Fitness Traits of Deoxynivalenol and Nivalenol-Producing Fusarium graminearum Species Complex Strains from Wheat.

Fusarium graminearum of the 15-acetyl-deoxynivalenol (15-ADON) chemotype is the main cause of Fusarium head blight (FHB) of wheat in southern Brazil. However, 3-ADON and nivalenol (NIV) chemotypes have been found in other members of the species complex causing FHB in wheat. To improve our understanding of the pathogen biology and ecology, we assessed a range of fitness-related traits in a sample of 30 strains representatives of 15-ADON (F. graminearum), 3-ADON (F. cortaderiae and F. austroamericanum), and NIV (F. meridionale and F. cortaderiae). These included perithecia formation on three cereal-based substrates, mycelial growth at two suboptimal temperatures, sporulation and germination, pathogenicity toward a susceptible and a moderately resistant cultivar, and sensitivity to tebuconazole. The most important trait favoring F. graminearum was a two times higher sexual fertility (>40% perithecial production index [PPI]) than the other species (<30% PPI); PPI varied among substrates (maize > rice > wheat). In addition, sensitivity to tebuconazole appeared lower in F. graminearum, which had the only strain with effective fungicide concentration to reduce 50% of mycelial growth >1 ppm. In the pathogenicity assays, the deoxynivalenol producers were generally more aggressive (1.5 to 2× higher final severity) toward the two cultivars, with 3-ADON or 15-ADON leading to higher area under the severity curve than the NIV strains in the susceptible and moderately resistant cultivars, respectively. There was significant variation among strains of the same species with regards asexual fertility (mycelial growth, macroconidia production, and germination), which suggested a strain- rather than a species-specific difference. These results contribute new knowledge to improve our understanding of the pathogen-related traits that may explain the dominance of certain members of the species complex in specific wheat agroecosystems.

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 journal's 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 Lin's 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.

Is the emergence of fungal resistance to medical triazoles related to their use in the agroecosystems? A mini review

Abstract Triazole fungicides are used broadly for the control of infectious diseases of both humans and plants. The surge in resistance to triazoles among pathogenic populations is an emergent issue both in agriculture and medicine. The non-rational use of fungicides with site-specific modes of action, such as the triazoles, may increase the risk of antifungal resistance development. In the medical field, the surge of resistant fungal isolates has been related to the intensive and recurrent therapeutic use of a limited number of triazoles for the treatment and prophylaxis of many mycoses. Similarities in the mode of action of triazole fungicides used in these two fields may lead to cross-resistance, thus expanding the spectrum of resistance to multiple fungicides and contributing to the perpetuation of resistant strains in the environment. The emergence of fungicide-resistant isolates of human pathogens has been related to the exposure to fungicides used in agroecosystems. Examples include species of cosmopolitan occurrence, such as Fusarium and Aspergillus, which cause diseases in both plants and humans. This review summarizes the information about the most important triazole fungicides that are largely used in human clinical therapy and agriculture. We aim to discuss the issues related to fungicide resistance and the recommended strategies for preventing the emergence of triazole-resistant fungal populations capable of spreading across environments.

Is the emergence of fungal resistance to medical triazoles related to their use in the agroecosystems? A mini review.

Triazole fungicides are used broadly for the control of infectious diseases of both humans and plants. The surge in resistance to triazoles among pathogenic populations is an emergent issue both in agriculture and medicine. The non-rational use of fungicides with site-specific modes of action, such as the triazoles, may increase the risk of antifungal resistance development. In the medical field, the surge of resistant fungal isolates has been related to the intensive and recurrent therapeutic use of a limited number of triazoles for the treatment and prophylaxis of many mycoses. Similarities in the mode of action of triazole fungicides used in these two fields may lead to cross-resistance, thus expanding the spectrum of resistance to multiple fungicides and contributing to the perpetuation of resistant strains in the environment. The emergence of fungicide-resistant isolates of human pathogens has been related to the exposure to fungicides used in agroecosystems. Examples include species of cosmopolitan occurrence, such as Fusarium and Aspergillus, which cause diseases in both plants and humans. This review summarizes the information about the most important triazole fungicides that are largely used in human clinical therapy and agriculture. We aim to discuss the issues related to fungicide resistance and the recommended strategies for preventing the emergence of triazole-resistant fungal populations capable of spreading across environments.

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.

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.

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.

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.

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.

Agressividade diferencial de espécies do complexo Fusarium graminearum em interação com o fungicida tebuconazole na redução do rendimento de trigo / Differential aggressiveness of species of the Fusarium graminearum complex in interaction with tebuconazole fungicide on the reduction of wheat grain yield

No Brasil, a giberela em trigo é causada por espécies do complexo Fusarium graminearum, especialmente F. graminearum sensu stricto (Fgss) e F. meridionale (Fmer), as quais variam quanto ao potencial toxigênico. O objetivo deste trabalho foi avaliar a relação entre características fenotípicas e agressividade dessas duas espécies associadas ao uso do fungicida tebuconazole, da classe dos triazóis, com a redução de rendimento do trigo. Em dez isolados Fgss e nove Fmer, foram avaliados: esporulação total, taxa de germinação e sensibilidade ao fungicida tebuconazole. A inoculação, para cada isolado, foi feita por aspersão em espigas e as variáveis severidade da doença, incidência de grãos giberelados e o peso de grãos foram avaliados. O efeito de tebuconazole na redução da doença foi avaliado em ação protetora, seguida de inoculação na espigueta central de plantas da cultivar 'BRS Guamirim', com os seguintes tipos de inóculo: somente Fgss, somente Fmer ou a mistura de ambos (1:1). Isolados Fgss apresentaram maior esporulação total, maior taxa de germinação e foram menos sensíveis ao tebuconazole em comparação a Fmer. Grãos giberelados por isolados Fmer apresentaram 50% maior peso do que aqueles provenientes de inoculações com Fgss. O tebuconazole apresentou efeito fungistático e os grãos de espigas tratadas com o fungicida apresentaram peso 25% superior aos não tratados. Sugere-se que diferenças no potencial de dano aos grãos pelas duas espécies, assim como o efeito fungistático de triazóis, podem ajudar a explicar a co-ocorrência de diferentes micotoxinas, o que ainda necessita ser confirmado com dados de campo.

