Long-chain chitooligosaccharides (COS) with degree of polymerization (DP) more than 4 are known to have potential biological activities. A hyper-transglycosylating mutant of an endo-chitinase from Serratia proteamaculans (SpChiD-Y28A) was used to synthesize COS with DP6 and DP7 using COS DP5 as substrate. Purified COS with DP5-7 were tested to elicit the defense response in rice seedlings. Among the COS used, DP7 strongly induced oxidative burst response as well as peroxidase, and phenylalanine ammonia lyase activites. A few selected marker genes in salicylic acid (SA)- and jasmonic acid-dependent pathways were evaluated by real-time PCR. The expression levels of pathogenesis-related (PR) genes PR1a and PR10 and defense response genes (chitinase1, peroxidase and β -1,3-glucanase) were up regulated upon COS treatment in rice seedlings. The DP7 induced Phenylalanine ammonia lyase and Isochorismate synthase 1 genes, with concomitant increase of Mitogen-activated protein kinase 6 and WRKY45 transcription factor genes indicated the possible role of phosphorylation in the transmission of a signal to induce SA-mediated defense response in rice.
Keywords: Chitinase; Chitooligosaccharides; Defense mechanism; Elicitor activity; Transglycosylation.
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