Purification and characterization of cassiicolin, the toxin produced by Corynespora cassiicola, causal agent of the leaf fall disease of rubber tree

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 849; no. 1; pp. 357 - 362
• Main Authors: de Lamotte, Frédéric, Duviau, Marie-Pierre, Sanier, Christine, Thai, Robert, Poncet, Joël, Bieysse, Daniel, Breton, Frédéric, Pujade-Renaud, Valérie
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.04.2007; Elsevier Science; Elsevier
• Subjects: Amino Acid Sequence; Analysis; Analytical, structural and metabolic biochemistry; Ascomycota - metabolism; Biological and medical sciences; Cassiicolin; Chromatography; Chromatography, Liquid - methods; Corynespora cassiicola; Electrophoresis - methods; Fundamental and applied biological sciences. Psychology; General pharmacology; Hevea - drug effects; Hevea - microbiology; Hevea brasiliensis; Life Sciences; Mass spectrometry; Medical sciences; Molecular Sequence Data; Molecular Weight; Mycotoxins - chemistry; Mycotoxins - isolation & purification; Mycotoxins - toxicity; Pharmacology. Drug treatments; Plant Leaves - drug effects; Plant Leaves - microbiology; Purification; Reproducibility of Results; Sequence Analysis, Protein; Spectrometry, Mass, Electrospray Ionization - methods; Toxin; Cassiicolin; Corynespora cassiicola; Hevea brasiliensis; Toxin; Purification; Amino acid sequence; Mass spectrometry; Chromatography; Laticiferous plant; Plant leaf; Fungi; Characterization; Dicotyledones; Aminoacid; Angiospermae; Plant origin; Euphorbiaceae; Tree; Spermatophyta; Fungi Imperfecti; Rubber; Thallophyta; TOXIN; AMINO ACID SEQUENCE; CORYNESPORA CASSIICOLA; PURIFICATION; MASS SPECTROMETRY; CASSIICOLIN; CHROMATOGRAPHY; BIOLOGIE DU DEVELOPPEMENT
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2006.10.051

Cassiicolin, a phytotoxin produced by the necrotrophic fungus Corynespora cassiicola, was purified to homogeneity from a rubber tree isolate. The optimized protocol involves reverse phase chromatography followed by size exclusion chromatography, with monitoring of the toxicity on detached rubber tree leaves. Cassiicolin appeared to be a peptide composed of 27 amino acids, glycosylated on the second residue, with a N-terminal pyroglutamic acid and 6 cysteines involved in disulfide bonds. Its molecular mass was estimated to be 2885 Da. No significant sequence homology with other proteins could be found. The availability of pure toxin in sufficient amount is a prerequisite for its structure determination, which is a key step in the understanding of the aggression mechanism.