Circadian Clock Contributes to Modulate Salinity Stress-Responsive Antioxidative Mechanisms and Chloroplast Proteome in Spinacia oleracea

Extreme abiotic stresses such as drought, salinity, and temperature reduce crop productivity significantly and pose a serious threat to the area of land used for agriculture. Therefore, there is a pressing need to create crops that can thrive in these circumstances. It has been noted that plants can maintain defense mechanisms during any environmental changes and anticipate diurnal patterns correct to a circadian-based clock. Therefore, the main aim of this study was to investigate the role of circadian core oscillators in response to salinity stress in an important vegetable crop, spinach, and obtain evidence to better understand salinity stress adaptation for crop productivity. Therefore, the current study was carried out to examine the circadian clock-based (morning–evening loop) salinity stress defense mechanism in spinach (Spinacia oleracea), a leafy vegetable crop with significant economic importance and health benefits. In the presence of dawn and dusk, the circadian clock-based defense mechanism was observed using the genotypes “Delhi Green” and “Malav Jyoti.” A photoperiodic rhythm consists of 4-h intervals for 12 h (morning–evening loop) in spinach was demonstrated under the salinity stress treatments (20 mM and 50 mM). The clock-controlled a large fraction of growth parameters such as plant height, biomass, and root-shoot ratio under salinity stress. Conversely, salinity stress resulted in upregulation of antioxidative parameters such as superoxide dismutase, ascorbate peroxidase, catalase, and other stress markers such as thiobarbituric acid reactive substances, proline content, and localizations of H2O2 and O2−1 but was altered and maintained at a certain photoperiodic time interval of the circadian clock. In distinction to results observed from antioxidative measurements performed with an early and late circadian duration of salt-treated plants, 10 am and 2 pm were revealed to be the rhythmic times for controlling salinity stress. Likewise, comprehensive measurements of the photosynthetic system under salinity stress at specific photoperiodic circadian time intervals, including net-photosynthetic rate, transpiration, stomatal conductance, PSII quantum yield, and stomata structure, were made at 10 am and 2 pm. The salinity stress response was down-streamed and the clock also regulated chloroplastic protein expression. Thus, according to our findings, photoperiodic circadian rhythms, particularly the morning–evening loop, enhanced plant survival rates by modulating cellular antioxidant mechanisms and chloroplastic proteins that further helped to reduce the effects of salinity stress.