoparium Morgan, Triscelophorus monosporus Ingold and Polychytrium aggregatum Ajello. Although the monthly obtained mycota had been composed by species of different taxonomic groups, the fungal succession was defined by the initial presence of typical terrestrial leaf inhabiting fungi (mostly Deuteromycotina), followed by species of Mastigomycotina and Zygomycotina. Combining culture methods and baiting techniques, it was possible to verify the presence of terrestrial fungi on the decomposition of submerged leaves and the importance of zoosporic fungi in the fungal succession. This is the first paper about the fungal succession on the decomposition of leaves submerged in a lotic ecosystem in Brazil.

Atividade da celulase de fungos isolados do solo da Estação Ecológica de Juréia-Itatins, São Paulo, Brasil

Oitenta fungos filamentosos isolados do solo da Mata Atlântica da região conhecida como Banhado Grande, Estação Ecológica de Juréia-Itatins, São Paulo, Brasil, foram analisados para avaliar seus potenciais quanto a produção de celulases em resposta à presença de celulose, como única fonte de carbono, em meio de cultura. Foi utilizada a técnica de coloração com vermelho congo e determinada a atividades da celulase em papel de filtro (FPase) e em carboximetilcelulose (CMCase). Os fungos foram diferenciados quanto à atividade dessas enzimas, pois tais atividades variaram em relação ao tipo de substrato e à metodologia aqui utilizados. A melhor atividade CMCase (1,64 U) foi obtida com o cultivo de Trichoderma harzianum (V) em meio de farelo de trigo após cultivo por 4 dias, a 25 ºC. Os resultados obtidos não forneceram evidências para diferenciar qualquer linhagem que tivesse melhor atividade da celulase em relação às demais. Contudo, sugerem que estudos mais detalhados com as linhagens de Trichoderma: T. harzianum III e V, T. inhamatum I, T. longibrachiatum, T. pseudokoningii II e T. viride I, serão necessários para avaliar se estas são potencialmente boas produtoras de celulase, sob condições adequadas de cultivo.

Mixobiota de Floresta Atlântica: novas referências de Physarales para o Estado da Paraíba, Nordeste do Brasil

(Mixobiota de Floresta Atlântica: novas referências de Physarales para o Estado da Paraíba, Nordeste do Brasil). A ordem Physarales engloba as famílias Elaeomyxaceae, Didymiaceae e Physaraceae, reunindo 16 gêneros e aproximadamente 371 espécies, muitas bem representadas nos Neotrópicos. Um estudo sobre a presença de representantes desta ordem na mixobiota da Floresta Atlântica foi desenvolvido na Área de Preservação Permanente Mata do Pau Ferro, localizada na Mesorregião do Brejo Paraibano (6°58'12' S e 35°42'15' W, 400-650 m.s.m., 600 ha). As coletas foram realizadas entre junho e dezembro de 2005, abrangendo o período chuvoso e o de estiagem, explorando diferentes substratos. Exsicatas representativas do material estudado estão depositadas no Herbário UFP. Dentre as Physarales assinaladas, constituem nova referência para a Paraíba o gênero Fuligo e as seguintes espécies: Diderma hemisphaericum (Bull.) Hornem., Didymium clavus (Alb. & Schwein.) Rabenh., D. nigripes (Link) Fr. (Didymiaceae), Fuligo septica (L.) F. H. Wigg., Physarum echinosporum Lister, P. pulcherrimum Berk. & Ravenel e P. viride (Bull.) Pers. (Physaraceae). São apresentadas descrições de cada espécie, baseadas no material estudado, acompanhadas de comentários e distribuição geográfica no Brasil.

