Multidimensional evaluation of salt tolerance in groundnut genotypes through biochemical responses

The manuscript explores the complex interplay between groundnut genotypes,salt tolerance and hormonal influence,shedding light on the dynamic responses of three specific groundnut genotypes,KDG-128,TG-37 A and GG-20,to salt treatments and gibberellic acid(GA3).The study encompasses germination,plant growth,total protein content and oil content as key parameters.Through comprehensive analysis,it identifies TG-37 A and KDG-128 as salt-tolerant genotypes,and GG-20 as salt-susceptible genotypes,which highlighting the potential for targeted breeding efforts to develop more resilient groundnut varieties.Moreover,the quantification of protein and oil content under different treatments provides vital data for optimizing nutritional profiles in groundnut cultivars.Principal Component Analysis(PCA) underscores the significance of the first principal component(PC1)in explaining the majority of variance,capturing primary trends and differences in plant length.Analysis of Variance(ANOVA) and hierarchical analysis confirm the presence of statistically significant differences in protein and oil content among the genotypes.Pearson's correlation coefficient matrix analysis reveals strong positive correlations between plant length and protein content,plant length and oil content,and a moderately positive correlation between protein content and oil content.These findings provide valuable insights into groundnut physiology,salt tolerance,and nutritional composition,with implications for future research in sustainable agriculture and crop improvement.

Advances in deciphering salt tolerance mechanism in maize

Maize(Zea mays L.)is a global cereal crop whose demand is projected to double by 2050.Along with worsening of farmland salinization,salt stress has become a major environmental threat to the sustainability of maize production worldwide.Accordingly,there is an urgent need to decipher salt-tolerant mechanisms and facilitate the breeding of salt-tolerant maize.As salt tolerance is a complex trait regulated by multiple genes,and maize germplasm varies widely in salt tolerance,efforts have been devoted to the identification and application of quantitative-trait loci(QTL)for salt tolerance.QTL associated with ion regulation,osmotic tolerance,and other aspects of salt tolerance have been discovered using genomewide association studies(GWAS),linkage mapping,and omics-based approaches.This review highlights recent advances in the molecular-level understanding of salt stress response in maize,in particular in(a)the discovery of salt-tolerance QTL,(b)the mechanisms of salt tolerance,(c)the development of salttolerant maize cultivars,and(d)current challenges and future prospects.

A cluster of mutagenesis revealed an osmotic regulatory role of the OsPIP1 genes in enhancing rice salt tolerance

Aquaporins play important regulatory roles in improving plant abiotic stress tolerance.To better understand whether the Os PIP1 genes collectively dominate the osmotic regulation in rice under salt stress,a cluster editing of the Os PIP1;1,Os PIP1;2 and Os PIP1;3 genes in rice was performed by CRISPR/Cas9 system.Sequencing showed that two mutants with Cas9-free,line 14 and line 18 were successfully edited.Briefly,line 14 deleted a single C base in both the Os PIP1;1 and Os PIP1;3 genes,and inserted a single T base in the Os PIP1;2 gene,respectively.While line 18 demonstrated an insertion of a single A base in the Os PIP1;1gene and a single T base in both the Os PIP1;2 and Os PIP1;3 genes,respectively.Multiplex editing of the Os PIP1 genes significantly inhibited photosynthetic rate and accumulation of compatible metabolites,but increased MDA contents and osmotic potentials in the mutants,thus delaying rice growth under salt stress.Functional loss of the Os PIP1 genes obviously suppressed the expressions of the Os PIP1,Os SOS1,Os CIPK24 and Os CBL4 genes,and increased the influxes of Na+and effluxes of K^(+)/H^(+)in the roots,thus accumulating more Na+in rice mutants under salt stress.This study suggests that the Os PIP1 genes are essential modulators collectively contributing to the enhancement of rice salt stress tolerance,and multiplex editing of the Os PIP1 genes provides insight into the osmotic regulation of the PIP genes.

