Proteomic Study of Biocontrol Mechanisms of Trichoderma harzianum ETS 323 in Response to Rhizoctonia solani

• Published in: Journal of agricultural and food chemistry Vol. 56; no. 16; pp. 6914 - 6922
• Main Authors: Tseng, Shih-Chi, Liu, Shu-Ying, Yang, Hsueh-Hui, Lo, Chaur-Tsuen, Peng, Kou-Cheng
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 27.08.2008
• Subjects: amino acid oxidase; beta-glucanase; beta-glucosidase; beta-mannosidase; Bioactive Constituents; biocontrol; Biological and medical sciences; biological control; biological control agents; Cell Wall - metabolism; cell wall-degrading enzymes; cell walls; cellulases; chitinase; endochitinase; enzyme activity; enzymes; Food industries; Fundamental and applied biological sciences. Psychology; fungal proteins; Glycoside Hydrolases - metabolism; Hypocrea lixii; O-glycoside hydrolases; Peptide Hydrolases - metabolism; Pest Control, Biological; Plant Diseases - microbiology; plant pathogenic fungi; proteinases; Proteome; Proteomics; Rhizoctonia - metabolism; Rhizoctonia solani; Thanatephorus cucumeris; Trichoderma - enzymology; Trichoderma - physiology; Trichoderma harzianum; xylanases; Trichoderma harzianum; Proteome; Rhizoctonia solani; biocontrol; cell wall-degrading enzymes; Enzyme; Biological control; Mechanism; Cell wall
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf703626j

To elucidate the entire range of proteins that are secreted by Trichoderma harzianum ETS 323 in its antagonism with Rhizoctonia solani, an in vivo interaction between them was mimicked and not only the secreted cell wall-degrading enzymes (CWDEs) but also all of the proteome were investigated. Seven CWDEs, chitinase, cellulase, xylanase, β-1,3-glucanase, β-1,6-glucanase, mannanase, and protease,were revealed by activity assay, in-gel activity stain, 2-DE, and LC-MS/MS analysis. Extracellular protein extracts from media that contained R. solani exhibited much higher CWDE activities than media that did not contain R. solani. Cellulase and mannanase activity, however, were insignificant. Activity stain also revealed that β-1,3-glucanase, β-1,6-glucanase, and xylanase activity occurred exclusively in media that contained R. solani. Furthermore, 35 of the 43 excised spots on the 2-DE gel were successfully analyzed by LC-MS/MS, and eight proteins were identified. They were two glycoside hydrolases, two proteases, two β-glucosidases, one endochitinase and, interestingly, one amino acid oxidase. Additionally, a possible mechanism was proposed to elucidate how the cell walls of R. solani are systematically enveloped and disintegrated.