Sclerotinia sclerotiorum, causal agent of white mould, is the most destructive and widely distributed soilborne pathogen of common bean during the autumn–winter season in Brazil. Nevertheless, little is known about the genetic structure of the pathogen population. Microsatellite (SSR) markers and mycelial compatibility groups (MCGs) were used to characterize 118 isolates collected from 20 bean fields located in the most important growing regions of Minas Gerais State (MG). Additionally, the genetic variability among 10 isolates obtained from a single sclerotium was investigated in 10 different sclerotia. Seventy SSR haplotypes and 14 MCGs were identified among the 118 isolates. The genetic differences within bean growing areas accounted for most of the genetic variation (72%). Despite the relatively high genotypic diversity, the SSR loci were at linkage disequilibrium. Moreover, 70% of the isolates were assigned to only two MCGs, and haplotypes of a given MCG were closely related. The discriminant analysis of principal components revealed five groups. There was strong genetic differentiation between isolates collected in one municipality in southern MG when compared to other regions. Common bean resistance to white mould should be assessed with representative isolates of the five genetic groups and, if possible, of the different MCGs detected in the present study. One to five haplotypes were detected among the 10 isolates obtained from a single sclerotium. Therefore, in order to ensure genetic identity of an isolate, hyphal tip or monoascosporic isolates should be used.
White mould (Sclerotinia sclerotiorum) is a destructive disease of soybean worldwide. However, little is known of its impact on soybean production in Brazil. A meta‐analytic approach was used to assess the relationship between disease incidence and soybean yield (35 trials) and between incidence and sclerotia production (29 trials) in experiments conducted in 14 locations across four seasons. Region, site elevation and season included as moderators in random‐effects and random‐coefficients models did not significantly explain the variability in the slopes of the incidence–yield relationship. The Pearson's r, obtained from back‐transforming the Fisher's Z estimated by an overall random‐effects model, showed that incidence of white mould was moderately and negatively correlated with yield (r = −0.76, P < 0.0001). A random‐coefficients model estimated a slope of −17.2 kg ha−1%−1, for a mean attainable yield of 3455 kg ha−1, indicating that a 10% increase in white mould incidence would result in a mean yield reduction of 172 kg ha−1. White mould incidence and production of sclerotia were strongly and positively correlated (r = 0.85, P < 0.0001). For every 10% increase in white mould incidence, 1 kg ha−1 of sclerotia was produced. The relationship between disease incidence and production of sclerotia was stronger in southern regions and at higher elevation. In the absence of management, economic losses associated with white mould epidemics, assuming 43% incidence in 22% of the soybean area, were estimated at approximately US $1.47 billion annually within Brazil.
Além do valor como recurso terapêutico, plantas medicinais também possuem potencial para serem utilizadas como fonte de princípios ativos contra fitopatógenos. O objetivo deste trabalho foi avaliar o efeito de óleos essenciais das espécies medicinais Baccharis dracunculifolia (alecrim-do-campo), Schinus terebinthifolius (aroeirinha) e Porophyllum ruderale (arnica-brasileira) sobre o crescimento dos fungos fitopatogênicos Fusarium oxysporum f. sp. phaseoli (Fop), F. solani f. sp. phaseoli (Fsp), Sclerotinia sclerotiorum (Ss), S. minor (Sm), Rhizoctonia solani (Rs), Sclerotium rolfsii (Sr) e Macrophomina phaseolina (Mp). Avaliou-se em placas de Petri o crescimento radial desses fungos em meio batata-dextrose-ágar (BDA) com cinco concentrações (0, 250, 500, 1000 e 3000 mg L-1) dos óleos essenciais. Discos de micélio (5 mm de diâmetro) de cada fungo em crescimento foram transferidos para placas de Petri que foram mantidas a 23°C no escuro por 48 horas. O óleo essencial de alecrim-do-campo foi o mais eficiente na redução do crescimento micelial de todos os fungos, com inibição completa quando se utilizou a concentração de 3000 mg L-1. A redução de crescimento variou de 29% (Fs) a 80% (Rs) a 250 mg L-1 do óleo essencial de alecrim-do-campo; a 500 mg L-1, variou de 29% (Fs) a 98% (Sr); e a 1000 mg L-1, de 41% (Fs) a 100% (Sr). A redução do crescimento dos fungos pelo óleo de aroeirinha na concentração de 3000 mg L-1 variou de 27% (Fsp) a 74% (Rs). Nessa concentração, o óleo de arnica-brasileira reduziu o crecimento micelial de Ss em 72%, o de Rs em 80% e o de Mp em 82%, sem efeitos significativos sobre o crescimento micelial de Fsp e Fop. Conclui-se que os óleos essenciais de alecrim-do-campo, aroeirinha e arnica-brasileira possuem potencial para o controle dos fungos fitopatogênicos estudados, com destaque para o óleo de alecrim-do-campo.
