Synergistic effects of carbon cycle metabolism and photosynthesis in Chinese cabbage under salt stress

Chinese cabbage(Brassica rapa ssp. pekinensis) has a long cultivation history and is one of the vegetable crops with the largest cultivation area in China. However, salt stress severely damages photosynthesis and hormone metabolism, nutritional balances, and results in ion toxicity in plants. To better understand the mechanisms of salt-induced growth inhibition in Chinese cabbage, RNA-seq and physiological index determination were conducted to explore the impacts of salt stress on carbon cycle metabolism and photosynthesis in Chinese cabbage. Here, we found that the number of thylakoids and grana lamellae and the content of starch granules and chlorophyll in the leaves of Chinese cabbage under salt stress showed a time-dependent response, first increasing and then decreasing. Chinese cabbage increased the transcript levels of genes related to the photosynthetic apparatus and carbon metabolism under salt stress, probably in an attempt to alleviate damage to the photosynthetic system and enhance CO_(2) fixation and energy metabolism. The transcription of genes related to starch and sucrose synthesis and degradation were also enhanced;this might have been an attempt to maintain intracellular osmotic pressure by increasing soluble sugar concentrations. Soluble sugars could also be used as potential reactive oxygen species(ROS) scavengers, in concert with peroxidase(POD)enzymes, to eliminate ROS that accumulate during metabolic processes. Our study characterizes the synergistic response network of carbon metabolism and photosynthesis under salt stress.

Genome-Wide Exploration of the Grape GLR Gene Family and Differential Responses of VvGLR3.1 and VvGLR3.2 to Low Temperature and Salt Stress

Grapes,one of the oldest tree species globally,are rich in vitamins.However,environmental conditions such as low temperature and soil salinization significantly affect grape yield and quality.The glutamate receptor(GLR)family,comprising highly conserved ligand-gated ion channels,regulates plant growth and development in response to stress.In this study,11 members of the VvGLR gene family in grapes were identified using whole-genome sequence analysis.Bioinformatic methods were employed to analyze the basic physical and chemical properties,phylogenetic trees,conserved domains,motifs,expression patterns,and evolutionary relationships.Phylogenetic and collinear analyses revealed that the VvGLRs were divided into three subgroups,showing the high conservation of the grape GLR family.These members exhibited 2 glutamate receptor binding regions(GABAb and GluR)and 3-4 transmembrane regions(M1,M2,M3,and M4).Real-time quantitative PCR analysis demonstrated the sensitivity of all VvGLRs to low temperature and salt stress.Subsequent localization studies in Nicotiana tabacum verified that VvGLR3.1 and VvGLR3.2 proteins were located on the cell membrane and cell nucleus.Additionally,yeast transformation experiments confirmed the functionality of VvGLR3.1 and VvGLR3.2 in response to low temperature and salt stress.Thesefindings highlight the significant role of the GLR family,a highly conserved group of ion channels,in enhancing grape stress resistance.This study offers new insights into the grape GLR gene family,providing fundamental knowledge for further functional analysis and breeding of stress-resistant grapevines.

Effects of Different Arbuscular Mycorrhizal Fungi on Physiology of Viola prionantha under Salt Stress

Arbuscular mycorrhizal(AM)fungi distribute widely in natural habits and play a variety of ecological functions.In order to test the physiological response to salt stress mediated by different AM fungi,Viola prionantha was selected as the host,the dominant AM fungus in the rhizosphere of V.philippica growing in Songnen saline-alkali grassland,Rhizophagus irregularis,and their mixtures were used as inoculants,and NaCl stress was applied after the roots were colonized.The results showed that V.philippica could be colonized by AM fungi in the field and the colonization rate ranged from 73.33%to 96.67%,and Claroideoglomus etunicatum was identified as the dominant AM fungi species in the rhizosphere of V.philippica by morphology combined with sequencing for AM fungal AML1/AML2 target.Inoculation with both the species resulted in the formation of mycorrhizal symbiosis(the colonization rate was more than 70%)and AM fungi significantly enhanced plants’tolerance to salt stress of varying magnitude.Higher activity of antioxidant enzymes and augmented levels of proline and other osmoregulators were observed in AM plants.The content of MDA in CK was higher than that in the inoculations with the stress of 100,200,and 250 mM.All indices except soluble protein content and MDA content were significantly correlated with AM fungal colonization indices.The analysis for different AM fungal effects showed that the mixtures and R.irregularis worked even better than C.etunicatum.These results will provide theoretical support for the exploration and screening of salt-tolerant AM fungi species and also for the application of AM-ornamental plants in saline-alkali urban greening.

