Giberelinas : efecto de la fuente de carbono sobre su producción en gibberella fujikuroi ATCC-12616 utilizando un sistema de fermentación sólido por lotes preparado a basede residuos agroindustriales (por) Martha Alicia Suárez Herrera

Tesis (Maestría en Ciencias, con especialidad en Microbiología Industrial) U.A.N.L.

Comparison of lipase production on crambe oil and meal by Fusarium sp (Gibberella fujikuroi complex)

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

Chicken fat and inorganic nitrogen source for lipase production by Fusarium sp. (Gibberella fujikuroi complex)

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

Structure Variation And Dynamics of the Nuclear Ribosomal Intergenic Spacer Region in Key Gibberella Fujikuroi Complex Species

The intergenic spacer (IGS) region of the ribosomal DNA was cloned and sequenced in eight species within the Gibberella fujikuroi species complex with anamorphs in the genus Fusarium , a group that includes the most relevant toxigenic species. DNA sequence analyses revealed two categories of repeated elements: long repeats and short repeats of 125 and 8 bp, respectively. Long repeats were present in two copies and were conserved in all the species analyzed, whereas different numbers of short repeat elements were observed, leading to species-specific IGS sequences with different length. In Fusarium subglutinans and Fusarium nygamai , these differences seemed to be the result of duplication and deletion events. Here, we propose a model based on unequal crossing over that can explain these processes. The partial IGS sequence of 22 Fusarium proliferatum isolates was also obtained to study variation at the intraspecific level. The results revealed no differences in terms of number or pattern of repeated elements and detected frequent gene conversion events. These results suggest that the homogenization observed at the intraspecific level might not be achieved primarily by unequal crossing-over events but rather by processes associated with recombination such as gene conversion events.

The P450–1 gene of Gibberella fujikuroi encodes a multifunctional enzyme in gibberellin biosynthesis

Recent studies have shown that the genes of the gibberellin (GA) biosynthesis pathway in the fungus Gibberella fujikuroi are organized in a cluster of at least seven genes. P450–1 is one of four cytochrome P450 monooxygenase genes in this cluster. Disruption of the P450–1 gene in the GA-producing wild-type strain IMI 58289 led to total loss of GA production. Analysis of the P450–1-disrupted mutants indicated that GA biosynthesis was blocked immediately after ent-kaurenoic acid. The function of the P450–1 gene product was investigated further by inserting the gene into mutants of G. fujikuroi that lack the entire GA gene cluster; the gene was highly expressed under GA production conditions in the absence of the other GA-biosynthesis genes. Cultures of transformants containing P450–1 converted ent-[14C]kaurenoic acid efficiently into [14C]GA14, indicating that P450–1 catalyzes four sequential steps in the GA-biosynthetic pathway: 7β-hydroxylation, contraction of ring B by oxidation at C-6, 3β-hydroxylation, and oxidation at C-7. The GA precursors ent-7α-hydroxy[14C]kaurenoic acid, [14C]GA12-aldehyde, and [14C]GA12 were also converted to [14C]GA14. In addition, there is an indication that P450–1 may also be involved in the formation of the kaurenolides and fujenoic acids, which are by-products of GA biosynthesis in G. fujikuroi. Thus, P450–1 displays remarkable multifunctionality and may be responsible for the formation of 12 products.

Evaluation of ent-Kaurenoic Acid Derivatives for their Anticariogenic Activity

Ent-kaur-16(17)-en-19-oic acid (kaurenoic acid, KA) is a tetracyclic diterpene prototype for natural anticaries agents. Six KA derivatives were prepared and their antimicrobial activity against the main microorganisms involved in the caries process evaluated. The sodium salt of KA (KA-Na) was the most active, displaying very promising MIC values for most pathogens. Time-kill assays against the primary causative agent of caries (Streptococcus mutans) indicated that KA and KA-Na only inhibited growth in the first 12 h, suggesting a bacteriostatic effect. After this period (12-24 h), their bactericidal effect was clearly noted. KA and KA-Na showed no synergy when combined with the gold standard anticariogenic (chlorhexidine dihydrochloride, CHD) in the checkerboard assays against S. mutans.

