9,10-KODA,an α-ketol produced by the tonoplastlocalized 9-lipoxygenase ZmLOX5,plays a signaling role in maize defense against insect herbivory

13-Lipoxygenases(LOXs)initiate the synthesis of jasmonic acid(JA),the best-understood oxylipin hormone in herbivory defense.However,the roles of 9-LOX-derived oxylipins in insect resistance remain unclear.Here,we report a novel anti-herbivory mechanism mediated by a tonoplast-localized 9-LOX,ZmLOX5,and its linolenic acid-derived product,9-hydroxy-10-oxo-12(Z),15(Z)-octadecadienoic acid(9,10-KODA).Transposon-insertional disruption of ZmLOX5 resulted in the loss of resistance to insect herbivory.lox5 knockout mutants displayed greatly reduced wound-induced accumulation of multiple oxylipins and defense metabolites,including benzoxazinoids,abscisic acid(ABA),and JA-isoleucine(JA-Ile).However,exogenous JA-Ile failed to rescue insect defense in lox5 mutants,while applications of 1 mM 9,10-KODA or the JA precursor,12-oxo-phytodienoic acid(12-OPDA),restored wild-type resistance levels.Metabolite profiling revealed that exogenous 9,10-KODA primed the plants for increased production of ABA and 12-OPDA,but not JA-Ile.While none of the 9-oxylipins were able to rescue JA-Ile induction,the lox5 mutant accumulated lower wound-induced levels of Ca^(2+),suggesting this as a potential explanation for lower wound-induced JA.Seedlings pretreated with 9,10-KODA exhibited rapid or more robust woundinduced defense gene expression.In addition,an artificial diet supplemented with 9,10-KODA arrested fall armyworm larvae growth.Finally,analysis of single and double lox5 and lox10 mutants showed that ZmLOX5 also contributed to insect defense by modulating ZmLOX10-mediated green leaf volatile signaling.Collectively,our study uncovered a previously unknown anti-herbivore defense and hormonelike signaling activity for a major 9-oxylipin α-ketol.