The Improvement of Soybean Salt Tolerance by Overexpressed GmPAO1

Polyamines play an important regulatory role during plant growth and development and adversity stress,and polyamine oxidase(PAO)is involved in polyamine catabolism.In this study,an up-regulated polyamine oxidase gene GmPAO1 was obtained by transcriptome sequencing analysis and screening at soybean seedling stages.Also,its expression pattern and function were analyzed.The identification results of transgenic GmPAO1 soybean positive lines showed that the relative expression level of GmPAO1 in the overexpressed lines was increased under salt stress.With increasing stress concentration,the seed germination rate decreased.However,the seed germination rate of the overexpressed lines was significantly higher than that of the control lines,and the phenotypic character of the root systems was also better than that of the control lines.The measurement of superoxide dismutase(SOD)and peroxidase(POD)activities and malondialdehyde and hydrogen peroxide contents revealed that the overexpressed soybean lines significantly increased the SOD and POD activities,significantly reducing the malondialdehyde content.Although the hydrogen peroxide content in the transformed plants gradually increased,the hydrogen peroxide content in the overexpression lines was still lower than that in the gene editing lines.Based on this,it was preliminarily judged that GmPAO1 can improve soybean salt tolerance.

Temperature/salt tolerance and oil recovery of xanthan gum solution enhanced by surface-modified nanosilicas

Amide-and alkyl-modified nanosilicas(AANPs)were synthesized and introduced into Xanthan gum(XG)solution,aiming to improve the temperature/salt tolerance and oil recovery.The rheological behaviors of XG/AANP hybrid dispersions were systematically studied at different concentrations,temperatures and inorganic salts.At high temperature(75C)and high salinity(10,000 mg,L1 NaCl),AANPs increase the apparent viscosity and dynamic modulus of the XG solution,and XG/AANP hybrid dispersion exhibits elastic-dominant properties.The most effective concentrations of XG and AANP interacting with each other are 1750 mg·L^(-1) and 0.74 wt%,respectively.The temperature tolerance of XG solution is not satisfactory,and high temperature further weakens the salt tolerance of XG.However,the AANPs significantly enhance the viscoelasticity the XG solution through hydrogen bonds and hydrophobic effect.Under reservoir conditions,XG/AANP hybrid recovers approximately 18.5%more OOIP(original oil in place)than AANP and 11.3%more OOIP than XG.The enhanced oil recovery mechanism of the XG/AANP hybrid is mainly increasing the sweep coefficient,the contribution from the reduction of oil-water interfacial tension is less.

Eco-friendly aqueous foam stabilized by cellulose microfibers with great salt tolerance and high temperature resistance

A low-cost eco-friendly aqueous foam,especially the robust foam with great tolerance to high salinity and high temperature,is in great demand in the oil industry,e.g.,oil and gas well or geothermal well drilling.Herein,an ultra-stable aqueous foam was developed using the biodegradable cellulose microfiber(CMF)as a foam stabilizer.The foam stabilized by CMF shows excellent tolerance to the high concentration of NaCl(6.0 wt%)and CaCl_(2)(0.25 wt%)and the related drainage half-life times(T_(0.5))reach 1750 and 2340 s respectively.By contrast,the foams without CMF are completely drained(T_(0.5)=0 s)when NaCl concentration is greater than 6.0 wt%or CaCl_(2) concentration is greater than 0.20 wt%.Notably,T0.5 of the foams stabilized by CMF at these saline concentrations still can maintain above 1000 s even after aging at 120℃ for 16 h,exhibiting an outstanding foam-stabilizing performance at high temperature.Experimental results suggest that the salt and high-temperature tolerance of CMF in foam stabilization is attributed to the electrically uncharged surfaces,the formation of a gel-like structure and the excellent thermal stability.This work not only provides a promising candidate of aqueous foam stabilizer to deal with high temperature and high salinity but also presents a natural-based solution for an environmentally friendly drilling industry in the future.

SlTPP4 participates in ABA-mediated salt tolerance by enhancing root architecture in tomato

Salinity tolerance is an important physiological index for crop breeding.Roots are typically the first plant tissue to withstand salt stress.In this study,we found that the tomato(Solanum lycopersicum)trehalose-6-phosphate phosphatase(SlTPP4)gene is induced by abscisic acid(ABA)and salt,and is mainly expressed in roots.Overexpression of SlTPP4 in tomato enhanced tolerance to salt stress,resulting in better growth performance.Under saline conditions,SlTPP4 overexpression plants demonstrated enhanced sucrose metabolism,as well as increased expression of genes related to salt tolerance.At the same time,expression of genes related to ABA biosynthesis and signal transduction was enhanced or altered,respectively.In-depth exploration demonstrated that SlTPP4 enhances Casparian band development in roots to restrict the intake of Na^(+).Our study thus clarifies the mechanism of SlTPP4-mediated salt tolerance,which will be of great importance for the breeding of salt-tolerant tomato crops.

