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One of the strategies that plants adopt to cope with an unfavorable environment is to sacrifice their growth for tolerance. Although moderate salt stress can induce root growth inhibition, the molecular mechanisms regulating this process have yet to be elucidated. Here, we found that overexpression of a zinc finger-homeodomain family transcription factor, HOMEOBOX PROTEIN 24 (HB24), led to longer primary roots than in the wild-type in the presence of 125 mM NaCl, whereas this phenotype was reversed for the hb24 loss-of-function mutant, indicating a negative impact of HB24 on salt-induced root growth inhibition. We then found that salt stress triggered the degradation of HB24 via the ubiquitin-proteasome pathway, as mediated by a plant U-box type E3 ubiquitin ligase 30 (PUB30) that directly targets HB24. We verified that HB24 is able to directly bind to the promoters of Sugars Will Eventually be Exported Transporter 11/12 (SWEET11/12) to regulate their expression in roots. Through genetic and biochemical assays, we further demonstrated that the HB24-SWEET11 module plays a negative role in salt-induced root growth inhibition. Therefore, we propose that under salt stress, PUB30 mediates HB24's degradation, thereby downregulating the expression of SWEET11, resulting in reduced sucrose supply and root growth inhibition., (© 2022 The Authors New Phytologist © 2022 New Phytologist Foundation.)

Oil palm is the most productive oil producing plant. Salt stress leads to growth damage and a decrease in yield of oil palm. However, the physiological responses of oil palm to salt stress and their underlying mechanisms are not clear. RNA-Seq was conducted on control and leaf samples from young palms challenged under three levels of salts (100, 250, and 500 mM NaCl) for 14 days. All three levels of salt stress activated EgSPCH expression and increased stomatal density of oil palm. Around 41% of differential expressed genes (DEGs) were putative EgSPCH binding target and were involved in multiple bioprocesses related to salt response. Overexpression of EgSPCH in Arabidopsis increased the stomatal production and lowered the salt tolerance. These data indicate that, in oil palm, salt activates EgSPCH to generate more stomata in response to salt stress, which differs from herbaceous plants. Our results might mirror the difference of salt-induced stomatal development between ligneous and herbaceous crops.

Arabidopsis thaliana SYNAPTOTAGMIN 1 (AtSYT1) was shown to be involved in responses to different environmental and biotic stresses. We investigated gas exchange and chlorophyll a fluorescence in Arabidopsis wild-type (WT, ecotype Col-0) and atsyt1 mutant plants irrigated for 48 h with 150 mM NaCl. We found that salt stress significantly decreases net photosynthetic assimilation, effective photochemical quantum yield of photosystem II (Φ PSII ), stomatal conductance and transpiration rate in both genotypes. Salt stress has a more severe impact on atsyt1 plants with increasing effect at higher illumination. Dark respiration, photochemical quenching (qP), non-photochemical quenching and Φ PSII measured at 750 µmol m -2 s -1 photosynthetic photon flux density were significantly affected by salt in both genotypes. However, differences between mutant and WT plants were recorded only for qP and Φ PSII . Decreased photosynthetic efficiency in atsyt1 under salt stress was accompanied by reduced chlorophyll and carotenoid and increased flavonol content in atsyt1 leaves. No differences in the abundance of key proteins participating in photosynthesis (except PsaC and PsbQ) and chlorophyll biosynthesis were found regardless of genotype or salt treatment. Microscopic analysis showed that irrigating plants with salt caused a partial closure of the stomata, and this effect was more pronounced in the mutant than in WT plants. The localization pattern of AtSYT1 was also altered by salt stress.

