Dauri José Tessmann

Molecular survey of trichothecene genotypes of Fusarium graminearum species complex from barley in Southern Brazil

Fusarium head blight is a disease of primary concern to small-grain cereals of Brazil, including barley. Its main causal agent, Fusarium graminearum species complex (Fg complex)¸ is able to produce mycotoxins, especially deoxynivalenol (DON) and nivalenol (NIV), that usually contaminate grain. Strains that produce DON may also produce its acetylated derivatives: 3-acetyl-DON (3-ADON) and 15-acetyl-DON (15-ADON). Ninety two isolates were obtained from samplings of barley grain during three years (2007, 2008 and 2009) from several fields in both southern and northern production regions of Rio Grande do Sul state, Brazil. These isolates were examined for polymerase chain-reaction-based (PCR) trichothecene genotype based on the amplification of portions of Tri3 and Tri12. There was no effect of year or region on the proportion of trichothecene genotypes. Overall, 66% of the strains (61/92) were 15-ADON, 4.4% (4/92) were 3-ADON and 29.3% (27/92) were NIV. The overall NIV/DON ratio estimated (0.41) was five times higher than that found in previous studies with strains from wheat grown in the same region. Species identification of nine strains representing the trichothecene genotypes, based on comparisons of DNA sequences of portions of the PHO, RED and URA genes with sequences from curated reference isolates of Fusarium from GenBank, revealed that they belong to F. graminearum sensu stricto (four 15-ADON and one 3-ADON strain), F. meridionale (three NIV strains) and F. austroamericanum (one 3-ADON strain). These results add to the current regional knowledge of trichothecene genotypes and species within the Fg complex affecting barley in the region.

Regional and Field-Specific Factors Affect the Composition of Fusarium Head Blight Pathogens in Subtropical No-Till Wheat Agroecosystem of Brazil

A multiyear survey of >200 wheat fields in Paraná (PR) and Rio Grande do Sul (RS) states was conducted to assess the extent and distribution of Fusarium graminearum species complex (FGSC) diversity in the southern Brazilian wheat agroecosystem. Five species and three trichothecene genotypes were found among 671 FGSC isolates from Fusarium head blight (FHB)-infected wheat heads: F. graminearum (83%) of the 15-acetyldeoxynivalenol (15-ADON) genotype, F. meridionale (12.8%) and F. asiaticum (0.4%) of the nivalenol (NIV) genotype, and F. cortaderiae (2.5%) and F. austroamericanum (0.9%) with either the NIV or the 3-ADON genotype. Regional differences in FGSC composition were observed, with F. meridionale and the NIV type being significantly (P<0.001) more prevalent in PR (>28%) than in RS (≤9%). Within RS, F. graminearum was overrepresented in fields below 600 m in elevation and in fields with higher levels of FHB incidence (P<0.05). Species composition was not significantly influenced by previous crop or the stage of grain development at sampling. Habitat-specific differences in FGSC composition were evaluated in three fields by characterizing a total of 189 isolates collected from corn stubble, air above the wheat canopy, and symptomatic wheat kernels. Significant differences in FGSC composition were observed among these habitats (P<0.001). Most strikingly, F. meridionale and F. cortaderiae of the NIV genotype accounted for the vast majority (>96%) of isolates from corn stubble, whereas F. graminearum with the 15-ADON genotype was dominant (>84%) among isolates from diseased wheat kernels. Potential differences in pathogenic fitness on wheat were also suggested by a greenhouse competitiveness assay in which F. graminearum was recovered at much higher frequency (>90%) than F. meridionale from four wheat varieties inoculated with an equal mixture of F. graminearum and F. meridionale isolates. Taken together, the data presented here suggest that FGSC composition and, consequently, the trichothecene contamination in wheat grown in southern Brazil is influenced by host adaptation and pathogenic fitness. Evidence that F. meridionale and F. cortaderiae with the NIV genotype are regionally significant contributors to FHB may have significant implications for food safety and the economics of cereal production.