Qualidade sanitária e produção de fumonisina B1 em grãos de milho na fase de pré-colheita

Trinta e seis (36) cultivares de milho foram avaliadas em relação à incidência de grãos ardidos, mofados e produção de fumonisina B1. Amostras de 1,2 kg de grãos foram analisadas visualmente para a quantificação de grãos ardidos (Fusarium subglutinans), mofados (Penicillium oxalicum) e para a análise de fumonisina B1. Os grãos ardidos foram submetidos à análise de sanidade (papel de filtro com congelamento) visando identificar os fungos a eles associados. A cultivar Hatã 3052 apresentou 7,6% de grãos ardidos, ultrapassando o limite de tolerância que é de 6,0%. As cultivares AG 5011, HT 7105-3, Dina 1000 e C 701 apresentaram 16,8% , 3,4%, 3,2% e 3,1% de grãos mofados, respectivamente, acima do limite de tolerância que é de 3,0%. O fungo Fusarium subglutinans (Gibberella fujikuroi var. subglutinans) foi o causador de grãos ardidos, cuja detecção variou de 50,0 a 99,0%. A análise de variância mostrou diferenças significativas entre as cultivares com relação às incidências de grãos ardidos e de grãos mofados. Com relação à produção de fumonisina B1, as cultivares Hatã 3052, NB 6077 e 983 P produziram 7,0; 6,1 e 5,9 µg.g-1 de grãos, respectivamente, diferindo significativamente da cultivar P3071 (2,2 µg.g-1 de grãos). Conclui-se que há diferenças significativas entre as cultivares de milho em relação à produção de grãos ardidos e mofados, bem como acentuada interação entre as cultivares e o fungo toxigênico Fusarium subglutinans (Gibberella fujikuroi var. subglutinans) quanto à biossíntese de fumonisina B1 em grãos de milho.

Mycoflora and fumonisin contamination in Brazilian sorghum from sowing to harvest

BACKGROUND: The aim of this study was to characterise the mycoflora and the presence of fumonisin in sorghum grains, correlating the results with the environment and abiotic factors. RESULTS: Fifty samples (five collections of ten samples each) of sorghum were analysed. All samples were found to be contaminated with fungi, with higher frequencies of Cladosporium spp. (61.8%) and Helminthosporium spp. (33.4%). Fusarium verticillioides was isolated from 15.1% of the samples, with 38% of them being contaminated with fumonisin B(1) (FB(1)) at levels ranging from 50 to 368.78 ng g(-1). Regarding abiotic factors, temperature, water activity and rainfall showed a positive correlation with the frequency of F. verticillioides and FB(1) production. There was a significant positive correlation between relative air humidity and FB(1) production. The results obtained from sexual crosses between standard F mating tester strains and the isolated strains confirmed that the strains isolated were F. verticillioides. CONCLUSION: It can be concluded that the decrease in F. verticillioides and fumonisin contamination occurred owing to atypical climatic factors during the period of sorghum cultivation, when there was any occurrence of rain and the level of water activity of grains did not reach 0.58. (C) 2010 Society of Chemical Industry

Unexpected homology between inducible cell wall protein QID74 of filamentous fungi and BR3 salivary protein of the insect Chironomus

