title: Separation and identification of candidate protein elicitors from the cultivation medium of Leptosphaeria maculans inducing resistance in Brassica napus.
abstract: The Dothideomycete Leptosphaeria maculans, a worldwide fungal pathogen of oilseed rape (Brassica napus), secretes a broad spectrum of molecules into the cultivation medium during growth in vitro. 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-related 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°C. Active components were further characterized by specific cleavage with the proteolytic enzymes trypsin and proteinase K and with glycosidases α-amylase and β-glucanase. The elicitor activity was eliminated by proteolytic digestion while glycosidases had no effect. The filtered medium was fractionated by either ion-exchange 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-specific secreted elicitors with a potential to be used as defense-activating agents in the protection of B. napus against L. maculans in agriculture. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:918-928, 2016.
authors.affiliation: Dept. of Biochemistry and Microbiology, University of Chemistry and Technology Prague, Technická 3, 166 28 Prague 6, Czech Republic.|Laboratory of Pathological Plant Physiology, Inst. of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojová 263, 165 02 Prague 6, Czech Republic.
authors.forename: Miroslava
authors.initials: M
authors.lastname: Nováková
authors.affiliation: Dept. of Biochemistry and Microbiology, University of Chemistry and Technology Prague, Technická 3, 166 28 Prague 6, Czech Republic.
authors.forename: Phuong Dinh
authors.initials: PD
authors.lastname: Kim
authors.affiliation: Laboratory of Pathological Plant Physiology, Inst. of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojová 263, 165 02 Prague 6, Czech Republic.
authors.forename: Vladimír
authors.initials: V
authors.lastname: Šašek
authors.affiliation: Laboratory of Pathological Plant Physiology, Inst. of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojová 263, 165 02 Prague 6, Czech Republic.
authors.forename: Lenka
authors.initials: L
authors.lastname: Burketová
authors.affiliation: Dept. of Biochemistry and Microbiology, University of Chemistry and Technology Prague, Technická 3, 166 28 Prague 6, Czech Republic.|Laboratory of Pathological Plant Physiology, Inst. of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojová 263, 165 02 Prague 6, Czech Republic.
authors.forename: Barbora
authors.initials: B
authors.lastname: Jindřichová
authors.affiliation: Dept. of Biochemistry and Microbiology, University of Chemistry and Technology Prague, Technická 3, 166 28 Prague 6, Czech Republic.
authors.forename: Jiří
authors.initials: J
authors.lastname: Šantrůček
authors.affiliation: Dept. of Biochemistry and Microbiology, University of Chemistry and Technology Prague, Technická 3, 166 28 Prague 6, Czech Republic.
authors.forename: Olga
authors.initials: O
authors.lastname: Valentová
mesh_terms: D000935:Antifungal Agents / Q000032:analysis / Q000378:metabolism / Q000494:pharmacology*; D001203:Ascomycota / Q000187:drug effects* / Q000378:metabolism; D029688:Brassica napus / Q000737:chemistry* / Q000378:metabolism / Q000382:microbiology; D002852:Chromatography, Ion Exchange; D003470:Culture Media / Q000737:chemistry*; D005656:Fungal Proteins / Q000032:analysis / Q000378:metabolism*; D013058:Mass Spectrometry
chemical_list: D000935:Antifungal Agents; D003470:Culture Media; D005656:Fungal Proteins
keywords: Brassica napus; Leptosphaeria maculans; elicitor; induced resistance; plant cell-wall-degrading enzymes