The lectin gene TRpL1 of tetraploid Robinia pseudoacacia L.response to salt stress

Lectins are natural proteins in animals,plants,and microorganisms and can be divided into 12 families.These lectins play important roles in various environmental stresses.Some polyploid plants show tolerance to environmental stresses and to insect pests.However,the mechanism of stress tolerance is unclear.Tetraploid Robinia pseudoacacia(4×)under salt stress showed higher tolerance than diploid R.pseudoacacia(2×).As lectin can improve stress tolerance,it was questioned whether the stress resistance of polyploid plants was related to the lectin protein.In this study,salt resistance of lectin gene TRpL1 was verified by its over-expression in plants.In addition,salt resistance of lectin protein by E.coli strains was detected.The data revealed that the over-expression transgenic plants of TRpL1 showed better salt tolerance than control plants under salt stress,and the TRpL1-expressing strain also grew better in the medium with added NaCl.Therefore,tetraploid plants can resist salt stress through TRpL1 protein regulation.

Identification of novel salt stress-responsive microRNAs through sequencing and bioinformatic analysis in a unique halophilic Dunaliella salina strain

Dunaliella salina is a classic halophilic alga.However,its molecular mechanisms in response to high salinity at the post transcriptional level remain unknown.A unique halophilic alga strain,DS-CN1,was screened from four D.salina strains via cell biological,physiological,and biochemical methods.High-throughput sequencing of small RNAs(sRNAs)of DS-CN1 in culture medium containing 3.42-mol/L NaCl(SS group)or 0.05-mol/L NaCl(CO group)was performed on the BGISEQ-500 platform.The annotation and sequences of D.salina sRNAs were profiled.Altogether,44 novel salt stress-responsive microRNAs(miRNAs)with a relatively high C content,with the majority of them being 24 nt in length,were identified and characterized in DS-CN1.Twenty-one differentially expressed miRNAs(DEMs)in SS and CO were screened via bioinformatic analysis.A total of 319 putative salt stress-related genes targeted(104 overlapping genes)by novel miRNAs in this alga were screened based on our previous transcriptome sequencing research.Furthermore,these target genes were classified and enriched by GO and KEGG pathway analysis.Moreover,5 novel DEMs(dsa-mir3,dsa-mir16,dsa-mir17,and dsa-mir26 were significantly upregulated,and dsa-mir40 was significantly downregulated)and their corresponding 10 target genes involved in the 6 significantly enriched metabolic pathways were verified by quantitative real-time PCR.Next,their regulatory relationships were comprehensively analyzed.Lastly,a unique salt stress response metabolic network was constructed based on the novel DEM-target gene pairs.Taken together,our results suggest that 44 novel salt stress-responsive microRNAs were identified,and 4 of them might play important roles in D.salina upon salinity stress and contribute to clarify its distinctive halophilic feature.Our study will shed light on the regulatory mechanisms of salt stress responses.

GhSCL4 Acts as a Positive Regulator in Both Transgenic Arabidopsis and Cotton during Salt Stress

GRAS transcription factors play important roles in plant abiotic stress response,but their characteristics and functions in cotton have not been fully investigated.A cotton SCL4/7 subgroup gene in the GRAS family,GhSCL4,was found to be induced by NaCl treatments.Nuclear localization and transactivation activity of GhSCL4 indicate its potential role in transcriptional regulation.Transgenic Arabidopsis thaliana over-expressing GhSCL4 showed enhanced resistance to salt and osmotic stress.What’s more,the transcript levels of salt stress-induced genes(AtNHX1 and AtSOS1)and oxidation-related genes(AtAPX3 and AtCSD2)were more highly induced in the GhSCL4 over-expression lines than in wild type after salt treatment.Furthermore,silencing of GhSCL4 resulted in reduced salt tolerance in cotton caused by reactive oxygen species(ROS)enrichment under salt treatment,and antioxidant enzyme activities were accordingly significantly reduced in GhSLC4-silenced lines.These results indicated that GhSCL4 enhances salt tolerance of cotton by detoxifying ROS.In addition,the transient expression assay confirmed an interactive relationship between GhSCL4 and GhCaM7,which indicated that salt tolerance conferred by GhSCL4 might be associated with salt-induced Ca^(2+)/CaM7-mediated signaling.Taken together,GhSCL4 acts as a positive regulator in cotton during salt stress that is potentially useful for engineering salt-tolerant cotton.

