Osmotic Stress Tolerance Research Articles

Coapplication of Effective Microorganisms and Nanomagnesium Boosts the Agronomic, Physio-Biochemical, Osmolytes, and Antioxidants Defenses Against Salt Stress in Ipomoea batatas

The application of bio- and nanofertilizers are undoubtedly opening new sustainable approaches toward enhancing abiotic stress tolerance in crops. In this study, we evaluated the application of effective microorganisms (EMs) of five groups belonging to photosynthetic bacteria, lactic acid bacteria, yeast, actinobacteria, and fermenting fungi combined with magnesium oxide (MgO) nanoparticles (MgO-NP) on the growth and productivity of sweet potato plants grown in salt-affected soils. In two field experiments carried out in 2020 and 2021, we tested the impacts of EMs using two treatments (with vs. without EMs as soil drench) coupled with three foliar applications of MgO-NP (0, 50, and 100 μg ml–1 of MgO, representing MgO-NP0, MgO-NP50, and MgO-NP100, respectively). In our efforts to investigate the EMs:MgO-NP effects, the performance (growth and yield), nutrient acquisition, and physio-biochemical attributes of sweet potatoes grown in salt-affected soil (7.56 dS m–1) were assessed. Our results revealed that salinity stress significantly reduced the growth parameters, yield traits, photosynthetic pigment content (chlorophylls a and b, and carotenoids), cell membrane stability, relative water content, and nutrient acquisition of sweet potatoes. However, the EMs+ and/or MgO-NP-treated plants showed high tolerance to salt stress, specifically with a relatively superior increase when any of the biostimulants were combined. The application of EMs and/or MgO-NP improved osmotic stress tolerance by increasing the relative water content and membrane integrity. These positive responses owed to increase the osmolytes level (proline, free amino acids, and soluble sugars) and antioxidative compounds (non-enzymatic concentration, enzymatic activities, phenolic acid, and carotenoids). We also noticed that soil salinity significantly increased the Na+ content, whereas EMS+ and/or MgO-NP-treated plants exhibited lower Na+ concentration and increased K+ concentration and K+/Na+ ratio. These improvements contributed to increasing the photosynthetic pigments, growth, and yield under salinity stress. The integrative application of EMs and MgO-NP showed higher efficacy bypassing all single treatments. Our findings indicated the potential of coapplying EMs and MgO-NP for future use in attenuating salt-induced damage beneficially promoting crop performance.

Improved osmotic stress tolerance in brewer's yeast induced by wheat gluten peptides.

The influences of three wheat gluten peptides (WGP-LL, WGP-LML, and WGP-LLL) on the osmotic stress tolerance and membrane lipid component in brewer's yeast were investigated. The results demonstrated that the growth and survival of yeast under osmotic stress were enhanced by WGP supplementation. The addition of WGP upregulated the expressions of OLE1 (encoded the delta-9 fatty acid desaturase) and ERG1 (encoded squalene epoxidase) genes under osmotic stress. At the same time, WGP addition enhanced palmitoleic acid (C16:1) content, unsaturated fatty acids/saturated fatty acids ratio, and the amount of ergosterol in yeast cells under osmotic stress. Furthermore, yeast cells in WGP-LL and WGP-LLL groups were more resistant to osmotic stress. WGP-LL and WGP-LLL addition caused 25.08% and 27.02% increase in membrane fluidity, 22.36% and 29.54% reduction in membrane permeability, 18.38% and 14.26% rise in membrane integrity in yeast cells, respectively. In addition, scanning electron microscopy analysis revealed that the addition of WGP was capable of maintaining yeast cell morphology and reducing cell membrane damage under osmotic stress. Thus, alteration of membrane lipid component by WGP was an effective approach for increasing the growth and survival of yeast cells under osmotic stress. KEY POINTS: •WGP addition enhanced cell growth and survival of yeast under osmotic stress. •WGP addition increased unsaturated fatty acids and ergosterol contents in yeast. •WGP supplementation improved membrane homeostasis in yeast at osmotic stress.

