Redox metabolism and cell wall modifications as global and local targets respectively, of cyanide induced dormancy release of walnut kernels.

The HCN-induced seed dormancy release necessitates alterations in reactive oxygen species (ROS) metabolism and radicle cell wall loosening. Little is known about the interaction of ROS metabolism with cell wall hydrolytic enzymes during HCN-induced seed dormancy release. Thus dormant walnut (Juglans regia L.) kernels were exposed to HCN (4 h) and studied for redox metabolism and cell wall-modifying enzymes during 10 days of incubation (DI) i.e. before radicle emergence. HCN increased ROS especially in the embryonic axes (EAs) but decreased ROS-generating NADPH oxidase and ROS scavenging superoxide dismutase (SOD) and peroxidase (POX) with no effects on catalase (CAT), ascorbate peroxidase (APX) and cell wall-modifying enzymes activities in short term up to 2 DI. In long term roughly from 4 DI onwards, HCN-exposed EA displayed greater superoxide anions and enhanced activities of POX, APX, NADPH oxidase, cell wall peroxidase (CW-POX), β- 1, 4-D glucanase, mannanase, polygacturonase and xylanase. Meanwhile HCN increased greater expression of POX and mannanase isoforms as revealed by in-gel activity assay. Except for higher activities of CAT, POX and APX, cotyledonary activities of CW-POX, mannanase and polygacturonase and to some extent β- 1, 4-D glucanase remained unaffected by HCN. Thus short term ROS accumulation in HCN-treated EA is due to declined SOD and POX activities. In long term the enhanced activities of both NADPH oxidase: CW-POX couple and cell wall-modifying enzymes in EA bring about wall loosening in preparation for radicle emergence. Evidences for the simultaneous operation of both mechanisms are provided in walnut EAs during dormancy release.