Proteome-wide identification of S-sulfenylated cysteines response to salt stress in Brassica napus root

Reactive oxygen species(ROS)play a key role in a variety of biological processes,such as the perception of abiotic stress,the integration of different environmental signals,and the activation of stress response networks.Salt stress could induce an increased ROS accumulation in plants,disrupting intracellular redox homeostasis,leading to posttranslational modifications(PTMs)of specific proteins,and eventually causing adaptive changes in metabolism.Here,we performed an iodoTMT-based proteomic approach to identify the sulfenylated proteins in B.napus root responsing to salt stress.Totally,1348 sulfenylated sites in 751 proteins were identified and these proteins were widely existed in different cell compartments and processes.Our study revealed that proteins with changed abundance and sulfenylation level in B.napus root under salt stress were mainly enriched in the biological processes of ion binding,glycolysis,ATP binding,and oxidative stress response.This study displays a landscape of sulfenylated proteins response to salt stress in B.napus root and provides some theoretical support for further understanding of the molecular mechanisms of redox regulation under salt stress in plants.

Chlorophyll Fluorescence Response of Persimmon Plants under Salt Stress

[Objectives]To study the photosynthetic response mechanism of persimmon seedlings to salt stress.[Methods]The chlorophyll fluorescence parameters of Diospyros virginiana and Diospyros lotus seedlings under 4%salt stress were studied by pot culture salt control method,including the minimal fluorescence(F_(0)),maximum fluorescence(F_(m)),potential activity of PS II(F_(v)/F_(0)),maximum photochemical efficiency of PS II(F_(v)/F_(m)),electron transport rate(ETR),actual photochemical efficiency of PS II(Y II),and photochemical quenching coefficient(q_(p)).[Results]Under 4%salt stress,the maximum fluorescence(F_(m)),maximum photochemical efficiency of PS II(F v/F m),and photochemical quenching coefficient(q_(p))of two persimmon plants decreased with time.The potential activity of PS II(F_(v)/F_(0)),actual photochemical efficiency of PS II(Y_(II)),and electron transport rate(ETR)decreased under salt stress.[Conclusions]This study indicates that the PS II reaction center in the persimmon leaves was damaged and the electron transport at the acceptor side was damaged under salt stress.It is expected to lay a foundation for the analysis of salt-tolerance mechanism of persimmon plants.

Effect of Exogenous Gibberellin on Wheat Seed Germination under Salt Stress

[Objectives]In this experiment,wheat seeds were treated with different concentrations of gibberellin and different concentrations of salt solution to study the change of germination index of wheat seeds.[Methods]The germination rate,germination potential and germination index of wheat seeds were measured by routine methods,and the effect of exogenous gibberellin on germination of wheat seeds under salt stress was observed.[Results]The germination rate,germination potential and germination index of wheat seeds under salt stress were significantly increased after exogenous treatment of 0.25 and 0.50 g/L gibberellin within the range of salt concentration gradient.However,when the concentration of gibberellin was too high,it would inhibit the germination of seeds.[Conclusions]Appropriate concentration of gibberellin can effectively alleviate the stress caused by salt on wheat seed germination.In this experiment,the best concentration of gibberellin to alleviate salt stress was 0.25 g/L.

