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 mold of common bean (Phaseolus vulgaris), caused by Sclerotinia sclerotiorum, is a major yield-limiting disease during the fall-winter season in Brazil. This study was conducted to evaluate the efficacy of decreasing within-row densities for an indeterminate growth habit (type IIIa) cultivar, keeping constant the between-row spacing of 0.5 m, to manage the disease. A modification of within-row plant distributions was also attempted in order to reduce white mold intensity. The study was conducted with sprinkler irrigation in two growing seasons (2000 and 2001) in Viçosa, State of Minas Gerais, Brazil, in an area naturally infested with sclerotia. In 2000, treatments were arranged as 3 × 2 × 2 factorial combination of within-row densities (15, 7.5, or 5 plants/m), within-row plant distributions (single and equidistant plants or equidistant groups of three plants), and fluazinam treatments (with or without). In 2001, 16, 12, 8, or 4 plants/m were combined with fluazinam treatments. Fluazinam was applied at both flowering onset and 10 or 13 days later. Average incidence was 92.6% in 2000 and 77.8% in 2001; severity index was 69.8% in 2000 and 40.2% in 2001; and yield was 1,656 kg/ha in 2000 and 2,542 kg/ha in 2001. White mold decreased and yield increased as within-row densities were reduced, regardless of fluazinam treatments. A distribution of equidistant groups of three plants was ineffective in reducing disease. The use of 4 equidistant plants/m in infested irrigated areas was an effective strategy in white mold management.
The objective of this study was to understand the infection process of Fusarium oxysporum f. sp. phaseoli (Fop) in bean cultivars classified as resistant (Manteigão Fosco 11), intermediate (VP8) and susceptible (Meia Noite). Plants of the three cultivars were inoculated at 10 days after emergence with a suspension of 1×10 6 conidia of Fop per mL. At 43 days after the inoculation, stem segments were observed with a scanner electronic microscope. The cultivars Manteigão Fosco 11 and VP8 presented an occluding material in the xylem vessels, which may have restricted tissue colonization by Fop. The resistance of bean cultivars to Fop seemed also to be explained by structural differences in the xylem tissue. Key words: Phaseolus vulgaris, Fusarium wilt, host defense.The fungus Fusarium oxysporum Schlecht. f. sp. phaseoli Kendrick & Snyder (Fop) is the causal agent of the Fusarium wilt on common beans (Phaseolus vulgaris L.) and is present in all the regions that produce beans in the world (Alves-Santos et al., 2002;Abawi & Pastor-Corrales, 1990;Schwartz & McMillan, 1989;Buruchara & Camacho, 2000). The inadequate rotation of cultures, especially in areas irrigated with central pivot, the lack of preventive measures of control of the pathogen dissemination and the increase of soil compaction made the Fusarium wilt one of the most important bean diseases in Brazil (Paula Júnior et al., 2006).The dark, thick-walled chlamydospores are the longterm survival structures in soil (Abawi & Pastor-Corrales, 1990). Management practices alone may not be enough to keep the disease in low levels of intensity. Thus, the most efficient and viable alternative for the control of this disease is the use of resistant cultivars (Abawi, 1989;Abawi & Pastor-Corrales, 1990;Paula Júnior et al., 2006).The pathogen is capable of penetrating intact root tissue, although penetration of older parts of root and hypocotyl tissue also occurs, usually through wounds or natural openings (Abawi, 1989;Dongo & Müller, 1969;Duque & Müller, 1969). After penetration, hyphae of Fop move inter-and intracellularly and invade the xylem vessels (Mace et al., 1981). The fungus is confined to xylem vessels until the later stages of disease development, although limited invasion of the adjacent tissues may occur; growth of hyphae and transportation of microconidia in the xylem vessels are observed in susceptible cultivars (Abawi, 1989). On the other hand, in resistant plants the colonization between adjacent xylem vessels is restricted, probably as a result of chemical and structural alterations (Mace et al., 1981), including vascular occlusion by the formation of gel plugs, tyloses, deposition of additional wall layers and infusion of these structures with phenols and other metabolites (Mace et al., 1981).Although there are some reports about the differences in the histopathology of stem tissues from resistant and susceptible bean cultivars infected by Fop, detailed studies that approach the infectious process in plant tissues are scarce, especially compari...
