Ionomic and Metabolomic Analyses Reveal Different Response Mechanisms to Saline–Alkali Stress Between Suaeda salsa Community and Puccinellia tenuiflora Community

Soil salinization imposes severe stress to plants, inhibits plant growth, and severely limits agricultural productivity and land utilization. The response of a single plant to saline-alkali stress has been well investigated. However, the plant community that usually works as a group to defend against saline–alkali stress was neglected. To determine the functions of plant community, in our current work, Suaeda salsa (S. salsa) community and Puccinellia tenuiflora (P. tenuiflora) community, two communities that are widely distributed in Hulun Buir Grassland in Northeastern China, were selected as research objects. Ionomic and metabolomic were applied to compare the differences between S. salsa community and P. tenuiflora community from the aspects of ion transport and phenolic compound accumulation, respectively. Ionomic studies demonstrated that many macroelements, including potassium (K) and calcium (Ca), were highly accumulated in S. salsa community whereas microelement manganese (Mn) was highly accumulated in P. tenuiflora community. In S. salsa community, transportation of K to aboveground parts of plants helps to maintain high K+ and low Na+ concentrations whereas the accumulation of Ca triggers the salt overly sensitive (SOS)-Na+ system to efflux Na+. In P. tenuiflora community, enrichment of Mn in roots elevates the level of Mn-superoxide dismutase (SOD) and increases the resistance to saline–alkali stress. Metabolomic studies revealed the high levels of C6C1-compounds and C6C3C6-compounds in S. salsa community and also the high levels of C6C3-compounds in P. tenuiflora community. C6C1-compounds function as signaling molecules to defend against stress and may stimulate the accumulation of C6C3C6-compounds. C6C3-compounds contribute to the elimination of free radicals and the maintenance of cell morphology. Collectively, our findings determine the abundance of phenolic compounds and various elements in S. salsa community and P. tenuiflora community in Hulun Buir Grassland and we explored different responses of S. salsa community and P. tenuiflora community to cope with saline–alkali stress. Understanding of plant response strategies from the perspective of community teamwork may provide a feasible and novel way to transform salinization land.

Positive Salt Tolerance Modulation via Vermicompost Regulation of SOS1 Gene Expression and Antioxidant Homeostasis in Viciafaba Plant

Strategic implementation of vermicompost as safe biofertilizer besides defensing saline soils offer dual function solving problems in developing countries. The current study aims to utilize vermicompost (VC) for amelioration of 200mM NaCl in Vicia faba Aspani cultivar and investigate the molecular role of salt overly sensitive pathway (SOS1). The experiment was conducted following a completely randomized design with three replicates. Treatments include 0; 2.5; 5; 10; 15% dried VC intermingled with soil mixture (clay: sand; 1:2) and/or 200 mM NaCl. The results show that salinity stress decreased broad bean fresh and dry weight; and K+/Na+. However, malonedialdehyde and H2O2 contents; increased. Application of 10% VC and salinity stress increases Ca2+ (41% and 50%), K+/Na+ (125% and 89%), Mg2+ (25% and 36%), N (8% and 11%), indole acetic acid (70% and 152%) and proteins (9% and 13%) for root and shoot, respectively, in comparison to salt treated pots. Moreover, all examined enzymatic antioxidants and their substrates increased, except glutathione reductase. A parallel decrease in abscisic acid (75% and 29%) and proline (59% and 58%) was also recorded for roots and leaves, respectively. Interestingly, the highly significant increase in gene expression of SOS1 (45-fold) could drive defense machinery of broad bean to counteract 200 mM NaCl.

Distinct Roles of N-Terminal Fatty Acid Acylation of the Salinity-Sensor Protein SOS3

The Salt-Overly-Sensitive (SOS) pathway controls the net uptake of sodium by roots and the xylematic transfer to shoots in vascular plants. SOS3/CBL4 is a core component of the SOS pathway that senses calcium signaling of salinity stress to activate and recruit the protein kinase SOS2/CIPK24 to the plasma membrane to trigger sodium efflux by the Na/H exchanger SOS1/NHX7. However, despite the well-established function of SOS3 at the plasma membrane, SOS3 displays a nucleo-cytoplasmic distribution whose physiological meaning is not understood. Here, we show that the N-terminal part of SOS3 encodes structural information for dual acylation with myristic and palmitic fatty acids, each of which commands a different location and function of SOS3. N-myristoylation at glycine-2 is essential for plasma membrane association and recruiting SOS2 to activate SOS1, whereas S-acylation at cysteine-3 redirects SOS3 toward the nucleus. Moreover, a poly-lysine track in positions 7–11 that is unique to SOS3 among other Arabidopsis CBLs appears to be essential for the correct positioning of the SOS2-SOS3 complex at the plasma membrane for the activation of SOS1. The nuclear-localized SOS3 protein had limited bearing on the salt tolerance of Arabidopsis. These results are evidence of a novel S-acylation dependent nuclear trafficking mechanism that contrasts with alternative subcellular targeting of other CBLs by S-acylation.