Response and adaptation to the accumulation and distribution of photosynthetic product in peanut under salt stress

To clarify the response and adaptability of peanut under salt stress,Huayu 25 was used as the material,and non-salt stress(CK),0.15% salt stress(S1),and 0.3% salt stress(S2) were applied as three treatments.The study analysed the effects of salt stress on photosynthetic characteristics,photosynthetic substances accumulation and distribution as well as the ecological adaptability of peanuts.The results showed that net photosynthetic rate(Pn),SPAD value,leaf area,and peanut yield were reduced under salt stress.Pn in CK was 13.71 and 28.72% higher than that in S1 and S2 at the 50 th day after planting,respectively.At the same growth period,the SPAD value among treatments was ranked as follows: CK>S1>S2.The 100-pod mass,100-kernel mass,kernel rate to pod,and pod mass per plant were reduced under salt stress,and the trend was CK>S1>S2.The distribution proportion of dry matter in different organs of peanut plant was changed to adapt to such stress.Roots under salt stress intensively distributed in a 0-40 cm soil layer for salt resistance.Dry mass proportion in stems and pods increased during the vegetative stage and early period of reproductive stage,respectively.The maximum growth rates of the pod volume,pod dry weight,and seed kernel dry weight all declined,and the pod and kernel volume at harvest were reduced,improving the seed plumpness under salt stress.This finding could be useful in growing peanut in saline soil.

Overaccumulation of glycine betaine makes the function of the thylakoid membrane better in wheat under salt stress

Wheat(Triticum aestivum L.) lines T1, T4, and T6 were genetically modified to increase glycine betaine(GB) synthesis by introduction of the BADH(betaine aldehyde dehydrogenase, BADH)gene from mountain spinach(Atriplex hortensis L.). These transgenic lines and WT of wheat(T. aestivum L.) were used to study the effect of increased GB synthesis on wheat tolerance to salt stress. Salt stress due to 200 mmol L-1Na Cl impaired the photosynthesis of the four wheat lines, as indicated by declines in net photosynthetic rate(Pn), stomatal conductance(Gs),maximum photochemical efficiency of PSII(Fv/Fm), and actual photochemical efficiency of PSII(ФPSII) and an increase in intercellular CO2concentration(Ci). In comparison with WT, the effect of salinity on the three transgenic lines was mild. Salt stress caused disadvantageous changes in lipids and their fatty acid compositions in the thylakoid membrane of the transgenic lines and WT. Under salt stress, the three transgenic lines showed slightly higher chlorophyll and carotenoid contents and higher Hill reaction activities and Ca2+-ATPase activity than WT. All the results suggest that overaccumulation of GB resulting from introduction of the BADH gene can enhance the salt tolerance of transgenic plants, especially in the protection of the components and function of thylakoid membranes, thereby making photosynthesis better. Changes in lipids and fatty acid compositions in the thylakoid membrane may be involved in the increased salt stress tolerance of the transgenic lines.

Physiological and Biochemical Mechanisms of Exogenous Calcium Chloride on Alleviating Salt Stress in Two Tartary Buckwheat(Fagopyrum tataricum)Varieties Differing in Salinity Tolerance

Salt stress is one of the most serious abiotic stresses limiting plant growth and development.Calcium as an essential nutrient element and important signaling molecule plays an important role in ameliorating the adverse effect of salinity on plants.This study aimed to investigate the impact of exogenous calcium on improving salt tolerance in Tartary buckwheat cultivars,cv.Xinong9920(salt-tolerant)and cv.Xinong9909(salt-sensitive).Four-week-old Tartary buckwheat seedlings under 100 mM NaCl stress were treated with and without exogenous calcium chloride(CaCl_(2)),Ca^(2+)chelator ethylene glycol tetraacetic acid(EGTA)and Ca^(2+)-channel blocker lanthanum chloride(LaCl_(3))for 10 days.Then,some important physiological and biochemical indexes were determined.The results showed that salt stress significantly reduced seedling growth,decreased photosynthetic pigments,inhibited antioxidants and antioxidant enzyme activities.However,it increased the reactive oxygen species(ROS)levels in the two Tartary buckwheat cultivars.Exogenous 10 mM CaCl_(2)application on salt-stressed Tartary buckwheat seedlings obviously mitigated the negative effects of NaCl stress and partially restored seedlings growth.Ca^(2+)-treated salt-stressed seedlings diplayed a suppressed accumulation of ROS,increased the contents of total chlorophyll,soluble protein,proline and antioxidants,and elevated the activities of antioxidant enzymes compared with salt stress alone.On the contrary,the addition of 0.5 mM LaCl_(3)and 5 mM EGTA on salt-stressed Tartary buckwheat seedlings exhibited the opposite effects to those with CaCl_(2)treatment.These results indicate that exogenous Ca^(2+)can enhance salt stress tolerance and Ca^(2+)supplementation may be an effective practice to cultivate Tartary buckwheat in saline soils.