High soil salinity negatively affects plant growth and development, leading to a severe decrease in crop production worldwide. Here, we report that a secreted peptide, PAMP-INDUCED SECRETED PEPTIDE 3 (PIP3), plays an essential role in plant salt tolerance through RECEPTOR-LIKE KINASE 7 (RLK7) in Arabidopsis (Arabidopsis thaliana). The gene encoding the PIP3 precursor, prePIP3, was significantly induced by salt stress. Plants overexpressing prePIP3 exhibited enhanced salt tolerance, whereas a prePIP3 knockout mutant had a salt-sensitive phenotype. PIP3 physically interacted with RLK7, a leucine-rich repeat RLK, and salt stress enhanced PIP3-RLK7 complex formation. Functional analyses revealed that PIP3-mediated salt tolerance is dependent on RLK7. Exogenous application of synthetic PIP3 peptide activated RLK7, and salt treatment significantly induced RLK7 phosphorylation in a PIP3-dependent manner. Notably, MITOGEN-ACTIVATED PROTEIN KINASE3 (MPK3) and MPK6 were downstream of the PIP3-RLK7 module in salt response signaling. Activation of MPK3/6 was attenuated in pip3 or rlk7 mutants under saline conditions. Therefore, MPK3/6 might amplify salt stress response signaling in plants for salt tolerance. Collectively, our work characterized a novel ligand-receptor signaling cascade that modulates plant salt tolerance in Arabidopsis. This study contributes to our understanding of how plants respond to salt stress., (© American Society of Plant Biologists 2021. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Various environmental stresses can induce production of reactive oxygen species (ROS) to turn on signaling for proper responses to those stresses. Plasma membrane (PM)-localized respiratory burst oxidase homologs (RBOHs), in particular RBOHD, produce ROS via the post-translational activation upon abiotic and biotic stresses. Although the mechanisms of RBOHD activation upon biotic stress have been elucidated in detail, it remains elusive how salinity stress activates RBOHD. Here, we present evidence that trafficking of PM-localized RBOHD to endosomes and then its recycling back to the PM is critical for ROS accumulation upon salinity stress. ateca4 plants that were defective in recycling of proteins from endosomes to the PM and clc2-1 and chc2-1 plants that were defective in endocytosis showed a defect in salinity stress-induced ROS production. In addition, ateca4 plants showed a defect in transient accumulation of GFP:RBOHD to the PM at the early stage of salinity stress. By contrast, ateca4 plants showed no defect in the increase in the ROS level and accumulation of RBOHD to the PM upon flg22 treatment as wild-type plants. Based on these observations, we propose that factors involved in the trafficking machinery such as AtECA4 and clathrin are important players in salt stress-induced, but not flg22-induced, ROS accumulation., (© 2021 Society for Experimental Biology and John Wiley & Sons Ltd.)

F-box protein genes have been shown to play vital roles in plant development and stress respones. In Arabidopsis, there are more than 600 F-box proteins, and most of their functions are unclear. The present study shows that the F-box (SKP1-Cullin/CDC53-F-box) gene At5g15710 (Salt and Drought Responsiveness, SDR) is involved in abiotic stress responses in Arabidopsis. SDR is expressed in all tissues of Arabidopsis and is upregulated by salt and heat stresses and ABA treatment but downregulated by drought stress. Subcellular localization analysis shows that the SDR protein colocalizes with the nucleus. 35S:AntiSDR plants are hypersensitive to salt stress, but 35S:SDR plants display a salt-tolerant phenotype. Furthermore, 35S:SDR plants are hypersensitive to drought stress, while 35S:AntiSDR plants are significantly more drought tolerant. Overall, our results suggest that SDR is involved in salt and drought stress responses in Arabidopsis., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Phytochromobilin (PΦB) participates in the regulation of plant growth and development as an important synthetase of photoreceptor phytochromes (phy). In addition, Arabidopsis long hypocotyl 2 (HY2) appropriately works as a key PΦB synthetase. However, whether HY2 takes part in the plant stress response signal network remains unknown. Here, we described the function of HY2 in NaCl signaling. The hy2 mutant was NaCl-insensitive, whereas HY2-overexpressing lines showed NaCl-hypersensitive phenotypes during seed germination. The exogenous NaCl induced the transcription and the protein level of HY2 , which positively mediated the expression of downstream stress-related genes of RD29A , RD29B , and DREB2A . Further quantitative proteomics showed the patterns of 7391 proteins under salt stress. HY2 was then found to specifically mediate 215 differentially regulated proteins (DRPs), which, according to GO enrichment analysis, were mainly involved in ion homeostasis, flavonoid biosynthetic and metabolic pathways, hormone response (SA, JA, ABA, ethylene), the reactive oxygen species (ROS) metabolic pathway, photosynthesis, and detoxification pathways to respond to salt stress. More importantly, ANNAT1-ANNAT2-ANNAT3-ANNAT4 and GSTU19-GSTF10-RPL5A-RPL5B-AT2G32060, two protein interaction networks specifically regulated by HY2, jointly participated in the salt stress response. These results direct the pathway of HY2 participating in salt stress, and provide new insights for the plant to resist salt stress.

Protein phosphorylation, especially serine/threonine and tyrosine phosphorylation, plays significant roles in signalling during plant growth and development as well as plant responses to biotic or abiotic stresses. Dual-specificity protein tyrosine phosphatases dephosphorylate components of these signalling pathways. Here, we report that an atypical dual-specificity protein tyrosine phosphatase, AtPFA-DSP3 (DSP3), negatively affects the response of plants to high-salt conditions. A DSP3 loss-of-function mutant showed reduced sensitivity to salt treatment. DSP3 was primarily localized in nuclei and was degraded during salt treatment. Compared to wild type, the level of ROS was lower in the dsp3 mutant and higher in plants ectopically expressing DSP3, indicating that higher DSP3 level was associated with increased ROS production. DSP3 interacted with and dephosphorylated MPK3 and MPK6. Genetic analyses of a dsp3mpk3 double mutant revealed that DSP3's effect on salt stress depends on MPK3. Moreover, the phosphatase activity of DSP3 was required for its role in salt signalling. These results indicate that DSP3 is a negative regulator of salt responses in Arabidopsis by directly modulating the accumulation of phosphorylated MPK3 and MPK6., (© 2021 John Wiley & Sons Ltd.)