Thellungiella halophila ST5 improves salt tolerance in cotton

Background: Salinity is a major abiotic stress to global agriculture which hampers crop growth and development, and eventually reduces yield. Transgenic technology is an e ective and e cient approach to improve crop salt tolerance but depending on the availability of e ective genes. We previously isolated Salt Tolerance5(ThST5) from the halophyte Thellungiella halophila, an ortholog of Arabidopsis SPT4-2 which encodes a transcription elongation factor. However, SPT4-2-confered salt tolerance has not been evaluated in crops yet. Here we report the evaluation of Th ST5-conferred salt tolerance in cotton(Gossypium hirsutum L.).Results: The ThST5 overexpression transgenic cotton plants displayed enhanced tolerance to salt stress during seed germination and seedling stage compared with wild type. Particularly, the transgenic plants showed improved salinity tolerance as well as yield under saline field conditions. Comparative transcriptomic analysis showed that ThST5 improved salt tolerance of transgenic cotton mainly by maintaining ion homeostasis. In addition, ThST5 also orchestrated the expression of genes encoding antioxidants and salt-responsive transcription factors.Conclusion: Our results demonstrate that ThST5 is a promising candidate to improve salt tolerance in cotton.

Plant salt tolerance and Na^+ sensing and transport

Salinity is a global challenge to agricultural production. Understanding Na^+ sensing and transport in plants under salt stress will be of benefit for breeding robustly salt-tolerant crop species. In this review, first, possible salt stress sensor candidates and the root meristem zone as a tissue harboring salt stress-sensing components are proposed. Then,the importance of Na^+ exclusion and vacuolar Na^+ sequestration in plant overall salt tolerance is highlighted. Other Na^+ regulation processes, including xylem Na^+ loading and unloading, phloem Na^+ recirculation, and Na^+ secretion, are discussed and summarized.Along with a summary of Na^+ transporters and channels, the molecular regulation of Na^+ transporters and channels in response to salt stress is discussed. Finally, some largely neglected issues in plant salt stress tolerance, including Na^+ concentration in cytosol and the role of Na^+ as a nutrient, are reviewed and discussed.

Salt tolerance in rice:Physiological responses and molecular mechanisms

Crop yield loss due to soil salinization is an increasing threat to agriculture worldwide.Salt stress drastically affects the growth,development,and grain productivity of rice(Oryza sativa L.),and the improvement of rice tolerance to salt stress is a desirable approach for meeting increasing food demand.The main contributors to salt toxicity at a global scale are Na^(+)and Cl^(-)ions,which affect up to 50%of irrigated soils.Plant responses to salt stress occur at the organismic,cellular,and molecular levels and are pleiotropic,involving(1)maintenance of ionic homeostasis,(2)osmotic adjustment,(3)ROS scavenging,and(4)nutritional balance.In this review,we discuss recent research progress on these four aspects of plant physiological response,with particular attention to hormonal and gene expression regulation and salt tolerance signaling pathways in rice.The information summarized here will be useful for accelerating the breeding of salt-tolerant rice.

Cloning and Characterization of a Salt Tolerance-Associated Gene Encoding Trehalose-6-Phosphate Synthase in Sweetpotato

