Background: There is an urgent need to discover biocontrol agents to control bacterial wilt. This study reports on a new lipopeptide-producing biocontrol strain FJAT-46737 and explores its lipopeptidic compounds, and this study investigates the antagonistic effects of these compounds. Results: Based on a whole genome sequence analysis, the new strain FJAT-46737 was identified as Bacillus velezensis, and seven gene clusters responsible for the synthesis of bioactive secondary metabolites in FJAT-46737 were predicted. The antimicrobial results demonstrated that FJAT-46737 exhibited broad-spectrum antimicrobial activities in vitro against three bacteria and three fungi. Pot experiments showed that the control efficiencies for tomato bacterial wilt of the whole cultures, the 2-fold diluted supernatants and the crude lipopeptide of FJAT-46737 were 66.2%, 82.0%, and 96.2%, respectively. The above results suggested that one of the antagonistic mechanisms of FJAT-46737 was the secretion of lipopeptides consisting of iturins, fengycins and surfactins. The crude lipopeptides had significant antagonistic activities against several pathogens (including Ralstonia solanacearum, Escherichia coli and Fusarium oxysporum) and fengycins were the major antibacterial components of the lipopeptides against R. solanacearum in vitro. Furthermore, the rich organic nitrogen sources (especially yeast extracts) in the media promoted the production of fengycin and surfactin by FJAT-46737. The secretion of these two lipopeptides was related to temperature fluctuations, with the fengycin content decreasing by 96.6% and the surfactins content increasing by 59.9% from 20°C to 40°C. The optimal temperature for lipopeptide production by FJAT-46737 varied between 20°C and 25°C. Conclusions: The B. velezensis strain FJAT-46737 and its secreted lipopeptides could be used as new sources of potential biocontrol agents against several plant pathogens, and especially the bacterial wilt pathogen R. solanacearum.

Fusarium wilt of banana (Musa spp.) caused by Fusarium oxysporum f. sp. cubense (Foc) is one of the most serious banana fungal diseases in the world. Understanding the infection process of Foc is important for development of effective ways in disease control. In order to follow infection and colonization of this pathogen from root to rhizome and pseudostem tissues of banana, a highly pathogenic strain FJAT-3076 of Foc race 4 (Foc4) was transformed with gene encoding green fluorescent protein (GFP) and the fungus carrying gfp (FJAT-3076-GFP) was used to inoculate banana plants (Cavendish cv. B.F.). After inoculation for 3 to 10 d, it was observed that the conidia and their germ-tubes had penetrated into epidermis of young roots. The hyphae were found inside the root xylem 10 d after inoculation in the rhizome and pseudostem xylem after inoculation for 17 d. All plants infected by Foc died in 24 d after inoculation. It was also observed that Foc had spread all over the xylem and part of hyphae reached the pseudostem surface. Hyphal population was found the highest in the pseudostem, lower in root and least in rhizome. Field survey confirmed that Foc4 were mostly present in the base of pseudostem and less in the rhizome. Thus, effective prevention of the Foc hyphae movement from the rhizome up to the pseudostem might delay or control banana wilt disease.

4'-Thiosemicarbazonegriseofulvin, a new thiosemicarbazide derivative of griseofulvin, was synthesized and evaluated for its potential in the control of enzymatic browning and postharvest disease of fruits. Browning on fruits is mainly due to the enzymatic oxidation of phenolic compounds catalyzed by tyrosinase. 4'-Thiosemicarbazonegriseofulvin could effectively inhibit the activity of tyrosinase, and its 50% inhibitory concentration (IC(50)) against tyrosinase was determined to be 37.8 μM. It was a reversible and noncompetitive inhibitor of tyrosinase, and its inhibition constant (K(I)) was determined to be 38.42 μM. The antifungal activity of 4'-thiosemicarbazonegriseofulvin was studied against four fungi (Fusarium oxysporum, Fusarium moniliforme, Fusarium solani, and Colletotrichum truncatum) that often cause postharvest diseases of fruits. The results showed that 4'-thiosemicarbazonegriseofulvin could also strongly inhibit the mycelial growth of the four target fungi; the 50% lethal concentration (LC(50)) values were 5.4, 7.0, 15.3, and 1.5 mM, respectively.

Aims: This study aimed to investigate the effect of metal ions on lipopeptide production by Bacillus subtilis strain FJAT-4 and the mechanism of negative regulation by Ca 2+ . Methods and Results:The quantitative measurement of lipopeptides in response to K + , Na + , Mg 2+ and Ca 2+ addition was carried out by LC-MS. The contents of fengycin and surfactin varied within the range of 116. 24-129.80 mg/L and 34.03-63.11 mg/L in the culture media containing K + , Na + and Mg 2+ , while the levels were 0.86 and 0.63 mg/L in the media containing Ca 2+ . Ca 2+ at a high concentration (45 mM) did not adversely affect the growth of strain FJAT-4, but caused significant downregulation of lipopeptide synthesis-related gene expression, corresponding to a decrease in lipopeptide production. This inhibition by Ca 2+ was further investigated by proteomic analysis. In total, 112 proteins were upregulated and 524 proteins were downregulated in the presence of additional Ca 2+ (45 mM). Among these differentially expressed proteins (DEPs), 28 were related to phosphotransferase activity, and 42 were related to kinase activity. The proteomics results suggested that altered levels of three two-component signal-transduction systems (ResD/ResE, PhoP/PhoR and DegU/DegS) might be involved in the control of expression of the fen and srfA operons of FJAT-4 under high calcium stress. Conclusions:The Ca 2+ at the high concentration (45 mM) triggers a decrease in lipopeptide production, which might be attributed to the regulation of three twocomponent signal-transduction systems ResD/ResE, PhoP/PhoR and DegU/DegS. Significance and Impact of the Study:The regulatory effect of calcium on the expression of genes encoding lipopeptide synthetases can be applied to optimize the production of lipopeptides.

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