Incorporação de microrganismos, aminoácidos, micronutrientes e reguladores de crescimento em sementes de alface pela técnica de peliculização

A peliculização é uma tecnologia que permite, dentre outros usos, a inoculação de microrganismos e a incorporação de aditivos às sementes, sem mudança no seu tamanho ou forma e é uma estratégia para garantir alto potencial de emergência em campo, ganhos de produtividade e melhor agregação de insumos agrícolas às sementes. O experimento foi conduzido na Universidade Federal de Lavras, com o objetivo de avaliar o efeito da incorporação de alguns microrganismos antagônicos, micorrizas, aminoácidos, micronutrientes e reguladores de crescimento, pela técnica de peliculização, na qualidade de sementes e mudas de alface. As sementes foram revestidas com dois tipos de polímeros, adicionando-se microrganismos antagônicos (Trichoderma viride, Trichoderma polysporhum, Trichoderma stromaticum, Beauveria bassiana, Metarhizium anisopliae), micorrizas, aminoácidos, micronutrientes e reguladores de crescimento com a metodologia e as dosagens recomendadas pelo fabricante dos produtos. Avaliou-se as porcentagens de germinação e emergência, o índice de velocidade de emergência, o número de plantas, a massa seca da parte aérea e da raiz e a altura de parte aérea das mudas. Concluiu-se a microbiolização de sementes com a mistura dos microrganismos Trichoderma viride, T. polysporhum, T. stromaticum, Beauveria bassiana, Metarhizium anisopliae e micorrizas reduz a germinação das sementes de alface. A inoculação de sementes de alface com Trichoderma viride e com reguladores de crescimento promovem aumento na emergência e no índice de veolicadade de emergência das plântulas.

Antagonistic activities of selected fungal isolates against Armillaria mellea

The antagonistic activities of six selected fungal isolates against Armilloria mellea were studied on two different concentrations of three media, on fungicides-amended malt extract agar (MEA) medium, and in glasshouse pots filled with John Innes No.2 compost and natural field soil. Trichoderma hamatum isolate Tham1 was found the most effective in reducing Armillaria growths on both the low and high concentrations of malt extract, potato dextrose and V-8 juice in MEA, potato dextrose agar (PDA) and V-8 juice agar (VJA), respectively, followed by T. harzianum isolate Th2 and T. viride isolate Tv3. Neither dose rate (200 or 2000 mg l(-1)) of fenpropidin allowed any growth of Armillaria on MEA, while that of the antagonists was also completely inhibited or greatly restricted. However, both dose rates of fosetyl-A1 allowed the growth of Armillaria and almost all the antagonists. Data on colony diameters of Armillaria showed Tham1 as the most effective antagonist along with Th2, Th23 and Tv3. Tham1 was also found the most effective in protecting hazel billets from colonization by Armillaria, followed by Th2 and Th23. Compared with 7.1 colonized billets in the inoculated controls, only 1.3, 2.6 and 2.7 billets (out of ten) were colonized, respectively, when protected with these antagonists. The results indicate that the Trichoderma isolates are able to maintain their antagonistic effects on A. mellea under a variety of nutritional, chemical and edaphic regimes. More investigations are needed to develop a system of control for the disease with these potential antagonists.

The effect of carrier substrate, dose rate and time of application on biocontrol efficacy of fungal antagonists against Armillaria root rot of strawberry plants.

In a glasshouse experiment using potted strawberry plants (cv. Cambridge Favourite) as hosts, the effect of selected fungal antagonists grown on 25 or 50 g of mushroom compost containing autoclaved mycelia of Agaricus bisporus, or wheat bran was evaluated against Armillaria mellea. Another glasshouse experiment tested the effect of application time of the antagonists in relation to inoculations with the pathogen. A significant interaction was found between the antagonists, substrates and dose rates. All the plants treated with Chaetomium olivaceum isolate Co on 50 g wheat bran survived until the end of the experiment which lasted 482 days, while none of them survived when this antagonist was added to the roots of the plants on 25 g wheat bran or 25 or 50 g mushroom compost. Dactylium dendroides isolate SP had a similar effect, although with a lower host survival rate of 33.3%. Trichoderma hamatum isolate Tham 1 and T. harzianum isolate Th23 protected 33.3% of the plants when added on 50 g and none when added on 25 g of either substrate, while 66.7% of the plants treated with T. harzianum isolate Th2 on 25 g, or T viride isolate TO on 50 g wheat bran, survived. Application of the antagonists on mushroom compost initially resulted in development of more leaves and healthier plants, but this effect was not sustained. Eventually, plants treated with the antagonists on wheat bran had significantly more leaves and higher health scores. The plants treated with isolate Th2 and inoculated with Armillaria at the same time had a survival rate of 66.7% for the duration of the experiment (475 days), while none of them survived that long when the antagonist and pathogen were applied with an interval of 85 days in either sequence. C. olivaceum isolate Co showed a protective effect only, as 66.7% of the plants survived when they were treated with the antagonist 85 days before inoculation with the pathogen, while none of them survived when the antagonist and pathogen were applied together or the infection preceded protection.