OsPP65 Negatively Regulates Osmotic and Salt Stress Responses Through Regulating Phytohormone and Raffinose Family Oligosaccharide Metabolic Pathways in Rice

Although type 2C protein phosphatases (PP2Cs) have been demonstrated to play important roles in regulating plant development and various stress responses, their specific roles in rice abiotic stress tolerance are still largely unknown. In this study, the functions of OsPP65 in rice osmotic and salt stress tolerance were investigated. Here, we report that OsPP65 is responsive to multiple stresses and is remarkably induced by osmotic and salt stress treatments. OsPP65 was highly expressed in rice seedlings and leaves and localized in the nucleus and cytoplasm. OsPP65 knockout rice plants showed enhanced tolerance to osmotic and salt stresses. Significantly higher induction of genes involved in jasmonic acid (JA) and abscisic acid (ABA) biosynthesis or signaling, as well as higher contents of endogenous JA and ABA, were observed in the OsPP65 knockout plants compared with the wild-type plants after osmotic stress treatment. Further analysis indicated that JA and ABA function independently in osmotic stress tolerance conferred by loss of OsPP65. Moreover, metabolomics analysis revealed higher endogenous levels of galactose and galactinol but a lower content of raffinose in the OsPP65 knockout plants than in the wild-type plants after osmotic stress treatment. These results together suggest that OsPP65 negatively regulates osmotic and salt stress tolerance through regulation of the JA and ABA signaling pathways and modulation of the raffinose family oligosaccharide metabolism pathway in rice. OsPP65 is a promising target for improvement of rice stress tolerance using gene editing.

Expression alteration analyses in the transgenic Arabidopsis carrying soybean Histidine-containing phosphotransmitter gene under salinity stress condition

Productivity of many crops is highly vulnerable to extreme external conditions. Environmental stress factors such as drought and salinity have become more and more serious due to climate change and appear in many areas worldwide with higher frequency. As both drought and salinity belong to osmotic stress, they have similar negative effects on plant growth, development, and productivity as well as trigger similar stress responses by plants. In a previous study analyzing the expression profile in two soybean (Glycine max) cultivars with contrasting drought-tolerant phenotypes, a member of two-component system (TCS) in soybean, GmHP08, was proposed to associate with the plant tolerance capacity to drought. Subsequent in planta study confirmed its action as a positive regulator under drought conditions, as the transgenic Arabidopsis plants ectopically expressing GmHP08 acquired better drought tolerance. Following this, the presented research further explored the possible function of GmHP08 in mediating plant response to salinity. The obtained data from RT-qPCR analyses suggested that GmHP08 might positively enhance the salt tolerance of the Arabidopsis transgenic plants by altering the transcriptional abundance of several stress-related genes, including RD29A, RD29B, ABI5, SAG13, and CSD1. Activities of these genes are known to be associated with osmoprotection, senescence process, and antioxidation, which contribute to salt-tolerance ability of the transgenic plants. These results provided the first line of molecular evidence regarding GmHP08 function in plant response to salinity conditions. Therefore, extensive studies should be conducted in future studies to elaborate on the mechanisms by which this TCS member could improve various types of osmotic stress tolerance in plants.

32Pi Labeled Transgenic Wheat Shows the Accumulation of Phosphatidylinositol 4,5-bisphosphate and Phosphatidic Acid Under Heat and Osmotic Stress.

The ensuing heat stress drastically affects wheat plant growth and development, consequently compromising its grain yield. There are many thermoregulatory processes/mechanisms mediated by ion channels, lipids, and lipid-modifying enzymes that occur in the plasma membrane and the chloroplast. With the onset of abiotic or biotic stresses, phosphoinositide-specific phospholipase C (PI-PLC), as a signaling enzyme, hydrolyzes phosphatidylinositol 4,5-bisphosphate (PIP2) to generate inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG) which is further phosphorylated into phosphatidic acid (PA) as a secondary messenger and is involved in multiple processes. In the current study, a phospholipase C (PLC) signaling pathway was investigated in spring wheat (Triticum aestivum L.) and evaluated its four AtPLC5 overexpressed (OE)/transgenic lines under heat and osmotic stresses through 32Pi radioactive labeling. Naturally, the wheat harbors only a small amount of PIP2. However, with the sudden increase in temperature (40°C), PIP2 levels start to rise within 7.5 min in a time-dependent manner in wild-type (Wt) wheat. While the Phosphatidic acid (PA) level also elevated up to 1.6-fold upon exposing wild-type wheat to heat stress (40°C). However, at the anthesis stage, a significant increase of ∼4.5-folds in PIP2 level was observed within 30 min at 40°C in AtPLC5 over-expressed wheat lines. Significant differences in PIP2 level were observed in Wt and AtPLC5-OE lines when treated with 1200 mM sorbitol solution. It is assumed that the phenomenon might be a result of the activation of PLC/DGK pathways. Together, these results indicate that heat stress and osmotic stress activate several lipid responses in wild-type and transgenic wheat and can explain heat and osmotic stress tolerance in the wheat plant.