Fungitoxicidade, atividade elicitora de fitoalexinas e proteção de pepino contra Colletotrichum lagenarium, pelo extrato aquoso de Eucalyptus citriodora

Secondary compounds present in medicinal plants can play important roles in plant-pathogen interactions, through immediate antimicrobial action or by inducing a defence mechanism in other plants that may be treated with such compounds. With the objective of verifying the potential of Eucalyptus citriodora in the alternative control of anthracnose in cucumber (Cucumus sativus), aqueous extract (AE) of this forest essence, autoclaved or unautoclaved, in concentrations of 0,1, 1, 5, 10, 15, 20 e 25% was used in the following experiments: induction of phytoalexins in sorghum (Sorghum bicolor) etiolated mesocotyls and soybean (Glycine max) cotyledons; fungitoxicity in vitro against Colletotrichum lagenarium conidia; local or systemic induction of resistance in cucumber plants. Water and Bion were used as control treatments. The results indicated that the autoclaved and unautoclaved AE induced phytoalexin synthesis in sorghum starting from the concentration of 1%. In soybean, gliceollin synthesis took place in 10% and 15% of the autoclaved and unautoclaved AEs, respectively. There was total inhibition in the spores germination and apressoria formation in concentrations of 20% and 1% of the autoclaved AE, respectively. As for the unautoclaved extract, there was 75% inhibition of the spores germination in 25% of AE and total inhibition in the apressoria formation in 15% of AE. Based on the parameter size of lesion, the unautoclaved aqueous extract of E. citriodora could potentially induce local resistance in cucumber against C. lagenarium.

Manejo de doenças de plantas em cultivo protegido

Plant disease management in greenhouse crops Crop production based on plasticulture has allowed substantial gains in yields for many horticultural crops. In the greenhouse, crop production may be more easily optimized and plants may be better exploited for their maximum yields. However, this may increase the impact of some biotic and abiotic diseases. Less important diseases may become highly destructive in crops cultivated in the greenhouse. Indeed, in some cases disease management may become more complex in the greenhouse than in the field. Disease management in greenhouse cropping systems requires flexibility and great efforts for integration of available control measures.

Production of mycotoxins by Fusarium graminearum isolated from small cereals (wheat, triticale and barley) affected with scab disease in Southern Brazil

Fusarium fungi are known to be pathogenic for plants and mycotoxin producers. The in vitro production of deoxynivalenol and zearalenone was qualitatively evaluated in 24 different isolates of Fusarium graminearum collected from small cereals associated with the scab disease, in southern Brazil. Isolates were cultivated in rice during 14 days at 28oC. Cultivates were extracted with methanol:water (40:60 v/v) and analyzed by thin layer chromatography. Other trichothecenes (diacetoxyscirpenol, fusarenon-X, neosolaniol and nivalenol) and zearalenol, often produced by Fusarium, were also analyzed. In the conditions used, it was possible to detect zearalenone and deoxynivalenol in 67% and 33% of the isolates, respectively. The presence of zearalenol, diacetoxyscirpenol and fusarenone was also detected. None of the isolates was found to produce nivalenol or neosolaniol.

Grape rust caused by Phakopsora euvitis, a new disease for Brazil

A ferrugem da videira foi constatada pela primeira vez no Brasil, ocorrendo em parreirais (Vitis spp.)comerciais no estado do Parana tendo recentemente atingido tambem o Estado de Sao Paulo. O fungo Phakopsora euvitis foi identificado como o agente causal da doenca. Esta ferrugem ocorre preferencialmente em folhas maduras causando a desfolha precoce das plantas infetadas.

Diagrammatic scale for assessment of grapevine rust

A diagrammatic scale with six levels of disease severity (1, 5, 12, 25, 50 and 75%) was developed to assess grapevine (Vitis spp.) rust severity, caused by the fungus Phakopsora euvitis. Two versions of the scale, in black-and-white and in color, were validated by seven raters without previous experience with grapevine rust, who estimated the severity of 30 grapevine leaves showing rust symptoms, with and without the use of the scales. Precision and accuracy were determined by linear regression, relating the assessments using the scales to actual severity. Actual severity was assessed with the software ASSESS®. Using the diagrammatic scales, some raters were able to improve precision and accuracy. The color scale provided slightly more precise and accurate estimates than the black-and-white scale.

