Alkaline Stress Research Articles

Advancements and prospects of CRISPR/Cas9 technologies for abiotic and biotic stresses in sugar beet.

Sugar beet is a crop with high sucrose content, known for sugar production and recently being considered as an emerging raw material for bioethanol production. This crop is also utilized as cattle feed, mainly when animal green fodder is scarce. Bioethanol and hydrogen gas production from this crop is an essential source of clean energy. Environmental stresses (abiotic/biotic) severely affect the productivity of this crop. Over the past few decades, the molecular mechanisms of biotic and abiotic stress responses in sugar beet have been investigated using next-generation sequencing, gene editing/silencing, and over-expression approaches. This information can be efficiently utilized through CRISPR/Cas 9 technology to mitigate the effects of abiotic and biotic stresses in sugar beet cultivation. This review highlights the potential use of CRISPR/Cas 9 technology for abiotic and biotic stress management in sugar beet. Beet genes known to be involved in response to alkaline, cold, and heavy metal stresses can be precisely modified via CRISPR/Cas 9 technology for enhancing sugar beet's resilience to abiotic stresses with minimal off-target effects. Similarly, CRISPR/Cas 9 technology can help generate insect-resistant sugar beet varieties by targeting susceptibility-related genes, whereas incorporating Cry1Ab and Cry1C genes may provide defense against lepidopteron insects. Overall, CRISPR/Cas 9 technology may help enhance sugar beet's adaptability to challenging environments, ensuring sustainable, high-yield production.

Exogenous Melatonin Alleviates the Inhibitory Effect of NaHCO3 on Tomato Growth by Regulating the Root pH Value and Promoting Plant Photosynthesis

Soil salinity is a severe threat to agricultural production. Most saline soils turn alkaline, increasing the soil pH and, in turn, hampering the growth and development of crops. In this study, the effects of a foliar spray of melatonin (MT; 100 μmol·L−1) on the pH of the root environment, growth of tomato seedlings, endogenous MT levels, rapid chlorophyll fluorescence induction kinetics, and key enzymes of the Calvin cycle under alkaline (60 mmol·L−1 NaHCO3) stress were studied in Riegel 87-5 tomatoes. The results revealed that the growth and photosynthesis of tomato seedlings were inhibited by increased pH in the root environment under alkali stress; however, the application of exogenous MT reduced the pH of the root environment, alleviated the inhibition of growth of tomato seedlings under alkali stress, increased the content of photosynthetic pigments, alleviated the damage of the donor and acceptor sides of the photosynthetic electron transport chain, increased the activity and efficiency of photosynthetic electron transport, and optimized the share of the light energy allocated to PSII reaction centers. Increased expression levels of Calvin-cycle enzymes, including fructose-1,6-bisphosphate aldolase (FBA), fructose-1,6-bisphosphate esterase (FBP), and phosphoglycerate kinase (PGK), led to enhanced photosynthetic performance in tomato seedlings. Exogenous MT boosted endogenous MT levels and stimulated the production and secretion of organic acids in the root system. This regulation of organic acid content reduced the environmental pH in the inter-root zone, alleviating the damage caused by alkali stress. This study indicated that the exogenous administration of MT may mediate an increase in endogenous MT levels, regulate the efficiency of photosynthesis and root pH levels, and play a crucial role in mitigating injury caused by alkali stress in tomato seedlings.

How does AT1 increase crop productivity under alkaline stress?

Alkalinity constrains crop production. Recently, Zhang et al. reported a negative regulator, Alkaline Tolerance 1 (AT1), attenuating phosphorylation of phosphatidylinositol 4,5-bisphosphate (PIP2) to block efflux of intracellular reactive oxygen species (ROS) under alkaline stress and boosting yield of cereal crops by 20-30%. However, further efforts are needed to exploit the application of AT1 in breeding alkaline-tolerant crops.