Characterization of the 19 Novel Cotton FLA Genes and Their Expression Profiling in Fiber Development and in Response to Phytohormones and Salt Stress

Fasciclin-like arabinogalactan proteins(FLAs),a subclass of arabinogalactan proteins(AGPs),are usually involved in cell development in plants.To investigate the expression profiling as well

Effect of Seed Soaking with Exogenous Proline on Seed Germination of Rice Under Salt Stress

To explore the germination mechanism of salt-stressed rice improved by exogenous proline, and provide a theoretical basis to rice direct sowing technology for salinized soil, the effects of soaking with proline on germination status, amylase activity and isoenzyme were studied in this paper. The results showed that germination status including germination energy(GE), germination rate(GR), relative germination energy(RGE) and relative germination rate(RGR) significantly decreased as the same as the activities of alpha-amylase, beta-amylase and the total amylase under salt stress. Soaking with exogenous proline improved the germination status of rice under salt stress. Moreover, GE and RGE of salt-stressed rice were improved with increasing of proline concentration at the range of 5-45 mmol ·L-1. Soaking with 15 mmol ·L-1 and 30 mmol ·L-1 proline significantly improved the amylase activities(e.g. alpha-amylase, beta-amylase and total amylase) of rice under salt stress. Salt stress inhibited the express of beta-amylase isoenzyme temporarily, but had few impacts on alpha-amylase isozyme. Soaking with 30 mmol ·L-1 proline brightened District I and increased the width of 'i' brand in District II of alpha-amylase isoenzyme, but had few impacts on beta-amylase isoenzyme. In a word, soaking with proline could effectively alleviate the inhibitory effects of salt stress on seed germination.

Overexpression of StCYS1 gene enhances tolerance to salt stress in the transgenic potato(Solanum tuberosum L.)plant

Salt stress seriously restricts the growth and yield of potatoes.Plant cystatins are vital players in biotic stress and development,however,their roles in salt stress resistance remain elusive.Here,we report that StCYS1 positively regulates salt tolerance in potato plants.An in vitro biochemical test demonstrated that StCYS1 is a bona fide cystatin.Overexpression of StCYS1 in both Escherichia coli and potato plants significantly increased their resistance to high salinity.Further analysis revealed that the transgenic plants accumulated more proline and chlorophyll under salt stress conditions.Moreover,the transgenic plants displayed higher H2O2 scavenging capability and cell membrane integrity compared with wild-type potato.These results demonstrate that StCYS1 is closely correlated with salt stress and its overaccumulation can substantially enhance salt stress resistance.

Transcriptome analysis reveals dynamic changes in the salt stress response in Salix

Soil salinization is a serious ecological problem worldwide and information regarding the salt tolerance mechanisms of Salix is scarce.To elucidate the dynamic changes in the molecular mechanisms of Salix under salt stress,we generated gene expression profiles and examined changes in the expression of those genes.RNA-Seq was used to produce six cDNA libraries constructed from the leaves of Salix ×jiangsuensis CL‘J2345’treated with NaCl for 0,2,6,12,24 and 48 h.In total,249 million clean reads were assembled into 12,739 unigenes,all of which were clustered into 10 profiles based on their temporal expression patterns.KEGG analysis revealed that as an early defense response,the biosynthesis pathways of cutin,suberin and wax,which are involved in cell wall structure,were activated beginning at 2 h.The expression of secondary metabolism genes,including those involved in the phenylpropanoid,flavonoid,stilbenoid,diarylheptanoid and gingerol pathways,peaked at 6 h and 24 h;the upregulated genes were mainly involved in plant hormone pathways and beta-alanine,galactose and betalain metabolism.We identified roles of key phytohormones and found ETH to be the major signaling molecule activating TFs at 12 h;ETH,ABA,IAA and SA were the key molecules at 24 h.Moreover,we found that the upregulated genes were associated with elevated levels of amino acids,sucrose,inositol,stress proteins and ROS-scavenging enzymes,contributing to the maintenance of water balance.This research constitutes the first detailed analysis of salt stress-related mechanisms in Salix and identifies potential targets for genetic manipulation to improve yields.

