Immunoengineered Chitosanase-Produced Chitosan Oligomers for Elevating Plant Resistance to Viral Infection

Abstract

Chitooligomers can act as plant biostimulants or biopesticides, but todays chitosan-based agro-biologics often lack sufficient efficacy. This is due to a lack of scalable production processes for structurally well-controlled chitosans combined with a limited understanding of structure-function relationships. Chitosans differ in their degree of polymerization (DP), fraction and pattern of acetylation (FA and PA). While the influence of DP and FA on antimicrobial and phytostimulatory properties is at least partially known, this is not yet the case for PA. PA can be partially controlled by using enzymatic rather than acid hydrolysis for oligomer production. We have used recombinant chitinases and chitosanases to hydrolyse a well-characterised chitosan polymer, and purified oligomers with different DP. We have structurally characterised the products and tested their abilities to protect tobacco from viral disease. Chitinase products were dominated by GlcNAc units at their reducing and non-reducing ends, with GlcN units dominating their centers, and v.v. for chitosanase products. While the chitinase-derived hydrolysates were inactive, the chitosanase-derived oligomers possessed elicitor and priming activities and protected plants from disease, and their activity increased with increasing DP. Clearly, the Bacillus chitosanase used is well-suited to set up a scalable production process for chitosan oligomers with promising agro-biologic properties. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=95 SRC="FIGDIR/small/731087v1_ufig1.gif" ALT="Figure 1"> View larger version (56K): org.highwire.dtl.DTLVardef@b50b59org.highwire.dtl.DTLVardef@1fc35c3org.highwire.dtl.DTLVardef@12db91aorg.highwire.dtl.DTLVardef@d3732f_HPS_FORMAT_FIGEXP M_FIG C_FIG