Trehalose plays an important role in metabolic regulation and abiotic stress tolerance in a variety of organisms. In plants, its biosynthesis is catalyzed by two key enzymes： trehalose-6-phosphate synthase（TPS） and trehalose-6-phosphate phosphatase（TPP）. In the present study, a TPS gene, named IbTPS, was first isolated from sweetpotato（Ipomoea batatas（L.） Lam.） cv. Lushu 3 by rapid amplification of cDNA ends（RACE）. The open reading frame（ORF） contained 2 580 nucleotides encoding 859 amino acids with a molecular weight of 97.433 kDa and an isoelectric point（pI） of 5.7. The deduced amino acid sequence showed high identities with TPS of other plants. Real-time quantitative PCR analysis revealed that the expression level of IbTPS gene was significantly higher in stems of Lushu 3 than in its leaves and roots. Subcellular localization analysis in onion epidermal cells indicated that IbTPS gene was located in the nucleus. Transgenic tobacco（cv. Wisconsin 38） plants over-expressing IbTPS gene exhibited significantly higher salt tolerance compared with the control plant. Trehalose and proline content was found to be significantly more accumulated in transgenic tobacco plants than in the wild-type and several stress tolerance related genes were up-regulated. These results suggest that IbTPS gene may enhance salt tolerance of plants by increasing the amount of treahalose and proline and regulating the expression of stress tolerance related genes.

Haplotype variations in QTL for salt tolerance in Chinese wheat accessions identified by marker-based and pedigree-based kinship analyses

Most modern wheat cultivars were selected on the basis of yield-related indices measured under optimal fertilizer and irrigation inputs.With climate change,land degradation and salinity caused by sea water encroachment,wheat is increasingly subjected to environmental stress.Moreover,expanding urbanization increasingly encroaches upon prime agricultural land in countries like China,and alternative cropping areas must be found.Some of these areas have moderate constraining factors,such as salinity.Therefore,it is important to investigate whether current genetic materials and breeding procedures are maintaining adequate variability to address future problems caused by abiotic stress.In this study,a panel of 307 wheat accessions,including local landraces,exotic cultivars used in Chinese breeding programs and Chinese cultivars released during different periods since1940,were subjected to a genome-wide association study to dissect the genetic basis of salinity tolerance.Both marker-based and pedigree-based kinship analyses revealed that favorable haplotypes were introduced in some exotic cultivars as well as a limited number of Chinese landraces from the 1940 s.However,improvements in salinity tolerance during modern breeding are not as obvious as that of yield.To broaden genetic diversity for increasing salt tolerance,there is a need to refocus attention on local landraces that have high degrees of salinity tolerance and carry rare favorable alleles that have not been exploited in breeding.

Molecular Cloning and Functional Characterization of a Salt Tolerance-Associated Gene IbNFU1 from Sweetpotato

Iron-sulfur cluster biosynthesis involving the nitrogen fixation（Nif） proteins has been proposed as a general mechanism acting in various organisms.NifU-like protein may play an important role in protecting plants against abiotic and biotic stresses.Based on the EST sequence selected from salt-stressed suppression subtractive hybridization（SSH） cDNA library constructed with a salt-tolerant mutant LM79,a NFU gene,termed IbNFU1,was cloned from sweetpotato（Ipomoea batatas（L.） Lam.） via rapid amplification of cDNA ends（RACE）.The cDNA sequence of 1 117 bp contained an 846 bp open reading frame encoding a 281 amino acids polypeptide with a molecular weight of 30.5 kDa and an isoelectric point（pI） of 5.12.IbNFU1 gene contained a conserved Cys-X-X-Cys motif in C-terminal of the iron-sulfur cluster domain.The deduced amino acid sequence had 66.08 to 71.99% sequence identity to NFU genes reported in Arabidopsis thaliana,Eucalyptus grandis and Vitis vinifera.Real-time quantitative PCR analysis revealed that the expression level of IbNFU1 gene was significantly higher in the roots of the mutant LM79 compared to the wild-type Lizixiang.Transgenic tobacco（cv.Wisconsin 38） plants expressing IbNFU1 gene exhibited significantly higher salt tolerance compared to the untransformed control plants.It is proposed that IbNFU1 gene has an important function for salt tolerance of plants.

Gm NAC15 overexpression in hairy roots enhances salt tolerance in soybean

The NAC （NAM, ATAF1/2 and CUC2） transcription factor family plays a key role in plant development and responses to abiotic stress. GmNAC15 （Glyma 15g40510.1）, a member of the NAC transcription factor family in soybean, was functionally characterized, especially with regard to its role in salt tolerance. In the present study, qRT-PCR （quantitative reverse transcription PCR） analysis indicated that GmNAC15 was induced by salt, drought, low temperature stress, and ABA treatment in roots and leaves. GmNAC15 overexpression in soybean （Glycine max） hairy roots enhanced salt tolerance. Transgenic hairy roots improved the survival of wild leaves; however, overexpression of GmNAC15 in hairy root couldn＇t influnce the expression level of GmNAC15 in leaf. GmNAC15 regulates the expression levels of genes responsive to salt stress. Altogether, these results provide experimental evidence of the positive effect of GmNAC15 on salt tolerance in soybean and the potential application of genetic manipulation to enhance the salt tolerance of important crops.

