Piérri Spolti

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.

Phenotypic and pathogenic traits of two species of the Fusarium graminearum complex possessing either 15-ADON or NIV genotype

Fusarium head blight (FHB) in Brazil is caused mainly by two members of the Fusarium graminearum species complex, each possessing either a DON/15-ADON chemotype (F. graminearum sensu stricto, Fgss) or a NIV chemotype (F. meridionale, Fmer). In this work, we aimed to characterize and compare isolates belonging to each species, obtained either from wheat or barley, in relation to phenotypic traits (mycelial growth, sporulation and germination) and pathogenicity (in vitro glume infection and in vivo central floret inoculation) to two Brazilian wheat cultivars, Guamirim (moderate susceptible) and BRS 194 (susceptible). Results showed significantly higher growth rates, greater spore production and quicker germination for the Fgss isolates compared to the Fmer isolates, which were also more sensitivity to tebuconazole than the Fgss isolates. All isolates were capable of infecting glume tissues of both varieties, with an overall higher infection frequency for Fgss than Fmer isolates when inoculated in cv. Guamirim than in cv. BRS194, which showed similar infection frequency between the species. Accordingly, in the central-floret inoculation assay, Fgss isolates were also more aggressive than Fmer isolates towards cv. Guamirim, but not towards BRS194, based on the mean area under disease progress curves. It is hypothesized that phenotypic traits and host resistance may play a role in the selection of more aggressive populations under field conditions, thus partially explaining the dominance of Fgss populations associated with FHB of wheat in Brazil.

Triazole Sensitivity in a Contemporary Population of Fusarium graminearum from New York Wheat and Competitiveness of a Tebuconazole-Resistant Isolate

A sample of 50 isolates, including 25 each of the 3-acetyldeoxynivalenol and the 15-acetyldeoxynivalenol trichothecene genotype, from a contemporary collection of Fusarium graminearum associated with Fusarium head blight (FHB) of wheat in New York varied in sensitivity to tebuconazole (effective concentration leading to a 50% reduction of mycelial growth [EC50] of 0.28 to 8.09 mg/liter; μ = 1.12 mg/liter) and metconazole (0.05 to 0.86 mg/liter; μ = 0.33). Mean sensitivity did not differ between the trichothecene genotype groups. Isolate Gz448NY11 from Steuben County is the first tebuconazole-resistant field isolate of F. graminearum reported in the Americas and has the lowest sensitivity to tebuconazole (EC50 = 8.09 mg/liter) documented for this species. Suppression of FHB and deoxynivalenol (DON) following application of a commercial rate of tebuconazole was significantly diminished in plants inoculated with the tebuconazole-resistant isolate compared with those inoculated with a tebuconazole-sensitive isolate well documented for its aggressiveness and toxigenicity on wheat. There was no diminution of FHB and DON suppression with either isolate following application of metconazole. Significantly more individuals of the tebuconazole-resistant isolate were recovered from spikes inoculated with an equal mixture of the two isolates and sprayed with tebuconazole. Future studies are needed on the epidemiology and monitoring of triazole-resistant isolates to understand the risk that fungicide resistance poses to disease management and food security.

Improving sooty blotch and flyspeck severity estimation on apple fruit with the aid of standard area diagrams

Sooty blotch and flyspeck is caused by numerous species of fungi that colonize the surface of apple fruit and thereby lower its market value. Although this disease poses a substantial threat to apple growers’ profitability in some regions, reliable and cost-effective methods for epidemiological and disease control studies have not been validated, nor are they widely available. We modified a standard area diagram to aid sooty blotch and flyspeck severity assessments and quantified its impact on accuracy and precision of visual estimates. Samples of ‘Fuji’ and ‘Mutsu’ fruit were photographed both from the top and laterally. Severity was assessed from a sub-sample of 160 images using image analysis software. Validation of the diagram was performed by eight raters who independently assessed severity in two series of selected images representing the lateral view and the top view, initially unaided and subsequently with the aid of the scale. Severity estimates ranged from 0.4% to 98% (most fruit had <10% severity). Accuracy and precision of the estimates were significantly improved when using the diagrammatic scale; concordance correlation coefficient values increased from 0.81 to 0.95. A strong tendency to underestimate severity for the mid-range to high levels was minimized when using the aid, which also improved reproducibility of the estimates among raters. In addition to strengthening evidence that a standard area diagram can be used reliably in sooty blotch and flyspeck studies, we expanded its application to disease assessment in the peduncle region, which enhances the usefulness of the method for evaluating efficacy of management practices.