The plasma membrane NADPH Oxidase-derived ROS as signaling molecules play crucial roles in salt stress response. As the motor organelle of cells, mitochondria are also important for salt tolerance. However, the possible interaction between NADPH Oxidase-derived ROS and mitochondria is not well studied. Here, a transgenic Arabidopsis expressing mitochondrial matrix-targeted pH-sensitive indicator cpYFP was used to monitor the pH dynamics in root cells under salt stress. A significant alkalization in mitochondria was observed when the root was exposed to NaCl or KCl, but not osmotic stress such as isotonic mannitol. Interestingly, when pretreated with the NADPH Oxidase inhibitor DPI, the mitochondrial alkalization in root cells was largely abolished. Genetic evidence further showed that salt-induced mitochondrial alkalization was significantly reduced in the loss of function mutant atrbohF . Pretreatment with endocytosis-related inhibitor PAO or TyrA23, which inhibited the ROS accumulation under salt treatment, almost abolished this effect. Furthermore, [Ca 2+ ] cyt increase might also play important roles by affecting ROS generation to mediate salt-induced mitochondrial alkalization as indicated by treatment with plasma membrane Ca 2+ channel inhibitor LaCl 3 and mitochondrial Ca 2+ uniporter inhibitor Ruthenium Red. Together, these results suggest that the plasma membrane NADPH Oxidase-derived ROS promote the mitochondrial alkalization under salt treatment, providing a possible link between different cellular compartments under salt stress.

Key Message: Overexpression of Bacillus amyloliquefaciens SN13-responsive OsNAM gene in Arabidopsis reveals its important role in beneficial plant and plant growth promoting rhizobacteria interaction by conferring stress tolerance and phytohormone modulation. Salinity is one of the major constraints that affect crop development and yield. Plants respond and adapt to salt stress via complex mechanisms that involve morpho-physiological, biochemical, and molecular changes. The expression of numerous genes is known to alter during various abiotic stresses and impart stress tolerance. Recently, some known rhizospheric microbes have also been used to mitigate the effects of abiotic stresses; however, the molecular basis of such interactions remains elusive. Therefore, the present investigation was aimed to elucidate the plant growth-promoting rhizobacteria (PGPR; Bacillus amyloliquefaciens-SN13) -induced crosstalk among salinity and phytohormones in OsNAM-overexpressed Arabidopsis plants. Transgenic plants showed increased germination percentage compared to wild-type (WT) seeds under 100 mM of NaCl. Phenotypic data showed increased root length, rosette diameter, leaf size, and biomass in transgenics than WT plants. Transgenic plants can also better maintain membrane integrity and osmolyte concentration under salinity as compared to WT. Further, gene expression analysis of AP2/ERF, GST, ERD4, and ARF2 genes showed differential expression and their positive modulation in transgenic Arabidopsis exposed to salt stress in the presence of SN13 as compared to uninoculated WT. Modulation in IAA, ABA, and GA content in inoculated plants showed the more pronounced positive effects of SN13 on transgenic plants that supported our findings on Arabidopsis-SN13 interaction. Overall, the study concludes that SN13 positively modulated expression of stress-responsive genes under salinity and alter phytohormones levels in OsNAM-overexpressed plants suggesting its extensive role in cross-talk among salinity and phytohormones in response to PGPR.