Comparisons between the in vitro and in vivo efficacies of potential fungal antagonists of Armillaria mellea

Seventeen fungal isolates were tested in vitro as potential antagonists of two isolates of the root rot pathogen, Armillaria mellea. Some of the isolates were also added on mushroom composts with living mycelia to the roots of Armillaria-inoculated potted strawberry plants in the glasshouse to find out if they had the same degree of efficacy against the disease. Dactylium dendroides isolate SP was the most effective in reducing mycelial growth of A. mellea isolate 1 (Am1), followed by Trichoderma harzianum isolate Th2 and T. viride isolate Tv4. Th2, Th22, Tv3 and SP grew extensively over Am1 colonies, disintegrating the rhizomorphs. Isolate Tham1 of T hamatum was the most effective in reducing mycelial growth of A. mellea isolate 2 (Am2), followed by Tv3. Th12, Th22, Tv1, Tv3 and SP inhibited the initiation and growth of rhizomorphs of Am2. Regeneration tests showed that both Am1 and Am2 attacked by Trichoderma isolates and SP were no longer viable. Th23 and SP were almost as effective in vivo as in vitro. But isolate Co of Chaetomium olivaceum, which was ineffective in vitro, was found effective in vivo. Conversely, Th2, which exhibited good antagonistic activity in vitro, performed poorly in vivo. These results show that the in vitro and in vivo efficacies of potential antagonists may not necessarily be closely correlated. Hence, there is a danger that potentially effective isolates may be discarded if decisions are made only on the basis of preliminary screening tests carried out under laboratory conditions.

Controle biológico de Colletotrichum acutatum, agente causal da queda prematura dos frutos cítricos

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

Comparative growth of trichoderma strains in different nutritional sources, using bioscreen c automated system

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Screening of culture condition for xylanase production by filamentous fungi

The objective of this research was to investigate xylanase production by filamentous fungi (Trichoderma viride) to determine the best cultivation conditions in the process, aiming toward optimization of enzyme production. The best temperature, as well as the best carbon source, for biomass production was determined through an automated turbidimetric method (Bioscreen-C). The enzyme activity of this fungus was separately evaluated in two solid substrates (wheat and soybean bran) and in Vogel medium, pure and by adding other carbon sources. Temperature effects, cultivation time, and spore concentrations were also tested. The best temperature and carbon source for enzyme and biomass production was 25 C and sorbitol, respectively. Maximum xylanase activity was achieved when the fungus was cultivated in wheat bran along with sorbitol (1%, w/v), using a spore concentration of 2 x 10(6) spores. mL(-1), pH 5.0, for 144 h cultivation. The study demonstrated not only the importance of the nature of the substrate in obtaining a system resistant to catabolic repression, but also the importance of the culture conditions for biosynthesis of this enzyme. T. viride showed a high potential for xylanase production under the conditions presented in these assays.