Separation and characterization of related substances of Lurasidone hydrochloride by LC-QTOF-MS techniques

In the present study, a reliable LC-QTOF-MS method was developed and employed for the separation and characterization of process-related substances and forced degradation products of Lurasidone hydrochloride. The chromatographic separation was carried out using an Agilent Poroshell 120 Bonus-RP C18 column (100 mm × 4.6 mm, 2.7 µm) and a mobile phase consisting of a gradient elution of 10 mM ammonium formate solution and methanol. The degradation studies followed the guidelines outlined in ICH Q1A (R2). It was observed that Lurasidone hydrochloride exhibited instability under photolytic, alkaline, and oxidative stress conditions, while remaining relatively stable under acidic and thermal stresses. Through positive ESI-QTOF mass spectrometric analysis, fourteen related compounds in total, including both process-related and stress degradation products, were identified based on the accurate masses of parent and product ions and calculated elemental compositions. Amongst these substances, nine had not been previously reported, and their formation mechanisms were speculated. The process-related substances were further confirmed by NMR spectra determination, and suggestions were proposed to eliminate them. This study highlights the potential for monitoring and controlling related substances during the manufacturing processes, providing valuable insights for process optimization and quality control of Lurasidone hydrochloride.

Comparative Study of the Priming Effect of Abscisic Acid on Tolerance to Saline and Alkaline Stresses in Rice Seedlings

The plant hormone abscisic acid (ABA) regulates the plant response to environmental stress; therefore, ABA priming is an effective strategy for enhancing stress tolerance in rice. In this study, we investigated the priming effects of 1 and 5 µM ABA on the biochemical and physiological traits associated with seedling growth performance in two rice cultivars exposed to saline (100 mM NaCl) and alkaline (15 mM Na2CO3) stress via root drenching. ABA pretreatment effectively reduced damage in rice seedlings by mitigating the increases in Na+/K+ ratio, membrane injury, contents of Na+, malondialdehyde, hydrogen peroxide, and superoxide anion radical, and prevented reductions in K+ and total chlorophyll contents, and ROS-related enzyme activities in both cultivars under saline and alkaline stresses. Rice seedlings with ABA pretreatment under alkaline stress had a stronger ability to maintain ion homeostasis, eliminate ROS, and induce changes in endogenous ABA levels via the upregulation of OsHKT1;5, OsSOS1, OsNHX5, OsPOX1, OsCATA, OsNCED3, OsSalT, and OsWsi18 and downregulation of OsRbohA than under saline stress. The saline–alkaline (SA)-sensitive cultivar demonstrated greater sensitivity to the priming effect of ABA than that of the SA-tolerant cultivar under both stress conditions. These findings have implications for rice adaptation to SA soils.

Genetic Variability and Association Analysis for Yield and its Components in Alkaline and Inland Saline Stress in Rice (Oryza sativa L.)

The experiment was conducted at Agricultural Research Station, Kampasagar, Nalgonda, Telangana, India during the rabi season (December, 2020 to May, 2021) to identify suitable lines tolerant to alkalinity and inland salinity stress in rice. Analysis of variance evinced plenteous quantum of variability in all the 36 genotypes investigated. Phenotypic coefficients of variation were higher than genotypic coefficients demonstrating the effect of the prevalent stress environment. High estimates of genotypic coefficient of variation, phenotypic coefficient of variation, heritability, and genetic advance as percent mean was observed for the traits, seedling mortality, number of filled grains panicle-1 and sterility %. These findings suggested the preponderance of additive gene action for these characters. The remaining traits were mostly under the influence of non-additive gene effects as they recorded low to moderate estimates of genetic advance. Studies from association analysis showed that number of productive tillers hill-1 and number of filled grains panicle-1 had a positive significant correlation with yield whereas, seedling mortality and sterility % had a negative significant association with yield. Path analysis revealed that traits number of productive tillers hill-1, number of grains panicle-1 and 1000 grain weight had a positive direct effect on yield. Hence primacy should be given to the above-mentioned traits to develop alkalinity and inland saline tolerant and high yielding rice cultivars. Lines CT 118911, Sahel 177, M 202 and KPS 10654 were found to be promising entries and can be used tolerant parents in the breeding programme. Three principal components combined could explain 68.86% of the total variation.

How do plants maintain pH and ion homeostasis under saline-alkali stress?

