Osmotic stresses, such as drought and high salinity, adversely affect plant growth and productivity. The phytohormone abscisic acid (ABA) accumulates in response to osmotic stress and enhances stress tolerance in plants by triggering multiple physiological responses through ABA signaling. Subclass III SNF1–related protein kinases 2 (SnRK2s) are key regulators of ABA signaling. Although SnRK2s have long been considered to be self-activated by autophosphorylation after release from PP2C-mediated inhibition, they were recently revealed to be activated by two independent subfamilies of group B Raf-like kinases, B2-RAFs and B3-RAFs, under osmotic stress conditions. However, the relationship between SnRK2 phosphorylation by these RAFs and SnRK2 autophosphorylation and the individual physiological roles of each RAF subfamily remain unknown. In this study, we indicated that B2-RAFs are constantly active and activate SnRK2s when released from PP2C-mediated inhibition by ABA-binding ABA receptors, whereas B3-RAFs are activated only under stress conditions in an ABA-independent manner and enhance SnRK2 activity. Autophosphorylation of subclass III SnRK2s is not sufficient for ABA responses, and B2-RAFs are needed to activate SnRK2s in an ABA-dependent manner. Using plants grown in soil, we found that B2-RAFs regulate subclass III SnRK2s at the early stage of drought stress, whereas B3-RAFs regulate SnRK2s at the later stage. Thus, B2-RAFs are essential kinases for the activation of subclass III SnRK2s in response to ABA under mild osmotic stress conditions, and B3-RAFs function as enhancers of SnRK2 activity under severe stress conditions.

Blossom-end rot (BER) in paprika (Capsicum annuum L.), as in tomato fruits, is a physiological disorder caused by calcium (Ca) deficiency in fruits. While studies have revealed that excessive nitrogen and potassium fertilization and high temperatures affect Ca transport to fruits and induce BER, few studies have investigated the effects of other elemental concentrations in paprika fruit and their association with BER occurrence. This study aimed to investigate the relationship between the changes in the elemental composition and incidence of BER in paprika fruits grown at different potassium nitrate (KNO 3 ) concentrations and different temperature conditions. Paprika plants were grown in rockwool blocks in a greenhouse at 25°C and 30°C for approximately four months and irrigated with liquid fertilizer and three different concentrations (0, 27.9, and 74.4 mM) of KNO 3 were added. Subsequently, the total fresh weights of ripe paprika fruits and the incidence of BER were measured, and the elemental compositions in the pericarp of the first and last sampled fruits were analyzed. The total fresh weights significantly decreased and the incidence of BER increased with increasing KNO 3 concentrations and temperature. The profiles of 11 mineral elements in the pericarps of paprika fruits revealed a significant positive or negative correlation between not only the concentration of Ca, but also that of several other elements including boron (which crosslinks pectin in the cell wall like Ca) and the incidence of BER. These results suggest that Ca deficiency may not be the only cause of BER occurrence, and that several elements may also be involved. The insights from this study will contribute to help predict the incidence of BER and stabilize crop production by improving fertilizer application and environmental control.