In plants, both hyperosmolality and salt stress induce cytosolic calcium increases within seconds, referred to as the hyperosmolality-induced [Ca 2+ ] cyt increases, OICI cyt , and salt stress-induced [Ca 2+ ] cyt increases, SICI cyt . Previous studies have shown that Arabidopsis reduced hyperosmolality-induced [Ca 2+ ] i increase 1 (OSCA1.1) encodes a hyperosmolality-gated calcium-permeable channel that mediates OICI cyt in guard cells and root cells. Multiple OSCA members exist in plants; for example, Oryza sativa has 11 OsOSCAs genes, indicating that OSCAs have diverse biological functions. Here, except for OsOSCA4.1, ten full-length OsOSCAs were separately subcloned, in which OsOSCA1.4 was exclusively localised to the plasma membrane and other nine OsOSCAs-eYFP co-localised with an endoplasmic reticulum marker in Arabidopsis mesophyll protoplasts. OsOSCA1.4 was further identified as a calcium-permeable ion channel that activates an inward current after receiving an osmotic signal exerted by hyperosmolality or salt stress, and mediates OICI cyt and SICI cyt in human embryonic kidney 293 (HEK293) cells. Moreover, overexpression of OsOSCA1.4 in Arabidopsis osca1 mutant complemented osmotic Ca 2+ signalling, root growth, and stomatal movement in response to hyperosmolality and salt stress. These results will facilitate further study of OsOSCA-mediated calcium signalling and its distinct roles in rice growth and development., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Main Conclusion: GhBASS5 is a member of the bile acid sodium symporter (BASS) gene family from cotton and a plastid-localized Na + transporter that negatively regulates salt tolerance of plants. Soil salinization is a major constraint on global cotton production, and Na + is the most dominant toxic ion in salinity stress. Hence, insights into the identities and properties of transporters that catalyze Na + movement between different tissues and within the cell compartments are vital to understand the salt-tolerant mechanisms of plants. Here, we identified the GhBASS5 gene, a member of the bile acid sodium symporter (BASS) gene family from cotton, served as a plastidic Na + transporter. GhBASS5 encodes a membrane protein localized in the plastid envelope. It was highly expressed in cotton roots and predominantly existed in the vascular cylinder. Heterogenous expression of GhBASS5 in Arabidopsis chloroplasts promoted Na + uptake into chloroplasts, which contributed to an increased cytoplasmic Na + concentration. And GhBASS5-overexpressed transgenic plants showed an increase in Na + translocation from roots to shoots and an elevated Na + content in both roots and shoots, but a dramatic decrease in the Na + efflux from root tissues and the K + /Na + ratio, especially under salt stress conditions. Furthermore, overexpressing GhBASS5 greatly damaged plastid functions and enhanced salt sensitivity in transgenic Arabidopsis when compared with wild-type plants under salt stress. Additionally, the salt-responsive transporter genes that regulate K + /Na + homeostasis were dramatically expressed in GhBASS5-overexpressed lines, especially under salt stress conditions. Taken together, our results suggest that GhBASS5 is a plastid-localized Na + transporter, and high expression of GhBASS5 impairs salt tolerance of plants via increasing Na + transportation and accumulation at both cell and tissue levels.

Crop yield is sensitive to salt stresses, for which Calcineurin B-like proteins (CBLs) are major response factors. This study shows that Arabidopsis CBL10, through protein S-acylation by protein S-acyl transferase10, targets to the vacuolar membrane to confer salt tolerance., (© 2019 Institute of Botany, Chinese Academy of Sciences.)

Phosphorus (P) availability and salinity stress are two major constraints for agriculture productivity. A combination of salinity and P starvation is known to be more deleterious to plant health. Plant growth promoting rhizobacteria are known to ameliorate abiotic stress in plants by increasing the availability of different nutrients. However, interaction mechanisms of plant grown under salinity and P stress condition and effect of beneficial microbe for stress alleviation is still obscure. Earlier we reported the molecular insight of auxin producing, phosphate solubilising Pseudomonas putida MTCC 5279 (RAR) mediated plant growth promotion in Arabidopsis thaliana. In present study new trait of proline and phosphatase production of RAR and its impact on modulation of physiological phenomenon under phosphate starved-salinity stress condition in A. thaliana has been investigated. Different physiological and molecular determinants under RAR- A. thaliana interaction showed that auxin producing RAR shows tryptophan dependence for growth and proline production in ATP dependant manner under salinity stress. However, under P deprived conditions growth and proline production are independent of tryptophan. RAR mediated lateral root branching and root hair density through modulation of abscisic acid signalling was observed. Acidic phosphatase activity under P starved and salinity stress condition was majorly modulated along with ROS metabolism and expression of stress responsive/phosphate transporter genes. A strong correlation of different morpho-physiological factor with RAR + salt conditions, showed We concluded that enhanced adverse effect of salinity with unavailability of P was dampened in presence of P. putida MTCC 5279 (RAR) in A. thaliana, though more efficiently salinity stress conditions. Therefore, alleviation of combined stress of salinity induced phosphate nutrient deficiency by inoculation of beneficial microbe, P. putida MTCC 5279 offer good opportunities for enhancing the agricultural productivity.

Previously, we reported that overexpression of AtRH17 , an Arabidopsis DEAD-box RNA helicase gene, confers salt stress-tolerance via a pathway other than the well-known salt stress-responsive pathways. To decipher the salt stress-responsive pathway in AtRH17 -overexpressing transgenic plants (OXs), we performed RNA-Sequencing and identified 397 differentially expressed genes between wild type (WT) and AtRH17 OXs. Among them, 286 genes were upregulated and 111 genes were downregulated in AtRH17 OXs relative to WT. Gene ontology annotation enrichment and KEGG pathway analysis showed that the 397 upregulated and downregulated genes are involved in various biological functions including secretion, signaling, detoxification, metabolic pathways, catabolic pathways, and biosynthesis of secondary metabolites as well as in stress responses. Genevestigator analysis of the upregulated genes showed that nine genes, namely, LEA4-5 , GSTF6 , DIN2 / BGLU30 , TSPO , GSTF7 , LEA18 , HAI1 , ABR , and LTI30 , were upregulated in Arabidopsis under salt, osmotic, and drought stress conditions. In particular, the expression levels of LEA4-5 , TSPO , and ABR were higher in AtRH17 OXs than in WT under salt stress condition. Taken together, our results suggest that a high AtRH17 expression confers salt stress-tolerance through a novel salt stress-responsive pathway involving nine genes, other than the well-known ABA-dependent and ABA-independent pathways.

