In this study, more than 150 bacteria showing antagonistic properties against bacterial and fungal pathogens of the tomato plant were isolated and characterized. The most efficient agents against these phytopathogenic microorganisms belong to the genus Bacillus: the best biocontrol isolates were representatives of Bacillus subtilis, B. mojavensis and B. amyloliquefaciens species. They intensively produced fengycin or/and surfactin depsipeptide antibiotics and also proved to be excellent protease secretors. It was proved, that the selected strains were able to use ethylenethiourea (ETU) as sole nitrogen source. These antagonistic and ETU-degrading Bacillus strains can be applied as biocontrol and also as bioremediation agents.
Surfactins are lipopeptide-type biosurfactants produced mainly by Bacillus species, consisting of a peptide loop of seven amino acids and a hydrophobic fatty acid chain (C12–C16). These molecules have been proven to exhibit various biological activities; thus, their therapeutic and environmental applications are considered. Within the surfactin lipopeptide family, there is a wide spectrum of different homologues and isomers; to date, more than 30 variants have been described. Since the newest members of these lipopeptides were described recently, there is no information that is available on their characteristic features, e.g., the dependence of their production from different cultivation parameters. This study examined the effects of both the different carbon sources and various metal ions on the surfactin production of a selected B. subtilis strain. Among the applied carbon sources, fructose and xylose had the highest impacts on the ratio of the different variants, regarding both the peptide sequences and the lengths of the fatty acids. Furthermore, the application of metal ions Mn2+, Cu2+ and Ni2+ in the media completely changed the surfactin variant compositions of the fermenting broths leading to the appearance of methyl esterified surfactin forms, and resulted in the appearance of novel surfactin variants with fatty acid chains containing no more than 11 carbon atoms.
Metal- and pesticide-tolerant biocontrol agents are preferred in integrated pest management, as such strains can be applied in combination with different pesticides. The Bacillus velezensis strain SZMC 6161J proved to be sensitive to copper, nickel, zinc, and cadmium, while manganese elevated its growth. At concentrations higher than 1 mmol L , zinc and iron inhibited the chymotrypsin-like activity of this strain. In addition, trypsin-like protease and palmitoyl esterase activities were insensitive to all tested heavy metals in the applied concentration range. We studied the effects of some widely used herbicides and fungicides on the growth of this strain. The presence of sulfonylurea herbicides, like bensulfuron-methyl, cinosulfuron, chlorsulfuron, ethoxysulfuron, triasulfuron, and primisulfuron-methyl strongly inhibited the biomass production of the strain even at the concentration of 6.25 mg L . Glyphosate also inhibited the growth above 30 mg L . Similarly, contact fungicides like captan, maneb, mancozeb, and thiram resulted in total inhibition at the concentration as low as 6.25 mg L . Interestingly, the sterol-biosynthesis-inhibiting fungicides imazalil, fenarimol, penconazole, and tebuconazole also proved to be potent inhibitors. Heavy metal- and fungicide-tolerant strains were isolated from the parental strain and their antagonistic abilities were evaluated. There was no substantial difference between the antagonism capability of wild-type strain and the resistant mutants.
(1) Background: Bacillus velezensis and Bacillus amyloliquefaciens are closely related members of the “operational group B. amyloliquefaciens”, a taxonomical unit above species level within the ”Bacillus subtilis species complex”. They have similar morphological, physiological, biochemical, phenotypic, and phylogenetic characteristics. Thus, separating these two taxa from each another has proven to be difficult to implement and could not be pushed easily into the line of routine analyses. (2) Methods: The aim of this study was to determine whether whole FAME profiling could be used to distinguish between these two species, using both type strains and environmental isolates. Initially, the classification was determined by partial sequences of the gyrA and rpoB genes and the classified isolates and type strains were considered as samples to develop the identification method, based on FAME profiles. (3) Results: The dissimilarities in 16:0, 17:0 iso, and 17:0 FA components have drawn a distinction between the two species and minor differences in FA 14:0, 15:0 iso, and 16:0 iso were also visible. The statistical analysis of the FA profiles confirmed that the two taxa can be distinguished into two separate groups, where the isolates are identified without misreading. (4) Conclusions: Our study proposes that the developed easy and fast-automated identification tool based on cellular FA profiles can be routinely applied to distinguish B. velezensis and B. amyloliquefaciens.
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