GhHAI2, GhAHG3, and GhABI2 Negatively Regulate Osmotic Stress Tolerance via ABA-Dependent Pathway in Cotton (Gossypium hirsutum L.).

The type 2C protein phosphatases (PP2Cs) are well known for their vital roles in plant drought stress responses, but their molecular mechanisms in cotton (Gossypium hirsutum L.) remain largely unknown. Here, we investigated the role of three clade A PP2C genes, namely, GhHAI2, GhAHG3, and GhABI2, in regulating the osmotic stress tolerance in cotton. The transcript levels of GhHAI2, GhAHG3, and GhABI2 were rapidly induced by exogenous abscisic acid (ABA) and polyethylene glycol (PEG) treatment. Silencing of GhHAI2, GhAHG3, and GhABI2 via virus-induced gene silencing (VIGS) improved osmotic tolerance in cotton due to decreased water loss, increase in both relative water content (RWC) and photosynthetic gas exchange, higher antioxidant enzyme activity, and lower malondialdehyde (MDA) content. The root analysis further showed that GhHAI2, GhAHG3, and GhABI2-silenced plants were more responsive to osmotic stress. Yeast two-hybrid (Y2H) and luciferase complementation imaging (LCI) assays further substantiated that GhHAI2, GhAHG3, and GhABI2 interact with the core receptors of ABA signaling, GhPYLs. The expression of several ABA-dependent stress-responsive genes was significantly upregulated in GhHAI2-, GhAHG3-, and GhABI2-silenced plants. Our findings suggest that GhHAI2, GhAHG3, and GhABI2 act as negative regulators in the osmotic stress response in cotton through ABA-mediated signaling.

Ectopic Expression of Kenaf (Hibiscus cannabinus L.) HcWRKY50 Improves Plants’ Tolerance to Drought Stress and Regulates ABA Signaling in Arabidopsis

Kenaf (Hibiscus cannabinus L.) is an environmentally friendly, multipurpose fiber crop suitable for osmotic stress tolerance studies. However, the mechanisms of tolerance remain largely unknown. Here, we identified a stress-responsive HcWRKY50 gene from kenaf (Hibiscus cannabinus L.) and studied its function and tolerance under drought stress. HcWRKY50 is a nuclear-localized protein. The overexpression of HcWRKY50 in Arabidopsis showed higher drought tolerance, exhibiting increased root length and lateral root number, and reduced stomatal density compared with the control lines. The seed germination and seedling growth of HcWRKY50 transgenic plants showed less sensitivity to ABA but they became more sensitive to ABA in their stomatal aperture. Furthermore, qRT-PCR analysis revealed that HcWRKY50 regulated ABA signaling by promoting the expression of several key ABA-responsive and stress-responsive genes such as RD29B and COR47 in transgenic lines. Taken together, this study demonstrated that the kenaf transcription factor HcWRKY50 regulates seed germination and seedling growth and improves drought stress tolerance via an ABA signaling pathway.

Gain-of-function of the cytokinin response activator ARR1 increases heat shock tolerance in Arabidopsis thaliana

ABSTRACT In addition to its well-established role in plant development, the hormone cytokinin regulates plant responses to biotic and abiotic stresses. It was previously shown that cytokinin signaling acts negatively upon drought and osmotic stress tolerance and that gain-of-function of the cytokinin response regulator ARR1 causes osmotic stress hypersensitivity. Here we show that increased ARR1 action increases tolerance to heat shock and that this is correlated with increased accumulation of the heat shock proteins Hsp17.6 and Hsp70. These results show that the heat shock tolerance of plants can be elevated by increasing the expression of a cytokinin response activator.