Fitness Attributes of Fusarium graminearum Isolates from Wheat in New York Possessing a 3-ADON or 15-ADON Trichothecene Genotype

In all, 50 isolates of Fusarium graminearum from wheat spikes in New York, including 25 isolates each of the 15-acetyl-deoxynivalenol (15-ADON) and 3-ADON genotype, were tested to determine whether 3-ADON isolates are more fit for saprophytic survival and pathogenicity on wheat spikes than are 15-ADON isolates. The isolates were characterized and compared for 14 different attributes of saprophytic fitness and pathogenic fitness on a susceptible wheat variety. Isolates of the two genotypes could not be differentiated for most of these traits. Three principle components-ascospore production on corn stalks, total trichothecene amount in wheat kernels, and incidence of diseased spikelets up from the point of inoculation-accounted for 29.4, 18.9, and 10.8% of the variation among the isolates, respectively. A bootstrapping procedure grouped the isolates into two distinct groups, with 27 and 23 isolates each, with isolates from both genotypes represented in similar proportions (15-ADON/3-ADON, n = 14/13 and 11/12). Within the contemporary population of F. graminearum causing wheat head blight in New York, isolates with a 3-ADON genotype did not possess any detectable advantage over isolates with a 15-ADON genotype in saprophytic fitness or in pathogenic fitness on a susceptible wheat cultivar.

Características patogênicas de isolados do complexo Fusarium graminearum e de Fusarium verticillioides em sementes e plântulas de milho

In Brazil, several Fusarium species are pathogenic to corn affecting seeds, seedlings, stalks and grains. This research aimed to compare isolates from F. verticillioides and from two species of the F. graminearum complex (F. graminearum sensu stricto and F. meridionale) prevalent on corn seeds in relation to mycelial growth rate, ability to colonize and reduce seed germination, reduction in seedling emergence and infection and colonization of corn stalk. Permutational test and contrast analysis was performed to discriminate the isolates and species. Principal component analysis was used to identify the variables leading to greatest variance among the isolates. With the exception of seed germination, F. verticillioides isolates showed lower values for mycelial growth rate, infection efficiency, reduction in seedling emergence and lesion size than F. graminearum complex isolates that were all similar. Seedling emergence and seed germination showed greater influence on the characterization of isolates, suggesting that seeds infected with F. graminearum isolates of any of the species tested represent a greater threat to crop stand than F. verticillioides isolates.

Controle do inóculo inicial para redução dos danos pela podridão: 'olho-de-boi' em macieiras

Aiming to evaluate strategies for the control of bulls eye rot (BER) based on the reduc- tion of initial inoculum the following fungicide treatments were applied in experimental plots established in 2006/07 and 2007/08 seasons in Vacaria - RS, southern of Brazil. During the plant dormancy period it was applied: lime sulfur (3.0%); copper oxychloride (0.5%); copper hydroxide (0.3%); and cuprous oxide (0.3%). The check treatment consisted of non sprayed plots. Branches of one and two years and floral buds were sampled and the associated inoculum recovered with a semi-selective media to evaluate the effect of the treatments on inoculum reduction. During harvest, BER incidence on fruits was evaluated in four cat- egories of fruits: i) fallen on the ground under the projection of the canopy; ii) in the plants; iii) harvested (for the detection of latent infections); and iv) harvested and stored for three months in a cold chamber. A linear relationship was observed between inoculum levels of Cryptosporiopsis sp. on buds and total losses by BER (R 2 = 0.948; P = 0.001). Applications of lime sulfur promoted the most reduction of inoculum on buds and 50% of reduction of total losses by bulls eye rot, differing significantly from the other treatments in 2006/07 season. In fruits, inoculum levels increased linearly during the season, being relatively higher in the 2007/08 season when the eradicated treatments led to a 40% control of losses, thus demonstrating the beneficial economic return by the reduction of initial inoculum of BER. Index terms: eradicate treatments, epyphite population,latent infections, fruit rots, Cryptosporiopsis sp..