ZmWRKY104 positively regulates salt tolerance by modulating ZmSOD4 expression in maize

Salinity impairs plant growth, limiting agricultural development. It is desirable to identify genes responding to salt stress and their mechanism of action. We identified a function of the Zea mays WRKY transcription factor, Zm WRKY104, in salt stress response. Zm WRKY104 was localized in the nucleus and showed transcriptional activation activity. Phenotypic and physiological analysis showed that overexpression of Zm WRKY104 in maize increased the tolerance of maize to salt stress and alleviated salt-induced increases in O;accumulation, malondialdehyde(MDA) content, and percent of electrolyte leakage. Further investigation showed that Zm WRKY104 increased SOD activity by regulating Zm SOD4 expression. Yeast onehybrid, electrophoretic mobility shift test, and chromatin immunoprecipitation–quantitative PCR assay showed that Zm WRKY104 bound directly to the promoter of Zm SOD4 by recognizing the W-box motif in vivo and in vitro. Phenotypic, physiological, and biochemical analysis showed that Zm SOD4 increased salt tolerance by alleviating salt-induced increases in O;accumulation, MDA content, and percent of electrolyte leakage under salt stress. Taken together, our results indicate that Zm WRKY104 positively regulates Zm SOD4 expression to modulate salt-induced O;accumulation, MDA content, and percent of electrolyte leakage, thus affecting salt stress response in maize.

Advanced Backcross QTL Analysis for the Whole Plant Growth Duration Salt Tolerance in Rice(Oryza sativa L.)

Salinity is a major factor limiting rice yield in coastal areas of Asia. To facilitate breeding salt tolerant rice varieties, the wholeplant growth duration salt tolerance（ST） was genetically dissected by phenotyping two sets of BC2F5 introgression lines（ILs） for four yield traits under severe natural salt stress and non-stress filed conditions using SSR markers and the methods of advanced backcross QTL（AB-QTL） analysis and selective introgression. Many QTLs affecting four yield traits under salt stress and nonstress conditions were identified, most（〉90%） of which were clustered in 13 genomic regions of the rice genome and involved in complex epistasis. Most QTLs affecting yield traits were differentially expressed under salt stress and non-stress conditions. Our results suggested that genetics complementarily provides an adequate explanation for the hidden genetic diversity for ST observed in both IL populations. Some promising Huanghuazhan（HHZ） ILs with favorable donor alleles at multiple QTLs and significantly improved yield traits under salt stress and non-stress conditions were identified, providing excellent materials and relevant genetic information for improving rice ST by marker-assisted selection（MAS） or genome selection.

Overexpression of SOS Genes Enhanced Salt Tolerance in Sweetpotato

The production of transgenic sweetpotato （cv.Xushu 18） plants exhibiting enhanced salt tolerance using salt overly sensitive （SOS） genes was achieved through Agrobacterium tumefaciens-mediated transformation.A.tumefaciens strain EHA105 harbors a binary vector pCAMBIA3301 with SOS genes （SOS1,SOS2 and SOS3） and bar gene.Selection culture was conducted using 0.3 mg L^-1 phosphinothricin （PPT）.A total of 40 plants were produced from the inoculated 170 cell aggregates via somatic embryogenesis.PCR analysis showed that 37 of the 40 regenerated plants were transgenic plants.The in vitro assay demonstrated that superoxide dismutase （SOD） and proline were significantly more accumulated and malonaldehyde （MDA） was significantly less accumulated in 21 transgenic plants than in control plants when they were exposed to 86 mmol L^-1 NaCl.Salt tolerance of these 21 plants was further evaluated with Hoagland solution containing 0,51,86,and 120 mmol L^-1 NaCl in the greenhouse.The results indicated that 6 of them had significantly better growth and rooting ability than the remaining 15 transgenic plants and control plants.Expression of SOS genes in the 6 salt-tolerant transgenic plants was demonstrated by RT-PCR analysis.This study provides an alternative approach for improving salt tolerance of sweetpotato.

