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Here, candidate elicitor molecules, which induce resistance in B. napus to L. maculans, were identified in the cultivation medium. The elicitation activity was indicated by increased transcription of pathogenesis\u2010related gene 1 (PR1) and enhanced resistance of B. napus plants to the invasion of L. maculans. The elicitation activity was significantly lowered when the cultivation medium was heated to 80\u00b0C. Active components were further characterized by specific cleavage with the proteolytic enzymes trypsin and proteinase K and with glycosidases \u03b1\u2010amylase and \u03b2\u2010glucanase. The elicitor activity was eliminated by proteolytic digestion while glycosidases had no effect. The filtered medium was fractionated by either ion\u2010exchange chromatography or isoelectric focusing. Mass spectrometry analysis of the most active fractions obtained by both separation procedures revealed predominantly enzymes that can be involved in the degradation of plant cell wall polysaccharides. This is the first study searching for L. maculans\u2010specific secreted elicitors with a potential to be used as defense\u2010activating agents in the protection of B. napus against L. maculans in agriculture. \u00a9 2016 American Institute of Chemical Engineers <jats:italic>Biotechnol. 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