Chloroplast biogenesis depends on an extensive interplay between the nucleus, cytosol, and chloroplasts, involving regulatory nucleus-encoded chloroplast proteins, as well as nucleocytosolic photoreceptors such as phytochromes (phys) and other extrachloroplastic factors. However, this whole process is only partially understood. Here, we describe the role of VENOSA4 (VEN4) in chloroplast development and acclimation to adverse growth conditions. A 35S : VEN4 - eGFP fusion protein localizes to the nucleus in Arabidopsis ( Arabidopsis thaliana ) protoplasts, and VEN4 homologs are present in a wide range of eukaryotes including humans, where the corresponding homolog (SAMHD1) cleaves dNTPs. Defective photosynthesis in ven4 seedlings results from reduced accumulation of photosynthetic proteins and appears to be caused by a reduction in the translational capacity of chloroplasts. The negative effect of the ven4 mutation on photosynthesis can be phenotypically suppressed by germinating seeds in the presence of excess dCTP or a pool of dNTPs, implying that VEN4, like human SAMHD1, is involved in dNTP catabolism. Moreover, VEN4 activity is also required for optimal responses to cold and salt stresses. In conclusion, our work emphasizes the importance of the nucleocytosolic compartment and the fine-tuning of dNTP levels for chloroplast translation and development., (© 2020 American Society of Plant Biologists. All Rights Reserved.)

Among many glycoproteins within the plant secretory system, KORRIGAN1 (KOR1), a membrane-anchored endo-β-1,4-glucanase involved in cellulose biosynthesis, provides a link between N- glycosylation, cell wall biosynthesis, and abiotic stress tolerance. After insertion into the endoplasmic reticulum, KOR1 cycles between the trans -Golgi network (TGN) and the plasma membrane (PM). From the TGN, the protein is targeted to growing cell plates during cell division. These processes are governed by multiple sequence motifs and also host genotypes. Here, we investigated the interaction and hierarchy of known and newly identified sorting signals in KOR1 and how they affect KOR1 transport at various stages in the secretory pathway. Conventional steady-state localization showed that structurally compromised KOR1 variants were directed to tonoplasts. In addition, a tandem fluorescent timer technology allowed for differential visualization of young versus aged KOR1 proteins, enabling the analysis of single-pass transport through the secretory pathway. Observations suggest the presence of multiple checkpoints/branches during KOR1 trafficking, where the destination is determined based on KOR1's sequence motifs and folding status. Moreover, growth analyses of dominant PM-confined KOR1-L 48 L 49 →A 48 A 49 variants revealed the importance of active removal of KOR1 from the PM during salt stress, which otherwise interfered with stress acclimation., (© 2020 American Society of Plant Biologists. All rights reserved.)

SCO (synthesis of cytochrome c oxidase) proteins are involved in the insertion of copper during the assembly of cytochrome c oxidase (COX), the final enzyme of the mitochondrial respiratory chain. Two SCO proteins, namely, homolog of copper chaperone 1 and 2 (HCC1 and HCC2) are present in seed plants, but HCC2 lacks the residues involved in copper binding, leading to uncertainties about its function. In this study, we performed a transcriptomic and phenotypic analysis of Arabidopsis thaliana plants with reduced expression of HCC1 or HCC2. We observed that a deficiency in HCC1 causes a decrease in the expression of several stress-responsive genes, both under basal growth conditions and after applying a short-term high salinity treatment. In addition, HCC1 deficient plants show a faster decrease in chlorophyll content, photosystem II quantum efficiency and COX levels after salinity stress, as well as a faster increase in alternative oxidase capacity. Notably, HCC2 deficiency causes opposite changes in most of these parameters. Bimolecular fluorescence complementation analysis indicated that both proteins are able to interact. We postulate that HCC1 is a limiting factor for COX assembly during high salinity conditions and that HCC2 probably acts as a negative modulator of HCC1 activity through protein-protein interactions. In addition, a direct or indirect role of HCC1 and HCC2 in the gene expression response to stress is proposed., (� The Author(s) 2019. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Actin filament, also known as microfilament, is one of two major cytoskeletal elements in plants and plays important roles in various biological processes. Like in animal cells, actin filaments have been thought to participate in autophagy in plants. However, surprisingly, in this study we found that actin filaments are dispensable for the occurrence of autophagy in plants. Disruption of actin filaments by short term treatment with actin polymerization inhibitors, cytochalasin D and latrunculin B, or transient overexpression of Profilin 3 in Nicotiana benthamiana had no effect on basal autophagy as well as the upregulation of nocturnal autophagy and salt stress-induced autophagy. Furthermore, anti-microfilament drug treatment affected neither basal nor salt stress-induced autophagy in Arabidopsis . In addition, prolonged perturbation of actin filaments by silencing Actin7 or 24-h treatment with microfilament-disrupting agents in N. benthamiana caused endoplasmic reticulum (ER) disorganization and subsequent degradation via autophagy involving ATG2, 3, 5, 6 and 7. Our findings reveal that, unlike mammalian cells, actin filaments are unnecessary for bulk autophagy in plants. Abbreviations: ATG: autophagy-related; CD: cytochalasin D; Cvt pathway: cytoplasm to vacuole targeting pathway; DMSO: dimethyl sulfoxide; ER: endoplasmic reticulum; LatB: latrunculin B; Nb: Nicotiana benthamiana ; PAS: phagophore assembly site; PRF3: Profilin 3; RER: rough ER; SER: smooth ER; TEM: transmission electron microscopy; TRV: Tobacco rattle virus ; VIGS: virus-induced gene silencing; wpi: weeks post-agroinfiltration.