Inheritance and QTL Mapping of Salt Tolerance in Rice

An F2 population derived from the cross between Jiucaiqing (Japonica) and IR36 (indica) was used to analyze the inheritance of salt tolerance in rice by genetic model of major-genes plus polygenes, and to map the corresponding QTLs by SSR molecular markers. Rice plants of P1, P2, F1 and F2 at 5- to 6- leaf stage were treated under 140 mmol/LNaCI for 10 days. Three indices representing the ability of salt tolerance of rice seedlings were measured, including salt tolerance rating (STR), Na+/K+ ratio in roots and dry matter weight of shoots (DWS). STR, Na+/K+ and DWS were all controlled by two major genes with modification by polygenes. Heritability of these traits from major genes was 17.8, 53.3 and 52.3%, respectively. The linkage map constructed by 62 SSR molecular markers covered a total length of about 1 142 cM. There were three QTLs detected for STR located on chromosome 1, 5 and 9, two QTLs for DWS on chromosomes 8 and 9, and two QTLs for Na+/K+ on chromosomes 2 and 6, one on each chromosome respectively. Single QTL accounted for 6.7 to 19.3% of phenotypic variation. Identification method of salt tolerance in rice and breeding of rice varieties with salt tolerance based on molecular markers assisted selection had been discussed.

Gamma Irradiation of Embryogenic Callus Cultures and In vitro Selection for Salt Tolerance in Sugarcane (Saccharum officinarum L.)

Radiation induced mutagenesis followed by in vitro selection was employed for salt tolerance in popular Indian sugarcane （Saccharum officinarum L.） cv. CoC-671. Embryogenic calli were gamma irradiated and exposed to different levels of NaCl （42.8, 85.6, 128.3, 171.1,213.9, 256.7, 299.5, or 342.2 mM）. The relative growth rate （RGR） decreased progressively with increasing salt stress and was the least with a salt stress of 256.7 mM （0.25±0.009）, almost 10 fold lesser than the control. The RGR was significantly lower in 85.6 mM and higher salt stressed calli than the control. The survival percent also decreased, with an increase in NaCl concentration. In case of 10 and 20 Gy irradiated calli, regeneration was observed up to 85.6 mM NaCl selection, medium, whereas, higher treatments （128.3 mM and beyond） exhibited browning initially. However, in the subsequent subcultures, regeneration was obtained in the case of 10 and 20 Gy irradiated calli on 128.3 and 171.1 mM NaCl selections. Higher dose of gamma irradiation （40 Gy） also showed regeneration, but only with 85.6 mM NaCI selection. The unirradiated calli regenerated the highest number of plantlets followed by 10 and 20 Gy irradiated calli on salt selection. A total of 147 plantlets were selected from different salt levels. The salt selected plants are being tested for their field performance.

The osmolyte-producing endophyte Streptomyces albidoflavus OsiLf-2 induces drought and salt tolerance in rice via a multi-level mechanism

Drought and salinity are major environmental stresses that impair crop growth and productivity worldwide. Improving drought and salt tolerance of crops with microbial mutualists is an effective and environmentally sound strategy to meet the demands of the ever-growing world population. In the present study, we found that the Streptomyces albidoflavus Osi Lf-2, a moderately salt-tolerant endophytic actinomycete, produced abundant osmolytes, including proline, polysaccharides, and ectoine. Inoculation with Osi Lf-2 increased the osmotic-adjustment ability of the rice host by increasing the proline content(by250.3% and 49.4%) and soluble sugar(by 20.9% and 49.4%) in rice under drought and salt conditions, relative to the uninoculated control. Osi Lf-2 increased stress responses in the rice host at the physiological and biochemical levels(photosynthesis efficiency, osmolytes and antioxidant content), and the gene level(osmolytes synthesis, stress-responsive and ion-transport related genes), raising rice yields under both greenhouse and saline–alkaline soil conditions. The use of endophytic actinomycetes offers a promising biotechnological approach to developing stress-tolerant plants.