A new PCR approach for the identification of Fusarium graminearum

The main objective of this work was to develop a PCR protocol for the identification of Fusarium graminearum, based on a pair of primers targeted to a segment of the 3´coding region of the gaoA gene that codes for the enzyme galactose oxidase (GO). This region has low homology with the same region of GO genes from other fungi. Genomic DNA from 17 strains of Fusarium spp. isolated from diseased cereals, from several other Fusarium species, and from other fungi genera was analyzed in a PCR assay using this primer set. The 17 strains of Fusarium spp. were also analyzed for the GO enzyme production in submerse fermentation in a new formulated liquid medium. All strains that were morphologically and molecularly identified as F. graminearum were able to secrete the enzyme and had a positive result in the used PCR protocol. No DNA fragment was amplified using genomic DNA from other Fusarium species and species of other fungi genera. The results suggest that the proposed PCR protocol is specific and can be considered as a new molecular tool for the identification of F. graminearum. In addition, the new formulated medium is a cheap alternative for screening for GO screening production by F. graminearum.

Hentriacontane: a leaf hydrocarbon from Syzygium jambos with stimulatory effects on the germination of urediniospores of Puccinia psidii

A crude Sohxlet extract from leaves of Syzygium jambos was sequentially fractionated using a silica gel flash column. A bioassay based on the numbers of urediniospores of Puccinia psidii that germinated in 2% water agar detected an active stimulant of germination when the fraction eluted with 100% n-hexane was used. The active fraction induced up to 88% increase in germination when added to a spore suspension in mineral oil. The active fraction was characterized as a hydrocarbon by 1H nuclear magnetic resonance, 13C nuclear magnetic resonance, and infrared analysis. Gas chromatography-mass spectrometry analysis indicated that the fraction was a long-chain 436 MW hydrocarbon with corresponding to C31H64, namely hentriacontane. This is the first time such a compound proved to be involved with stimulation of fungal spore germination. These results may contribute to better understanding the infection process of rusts.

Fusarium species and fumonisins associated with maize kernels produced in Rio Grande do Sul State for the 2008/09 and 2009/10 growing seasons

Ear rots caused by Fusarium spp. are among the main fungal diseases that contribute to poor quality and the contamination of maize grains with mycotoxins. This study aimed to determine the visual incidence of fungal-damaged kernels (FDKs), the incidence of two main Gibberella (a teleomorph of Fusarium) complexes (G. fujikuroi and G. zeae) associated with maize using a seed health blotter test, and the fumonisin levels, using high performance liquid chromatography, in samples of maize grains grown across 23 municipalities during the 2008/09 and 2009/10 growing seasons. Additionally, 104 strains that were representative of all of the analysed samples were identified to species using PCR assays. The mean FDK was seven per cent, and only six of the samples had levels greater than six per cent. Fusarium spp. of the G. fujikuroi complex were present in 96% of the samples, and G. zeae was present in 18% of the samples (5/27). The mean incidence of G. fujikuroi was 58%, and the incidence of G. zeae varied from 2 to 6%. FB1 was found in 58.6%, FB2 in 37.9%, and both toxins in 37.9% of the samples. The FB1 and FB2 levels were below the quantification limits for 41.3% of the samples, and the mean FB1 levels (0.66 μg/g) were higher than the mean FB2 levels (0.42 μg/g). The PCR identification separated the 104 isolates into three of the G. fujikuroi complex: F. verticillioides (76%), F. subglutinans (4%) and F. proliferatum (2%); and G. zeae (anamorph = F. graminearum) (18%). Our results confirmed the dominance of F. verticillioides, similar to other regions of Brazil, but they differed due to the relatively higher incidence of F. graminearum. Total fumonisin levels were below the maximum limit determined by current Brazilian regulations.