AcCIPK5, a pineapple CBL-interacting protein kinase, confers salt, osmotic and cold stress tolerance in transgenic Arabidopsis

Plant-specific calcineurin B-like proteins (CBLs) and their interacting kinases, CBL-interacting protein kinases (CIPKs) module, are essential for dealing with various biotic and abiotic stress. The kinases (CIPKs) of this module have been well studied in several plants; however, the information about pineapple CIPKs remains limited. To understand how CIPKs function against environmental cues in pineapple, the CIPK5 gene of pineapple was cloned and characterized. The phylogenetic analyses revealed that AcCIPK5 is homologous to the CIPK12 of Arabidopsis and other plant species. Quantitative real-time PCR (qRT-PCR) analysis revealed that AcCIPK5 responds to multiple stresses, including osmotic, salt stress, heat and cold. Under optimal conditions, AcCIPK5 gets localized to the cytoplasm and cell membrane. The ectopic expression of AcCIPK5 in Arabidopsis improved the germination under osmotic and salt stress. Furthermore, AcCIPK5 positively regulated osmotic, drought, salt and cold tolerance and negatively regulated heat and fungal stress in Arabidopsis. Besides, the expression of AcCIPK impacted ABA-related genes and ROS homeostasis. Overall, the present study demonstrates that AcCIPK5 contributes to multiple stress tolerance and has the potential to be utilized in the development of stress-tolerant crops.

Expression profiling of TaARGOS homoeologous drought responsive genes in bread wheat

Drought tolerant germplasm is needed to increase crop production, since water scarcity is a critical bottleneck in crop productivity worldwide. Auxin Regulated Gene involved in Organ Size (ARGOS) is a large protein family of transcription factors that plays a vital role in organ size, plant growth, development, and abiotic stress responses in plants. Although, the ARGOS gene family has been discovered and functionalized in a variety of crop plants, but a comprehensive and systematic investigation of ARGOS genes in locally used commercial wheat cultivars is still yet to be reported. The relative expression of three highly conserved TaARGOS homoeologous genes (TaARGOS-A, TaARGOS-B, TaARGOS-D) was studied in three drought-tolerant (Pakistan-2013, NARC-2009 and NR-499) and three sensitive (Borlaug-2016, NR-514 and NR-516) wheat genotypes under osmotic stress, induced by PEG-6000 at 0 (exogenous control), 2, 4, 6, and 12 h. The normalization of target genes was done using β-actin as endogenous control, whereas DREB3, as a marker gene was also transcribed, reinforcing the prevalence of dehydration in all stress treatments. Real-time quantitative PCR revealed that osmotic stress induced expression of the three TaARGOS transcripts in different wheat seedlings at distinct timepoints. Overall, all genes exhibited significantly higher expression in the drought-tolerant genotypes as compared to the sensitive ones. For instance, the expression profile of TaARGOS-A and TaARGOS-D showed more than threefold increase at 2 h and six to sevenfold increase after 4 h of osmotic stress. However, after 6 h of osmotic stress these genes started to downregulate, and the lowest gene expression was noticed after 12 h of osmotic stress. Among all the homoeologous genes, TaARGOS-D, in particular, had a more significant influence on controlling plant growth and drought tolerance as it showed the highest expression. Altogether, TaARGOSs are involved in seedling establishment and overall plant growth. In addition, the tolerant group of genotypes had a much greater relative fold expression than the sensitive genotypes. Ultimately, Pakistan-2013 showed the highest relative expression of the studied genes than other genotypes which shows its proficiency to mitigate osmotic stress. Therefore, it could be cultivated in arid and semi-arid regions under moisture-deficient regimes. These findings advocated the molecular mechanism and regulatory roles of TaARGOS genes in plant growth and osmotic stress tolerance in contrasting groups of wheat genotypes, accompanied by the genetic nature of identified genotypes in terms of their potential for drought tolerance.

Maize Transcription Factor ZmARF4 Confers Phosphorus Tolerance by Promoting Root Morphological Development.

Plant growth and development are closely related to phosphate (Pi) and auxin. However, data regarding auxin response factors (ARFs) and their response to phosphate in maize are limited. Here, we isolated ZmARF4 in maize and dissected its biological function response to Pi stress. Overexpression of ZmARF4 in Arabidopsis confers tolerance of Pi deficiency with better root morphology than wild-type. Overexpressed ZmARF4 can partially restore the absence of lateral roots in mutant arf7 arf19. The ZmARF4 overexpression promoted Pi remobilization and up-regulated AtRNS1, under Pi limitation while it down-regulated the expression of the anthocyanin biosynthesis genes AtDFR and AtANS. A continuous detection revealed higher activity of promoter in the Pi-tolerant maize P178 line than in the sensitive 9782 line under low-Pi conditions. Meanwhile, GUS activity was specifically detected in new leaves and the stele of roots in transgenic offspring. ZmARF4 was localized to the nucleus and cytoplasm of the mesophyll protoplast and interacted with ZmILL4 and ZmChc5, which mediate lateral root initiation and defense response, respectively. ZmARF4 overexpression also conferred salinity and osmotic stress tolerance in Arabidopsis. Overall, our findings suggest that ZmARF4, a pleiotropic gene, modulates multiple stress signaling pathways, and thus, could be a candidate gene for engineering plants with multiple stress adaptation.