A gene, qid74, of mycoparasitic filamentous fungus Trichoderma harzianum and its allies encodes a cell wall protein that is induced by replacing glucose in the culture medium with chitin (simulated mycoparasitism conditions). Because no trace of this gene can be detected in related species such as Gibberella fujikuroi and Saccharomyces cerevisiae, the qid74 gene appears to have arisen de novo within the genus Trichoderma. Qid74 protein, 687 residues long, is now seen as highly conserved tandem repeats of the 59-residue-long unit. This unit itself, however, may have arisen as tandem repeats of the shorter 13-residue-long basic unit. Within the genus Trichoderma, the amino acid sequence of Qid74 proteins has been conserved in toto. The most striking is the fact that Qid74 shares 25.3% sequence identity with the carboxyl-terminal half of the 1,572-residue-long BR3 protein of the dipteran insect Chironomus tentans. BR3 protein is secreted by the salivary gland of each aquatic larva of Chironomus to form a tube to house itself. Furthermore, the consensus sequence derived from these 59-residue-long repeating units resembles those of epidermal growth factor-like domains found in divergent invertebrate and vertebrate proteins as to the positions of critical cysteine residues and homology of residues surrounding these cysteines.

Quantificação de propágulos de Gibberella zeae no ar e infecção de anteras em trigo

Devido ao aumento da ocorrência e da intensidade de epidemias nos últimos anos, a giberela tem sido apontada como uma das doenças que mais danos causa em cereais de inverno. O principal inóculo do fungo patógeno (Giberella zeae) são os ascosporos produzidos em peritécios sobre os restos culturais da maioria das espécies cultivadas. Os esporos são transportados a longas distâncias pelo vento e depositados sobre as anteras causando infecção. Os objetivos do presente trabalho foram quantificar a densidade de esporos no ar, a incidência da infecção em anteras de trigo (Triticum aestivum), a intensidade da doença no campo e elucidar o papel das anteras no processo infeccioso. Durante o período correspondente à antese do trigo, foram coletados, em média, 6,6 esporos/10 cm²/dia, em 1999 e 13,5 esporos/10 cm²/dia em 2000. No estádio de floração plena, a infecção por G. zeae foi de 11,8% nas anteras soltas e de 24,3% nas anteras presas. As condições ambientais após a antese influenciaram a intensidade da doença no campo. Os resultados reforçam que as anteras presas desempenham um papel importante no processo infeccioso e, portanto, devem ser os principais sítios de infecção a serem protegidos com fungicidas.

Danos causados pela infecção de Gibberella zeae em trigo

A giberela do trigo (Triticum aestivum), causada por Gibberella zeae, é uma doença de infecção floral com freqüente ocorrência em regiões onde, após o início da floração do trigo, ocorrem períodos prolongados de chuva (> 48 h) e temperaturas médias (> 20 ºC). Os danos na redução do rendimento de grãos, causados pela infecção natural de giberela no campo, foram quantificados em diferentes cultivares de trigo nas safras agrícolas de 2001 e 2002, no município de Passo Fundo, RS. Do estádio de grão leitoso até a maturação, foram identificadas e marcadas todas as espigas de trigo gibereladas, em uma área de 1 m², amostrando-se três repetições por área de trigo. As espigas gibereladas e sadias foram colhidas, secas, contadas e trilhadas separadamente. Os danos foram obtidos pela diferença entre o rendimento real e a estimativa do rendimento potencial, calculados com base no número total de espigas, no número de espigas sadias e no número de espigas gibereladas. O dano médio em 25 amostras de trigo coletadas em 2001, foi de 13,4%, variando de 6,4 à 23,1%. Na safra de 2002, em 18 amostras, o dano médio foi de 11,6%, variando de 3,1 a 20,5%. As reduções médias no rendimento de grãos foram de 394,4 Kg.ha-1 e 356,2 Kg.ha-1 para safras de 2001 e 2002, respectivamente. Nas duas safras, o dano médio da giberela, nos diferentes cultivares, foi de 375,3 kg.ha-1 ou 6,26 sacos de trigo/ha.

