Aims: This work was conducted to identify the antifungal compounds produced by two previously isolated Bacillus sp. strains: ARP23 and MEP218. Both strains were subjected to further analysis to determine their taxonomic position and to identify the compounds responsible for their antifungal activity as well as to evaluate the efficiency of these strains to control sclerotinia stem rot in soybean.
Methods and Results: The antifungal compounds were isolated by acid precipitation of cell‐free supernatants, purified by RP‐HPLC and then tested for antagonistic activity against Sclerotinia sclerotiorum. Mass spectra from RP‐HPLC eluted fractions showed the presence of surfactin C15, fengycins A (C16–C17) and B (C16) isoforms in supernatants from strain ARP23 cultures, whereas the major lipopeptide produced by strain MEP218 was iturin A C15. Alterations in mycelial morphology and sclerotial germination were observed in the presence of lipopeptides‐containing supernatants from Bacillus strains cultures. Foliar application of Bacillus amyloliquefaciens strains on soybean plants prior to S. sclerotiorum infection resulted in significant protection against sclerotinia stem rot compared with noninoculated plants or plants inoculated with a nonlipopeptide‐producing B. subtilis strain.
Conclusions: Both strains, renamed as B. amyloliquefaciens ARP23 and MEP218, were able to produce antifungal compounds belonging to the cyclic lipopeptide family. Our data suggest that the foliar application of lipopeptide‐producing B. amyloliquefaciens strains could be a promising strategy for the management of sclerotinia stem rot in soybean.
Significance and Impact of the Study: Sclerotinia stem rot was ranked as one of the most severe soybean disease in Argentina and worldwide. The results of this study showed the potential of B. amyloliquefaciens strains ARP23 and MEP218 to control plant diseases caused by S. sclerotiorum.
The development of alternatives for the use of chemical pesticides for plant disease control is the present-day and ongoing challenge for achieving sustainable agriculture. Pseudomonas fluorescens SF4c, native strain from wheat, produces tailocins (phage-tail-like bacteriocins) with antimicrobial activity against several phytopathogenic strains. We thus investigated the efficacy of foliar application of these bacteriocins to control the bacterial-spot disease in tomato caused by Xanthomonas vesicatoria Xcv Bv5-4a. The disease severity and incidence index were reduced by 44 and 36%, respectively; while the number of viable cells of X. vesicatoria Xcv Bv5-4a decreased after bacteriocin treatment. Furthermore, bacteriocin was effective in reducing bacterial-spot-disease symptoms on tomato fruits even when applied 12 h after infection. Tailocin activity was not affected by abiotic influences such as adjuvant, light and temperature and, biotic factors such as apoplastic-fluids. In contrast, no antibacterial activity of these tailocins was observed when the bacteriocin was exposed to extremely dry conditions. Finally, that no cytotoxic effects on mammalian cells were observed with this representative tailocins is highly significant and demonstrates the safety of such compounds in humans. All these findings indicate that the SF4c tailocins represent an attractive alternative to copper-containing bactericides for use in the control of bacterial spot.
A bacterial collection of approximately one thousand native strains, isolated from saline soils of Cordoba province (Argentina), was established. From this collection, a screening to identify those strains showing plant growth promotion and biocontrol activities, as well as salt tolerance, was performed. Eight native strains tolerant to 1 M: NaCl and displaying plant growth promotion and/or biocontrol features were selected for further characterization. Strains MEP(2 )18, MRP(2 )26, MEP(2 )11a, MEP(3 )1, and MEP(3 )3b significantly increased the growth of maize seedlings under normal and saline conditions, whereas isolates ARP(2 )3, AEP(1 )5, and ARP(2 )6 were able to increase the root dry weight of agropyre under saline conditions. On the other hand, strains MEP(2 )18 and ARP(2 )3 showed antagonistic activity against phytopathogenic fungi belonging to Sclerotinia and Fusarium genus. Antifungal activity was found in cell-free supernatants, and it was heat and protease resistant. Strains MEP(2)18 and ARP(2)3 were identified as Bacillus sp. and strains MEP(2)11a and MEP(3)3b as Ochrobactrum sp. according to the sequence analysis of 16S rRNA gene.
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