Sclerotinia sclerotiorum populations from tropical agricultural zones have been suggested to be more variable compared to those from temperate zones. However, no data were available comparing populations from both zones using the same set of markers. In this study, we compared S. sclerotiorum populations from the United States of America (USA, temperate) and southeast Brazil (tropical) using the frequency of mycelial compatibility groups (MCGs) and 13 microsatellite (SSR) markers. Populations were sourced from diseased plants within leguminous crops in New York, USA (NY; n = 78 isolates), and Minas Gerais State, Brazil (MG; n = 109). Twenty MCGs were identified in NY and 14 were previously reported in MG. The effective number of genotypes based on Hill’s number of order 0, which corresponded to the number of multilocus genotypes (MLGs) were 22 (95% CI = 15.6–28.4) and 24 (95% CI = 18.9–29.1) in NY and MG, respectively. Clonal fractions of MLGs were 71.8% (NY) and 78.0% (MG). The effective number of genotypes based on Hill’s number of orders 1 and 2 in NY were 8.9 (95% CI = 5.2–12.6) and 4.4 (95% CI = 2.6–6.1), respectively. For MG these indices were 11.4 (95% CI = 8.7–14.1) and 7.1 (95% CI = 5.1–9.0), respectively. There were no significant differences of allelic richness, private allelic richness, gene diversity, effective number of alleles and genotype evenness between the NY and MG populations. The populations were differentiated, with 29% of total variance attributed to differences between them and G''ST and Jost’s D indices higher than 0.50. Cluster analysis revealed dissimilarity higher than 80% among most MLGs from both populations. Different alleles segregated in the populations but both had similar levels of genotypic variability.
Thiophanate-methyl (TM), fluazinam, and procymidone are fungicides extensively used for white mold control of common bean in Brazil. We assessed the sensitivity of Brazilian isolates of Sclerotinia sclerotiorum to these three fungicides using discriminatory doses and concentration that results in 50% mycelial growth inhibition (EC50) values. In total, 282 isolates from the most important production areas were screened and none was resistant to fluazinam or procymidone. The EC50 values varied from 0.003 to 0.007 and from 0.11 to 0.72 μg/ml for fluazinam and procymidone, respectively. One isolate was resistant to TM. The EC50 of the TM-resistant isolate was greater than 100 μg/ml, whereas the EC50 of the sensitive isolates varied from 0.38 to 2.23 μg/ml. The TM-resistant isolate had a L240F mutation in the β-tubulin gene. This is the first report of mutation at codon 240 causing resistance to a benzimidazole fungicide in S. sclerotiorum. The high-resolution melting analysis allowed the distinction of TM-sensitive and -resistant isolates by specific melting peaks and curves. The TM-resistant isolate had mycelial growth, sclerotia production, and aggressiveness comparable with that of the sensitive isolates, indicating that this genotype will likely compete well against sensitive isolates in the field. This study demonstrates that resistance to TM, fluazinam, and procymidone is nonexistent or rare. Resistance management practices should be implemented, however, to delay the spread of TM-resistant genotypes.