Factors affecting density of airborne Gibberella zeae inoculum

Fusarium head blight (FHB) is a disease of increasing concern in the production of wheat (Triticum aestivum). This work studied some of the factors affecting the density of airborne Gibberella zeae inoculum. Spore samplers were placed at the edge of a field in order to observe spore deposition over a period of 45 days and nights in September and October, the period that coincides with wheat flowering. Gibberella zeae colonies were counted for each period and values transformed to relative density. A stepwise regression procedure was used to identify weather variables helpful in predicting spore cloud density. In general, a predominant night-time spore deposition was observed. Precipitation and daily mean relative humidity over 90% were the factors most hightly associated with peak events of spores in the air. Models for predicting spore cloud density simulated reasonably well with the fluctuation of airborne propagules during both night and day, with potential to be integrated into an FHB risk model framework.

Avaliação da deposição de calda em anteras de trigo, para o estudo do controle químico de giberela (Gibberella zeae)

A giberela, causada pelo fungo Gibberella zeae (Fusarium graminearum), é uma importante doença de infecção floral do trigo. Para controlar essa doença, o fungicida deve ser aplicado durante a antese, de modo a proteger as anteras. Os objetivos do presente trabalho foram de construir uma barra de pulverização que permitiu utilizar bicos em diferentes arranjos e quantificar a presença de corante nas anteras quando aplicado com diferentes pontas de pulverização, direções dos bicos na barra e com diferentes volumes de calda. Utilizaram-se pontas de jato plano (XR-11002), jato plano duplo (110DB2) e jato cônico vazio (JA-2), espaçadas em 0,50 m, em uma barra tradicional e numa barra modificada. Na barra modificada, foram utilizados corpos duplos giratórios, posicionando-se os bicos na vertical, 30º ou 45º para frente e para trás em relação à vertical. Os resultados mostraram que a utilização de dois bicos no mesmo ponto da barra aumentou significativamente o número de anteras que receberam o fungicida. As três pontas de pulverização comportaram-se de maneira semelhante. Dois bicos formando ângulo de 30º em relação à vertical proporcionaram a mesma quantidade de anteras cobertas com corante do que dois bicos posicionados na vertical. A angulação dos bicos em 45º com a vertical proporcionou maior número de anteras atingidas pela calda.

Molecular characterization and fumonisin production by Fusarium verticillioides isolated from corn grains of different geographic origins in Brazil

Gibberella moniliformis is most commonly associated with maize worldwide and produces high levels of fumonisins, some of the most agriculturally important mycotoxins. Studies demonstrate that molecular methods can be helpful for a rapid identification of Fusarium species and their levels of toxin production. The purpose of this research was to apply molecular methods (AFLP, TEF-1 alpha partial gene sequencing and PCR based on MAT alleles) for the identification of Fusarium species isolated from Brazilian corn and to verify if real time RT-PCR technique based on FUM1 and FUM19 genes is appropriated to estimate fumonisins B(1) and B(2) production levels. Among the isolated strains, 96 were identified as Fusarium verricillioides, and four as other Fusarium species. Concordant phylogenies were obtained by AFLP and TEF-1 alpha sequencing, permitting the classification of the different species into distinct clades. Concerning MAT alleles, 70% of the F. verricillioides isolates carried the MAT-1 and 30% MAT-2. A significant correlation was observed between the expression of the genes and toxin production r=0.95 and r=0.79 (correlation of FUM1 with FB(1) and FB(2), respectively, P < 0.0001): r=0.93 and r =0.78 (correlation of FUM19 with FB(1) and FB(2). respectively, P < 0.0001). Molecular methods used in this study were found to be useful for the rapid identification of Fusarium species. The high and significant correlation between FUM1 and FUM19 expression and fumonisins production suggests that real time RT-PCR is suitable for studies considering the influence of abiotic and biotic factors on expression of these genes. This is the first report concerning the expression of fumonisin biosynthetic genes in Fusarium strains isolated from Brazilian agricultural commodity. (c) 2010 Elsevier B.V. All rights reserved.

Flowchart for the production of perithecia of Gibberella zeae in wheat grains.

2016