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Fig. S1 Fractionation of proteins in mycelia from the fungus Alternaria brassicicola.

Fig. S2 Two-dimensional electrophoresis (2-DE) analyses of proteins in fraction S from control cultures of Alternaria brassicicola (A) and cultures treated with 0.10 mM brassinin (B). Total loading protein, 1 mg. Proteins were resolved on a gel using isoelectric focusing gradient 3–10 NL strips in the first dimension, and 10% polyacrylamide SDS-PAGE (sodium dodecylsulphate-polyacrylamide gel electrophoresis) gels in the second dimension. Protein spots were visualized by Coomassie blue staining of the gels. Each result is a compilation of three independent experiments and differentially expressed spots are labelled as ‘U’ for up-regulated proteins and ‘D’ for down-regulated proteins relative to control. Sixteen differentially expressed protein spots marked with arrows were identified using capillary liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) (Tables S2 and S4).

Fig. S3 Summary of metabolic pathways of Alternaria brassicicola affected by brassinin at 0.50 and 0.10 mM (Tables S1 and S2). Enzymes identified in this investigation are boxed: green represents up-regulated and yellow represents down-regulated proteins (red type, from 0.50 mM brassinin; blue type, from 0.10 mM brassinin; black underline, present in both 0.50 and 0.10 mM brassinin). P, phospho or phosphate; PP, diphospho or diphosphate.

Fig. S4 Confocal laser scanning microscopy images of mycelial cells from Alternaria brassicicola treated with rhodamine 123 and 3,3′-dihexyloxacarbocyanine iodide [DIOC6(3)]. Cells were incubated for 30 min in potato dextrose broth (PDB) medium containing 20 mM rhodamine 123 without (A) or with 0.10 mM brassinin (B) or 0.50 mM brassinin (C). Hyphae stained with 10 mM DIOC6(3) without (D) or with 0.10 mM brassinin (E) or 0.50 mM brassinin (F). Scale bar, 10 µm.

Table S1 Proteins differentially expressed in cultures of Alternaria brassicicola (controls) and cultures induced with brassinin at 0.50 mM. Proteins were identified using capillary liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS).

Table S2 Proteins differentially expressed in cultures of Alternaria brassicicola (controls) and cultures induced with brassinin at 0.10 mM. Proteins were identified using capillary liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS).

Table S3 Induction with 0.50 mM brassinin.

Table S4 Induction with 0.10 mM brassinin.

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MPP_765_sm_figureS1-4.pdf1286KSupporting info item
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MPP_765_sm_tableS3-4.xls615KSupporting info item

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