Common bean breeding programs for white mold (WM) resistance are in their initial stages in Brazil. Sources of partial resistance to WM are available abroad but their performance in Brazil is unknown. In two greenhouse (straw test) and three field experiments conducted in three districts in the state of Minas Gerais, Brazil, we evaluated a total of 23 lines with putative WM resistance with the objective to select lines with resistance to WM and other diseases associated with high yield potential. Two field-resistant local lines, two susceptible local cultivars, and two susceptible international lines were also included in the study. In the greenhouse, Cornell 605, A 195, and G122 were among the lines with the highest partial resistance to WM. In the field, these three lines were highly resistant to WM and had intermediate resistance or were resistant to anthracnose, angular leaf spot, rust, and Fusarium wilt. Cornell 605 and A 195 had high-yield potential but G122 yielded 47% less than the local lines under WM pressure. Our results suggest that Cornell 605 and A 195 are the most useful sources of resistance to WM for use in common bean breeding programs in Brazil.
White mold (Sclerotinia sclerotiorum) of leguminous crops in New York is generally managed with preventive applications of fungicides. However, no research has been conducted during the last decade to assess the sensitivity of the S. sclerotiorum population to fungicides or compare their performance under field conditions. The sensitivity of S. sclerotiorum to boscalid, fluazinam, and thiophanate-methyl was assessed in 151 isolates from 15 fields across New York using an agar dilution method with discriminatory concentrations. In addition, the effective concentration at which mycelial growth is reduced by 50% (EC50) was estimated for one representative isolate from each field. The efficacy of commercial formulations of each fungicide on white mold incidence in plants and pods was also tested in two field trials (2015 and 2016). The EC50 values ranged from 0.068 to 0.219, 0.001 to 0.002, and 1.23 to 2.15 µg/ml for boscalid, fluazinam, and thiophanate-methyl, respectively. Evidence of resistance was not found using the discriminatory concentration tests. The mycelial growth inhibition relative to the control ranged from 56 to 83%, 66 to 84%, and 53 to 83% at discriminatory concentrations of boscalid (5 µg a.i./ml), fluazinam (0.05 µg a.i./ml), and thiophanate-methyl (5 µg a.i./ml), respectively. Fourteen isolates with mycelial growth inhibition lower than 60% at 5 µg/ml of thiophanate-methyl, did not exhibit point mutations within a partial sequence of the β-tubulin gene. In the field trials, fungicides effectively reduced white mold incidence on plants by 75% (2015) and 93% (2016) and on pods by 81% (2015) and 87% (2016), both relative to the nontreated plots. However, fungicide applications led to significant increases in pod yield, relative to the nontreated plots, only in 2015 when the incidence of white mold on plants and pods were higher (85 and 49.2%) than in 2016 (31.3 and 10.3%). Although fungicide resistance was not detected, and thus control failures reported by New York snap bean growers may be due to other factors, further monitoring of sensitivity within the S. sclerotiorum population is encouraged as well as the use of rational systems to base their judicious and economic use.
White mold caused by the fungus Sclerotinia sclerotiorum is the most yield-limiting disease of common bean in Brazil. To date, there has been no commercial cultivar resistant to this disease. In a greenhouse we evaluated white mold resistance sources (Cornell 605, A195 and G122) against eight isolates of S. sclerotiorum from five Brazilian states. A Brazilian cultivar (BRSMG Madrepérola) and a susceptible check (Beryl) were used as control. Treatments were arranged in factorial combinations (5 × 8) in a completely random design with four replicates.Disease severity was assessed on a rating scale of 1-to-9 together with lesion length, which was used to determine an area under the disease progress curve (AUDPC). Polymorphisms detected in ten microsatellite loci were used to assess variability between the isolates. Each isolate was a distinct haplotype; they formed a genetic tree with two clusters. One cluster was formed by three isolates collected from the states of Minas Gerais and São Paulo (southeastern); the others, by isolates from Paraná, Santa Catarina (southern), Goiás (Mid-western), and again, Minas Gerais.Genotype × isolate interaction was significant. In general, Beryl was more susceptible than BRSMG Madrepérola. Considering the AUDPC and/or the white mold reaction score, Cornell 605 exhibited more physiological resistance than BRSMG Madrepérola to seven isolates, A195 to five isolates, and G122 to two isolates. Our results suggest that Cornell 605 is the best source of resistance to white mold for the southern region, whereas Cornell 605 and A195 are somewhat superior to G122 for the southeastern and mid-western regions.
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