Foraging activity and resource use by three parrotfish species at Fernando de Noronha Archipelago, tropical West Atlantic

Parrotfishes are important components of the herbivore and detritivore guilds of tropical and subtropical reefs. Most of parrotfish species are protogynous hermaphrodites that change colour and sex, from initial phase females or males (IP) to terminal phase males (TP). We studied the foraging behaviour of Sparisoma amplum, S. axillare and S. frondosum, three syntopic scarids on the rocky reefs of Fernando de Noronha Archipelago, Tropical West Atlantic. The three parrotfish species differed in food selection and preference, but IP and TP individuals of the same species preferred the same food types, except for S. amplum. Feeding rates of IP individuals were higher than those of TP individuals, but the distribution of feeding frequencies throughout the day of IP and TP individuals of the same species was similar. IP individuals had higher feeding rates than TP ones, which seems related to the fact that TP individuals spend a large amount of time patrolling their territories and chasing away conspecific individuals at the study site. The general foraging pattern we found for S. amplum, S. axillare and S. frondosum is similar to patterns found for other parrotfish species in the Western Atlantic.

New aerophytic morphospecies of Nostoc (Cyanobacteria) from São Paulo State, Brazil

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

Asymmetric microbial reduction of ketones: absolute configuration of trans-4-ethyl-1-(1S-hydroxyethyl)cyclohexanol

A set of five fungal species, Botrytis cinerea, Trichoderma viride and Eutypa lata, and the endophytic fungi Colletotrichum crassipes and Xylaria sp., was used in screening for microbial biocatalysts to detect monooxygenase and alcohol dehydrogenase activities (for the stereoselective reduction of carbonyl compounds). 4-Ethylcyclohexanone and acetophenone were biotransformed by the fungal set. The main reaction pathways involved reduction and hydroxylations at several positions including tertiary carbons. B. cinerea was very effective in the bioreduction of both substrates leading to the chiral alcohol (S)-1-phenylethanol in up to 90% enantiomeric excess, and the cis-trans ratio for 4-ethylcyclohexanol was 0:100. trans-4-Ethyl-1-(1S-hydroxyethyl)cyclohexanol, obtained from biotransformation by means of an acyloin-type reaction, is reported here for the first time. The absolute configurations of the compounds trans-4-ethyl-1-(1S-hydroxyethyl)cyclohexanol and 4-(1S- and 4-(1R-hydroxyethyl)cyclohexanone were determined by NMR analysis of the corresponding Mosher's esters. (C) 2009 Elsevier Ltd. All rights reserved.

SEGREGAÇÃO NATURAL DE VIRÓIDES DOS CITROS DETECTADA NA INDEXAÇÃO BIOLÓGICA E BIOQUÍMICA E CONFIRMADA PELOS SINTOMAS DE PLANTAS DE CAMPO

Estudaram-se os procedimentos de indexação biológica e bioquímica em um experimento de avaliação do uso de viróides como agentes ananicantes para pomeleiro 'Marsh Seedless'. em algumas parcelas, houve segregação de viróides, sendo que, em duas, a segregação foi de CEVd (viróide da exocorte dos citros) e, em outras duas, não foi detectada a presença de CVd-II (viróide dos citros II). Como conseqüência desta segregação, não ocorreram sintomas típicos da exocorte em cidra nas parcelas nas quais houve segregação de CEVd, o que ocorreu nas parcelas em que a segregação foi de CVd-II, indicando que é necessária a presença de CEV para a manifestação de sintomas típicos de exocorte em cidra. Numa quinta parcela, não se detectaram viróides, provavelmente, devido à ausência dos mesmos no material de inoculação do teste biológico (escape), já que a planta de campo teve desenvolvimento vegetativo normal e ausência de sintomas no tronco. Finalmente, em uma sexta parcela, o que ocorreu, foi uma segregação drástica, não relatada na literatura, já que a planta de campo mostrou redução de porte e sintomas no tronco. A coleta de varetas para indexação deve ser mais rigorosa em função da ocorrência de segregação.

Atividade da celulase de fungos isolados do solo da Estação Ecológica de Juréia-Itatins, São Paulo, Brasil

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