Fusarium head blight of wheat in Brazil is caused mainly by two species of the Fusarium graminearum species complex: F. graminearum sensu stricto (Fgss) and F. meridionale (Fmer), which vary in relation to toxigenic potential. The aim of this study was to evaluate the effect of phenotypic traits and aggressiveness of these two species, associated with the tebuconazole fungicide, of the triazole group, on the reduction of wheat yield. Ten Fgss strains and Fmer strains were evaluated with regards to: total sporulation, germination rate and tebuconazole sensitivity. The strains were spray-inoculated onto BRS Guamirim cv. 'plants' at full flowering and disease severity, Fusarium-damaged kernels, and kernel weight were evaluated. The effect of tebuconazole in disease control was evaluated through spraying the fungicide and then inoculating the central-floret of the wheat head with the following inoculum treatments: a mixture of all Fgss strains, a mixture of all Fmer strains or a mixture of all isolates of both species (1:1). Strains of Fgss showed higher total sporulation and germination rates and were less sensitivity to tebuconazole. The weight of Fusarium-damaged kernels from inoculations with Fmer strains was 50% higher than those inoculated with Fgss. A fungistatic effect on the disease was found for tebuconazole application and harvested kernels showed 25% higher grain weight than the untreated kernels. It is suggested that the distinct yield loss potential by these two species, in association with the fungistatic effect of triazoles could explain the co-occurrence of different mycotoxins in the harvested kernels, which needs to be proven with field data.

Características patogênicas de isolados do complexo Fusarium graminearum e de Fusarium verticillioides em sementes e plântulas de milho / Pathogenic traits of Fusarium graminearum complex and Fusarium verticillioides isolates on seeds and seedlings of maize

No Brasil, diversas espécies de Fusarium são patogênicas ao milho, afetando sementes, plântulas, colmos e grãos. Objetivou-se comparar isolados de F. verticillioides e de duas espécies do complexo F. graminearum (F. graminearum sensu stricto e F. meridionale), de ocorrência predominante em sementes de milho, quanto à taxa de crescimento micelial, capacidade de colonização e redução da germinação da semente, redução da emergência de plântulas e infecção e colonização de colmos de milho.Testes permutacionais e análise de contraste foram usados para discriminar os isolados. Análise de componentes principais foi utilizada para identificar as variáveis responsáveis pela maior variância entre os isolados. Com exceção da redução da germinação, os isolados F. verticillioides apresentaram menores valores de taxa de crescimento micelial, eficiência de infecção, redução na emergência de plântulas e comprimento de lesão em colmos, quando comparados aos demais, que não se diferenciaram. As variáveis redução da emergência e redução da germinação apresentaram maior influência na caracterização dos isolados, sugerindo que sementes infectadas com isolados de qualquer uma das espécies do complexo F. graminearum testadas representam maior risco ao estabelecimento da cultura do que isolados de F. verticillioides.

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.

Meio semiseletivo para recuperação e quantificação de Cryptosporiopsis perennans em maçãs / Semi-selective medium for recovery and quantification of Cryptosporiopsis perennans from apple fruits

A podridão "olho de boi", causada pelo fungo Cryptosporiopsis perennans, é uma das doenças de pós-colheita da macieira mais importantes no Brasil. Os objetivos deste trabalho foram desenvolver e avaliar um meio semiseletivo para a recuperação e mensuração da população epífita de C. perennans em maçãs "Fuji". A suplementação de BDA com tetraciclina, nistatina e iprodiona, nas respectivas concentrações de 12,5; 1 e 1µg p.a mL-1, proporcionou o desenvolvimento de colônias de C. perennans estatisticamente igual (P>0,05) ao observado no meio BDA. Em condições de infecção natural e em amostras pré-inoculadas com C. perennans, o meio semiseletivo permitiu detectar e quantificar a população do fungo na superfície dos frutos, além de ter inibido o crescimento de fungos leveduriformes, dematiáceos e leveduras. Em lavagens de frutos previamente inoculados, o número de conídios recuperados com o meio semiseletivo apresentou correlação (r=0,88; P<0,05) com a concentração do inóculo aspergido sobre os frutos.

Bull's eye rot, caused by Cryptosporiopsis perennans, is one of the most important apple's posharvest diseases in Brazil. The aim of this research was to develop and evaluate a semi-selective media to recovery and quantification of the epiphytic population of C. perennans on the apple fruit surface. A PDA medium amended with tetracycline, nistatine and iprodione in concentrations of 12.5; 1 and 1µg i.a.mL-1, respectively, allowed the development of C. perennans colonies statistically similar (P>0.05) to those in PDA media. Nistatine concentrations higher than 1µg.mL-1 inhibited the growth of the pathogen. Under natural infection conditions and pre-inoculation with C. perennans, the media allowed the detection and quantification of the fungal population on the fruit surface without affecting C. perennans development. Number of conidia observed in pre-inoculated fruits was correlated (r=0.88; P<0.05) to inoculum concentration sprayed on the fruit surface.