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.
Fusarium root rot (FRR) is a distributed disease of common beans in Brazil. Our main aim was to verify if there are genotypes, mainly advanced lines from the common bean breeding program with levels of resistance to FRR similar to those of the line A-300. We also compared three developmental stages for FRR assessment. Genotypes of six classes were evaluated in greenhouse and field experiments. In greenhouse, substrate was infested with chlamydospores. In field, genotypes were screened in area infested with Fusarium solani f. sp. phaseoli. Plants were rated for disease at V3, R5 and R7 stages. Correlation among area under the disease progress curve (AUDPC) in greenhouse and in the field experiments was significant. Genotypes were ranked into four groups based on AUDPC. Sixteen advanced lines were as resistant to FRR as A-300. Correlation between the disease rating at R5 stages and AUDPC was higher than those among AUDPC and either disease rating at V3 or R7 in all experiments. Our results indicate that there are advanced lines with levels of FRR resistance similar to those of A-300 and that the FRR assessment at the R5 stage is more appropriated than at either V3 or R7.
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.
The usefulness of sclerotia morphology traits and mycelial growth rate to infer phenotypic variability in populations of Sclerotinia sclerotiorum associated with common beans in Minas Gerais State (MG) was assessed in two experiments. Isolates (131) were collected in the Northwestern, Alto Paranaíba, Zona da Mata and Southern regions of MG and the mycelial growth rate, the number, weight and dimensions (length, width and thickness) of sclerotia and mycelial pigmentation were assessed. There was a high variability among isolates, even among colonies of the same isolate. The variance of the first experiment for the mycelial growth rate, number, weight, length, width and thickness of sclerotia, were 0. 01, 190.15, 0.01, 0.62, 0.12 and 0.05, whereas in the second, the variances were 0.01, 28.48, 0.002, 0.18, 0.06 and 0.03, respectively. Most correlations between traits were of low magnitude except that between length and width (r = 0.84, 'P < 0.001). Regarding colony pigmentation, non-pigmented, pigmented and highly pigmented isolates were identified, but for most isolates this characteristic varied between experiments. No groups based on phenotypic traits were detected. The morphological traits evaluated in the present study are not suitable for characterization of populations of S. sclerotiorum. Key words: Phaseolus vulgaris, sclerotium, phenotypic variability, white mold.Several biological features make the ascomycete Sclerotinia sclerotiorum (Lib.) de Bary a highly threatening fungal pathogen to important crops in Brazil. Sclerotinia sclerotiorum is a homothallic fungus that does not produce conidia, but abundant sclerotia (Bolton et al., 2006). Sclerotia are asexual resting structures that allow pathogen survival when no host is available (Adams & Ayers, 1979). The pathogen has a host range that encompasses over 400 plant species (Boland & Hall 1994). Furthermore, pathogen propagules (sclerotia) can be easily spread over long-distance by contaminated seeds. Sclerotinia sclerotiorum causes white mold in crops such as cotton, soybean, potato, sunflower etc., and is the main soilborne disease affecting common bean (Phaseolus vulgaris L.) in Minas Gerais State (MG), Brazil.In Brazil common bean is produced in the springsummer, summer-fall, and fall-winter growing seasons (Vieira et al., 2010). Considering the three growing seasons, MG is the second largest producer, but ranks first during the fall-winter, with an average yield of 2600 kg ha -1 (CONAB, 2013). In the fall-winter growing season, moderate temperatures and high humidity provided by irrigation create conducive environmental conditions for white mold epidemics (Paula Júnior et al., 2006). Planting white mold resistant cultivar would be attractive alternative for disease management. Nevertheless, to date there are no resistant varieties available to growers. Sound breeding programs require proper understanding of the pathogen variability. The variability of populations of S. sclerotiorum has been investigated for different crops around the world an...
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