Elucidating ROS signaling networks and physiological changes involved in nanoscale zero valent iron primed rice seed germination sensu stricto

Reactive oxygen species (ROS) play pivotal roles during seed dormancy and germination. Metabolically active cells of seeds generate ROS and successful germination is governed by internal ROS contents, maintained within an optimum "oxidative window" by several ROS scavengers. Although ROS was previously considered hazardous, optimum ROS generation in seeds can mediate early seed germination by acting as messengers for cell signaling involved in endosperm weakening, stored food mobilization, etc. Recent reports suggest that nano-priming can expedite seed germination rates and enhance seed quality and crop performances. However, nanoparticle-driven signal cascades involved during seed germination are still unknown. The present study is aimed to explore molecular mechanisms for promoting germination in nanoprimed seeds and to investigate the plausible role of nanoparticle-mediated ROS generation in this process. Here rice seeds were primed with 20 mg L-1 nanoscale zero valent iron (nZVI) for 72 h and several biochemical and physiological changes were monitored at different time points (5, 10, 20, 40, 60, and 80 h). To gain insight into roles of ROS in germination rate enhancement, intercellular ROS inhibitor, diphenyleneiodonium (DPI) was taken as another priming agent. Seed priming with DPI impaired seed germination percentage, hydrolytic enzyme activities due to ROS imbalance. On the contrary, seeds primed with both DPI and nZVI could recover from deleterious consequences of DPI treatment. Although DPI impaired intercellular ROS generation, nZVI can generate ROS independently which was confirmed from ROS localization assay. In both nZVI and the DPI and nZVI co-primed sets, significant upregulation in genes like OsGA3Ox2, OsGAMYB were observed which are responsible for regulating the activity of several hydrolases and mediates efficient mobilization of storage food reserves of seeds. Thus, nZVI priming has potential to regulate intracellular ROS levels and orchestrate all the metabolic activities which eventually upregulates seed germination rate and seed vigour.