The red seaweed Kappaphycus alvarezii antiporter gene (KaNa+/H+) confers abiotic stress tolerance in transgenic tobacco.

Plant establishment, growth, development and productivity are adversely affected by abiotic stresses that are dominant characteristics of environmentally challenged/degraded habitats created in the Anthropocene. Crop breeding for climate resilience properties is need of the hour to sustain the crop productivity. We report on the characterization of Kappaphycus alvarezii (a red seaweed) Na+/H+ antiporter gene (KaNa+/H+) for enhanced salt and osmotic stress tolerance. The KaNa+/H+ antiporter gene was cloned and over-expressed in tobacco under the control of CaMV35S promoter. Transgenic analysis was carried out to assess the stress tolerance ability of tobacco over-expressing KaNa+/H+ antiporter gene. Over-expression of KaNa+/H+ gene improved the seed germination and seed vigor index under stress. Transgenic plants grew better and exhibited delayed leaf senescence. Improved K+/Na+, carotenoid/total chlorophyll and relative water content; lower accumulation of reactive oxygen species (ROS), MDA and Na+; lower electrolyte leakage; better membrane stability index and accumulation of K+, photosynthetic pigment, starch, sugar, free amino acid, proline and polyphenol contents indicated better physiological health of the transgenic tobacco under stress. Transgenic tobacco exhibited higher photosynthesis, photosystem II efficiency, electron transfer rate, photochemical quenching and activity of water splitting complex. Compared with control tobacco, transgenic tobacco exhibited higher expression of stress-defence genes under stress and better recovery after long-term osmotic stress. Lower Na+ cytotoxicity, lower accumulation of ROS and maintenance of the membrane integrity helped transgenic tobacco to maintain the physiological functioning under stress. Present results established K. alvarezii as a potential gene resource and the KaNa+/H+ antiporter gene as a potential candidate gene in molecular breeding of cropsfor development of the degraded land.

Integrative genome and metabolome analysis reveal the potential mechanism of osmotic stress tolerance in Bifidobacterium bifidum

Osmotic stress affects the viability of probiotics, especially Bifidobacteria, which are less resistant to environmental stress. In this study, an adaptive evolution technique was applied to Bifidobacterium bifidum to select tolerant strains against high osmotic stress. B. bifidum CCFM16 was subjected to a gradually increasing penetration environment for 1000 generations. The generation time of the hyperosmotic tolerance mutant CCFM16m is 1/3 of its ancestor at an osmotic stress of 1300 mOsm/kg. Whole-genome comparison, candidate gene expression, and 3D protein structure simulation were performed and the results indicated that critical mutations were related to glutamate synthesis and transport. Untargeted metabolome analysis showed that compared to parent strain, the difference was mainly reflected in the increased accumulation of amino acids, especially proline, while the accumulation of central carbon decreased. Increase in the key enzyme F6PPK in the “bifidus” pathway indicated that cells transfer more energy from basal metabolism to resist osmotic stress. Since there was no proline transporter protein, excessive accumulation of proline showed that under long-term high osmotic stress, B. bifidum CCFM16 evolved a protective mechanism to synthesize proline from glutamic acid, an osmotic protection system with proline as the main compatible solute was formed.

Transcriptional and Physiological Analyses to Assess the Effects of a Novel Biostimulant in Tomato.