Effects of 1-aminobenzotriazole on the growth and physiological characteristics of Tamarix chinensis cuttings under salt stress

vegetation restoration is a main ecological remediation technology for greening saline and alkaline soils.The objectives of this study were to determine the effect of1-aminobenzotriazole(ABT-1) on the growth and physiology of Tamarix chinensis under salt stress and to determine a suitable ABT-1 concentration and soil salinity(Sc) for propagating T.chihehsis-cuttings.Cuttings were soaked in water and ABT-1 solutions at three concentrations(50,100,and 200 mg L^(-1)) and propagated in pots containing four soil salinity levels,mild(0.3%),moderate(0.6%),and severe(0.9% and 1.2%),and compared with a control.The cuttings were measured to determine growth indices and physiological and biochemical indices(e.g.,chlorophyll content,superoxide dismutase activity,peroxidase activity,and malondialdehyde content).ABT-1 was effective in improving survival,growth,and physiological processes of cuttings under salt stress.However,there was a threshold effect when using ABT-1 to facilitate propagation under salt stress.ABT-1 effects were insignificant when applied at low concentrations(<100 mg L^(-1)).At a high concentration(> 100 mg L^(-1)),ABT-1 limited growth and physiological activities.Under a salt stress level(Sc ≤0.9%),ABT applied at a 100 mg L^(-1)concentration increased chlorophyll content and superoxide dismutase and peroxidase activities in the leaves and reduced malondialdehyde accumulation and membrane lipid peroxidation effects.As a result,ABT-1 enhanced the resistance of T.chinensis to salt stress.However,under high salt stress(>0.9%) and ABT-1 concentration(> 100 mg L^(-1)),the physiological regulatory ability of T.chinensis seedlings weakened.T.chinensis grew well at a salt stress ≤0.9% and ABT ≤100 mg L^(-1) and exhibited relatively high physiological regulatory ability and high salt adaptability.

Functional characterization of the Arabidopsis SERRATE under salt stress

SERRATE(SE)plays critical roles in RNA metabolism and plant growth regulation.However,its function in stresseresponse processes remains largely unknown.Here,we examined the regulatory role of SE using the se-1 mutant and its complementation line under saline conditions.The expression of SE was repressed by salt treatment at both mRNA and protein levels.After treatment with different NaCl concentrations,the se-1 mutants showed increased sensitivity to salinity.This heightened sensitivity was evidenced by decreased germination,reduced root growth,more serious chlorosis,and increased conductivity of the mutants compared with the wild type.Further analysis revealed that SE regulates the pre-mRNA splicing of several well-characterized marker genes associated with salt stress tolerance.Our data thus imply that SE may function as a key component in plant response to salt stress by modulating the splicing of salt stress-associated genes.

Analysis of DNA Cytosine Methylation on Cotton under Salt Stress

DNA methylation,especially methylation of cytosine in eukaryotic organisms,has been implicated in gene regulation,genomic imprinting,the timing of DNA replication,and determination of chromatin structure.It was reported that 6.5% of the whole cytosine residues in the nuclear DNA in

Metabolic responses to combined water deficit and salt stress in maize primary roots

Soil water deficit and salt stress are major limiting factors of plant growth and agricultural productivity. The primary root is the first organ to perceive the stress signals for drought and salt stress. In this study, maize plant subjected to drought, salt and combined stresses displayed a significantly reduced primary root length relative to the control plants. GC-MS was used to determine changes in the metabolites of the primary root of maize in response to salt, drought and combined stresses. A total of 86 metabolites were measured, including 29 amino acids and amines, 21 organic acids, four fatty acids, six phosphoric acids, 10 sugars, 10 polyols, and six others. Among these, 53 metabolites with a significant change under different stresses were identified in the primary root, and the content of most metabolites showed down-accumulation. A total of four and 18 metabolites showed significant up-and down-accumulation to all three treatments, respectively. The levels of several compatible solutes, including sugars and polyols, were increased to help maintain the osmotic balance. The levels of metabolites involved in the TCA cycle, including citric acid, ketoglutaric acid, fumaric acid, and malic acid, were reduced in the primary root. The contents of metabolites in the shikimate pathway, such as quinic acid and shikimic acid, were significantly decreased. This study reveals the complex metabolic responses of the primary root to combined drought and salt stresses and extends our understanding of the mechanisms involved in root responses to abiotic tolerance in maize.