Proteomic study of biocontrol mechanisms of Trichoderma harzianum ETS 323 in response to Rhizoctonia solani.
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, beta-1,3-glucanase, beta-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 beta-1,3-glucanase, beta-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 beta-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.
Tseng
Shih-Chi
SC
Institute of Biotechnology, National Dong Hwa University, Hualien, 97401, Taiwan.
Liu
Shu-Ying
SY
Yang
Hsueh-Hui
HH
Lo
Chaur-Tsuen
CT
Peng
Kou-Cheng
KC
D002473:Cell Wall / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D010572:Pest Control, Biological; D010935:Plant Diseases / Q000382:microbiology; D040901:Proteomics; D012232:Rhizoctonia / Q000378:metabolism*; D014242:Trichoderma / Q000201:enzymology / Q000502:physiology*
D006026:Glycoside Hydrolases; D010447:Peptide Hydrolases