This work aimed to study the effects in tomato (Solanum lycopersicum L.) of foliar applications of a novel calcium-based biostimulant (SOB01) using an omics approach involving transcriptomics and physiological profiling. A calcium-chloride fertilizer (SOB02) was used as a product reference standard. Plants were grown under well-watered (WW) and water stress (WS) conditions in a growth chamber. We firstly compared the transcriptome profile of treated and untreated tomato plants using the software RStudio. Totally, 968 and 1,657 differentially expressed genes (DEGs) (adj-p-value < 0.1 and |log2(fold change)| ≥ 1) were identified after SOB01 and SOB02 leaf treatments, respectively. Expression patterns of 9 DEGs involved in nutrient metabolism and osmotic stress tolerance were validated by real-time quantitative reverse transcription PCR (RT-qPCR) analysis. Principal component analysis (PCA) on RT-qPCR results highlighted that the gene expression profiles after SOB01 treatment in different water regimes were clustering together, suggesting that the expression pattern of the analyzed genes in well water and water stress plants was similar in the presence of SOB01 treatment. Physiological analyses demonstrated that the biostimulant application increased the photosynthetic rate and the chlorophyll content under water deficiency compared to the standard fertilizer and led to a higher yield in terms of fruit dry matter and a reduction in the number of cracked fruits. In conclusion, transcriptome and physiological profiling provided comprehensive information on the biostimulant effects highlighting that SOB01 applications improved the ability of the tomato plants to mitigate the negative effects of water stress.

Diversity and bioprospecting for industrial hydrolytic enzymes of microbial communities isolated from deserted areas of south-east Morocco.

The current study aimed to analyze bacterial communities' diversity and abundance in three different deserted areas (Merzouga, Mhamid Elghizlane, and Erg lihoud) located in Moroccan Sahara, as well as to investigate osmotolerant microorganisms producing hydrolytic enzymes. The isolates were taxonomically affiliated using 16S rRNA gene sequencing. Four different hydrolase activities (amylase, lipase, cellulase, and protease) and osmotic stress tolerance were evaluated. The phylogenetic analysis of 364 screened isolates belonged to three phyla (Firmicutes 73%, Proteobacteria 26% and Actinobacteria 1%) and 18 different genera, from Bacillus, Ornithinibacillus, Paenibacillus, Geobacillus, Pseudomonas, Acinetobacter, Agrobacterium, Arthrobacter, Paenarthrobacter, Enterobacter, Staphylococcus, Erwinia, Herbasprillum, Ocuria, Massilia, Planomicrobium, Hodococcus, and Stenotrophomonas. The results detected a high proportion of osmotolerant and enzymes producing bacteria, many isolates can tolerate up to 55 °C (40%, 28%, and 30% in Merzouga, Mhamid Elghizlane, and Erg lihoudi, respectively). Meanwhile, the salinity tolerance reached 12% in some isolates with different proportions in each site, 29% in Merzouga, 24% in Mhamid Elghizlane, and 9% in Erg lihoudi. Furthermore, the enzymatic tests showed the presence of an amylolytic, lipolytic, cellulolytic, proteolytic activities in 20%, 31%, 63% and 72% of total strains, respectively.As a result, the present study is thus a preliminary yet critical step towards identifying the best bacterial candidates for further biotechnological applications.

SAPK9-OsNF-YA3-SLR1 Modulates the Balance between Plant Growth and Osmotic Stress Tolerance By Regulating GA and ABA Signaling in Rice

SAPK9-OsNF-YA3-SLR1 Modulates the Balance between Plant Growth and Osmotic Stress Tolerance By Regulating GA and ABA Signaling in Rice

Affinity Purification Followed by Liquid Chromatography-Tandem Mass Spectrometry to Identify Proteins Interacting with ABA Signaling Components.

Abscisic acid (ABA) is a key phytohormone involved in plant development, seed germination and responses to osmotic stresses, such as drought and high salinity. SNF1-related protein kinases (SnRK2s) play important roles in ABA-dependent and ABA-independent osmotic stress signaling. SnRK2s phosphorylate transcription factors and ion channels in response to ABA or osmotic stress to induce the expression of stress-responsive genes and stomatal closure, respectively, to confer osmotic stress tolerance. The activity of SnRK2s is directly or indirectly regulated by several protein factors. Identification of downstream substrates or upstream regulators of SnRK2s is very useful for elucidating protein components that regulate ABA and osmotic stress signaling. Here, we describe the use of affinity purification by coimmunoprecipitation and liquid chromatography-tandem mass spectrometry to identify protein complexes involved in ABA and osmotic stress signaling in plants. We previously identified several protein factors that regulate ABA and osmotic stress signaling by using this method.

Introgression of SbERD4 Gene Encodes an Early-Responsive Dehydration-Stress Protein That Confers Tolerance against Different Types of Abiotic Stresses in Transgenic Tobacco.

