Genes involved in cysteine metabolism are important for tellurite resistance in bacteria (Chasteen et al., 2009). We then decided to compare the tellurite sensitivity of strains BSIP1215 and BSIP1793 (ΔcymR). On plates containing methionine, the ΔcymR mutant was less resistant to tellurite than the wild-type strain with a growth inhibition area diameter of 47.7 and 30.3 mm, respectively (Fig. 4b). In contrast, on plates containing cystine, the same growth inhibition area diameter was obtained for both strains (40.2 mm for BSIP1793 and 40.6 mm for BSIP1215) (Fig. 4b). In addition, the black deposits were much more prevalent for the ΔcymR mutant than for

the wild-type strain and the this website blackening mostly surrounded the paper disk for strain BSIP1793

(Fig. 4a, left Selleckchem Bafilomycin A1 panel). Tellurite might be reduced by the H2S produced by bacteria. The significant amount of H2S produced in the ΔcymR mutant was probably responsible for the quantity of tellurium deposits observed with this mutant. The diffusion of H2S into the plate could also explain why tellurite reduction occurred even in the zone of growth inhibition. To confirm the possible role of H2S in this phenomenon, we repeated the same disk assay, but kept the lid of the plate open in a moisturized atmosphere, allowing H2S diffusion outside from the plate. The growth inhibition area diameter of the ΔcymR mutant then markedly increased in the open plates, reaching 52.7 mm instead of 38.1 mm for the wild-type strain. Simultaneously, the blackening around the paper disk disappeared

Immune system (Fig. 4a, right panel). A similar result was obtained when 5 mL of alkaline agar enriched with zinc acetate was poured on the lid to absorb H2S (data not shown). This indicated that H2S obtained from cysteine degradation probably participated in tellurite reduction, protecting the ΔcymR mutant from its toxicity. When H2S escaped from the plate, we observed a drastic increase in tellurite sensitivity for the ΔcymR strain similar to that obtained in the presence of methionine under conditions producing less H2S (Fig. 3a). We then tested the effect of CymR inactivation on the susceptibility of B. subtilis to other stress stimuli. We compared the sensitivity of strains BSIP1215 and BSIP1793 (ΔcymR) to paraquat, H2O2 and diamide using disk diffusion assays. The ΔcymR mutant was significantly more sensitive than the wild-type strain to diamide, a specific thiol oxidant that causes disulfide stress. This effect was observed with plates containing cystine or methionine (Table 1, data not shown). We further tested the effect of H2O2 and paraquat. On plates with methionine, the growth inhibition area in the presence of 10 μL of 2 M paraquat was 58.8 mm for the ΔcymR mutant and 49.3 mm for the wild-type strain. Under the same conditions, the zone of growth inhibition in the presence of 10 μL of 10 M H2O2 was 52.1 mm for the ΔcymR mutant and 41.4 mm for the wild-type strain.

Disease symptoms were measured including stem lesions after 10 weeks of planting. Stem lesions were evaluated using a scale of 1–5 as described previously by Sturz et al. (1995). After 3 months, the yielded tubercles (g), per pot

treatment, were recorded. Statistical analyses were used as described above. All fungal isolates were identified using ITS regions of rDNA and blast search. All isolates showed 100% homology with E. nigrum, A. longipes, R. solani, Lapatinib solubility dmso and T. atroviride (Table 1). One isolate showed 99.6% homology with Phomopsis subordinaria and was therefore named as Phomopsis sp. The blast scores are summarized in Table 1. The confrontation cultures between R. solani and isolates E1, E8, and E18 (identified as E. nigrum) showed clear inhibition zones and different patterns of interactions (Fig. 1). Isolates E2 and R24, identified as T. atroviride and Phomopsis sp., respectively, showed fast growth and covered the plate completely including the mycelium of R. solani. Isolate E13, identified as A. longipes, also showed an inhibition zone against the pathogenic fungus. Antagonistic isolates Selleck Anti-infection Compound Library showed different inhibition rates when confronted with R. solani (Table 1). The highest inhibition rate was observed

with T. atroviride, followed by Phomopsis sp., A. longipes, and E. nigrum. Nevertheless, these inhibition rates were statistically significant at P≤0.05. Figure 2 shows the different patterns of interactions between antagonistic isolates. The antagonist mycelium was easily distinguished from R. solani mycelium by hyphal morphology (Fig. 2f). Trichoderma atroviride hyphae established close contact with those of R. solani by coiling (Fig. 2e). The coils were usually very dense and appeared to tightly encircle the R. solani hyphae. After 7 days, T. atroviride hyphae penetrated R. solani hyphae and caused a loss of turgor. Phomopsis sp. invaded the R. solani colony and limited its growth (Fig. 2d). The hyphal density of Phomopsis sp. was higher than R. solani. Alternaria longipes also showed a denser hyphae than

