These authors contributed equally to this work.
Both extracellular chitinase and a new cyclic lipopeptide, chromobactomycin, contribute to the biocontrol activity of Chromobacterium sp. C61
Version of Record online: 11 SEP 2013
© 2013 BSPP AND JOHN WILEY & SONS LTD
Molecular Plant Pathology
Volume 15, Issue 2, pages 122–132, February 2014
How to Cite
Kim, H. J., Choi, H. S., Yang, S. Y., Kim, I. S., Yamaguchi, T., Sohng, J. K., Park, S. K., Kim, J.-C., Lee, C. H., Gardener, B. M. and Kim, Y. C. (2014), Both extracellular chitinase and a new cyclic lipopeptide, chromobactomycin, contribute to the biocontrol activity of Chromobacterium sp. C61. Molecular Plant Pathology, 15: 122–132. doi: 10.1111/mpp.12070
- Issue online: 6 JAN 2014
- Version of Record online: 11 SEP 2013
- Accepted manuscript online: 20 AUG 2013 05:09AM EST
- Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry, and Fisheries. Grant Number: 311019-03
- Ministry for Food, Agriculture, Forest, and Fisheries
- National Research Foundation of Korea. Grant Number: R32-2009-000-20047-0
Fig. S1 Effect of the purified Chi54 protein on the germination rate of phytopathogenic fungal spores. Fungal spores of Colletotrichum gloeosporioides (Cg), Botrytis cinerea (Bc), Cladosporium spharospermum (Cs) and Fusarium oxysporum (Fo) were treated with different concentrations of the purified Chi54 protein in 10 mm sodium acetate buffer (pH 6.0) or the buffer without the Chi54 protein. The germination rate was measured by counting the number of fungal spores that germinated versus the total number of spores using a haemocytometer under a phase contrast microscope 10 h after incubation at 28 °C. The data are the means of two independent experiments and the vertical bars indicate the standard deviations. Different letters indicate significant differences between fungal pathogens according to Duncan's multiple range test (P < 0.05).
Fig. S2 High-performance liquid chromatograms of ethyl acetate-extracted compounds from cell-free filtrates of Chromobacterium sp. C61. The solvent system was 85% (v/v) aqueous methanol supplemented with 0.1% trifluoroacetic acid at 2.0 mL/min, and the peaks were detected at 210 nm. The chromobactomycin peak from Chromobacterium sp. C61 is shown with an arrow, and this material was used to obtain the time-of-flight mass spectrum provided.
Fig. S3 Ion trap-time-of-flight mass spectrometry (IT-TOF MS), MS2 and MS3 data of the isolated antifungal compound.
Fig. S4 Correlation spectra of dihydroxybutyric acid (Dhb) in methanol-d6 of the isolated compound.
Fig. S5 Heteronuclear multiple-bond correlation spectra of dihydroxybutyric acid (Dhb) and Tyr-5 in methanol-d6 of the isolated compound.
Fig. S6 Heteronuclear multiple-quantum correlation spectra of β-hydroxymyristate in methanol-d6 of the isolated compound.
Fig. S7 Heteronuclear multiple-bond correlation spectra in methanol-d6 identifying all carbonyl groups in the amino acid residues of the isolated compound.
Fig. S8 Heteronuclear multiple-bond correlation spectra of Tyr-5 in methanol-d6 of the isolated compound.
Fig. S9 Connectivities observed in correlation and heteronuclear multiple-bond correlation spectra, leading to sequence assignment of the isolated compound.
Fig. S10 The proposed modifications of chromobactomycin structure identified by tandem mass spectrometry (MS2). Dhb, dihydroxybutyric acid; Gln, glutamine; Gly, glycine; His, histidine; HMS, β-hydroxymyristate; 3-HTDA, 3-hydroxytetradecanoic acid; Thr, threonine; Tyr, tyrosine.
Fig. S11 Effects of the chi54 mutations on aerobic growth in the presence of chitin. Cultures were grown aerobically in potato dextrose broth (PDB) with 0.2% chitin, and the growth of bacterial strains was determined by measuring the colony-forming units/mL (CFU/mL) of serial diluted cultures. Bacterial populations were assessed at each time point using analysis of variance (ANOVA), and mean CFU/mL values were compared according to Duncan's multiple range test (*P < 0.05 and **P < 0.01). The data are the means of three independent experiments and vertical bars indicate standard deviations.
Table S1 1H chemical shift assignments of chromobactomycin in methanol-d4 (A) and dimethylsulphoxide (DMSO)-d6 (B).
Table S2 13C chemical shift assignments of chromobactomycin in methanol-d4 (A) and dimethylsulphoxide (DMSO)-d6 (B).
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