Salicornia brachiata is an extreme halophyte that commonly grows on marsh conditions and is also considered a promising resource for drought and salt-responsive genes. To unveil a glimpse of stress endurance by plants, it is of the utmost importance to develop an understanding of stress tolerance mechanisms. ‘Early Responsive to Dehydration’ (ERD) genes are defined as a group of genes involved in stress tolerance and the development of plants. To increase this understanding, parallel to this expedited thought, a novel SbERD4 gene was cloned from S. brachiata, characterized, and functionally validated in the model plant tobacco. The study showed that SbERD4 is a plasma-membrane bound protein, and its overexpression in tobacco plants improved salinity and osmotic stress tolerance. Transgenic plants showed high relative water, chlorophylls, sugars, starch, polyphenols, proline, free amino acids, and low electrolyte leakage and H2O2 content compared to control plants (wild type and vector control) under different abiotic stress conditions. Furthermore, the transcript expression of antioxidant enzyme encoding genes NtCAT, NtSOD, NtGR, and NtAPX showed higher expression in transgenic compared to wild-type and vector controls under varying stress conditions. Overall, the overexpression of a novel early responsive to dehydration stress protein 4-encoding gene (SbERD4) enhanced the tolerance of the plant against multiple abiotic stresses. In conclusion, the overexpression of the SbERD4 gene mitigates plant physiology by enduring stress tolerance and might be considered as a promising key gene for engineering salinity and drought stress tolerance in crops.

Relative contribution of different members of OsDREB gene family to osmotic stress tolerance in indica and japonica ecotypes of rice.

Drought/osmotic stress is the single largest production constraint in rain-fed rice cultivation. Different members of the DREB gene family are known to contribute to osmotic stress tolerance. In this study, an attempt was made to understand their relative contribution towards osmotic stress tolerance in indica and japonica ecotypes of rice. Two genotypes (one tolerant and one susceptible) from each ecotype were grown hydroponically, and 21-day-old seedlings were subjected to polyethylene glycol-induced osmotic stress (15% PEG-6000, equivalent to -3.0 bars osmotic potential). The tolerant genotypes CR143 and Moroberekan were found to have superior root traits (total root length, surface area and volume), better plant water status and increased total dry biomass as compared to their susceptible counterparts after 10 days of osmotic stress. Different members of the DREB gene family were differentially induced in response to osmotic shock (1 h after stress) and osmotic stress (24 h after stress), which also differed between the two rice ecotypes. From the gene expression profiles of 10 DREB genes (both DREB1 and DREB2 families), in indica two DREB genes, DREB1B and DREB1G, were significantly correlated with stress tolerance indices, whereas in japonica significant correlations with five DREB genes (DREB1A, DREB1B, DREB1D, DREB1E and DREB2B) were observed. We found that only one member, i.e. DREB1B, showed a significant correlation with drought tolerance indices in both indica and japonica ecotypes. This study provides an overview of the relative contribution of different members of the DREB gene family and their association with drought/osmotic stress tolerance in rice.

Maize PHYTOMELATONIN RECEPTOR1 functions in plant tolerance to osmotic and drought stress.

Phytomelatonin is a universal signal molecule that regulates plant growth and stress responses; however, only one receptor that can directly bind with and perceive melatonin signaling has been identified so far, namely AtPMTR1/CAND2 in Arabidopsis. Whether other plants contain a similar receptor and, if so, how it functions is still unknown. In this study, we identified a new phytomelatonin receptor in the monocot maize (Zea mays), and investigated its role in plant responses to osmotic and drought stress. Using homology searching, we identified a plasma membrane-localized protein, Zm00001eb214610/ZmPMTR1, with strong binding activity to melatonin as a potential phytomelatonin receptor in maize. Overexpressing ZmPMTR1 in Arabidopsis Col-0 promoted osmotic stress tolerance, and rescued osmotic stress sensitivity of the Arabidopsis cand2-1 mutant. Furthermore, ZmPMTR1 also largely rescued defects in melatonin-induced stomatal closure in the cand2-1 mutant, thereby reducing water loss rate and increasing tolerance to drought stress. In addition, we identified a maize mutant of ZmPMTR1, EMS4-06e2fl, with a point-mutation causing premature termination of protein translation, and found that this mutant had lower leaf temperatures, increased rate of water loss, and enhanced drought stress sensitivity. Thus, we present ZmPMTR1 as the first phytomelatonin receptor to be identified and examined in a monocot plant, and our results indicate that it plays an important function in the response of maize to drought stress.