Identification and Characterization of Brassica oleracea var. capitata Chlorophyll a/b Binding Protein Bound to Trichoderma harzianum ETS 323 L-Amino Acid Oxidase

• Main Authors: Chong-Fu Liao, 廖崇富
• Other Authors: Kou-Cheng Peng
• Format: Others
• Language: zh-TW
• Published: 2019
• Online Access: http://ndltd.ncl.edu.tw/handle/x8az9s

碩士 === 國立東華大學 === 生命科學系 === 107 === T. harzianum ETS 323 is a well-known environmental biological control agent. Its mechanisms mainly contributes to production of extracellular proteins, secondary metabolites and resistance induction. Among these, L-amino acid oxidase and chrysophanol have been proven antibiotic activities and resistance induction of Brassica oleracea. The further interest would be to investigating binding proteins of the B. oleracea to ThLAAO. By using ThLAAO affiliated affinity chromatography, a 28.3 kDa ThLAAO binding membrane protein of B. oleracea was isolated and identified as Chlorophyll a/b binding protein. In the present of the ThLAAO, later infected with Botrytis cinerea, cab expression of B. oleracea leaf increase 2.5 folds, the cab of B. oleracea leaf that treated with His6-CAB later infection B. cinerea decreasing 7 folds. A mole ratio 1:1 of ThLAAO and His6-CAB mixture was applied to the B. oleracea leaf later infected with B. cinerea showing that His6-CAB can antagonize the ThLAAO resistance induction capability corresponding to decreasing cab expression 10 folds. The percentage of infection area of B. oleracea leaf were ThLAAO is 0 to 15 ％ His6-CAB is 50 to 100 ％ and ThLAAO mixture with His6-CAB is 70 to 100 ％accordingly decrease cab expression, respectively. ThLAAO-Cy5.5 treated B. oleracea root showing that ThLAAO-Cy5.5 enter root and transport to the leaves where have been proven in chloroplast in other investigation. This study showed that B. oleracea chlorophyll a/b binding protein is the binding protein of T. harzianum ETS 323 L-amino acid oxidase that induces pathogen resistance.