Soil salinity is one of the serious environmental issues worldwide. In the present study, we made an attempt to isolate endophytic actinobacteria from halophyte and evaluate their growth promoting ability in Arabidopsis thaliana under salt stress through transcriptomic analysis. Two endophytic strains SYSU 333322 and SYSU 333140 were isolated and 16S rRNA gene sequence analysis suggests that these strains belong to Arthrobacter endophyticus and Nocardiopsis alba, respectively. To evaluate the growth promoting ability of two strains in Arabidopsis thaliana four experimental set up were designed. Set up designated s322 and s140 includes strains SYSU 333322 and SYSU 333140, respectively inoculated with A. thaliana under salt stress; set up designated MS322 and MS140 includes strains SYSU 333322 and SYSU 333140, respectively inoculated with A. thaliana without salt stress; MS includes seedlings without bacterial strains and salt stress; C150 includes seedlings grown in 150 mmol L -1 NaCl. A. endophyticus strain SYSU 333322 and N. alba strain SYSU 333140 were efficient to promote A. thaliana growth under salt stress A. endophyticus strain SYSU 333322 was more efficient than N. alba strain SYSU 333140 for growth promotion. Although A. endophyticus strain SYSU 333322 and N. alba strain SYSU 333140 were isolated from the same host, their mechanism of growth promotion in A. thaliana under salt stress was different. Gene encoding for chlorophyll a reductase, peptide-methionine (R)-S-oxide reductase, and potassium ion uptake were up-regulated when A. thaliana inoculated with strain SYSU 333322 and SYSU 333140 under salt stress. Pathways such as carotenoid biosynthesis, phenylalanine metabolism, phenylpropanoid biosynthesis, glycerolipid metabolism, and nitrogen metabolism played a crucial role in enhancing the salt stress tolerance of A. thaliana. Our results suggest that different bacteria have a different mechanism to promote plant growth under salt stress and hence it is necessary to understand the mechanism to overcome soil salinity problem., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Vitamin B 6 (pyridoxine) is vital for key metabolic reactions and reported to have antioxidant properties in planta. Therefore, enhancement of vitamin B 6 content has been hypothesized to be a route to improve resistance to biotic and abiotic stresses. Most of the current studies on vitamin B 6 in plants are on eudicot species, with monocots remaining largely unexplored. In this study, we investigated vitamin B 6 biosynthesis in rice, with a view to examining the feasibility and impact of enhancing vitamin B 6 levels. Constitutive expression in rice of two Arabidopsis thaliana genes from the vitamin B 6 biosynthesis de novo pathway, AtPDX1.1 and AtPDX2, resulted in a considerable increase in vitamin B 6 in leaves (up to 28.3-fold) and roots (up to 12-fold), with minimal impact on general growth. Rice lines accumulating high levels of vitamin B 6 did not display enhanced tolerance to abiotic stress (salt) or biotic stress (resistance to Xanthomonas oryzae infection). While a significant increase in vitamin B 6 content could also be achieved in rice seeds (up to 3.1-fold), the increase was largely due to its accumulation in seed coat and embryo tissues, with little enhancement observed in the endosperm. However, seed yield was affected in some vitamin B 6 -enhanced lines. Notably, expression of the transgenes did not affect the expression of the endogenous rice PDX genes. Intriguingly, despite transgene expression in leaves and seeds, the corresponding proteins were only detectable in leaves and could not be observed in seeds, possibly pointing to a mode of regulation in this organ., (© 2019 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)

Salinity is detrimental to plant growth, crop production and food security worldwide. Excess salt triggers increases in cytosolic Ca 2+ concentration, which activate Ca 2+ -binding proteins and upregulate the Na + /H + antiporter in order to remove Na + . Salt-induced increases in Ca 2+ have long been thought to be involved in the detection of salt stress, but the molecular components of the sensing machinery remain unknown. Here, using Ca 2+ -imaging-based forward genetic screens, we isolated the Arabidopsis thaliana mutant monocation-induced [Ca 2+] i increases 1 (moca1), and identified MOCA1 as a glucuronosyltransferase for glycosyl inositol phosphorylceramide (GIPC) sphingolipids in the plasma membrane. MOCA1 is required for salt-induced depolarization of the cell-surface potential, Ca 2+ spikes and waves, Na + /H + antiporter activation, and regulation of growth. Na + binds to GIPCs to gate Ca 2+ influx channels. This salt-sensing mechanism might imply that plasma-membrane lipids are involved in adaption to various environmental salt levels, and could be used to improve salt resistance in crops.