Overexpression of NtSOS2 From Halophyte Plant N. tangutorum Enhances Tolerance to Salt Stress in Arabidopsis

The Salt Overly Sensitive (SOS) signaling pathway is key in responding to salt stress in plants. SOS2, a central factor in this pathway, has been studied in non-halophytes such as Arabidopsis and rice, but has so far not been reported in the halophyte Nitraria tangutorum. In order to better understand how Nitraria tangutorum acquires its tolerance for a high salt environment, here, the NtSOS2 was cloned from Nitraria tangutorum, phylogenetic analyses showed that NtSOS2 is homologous to the SOS2 of Arabidopsis and rice. Gene expression profile analysis showed that NtSOS2 localizes to the cytoplasm and cell membrane and it can be induced by salt stress. Transgenesis experiments showed that exogenous expression of NtSOS2 reduces leaf mortality and improves the germination rate, biomass and root growth of Arabidopsis under salt stress. Also, exogenous expression of NtSOS2 affected the expression of ion transporter-related genes and can rescue the phenotype of sos2-1 under salt stress. All these results revealed that NtSOS2 plays an important role in plant salt stress tolerance. Our findings will be of great significance to further understand the mechanism of salt tolerance and to develop and utilize molecular knowledge gained from halophytes to improve the ecological environment.

Morphological Analysis, Protein Profiling and Expression Analysis of Auxin Homeostasis Genes of Roots of Two Contrasting Cultivars of Rice Provide Inputs on Mechanisms Involved in Rice Adaptation towards Salinity Stress

Plants remodel their root architecture in response to a salinity stress stimulus. This process is regulated by an array of factors including phytohormones, particularly auxin. In the present study, in order to better understand the mechanisms involved in salinity stress adaptation in rice, we compared two contrasting rice cultivars—Luna Suvarna, a salt tolerant, and IR64, a salt sensitive cultivar. Phenotypic investigations suggested that Luna Suvarna in comparison with IR64 presented stress adaptive root traits which correlated with a higher accumulation of auxin in its roots. The expression level investigation of auxin signaling pathway genes revealed an increase in several auxin homeostasis genes transcript levels in Luna Suvarna compared with IR64 under salinity stress. Furthermore, protein profiling showed 18 proteins that were differentially regulated between the roots of two cultivars, and some of them were salinity stress responsive proteins found exclusively in the proteome of Luna Suvarna roots, revealing the critical role of these proteins in imparting salinity stress tolerance. This included proteins related to the salt overly sensitive pathway, root growth, the reactive oxygen species scavenging system, and abscisic acid activation. Taken together, our results highlight that Luna Suvarna involves a combination of morphological and molecular traits of the root system that could prime the plant to better tolerate salinity stress.

Salinity Stress-Mediated Suppression of Expression of Salt Overly Sensitive Signaling Pathway Genes Suggests Negative Regulation by AtbZIP62 Transcription Factor in Arabidopsis thaliana

Salt stress is one of the most serious threats in plants, reducing crop yield and production. The salt overly sensitive (SOS) pathway in plants is a salt-responsive pathway that acts as a janitor of the cell to sweep out Na+ ions. Transcription factors (TFs) are key regulators of expression and/or repression of genes. The basic leucine zipper (bZIP) TF is a large family of TFs regulating various cellular processes in plants. In the current study, we investigated the role of the Arabidopsis thaliana bZIP62 TF in the regulation of SOS signaling pathway by measuring the transcript accumulation of its key genes such as SOS1, 2, and 3, in both wild-type (WT) and atbzip62 knock-out (KO) mutants under salinity stress. We further observed the activation of enzymatic and non-enzymatic antioxidant systems in the wild-type, atbzip62, atcat2 (lacking catalase activity), and atnced3 (lacking 9-cis-epoxycarotenoid dioxygenase involved in the ABA pathway) KO mutants. Our findings revealed that atbzip62 plants exhibited an enhanced salt-sensitive phenotypic response similar to atnced3 and atcat2 compared to WT, 10 days after 150 mM NaCl treatment. Interestingly, the transcriptional levels of SOS1, SOS2, and SOS3 increased significantly over time in the atbzip62 upon NaCl application, while they were downregulated in the wild type. We also measured chlorophyll a and b, pheophytin a and b, total pheophytin, and total carotenoids. We observed that the atbzip62 exhibited an increase in chlorophyll and total carotenoid contents, as well as proline contents, while it exhibited a non-significant increase in catalase activity. Our results suggest that AtbZIP62 negatively regulates the transcriptional events of SOS pathway genes, AtbZIP18 and AtbZIP69 while modulating the antioxidant response to salt tolerance in Arabidopsis.

SOS Yolağından Sorumlu Arabidopsis Mutantlarının Tuz Stresi Altındaki Hassasiyetlerinin Karşılaştırılması

Salinity stress is one of the most important and common abiotic stress factors that cause significant physiological and metabolic changes in plants, negatively affecting plant growth and development, and causing decrease in product quality and quantity. The elucidation of the molecular control mechanisms associated with salt stress tolerance is based on the activation and /or inactivation of various stress-related genes. Salt Overly Sensitive (SOS) tolerance mechanism under salt stress is of great importance in terms of salt tolerance of the plants. Although this mechanism has been studied for many years, the physiological changes that the plants give as a result of mutation of the genes in the pathway under different levels of sodium chloride (NaCl) during development have not been examined comparatively. In this study, we found that the Arabidopsis thaliana sos1-1 mutant plant showed sensitivity to 10 mM NaCl while the sos3-1 and hkt1-1 mutants showed tolerance. The sos1-1, sos3-1 and hkt1-1 mutants showed increasing sensitivity when NaCl was applied beyon 50 mM of concentration. In addition, plants did not show significant sensitivity for 1 day of stress application, while significant effects were observed in plant root length when exposed to salinity for 3 to 4 days. Col-0, hkt1-1 and sos3-1 roots treated with low levels of NaCl for a short term were positively affected in length. In the light of these results, the amount and duration of salt stress is very critical in Arabidopsis thaliana's responses to the stress and determination of molecular tolerance pathways.