Programmed cell death (PCD) is a process intended for the maintenance of cellular homeostasis by eliminating old, damaged, or unwanted cells. In plants, PCD takes place during developmental processes and in response to biotic and abiotic stresses. In contrast to the field of animal studies, PCD is not well understood in plants. Calcium (Ca²⁺) is a universal cell signaling entity and regulates numerous physiological activities across all the kingdoms of life. The cytosolic increase in Ca²⁺ is a prerequisite for the induction of PCD in plants. Although over the past years, we have witnessed significant progress in understanding the role of Ca²⁺ in the regulation of PCD, it is still unclear how the upstream stress perception leads to the Ca²⁺ elevation and how the signal is further propagated to result in the onset of PCD. In this review article, we discuss recent advancements in the field, and compare the role of Ca²⁺ signaling in PCD in biotic and abiotic stresses. Moreover, we discuss the upstream and downstream components of Ca²⁺ signaling and its crosstalk with other signaling pathways in PCD. The review is expected to provide new insights into the role of Ca²⁺ signaling in PCD and to identify gaps for future research efforts.

Background: Salinity is one of the most lethal abiotic stresses which affect multiple aspects of plant physiology. Natural variations in plant germplasm are a great resource that could be exploited for improvement in salt tolerance. Eruca sativa (E. sativa) exhibits tolerance to abiotic stresses. However, thorough evaluation of its salt stress tolerance and screening for traits that could be reliably applied for salt tolerance needs to be studied. The current study was designed to characterize 25 E. sativa accessions, originating from diverse geographical regions of Pakistan, for the salt stress tolerance. Methods: Salt stress (150 mM NaCl) was applied for 2 weeks to the plants at four leaf stage in hydroponics. Data of the following morpho-physiological traits were collected from control and treated plants of all the accessions: root length (RL), shoot length (SL), plant height (PH), leaf number (LN), leaf area (LA), fresh weight (FW), dry weight (DW), chlorophyl content (SPAD), electrolyte leakage (EL), relative water content (RWC), gas exchange parameters and mineral ion content. Salt tolerance was determined based on membership function value (MFV) of the tested traits. Results: Compared with control, the salt-stressed group had significantly reduced mean SL, RL, PH, LN, LA, FW, DW and SPAD. NaCl treatment triggered a slight increase in EL in few accessions. Mean RWC of control and treated groups were not significantly different although few accessions exhibited variation in this trait. Salt stress caused a significant reduction in photosynthesis rate (PR), transpiration rate (TR) and stomatal conductance (SC) but intercellular CO₂ (Ci) was not significantly different between control and treated groups. Compared with control, the salt-stressed plants accumulated significantly higher Na⁺, K⁺ and Ca²⁺ while significantly lower Mg²⁺. K⁺/Na⁺ ratio was significantly decreased in salt-stressed plants compared with control. Importantly, significant inter-accession variations were found for all the tested traits. The principal component analysis identified SL, RL, PH, LN, LA, FW, DW and PR as the most significant traits for resolving inter-accession variability. Based on MFV of the tested traits, accessions were categorized into five standard groups. Among 25 accessions, one accession was ranked as highly tolerant, four as tolerant while 15 accessions were ranked as moderately tolerant. Of the remaining five accessions, four were ranked as sensitive while one accession as highly sensitive. Conclusion: E. sativa accessions were found to exhibit significant genetic diversity in all the tested traits. A few most significant traits for dissecting the genetic variability were identified that could be used for future large-scale germplasm screening in E. sativa. Salt tolerant accessions could be a good resource for future breeding programs aiming to improve salt stress tolerance.

Drought stress causes lower crop production globally. Plants have acquired many adaptations to overcome drought stress. Mungbean (Vigna radiata (L.) R.Wilczek) is a legume crop widely cultivated in South, East and Southeast Asia. It is grown in high-temperature areas where drought is the main cause of reduced plant growth and productivity. Plants cope with drought stress by activating different signalling mechanisms. The sucrose non-fermenting-1-related protein kinase 2 family (SnRK2s) is known to play vital roles in osmotic stress and in abscisic acid (ABA) signalling pathways by phosphorylating downstream targets. The genes encoding SnRK2s in mungbean and their detailed characterisation remain unexplored. We have conducted extensive genome-wide analysis for gene prediction, in silico gene analysis, evolutionary analysis and gene-expression profiling under drought-stress conditions by quantitative real-time PCR. Through genome-wide analysis, eight SnRK2 genes were predicted in the mungbean genome and were assigned the names VrSnRK2.1–VrSnRK2.8, according to their order on the chromosomes. The VrSnRK2 genes identified were classified into three clusters based on their phylogenetic relationship with those of Arabidopsis thaliana. Drought stress was imposed on 11-day-old mungbean plants by completely withholding water for 3 days. According to real-time qPCR data, the expression of most of the VrSnRK2 genes was induced by drought stress, indicating their role in the drought-stress response. One of the genes, namely SnRK2.6c, showed highest expression level (12-fold) under drought stress, possibly indicating a critical role under water-deficit conditions. These data provide important information about the VrSnRK2 gene family in mungbean. The results will help in future functional characterisation of VrSnRK2 genes.

The cyclic nucleotide cyclic guanosine monophosphate (cGMP) is a powerful cell signaling molecule involved in biotic and abiotic stress perception and signal transduction. In the model plant Arabidopsis thaliana, salt and osmotic stress rapidly induce increase in cGMP which plays role by modulating the activity of monovalent cation transporters, possibly by direct binding to these proteins and by altering the expression of many abiotic stress responsive genes. In a recent study, a membrane permeable analogue of cGMP (8-bromo-cGMP) was found to have a promotive effect on soluble sugar, flavonoids and lignin content, and membrane integrity in Solanum lycopersicum seedlings under salt stress. However, it remains to be elucidated how salt stress affects the endogenous cGMP level in S. lycopersicum and if Br-cGMP-induced improvement in salt tolerance in S. lycopersicum involves altered cation fluxes. The current study was conducted to answer these questions. A rapid increase (within 30 s) in endogenous cGMP level was determined in S. lycopersicum roots after treatment with 100 mM NaCl. Addition of membrane permeable Br-cGMP in growth medium remarkably ameliorated the inhibitory effects of NaCl on seedlings’ growth parameters, chlorophyll content and net photosynthesis rate. In salt stressed plants, Br-cGMP significantly decreased Na⁺ content by reducing its influx and increasing efflux while it improved plants K⁺ content by reducing its efflux and enhancing influx. Furthermore, supplementation with Br-cGMP improved plant’s proline content and total antioxidant capacity, resulting in markedly decreased electrolyte leakage under salt stress. Br-cGMP increased the expression of Na⁺/H⁺ antiporter genes in roots and shoots of S. lycopersicum growing under salt stress, potentially enhancing plant’s ability to sequester Na⁺ into the vacuole. The findings of this study provide insights into the mechanism of cGMP-induced salt stress tolerance in S. lycopersicum.