Antifungal activity of a novel endochitinase gene (chit36) from Trichoderma harzianum Rifai TM.
A novel 36-kDa endochitinase named chit36 has been isolated and characterized from Trichoderma harzianum Rifai TM. Partial amino acid sequences from the purified protein were used to clone the fungal cDNA, based on polymerase chain reaction with degenerate primers. The complete open reading frame encodes a 344-amino acid protein which shows 84% similarity to a putative chitinase from Streptomyces coelicolor. Chit36 was overexpressed under the pki1 constitutive promoter from Trichoderma reesei via biolistic transformation of T. harzianum TM. Stable transformants showed expression and endochitinase activity of chit36 in glucose-rich medium. Culture filtrates containing secreted CHIT36 as the sole chitinolytic enzyme completely inhibited the germination of Botrytis cinerea conidia. Growth of Fusarium oxysporum f. sp. melonis and Sclerotium rolfsii were significantly inhibited on agar plates on which the Trichoderma transformants had previously been grown.
Viterbo
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Otto-Warburg-Center for Agricultural Biotechnology, The Hebrew University of Jerusalem, Faculty of Agriculture, P.O. Box 12, 76100, Rehovot, Israel. viterbo@agri.huji.il
Haran
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S
Friesem
D
D
Ramot
O
O
Chet
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I
D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000302:isolation & purification / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D002688:Chitinases / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D005656:Fungal Proteins; D005670:Fusarium / Q000187:drug effects; D008969:Molecular Sequence Data; D010572:Pest Control, Biological; D010935:Plant Diseases / Q000382:microbiology; D017386:Sequence Homology, Amino Acid; D013172:Spores, Fungal / Q000187:drug effects; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics
D000935:Antifungal Agents; D005656:Fungal Proteins; C434315:Chit36 protein, Hypocrea lixii; D002688:Chitinases