R. solani, but no evidence of any hyphal penetration was observed. However, Fossariinae these cocultured R. solani hyphae showed an abnormal morphology in comparison with hyphae of R. solani grown alone (Fig. 2f). This may be due to a reduction in cell turgor. Epicoccum nigrum isolates grow alongside of R. solani hyphae and then wind around it, causing lysis of its hyphae (Fig. 2a and b). Epicoccum nigrum did not show any evidence of penetration, although clear inhibition zones were observed where R. solani mycelia were almost dead. All antagonistic fungal isolates are capable of producing volatile compounds when grown on PDA media. Table 2 shows a significant difference between various antagonist isolates. The highest inhibition was recorded by T. atrovirde (81.81%), followed by Phomopsis sp. (38.63%), A. longipes (21.02%), and E. nigrum E18 (20.73%), E1 (11.36%), and E8 (10.22%), respectively.

Disease symptoms were measured including stem lesions after 10 weeks of planting. Stem lesions were evaluated using a scale of 1–5 as described previously by Sturz et al. (1995). After 3 months, the yielded tubercles (g), per pot

treatment, were recorded. Statistical analyses were used as described above. All fungal isolates were identified using ITS regions of rDNA and blast search. All isolates showed 100% homology with E. nigrum, A. longipes, R. solani, www.selleckchem.com/products/bmn-673.html and T. atroviride (Table 1). One isolate showed 99.6% homology with Phomopsis subordinaria and was therefore named as Phomopsis sp. The blast scores are summarized in Table 1. The confrontation cultures between R. solani and isolates E1, E8, and E18 (identified as E. nigrum) showed clear inhibition zones and different patterns of interactions (Fig. 1). Isolates E2 and R24, identified as T. atroviride and Phomopsis sp., respectively, showed fast growth and covered the plate completely including the mycelium of R. solani. Isolate E13, identified as A. longipes, also showed an inhibition zone against the pathogenic fungus. Antagonistic isolates Idasanutlin cell line showed different inhibition rates when confronted with R. solani (Table 1). The highest inhibition rate was observed

with T. atroviride, followed by Phomopsis sp., A. longipes, and E. nigrum. Nevertheless, these inhibition rates were statistically significant at P≤0.05. Figure 2 shows the different patterns of interactions between antagonistic isolates. The antagonist mycelium was easily distinguished from R. solani mycelium by hyphal morphology (Fig. 2f). Trichoderma atroviride hyphae established close contact with those of R. solani by coiling (Fig. 2e). The coils were usually very dense and appeared to tightly encircle the R. solani hyphae. After 7 days, T. atroviride hyphae penetrated R. solani hyphae and caused a loss of turgor. Phomopsis sp. invaded the R. solani colony and limited its growth (Fig. 2d). The hyphal density of Phomopsis sp. was higher than R. solani. Alternaria longipes also showed a denser hyphae than

R. solani, but no evidence of any hyphal penetration was observed. However, Methocarbamol these cocultured R. solani hyphae showed an abnormal morphology in comparison with hyphae of R. solani grown alone (Fig. 2f). This may be due to a reduction in cell turgor. Epicoccum nigrum isolates grow alongside of R. solani hyphae and then wind around it, causing lysis of its hyphae (Fig. 2a and b). Epicoccum nigrum did not show any evidence of penetration, although clear inhibition zones were observed where R. solani mycelia were almost dead. All antagonistic fungal isolates are capable of producing volatile compounds when grown on PDA media. Table 2 shows a significant difference between various antagonist isolates. The highest inhibition was recorded by T. atrovirde (81.81%), followed by Phomopsis sp. (38.63%), A. longipes (21.02%), and E. nigrum E18 (20.73%), E1 (11.36%), and E8 (10.22%), respectively.