In Arabidopsis , the salt overly sensitive (SOS) pathway, consisting of calcineurin B-like protein 4 (CBL4/SOS3), CBL-interacting protein kinase 24 (CIPK24/SOS2) and SOS1, has been well defined as a crucial mechanism to control cellular ion homoeostasis by extruding Na + to the extracellular space, thus conferring salt tolerance in plants. CBL10 also plays a critical role in salt tolerance possibly by the activation of Na + compartmentation into the vacuole. However, the functional relationship of the SOS and CBL10-regulated processes remains unclear. Here, we analyzed the genetic interaction between CBL4 and CBL10 and found that the cbl4 cbl10 double mutant was dramatically more sensitive to salt as compared to the cbl4 and cbl10 single mutants, suggesting that CBL4 and CBL10 each directs a different salt-tolerance pathway. Furthermore, the cbl4 cbl10 and cipk24 cbl10 double mutants were more sensitive than the cipk24 single mutant, suggesting that CBL10 directs a process involving CIPK24 and other partners different from the SOS pathway. Although the cbl4 cbl10, cipk24 cbl10, and sos1 cbl10 double mutants showed comparable salt-sensitive phenotype to sos1 at the whole plant level, they all accumulated much lower Na + as compared to sos1 under high salt conditions, suggesting that CBL10 regulates additional unknown transport processes that play distinct roles from the SOS1 in Na + homeostasis.

MYB proteins are major transcription factors that play significant roles in plant defenses against various stresses. However, available information regarding stress-related MYB genes in maize is minimal. Herein, a maize MYB gene, ZmMYB3R, was cloned and functionally characterized. Subcellular localisation analysis showed that ZmMYB3R is localised to the nucleus. Yeast one-hybrid results revealed that ZmMYB3R has trans-activation activity, and a minimal activation domain at the C-terminus spanning residues 217-563. Gene expression analysis suggested that ZmMYB3R was induced by drought, salt and abscisic acid (ABA). Transgenic Arabidopsis plants overexpressing ZmMYB3R displayed enhanced growth performance and higher survival rates, elevated catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD) enzyme activities, increased sensitivity to ABA, and regulation of the stomatal aperture, suggesting that ZmMYB3R enhances tolerance to drought and salt stress. qRT-PCR assays revealed elevated expression levels of stress/ABA genes in transgenic plants following stress treatments. Moreover, transgenic plants accumulated higher ABA content than wild-type plants under drought and salt stress conditions. Collectively, these results indicate that ZmMYB3R is a positive transcription factor that enhances tolerance to drought and salt stress via an ABA-dependent pathway. The findings may prove useful for engineering economically important crops., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

Calcium is a universal secondary messenger that triggers many cellular responses. However, it is unclear how a calcium signal is coordinately decoded by different calcium sensors, which in turn regulate downstream targets to fulfill a specific physiological function. Here we show that SOS2-LIKE PROTEIN KINASE5 (PKS5) can negatively regulate the Salt-Overly-Sensitive signaling pathway in Arabidopsis. PKS5 can interact with and phosphorylate SOS2 at Ser 294 , promote the interaction between SOS2 and 14-3-3 proteins, and repress SOS2 activity. However, salt stress promotes an interaction between 14-3-3 proteins and PKS5, repressing its kinase activity and releasing inhibition of SOS2. We provide evidence that 14-3-3 proteins bind to Ca 2+ , and that Ca 2+ modulates 14-3-3-dependent regulation of SOS2 and PKS5 kinase activity. Our results suggest that a salt-induced calcium signal is decoded by 14-3-3 and SOS3/SCaBP8 proteins, which selectively activate/inactivate the downstream protein kinases SOS2 and PKS5 to regulate Na + homeostasis by coordinately mediating plasma membrane Na + /H + antiporter and H + -ATPase activity.

