Early axis growth during seed germination is gravitropic and mediated by ROS and calcium.

In plant establishment, seed germination is characterized by the emergence of a radicle for secured anchorage to the soil and nutrient and water uptake. Early growth of germinating axes appears to be gravisensitive, and the regulation of this process is largely uncharacterized, particularly in case of epigeally germinating species. Our previous work on the germination of Vigna radiata seeds demonstrated the role of apoplastic reactive oxygen species (ROS) in germination-associated axis growth. This study attempts to explore a possibly similar role of ROS in the gravitropic bending of germinating axes. Pharmacological and histological studies correlated the curvature growth of the axis (due to cell elongation in the cortical region of the upper side) with apoplastic superoxide accumulation. The superoxide was produced by diphenylene iodonium chloride (DPI)-insensitive NADH oxidase, which was different from the DPI-sensitive NADPH oxidase active in the apical elongation zone of the radicle. This NADH oxidase was differentially controlled by IAA, and its activation required influx of apoplastic Ca2+. This study shows that the early axis growth in germinating seeds is gravisensitive, which is distinct spatially as well as temporally from the elongation growth of the axis (radicle) and controlled by auxin and cytosolic Ca2+ through NADH oxidase-dependent ROS production.

Superoxide and its metabolism during germination and axis growth of Vigna radiata (L.) Wilczek seeds.

Involvement of reactive oxygen species in regulation of plant growth and development is recently being demonstrated with various results depending on the experimental system and plant species. Role of superoxide and its metabolism in germination and axis growth was investigated in case of Vigna radiata seeds, a non-endospermous leguminous species having epigeal germination, by studying the effect of different reactive oxygen species (ROS) inhibitors, distribution of O2(•)- and H2O2 and ROS enzyme profile in axes. Germination percentage and axis growth were determined under treatment with ROS inhibitors and scavengers. Localization of O2(•)- and H2O2 was done using nitroblue tetrazolium (NBT) and 3,3',5,5'-tetramethyl benzidine dihydrochloride hydrate (TMB), respectively. Apoplastic level of O2(•)- was monitored by spectrophotometric analysis of bathing medium of axes. Profiles of NADPH oxidase and superoxide dismutase (SOD) were studied by in-gel assay. Germination was retarded by treatments affecting ROS level except H2O2 scavengers, while axis growth was retarded by all. Superoxide synthesis inhibitor and scavenger prevented H2O2 accumulation in axes in later phase as revealed from TMB staining. Activity of Cu/Zn SOD1 was initially high and declined thereafter. Superoxide being produced in apoplast possibly by NADPH oxidase activity is further metabolized to (•)OH via H2O2. Germination process depends possibly on (•)OH production in the axes. Post-germinative axis growth requires O2(•)- while the differentiating zone of axis (radicle) requires H2O2 for cell wall stiffening.