Disease Risk, Spatial Patterns, and Incidence-Severity Relationships of Fusarium Head Blight in No-till Spring Wheat Following Maize or Soybean

The first large-scale survey of Fusarium head blight (FHB) in commercial wheat fields in southern Brazil was conducted over three years (2009 to 2011). The objectives were to: (i) evaluate whether increased FHB risk is associated with within-field maize residue; (ii) determine the spatial pattern of FHB incidence; and (iii) quantify the relationship between FHB incidence and severity. FHB was assessed in a total of 160 fields between early milk and dough. Incidence ranged from 1.0 to 89.9% (median = 25%) and severity from 0.02 to 18.6% (median = 1.3%). FHB risk was neither lower nor higher in wheat following maize than in wheat following soybean. Only 18% of fields were classified as having aggregated patterns of FHB-symptomatic spikes. A binary power law description of the variances was consistent with an overall random pattern of the disease. These results conform with the hypothesis that FHB epidemics in southern Brazil are driven by sufficient atmospherically-transported inoculum from regional sources. The incidence-severity relationship was coherent across growing season, growth stage, and previous crop; one common fitted curve described the relationship across all observations. Estimating severity from incidence may be useful in reducing the workload in epidemiological surveys.

Trichothecene Genotype Composition of Fusarium graminearum Not Differentiated Among Isolates from Maize Stubble, Maize Ears, Wheat Spikes, and the Atmosphere in New York

In order to test the hypothesis that the trichothecene genotype composition of local populations of Fusarium graminearum is structured by specific habitats, a collection of 1,407 isolates was obtained from overwintered maize stubble, mature maize ears and wheat spikes, and the atmosphere 1.5 m aboveground during the flowering stage of these crops. These isolates were sampled at three diverse agricultural locations in New York State: namely, Aurora (sampled in 2012 and 2013) in central New York, Belmont (sampled in 2013) in southwestern New York, and Willsboro (sampled in 2013) in northeastern New York. Approximately 100 isolates of F. graminearum from each habitat were collected within a 10-mile2 area in each location. Polymerase chain reaction assays were used to identify three main B-trichothecene genotypes--3-acetyldeoxynivalenol (3-ADON), 15-ADON, or nivalenol (NIV)--based on amplification of portions of Tri3 and Tri12 genes. All but the NIV genotype were detected. The 15-ADON genotype predominated in most locations; frequencies were 92% (652/709) at Aurora, 78% (332/379) at Belmont, and 53% (167/319) at Willsboro. Frequencies of any genotype did not differ in general among the four habits in each location. An exception was in Aurora 2012, where only 5 in 24 3-ADON isolates were found in samplings from the air and grains of both crops. As viewed by the composition of trichothecene genotypes, local populations of F. graminearum appear not to be structured by these four habitats inclusive of pathogenic and saprophytic phases of the fungus life cycle. The similar frequency of 3-ADON and 15-ADON in eastern New York (Willsboro), which is less than 400 km away from the Aurora sampling location in the central area of the state, suggests that regional populations may be differentiated based on selection associated with climatic or landscape features not currently identified.

Standard Area Diagrams for Aiding Severity Estimation: Scientometrics, Pathosystems, and Methodological Trends in the Last 25 Years

Standard area diagrams (SAD) have long been used as a tool to aid the estimation of plant disease severity, an essential variable in phytopathometry. Formal validation of SAD was not considered prior to the early 1990s, when considerable effort began to be invested developing SAD and assessing their value for improving accuracy of estimates of disease severity in many pathosystems. Peer-reviewed literature post-1990 was identified, selected, and cataloged in bibliographic software for further scrutiny and extraction of scientometric, pathosystem-related, and methodological-related data. In total, 105 studies (127 SAD) were found and authored by 327 researchers from 10 countries, mainly from Brazil. The six most prolific authors published at least seven studies. The scientific impact of a SAD article, based on annual citations after publication year, was affected by disease significance, the journals impact factor, and methodological innovation. The reviewed SAD encompassed 48 crops and 103 unique diseases across a range of plant organs. Severity was quantified largely by image analysis software such as QUANT, APS-Assess, or a LI-COR leaf area meter. The most typical SAD comprised five to eight black-and-white drawings of leaf diagrams, with severity increasing nonlinearly. However, there was a trend toward using true-color photographs or stylized representations in a range of color combinations and more linear (equally spaced) increments of severity. A two-step SAD validation approach was used in 78 of 105 studies for which linear regression was the preferred method but a trend toward using Lins correlation concordance analysis and hypothesis tests to detect the effect of SAD on accuracy was apparent. Reliability measures, when obtained, mainly considered variation among rather than within raters. The implications of the findings and knowledge gaps are discussed. A list of best practices for designing and implementing SAD and a website called SADBank for hosting SAD research data are proposed.