Sessile plants reprogram their metabolic and developmental processes during adaptation to prolonged environmental stresses. To understand the molecular mechanisms underlying adaptation of plant cells to saline stress, we established callus suspension cell cultures from Arabidopsis roots adapted to high salt for an extended period of time. Adapted cells exhibit enhanced salt tolerance compared with control cells. Moreover, acquired salt tolerance is maintained even after the stress is relieved, indicating the existence of a memory of acquired salt tolerance during mitotic cell divisions, known as mitotic stress memory. Metabolite profiling using 1H-nuclear magnetic resonance (NMR) spectroscopy revealed metabolic discrimination between control, salt-adapted and stress-memory cells. Compared with control cells, salt-adapted cells accumulated higher levels of sugars, amino acids and intermediary metabolites in the shikimate pathway, such as coniferin. Moreover, adapted cells acquired thicker cell walls with higher lignin contents, suggesting the importance of adjustments of physical properties during adaptation to elevated saline conditions. When stress-memory cells were reverted to normal growth conditions, the levels of metabolites again readjusted. Whereas most of the metabolic changes reverted to levels intermediate between salt-adapted and control cells, the amounts of sugars, alanine, γ-aminobutyric acid and acetate further increased in stress-memory cells, supporting a view of their roles in mitotic stress memory. Our results provide insights into the metabolic adjustment of plant root cells during adaptation to saline conditions as well as pointing to the function of mitotic memory in acquired salt tolerance., (� The Author 2018. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Cells maintain integrity despite changes in their mechanical properties elicited during growth and environmental stress. How cells sense their physical state and compensate for cell-wall damage is poorly understood, particularly in plants. Here we report that FERONIA (FER), a plasma-membrane-localized receptor kinase from Arabidopsis, is necessary for the recovery of root growth after exposure to high salinity, a widespread soil stress. The extracellular domain of FER displays tandem regions of homology with malectin, an animal protein known to bind di-glucose in vitro and important for protein quality control in the endoplasmic reticulum. The presence of malectin-like domains in FER and related receptor kinases has led to widespread speculation that they interact with cell-wall polysaccharides and can potentially serve a wall-sensing function. Results reported here show that salinity causes softening of the cell wall and that FER is necessary to sense these defects. When this function is disrupted in the fer mutant, root cells explode dramatically during growth recovery. Similar defects are observed in the mur1 mutant, which disrupts pectin cross-linking. Furthermore, fer cell-wall integrity defects can be rescued by treatment with calcium and borate, which also facilitate pectin cross-linking. Sensing of these salinity-induced wall defects might therefore be a direct consequence of physical interaction between the extracellular domain of FER and pectin. FER-dependent signaling elicits cell-specific calcium transients that maintain cell-wall integrity during salt stress. These results reveal a novel extracellular toxicity of salinity, and identify FER as a sensor of damage to the pectin-associated wall., (Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Regulation of cortical microtubule reorganization is essential for plant cell survival under high salinity conditions. In response to salt stress, microtubules undergo rapid depolymerization followed by reassembly to form a new microtubule network that promotes cell survival; however, the upstream regulatory mechanisms for this recovery response are largely unknown. In this study, we demonstrate that ethylene signaling facilitates salt stress-induced reassembly of cortical microtubules in Arabidopsis ( Arabidopsis thaliana ). Microtubule depolymerization was not affected under salt stress following the suppression of ethylene signaling with Ag + or in ethylene-insensitive mutants, whereas microtubule reassembly was significantly inhibited. ETHYLENE-INSENSITIVE3, a key transcription factor in the ethylene signaling pathway, was shown to play a central role in microtubule reassembly under salt stress. In addition, we performed functional characterization of the microtubule-stabilizing protein WAVE-DAMPENED2-LIKE5 (WDL5), which was found to promote ethylene-associated microtubule reassembly and plant salt stress tolerance. These findings indicate that ethylene signaling regulates microtubule reassembly by up-regulating WDL5 expression in response to salt stress, thereby implicating ethylene signaling in salt-stress tolerance in plants., (© 2018 American Society of Plant Biologists. All Rights Reserved.)

Phosphorylation of the RNA polymerase II (Pol II) C-terminal domain (CTD) regulates transcription of protein-coding mRNAs and noncoding RNAs. CTD function in transcription of protein-coding RNAs has been studied extensively, but its role in plant noncoding RNA transcription remains obscure. Here, using Arabidopsis thaliana CTD PHOSPHATASE-LIKE4 knockdown lines ( CPL4 RNAi ), we showed that CPL4 functions in genome-wide, conditional production of 3'-extensions of small nuclear RNAs (snRNAs) and biogenesis of novel transcripts from protein-coding genes downstream of the snRNAs (snRNA-downstream protein-coding genes [snR-DPGs]). Production of snR-DPGs required the Pol II snRNA promoter (PIIsnR), and CPL4 RNAi plants showed increased read-through of the snRNA 3'-end processing signal, leading to continuation of transcription downstream of the snRNA gene. We also discovered an unstable, intermediate-length RNA from the SMALL SCP1-LIKE PHOSPHATASE14 locus ( imRNA SSP14 ), whose expression originated from the 5' region of a protein-coding gene. Expression of the imRNA SSP14 was driven by a PIIsnR and was conditionally 3'-extended to produce an mRNA. In the wild type, salt stress induced the snRNA-to-snR-DPG switch, which was associated with alterations of Pol II-CTD phosphorylation at the target loci. The snR-DPG transcripts occur widely in plants, suggesting that the transcriptional snRNA-to-snR-DPG switch may be a ubiquitous mechanism to regulate plant gene expression in response to environmental stresses., (© 2017 American Society of Plant Biologists. All rights reserved.)