Salt and alkaline stresses often occur together, severely threatening plant growth and crop yields. Salt stress induces osmotic stress, ionic stress, and secondary stresses, such as oxidative stress. Plants under saline-alkali stress must develop suitable mechanisms for adapting to the combined stress. Sustained plant growth requires maintenance of ion and pH homeostasis. In this review, we focus on the mechanisms of ion and pH homeostasis in plant cells under saline-alkali stress, including regulation of ion sensing, ion uptake, ion exclusion, ion sequestration, and ion redistribution among organs by long-distance transport. We also discuss outstanding questions in this field.

An insight into variability and association among yield and its components under alkalinity and inland salinity stress in rice (Oryza sativa L.)

An insight into variability and association among yield and its components under alkalinity and inland salinity stress in rice (Oryza sativa L.)

Characterization and phylogenetic analysis of the complete mitochondrial genome of a unique economic plant <i>Lycium barbarum</i> L.

<i>Lycium barbarum</i> L. (<i>L. barbarum</i>), an important medicinal and food crop from the Solanaceae family, is known for excellent tolerance to saline–alkaline and drought stresses. Therefore, it has significant research and application value. The nuclear and chloroplast genomes of <i>L. barbarum</i> have been previously reported; however, its mitochondrial genome is still unexplored. In this study, mitochondrial DNA sequencing and assembly in <i>L. barbarum</i> were studied via second- and third-generation sequencing technologies using the anthers collected from the main cultivars “Ningqi No.1” (fertile line) and its male-sterile natural mutant “Ningqi No.5” from Ningxia, China. The results revealed that “Ningqi No.1” and “Ningqi No.5” have the same mitochondrial circular genome structure with a total length of 413,881 bp. In total, 61 genes were annotated, including 35 protein-coding genes, four rRNA genes, 22 tRNA genes, and 2,242 open reading frames with unknown functions. The repeat sequences in mtDNA were analyzed, in which short repeats (30–59 bp) exhibited the highest number. Overall, 23 protein-coding genes in “Ningqi No.1” and “Ningqi No.5” exhibited RNA-editing phenomenon, with different RNA-editing site number and distribution position. Phylogenetic analysis demonstrated that <i>L. barbarum</i> has the closest relationship with <i>Hyoscyamus niger</i>. These results provided a basis for a comprehensive understanding of the mitochondrial genome of <i>L. barbarum</i> and the relevant data for omics studies on <i>L. barbarum</i>.

Paclobutrazol Improves the Chlorophyll Content and Antioxidant Activities of Red Rice in Response to Alkaline Stress

Paclobutrazol Improves the Chlorophyll Content and Antioxidant Activities of Red Rice in Response to Alkaline Stress

Enhancement of sulfur metabolism and antioxidant machinery confers Bacillus sp. Jrh14-10–induced alkaline stress tolerance in plant

Alkaline stress is a major environmental challenge that restricts plant growth and agricultural productivity worldwide. Plant growth-promoting rhizobacteria (PGPR) can be used to effectively enhance plant abiotic stress in an environment-friendly manner. However, PGPR that can enhance alkalinity tolerance are not well-studied and the mechanisms by which they exert beneficial effects remain elusive. In this study, we isolated Jrh14-10from the rhizosphere soil of halophyte Halerpestes cymbalaria (Pursh) Green and found that it can produce indole-3-acetic acid (IAA) and siderophore. By 16S rRNA gene sequencing, it was classified as Bacillus licheniformis. Inoculation Arabidopsis seedlings with Jrh14-10 significantly increased the total fresh weight (by 148.1%), primary root elongation (by 1121.7%), and lateral root number (by 108.8%) under alkaline stress. RNA-Seq analysis showed that 3389 genes were up-regulated by inoculation under alkaline stress and they were associated with sulfur metabolism, photosynthetic system, and oxidative stress response. Significantly, the levels of Cys and GSH were increased by 144.3% and 48.7%, respectively, in the inoculation group compared to the control under alkaline stress. Furthermore, Jrh14-10 markedly enhanced the activities of antioxidant enzymes, resulting in lower levels of O2•–, H2O2, and MDA as well as higher levels of Fv/Fm in alkaline-treated seedlings. In summary, Jrh14-10 can improve alkaline stress resistance in seedlings which was accompanied by an increase in sulfur metabolism-mediated GSH synthesis and antioxidant enzyme activities. These results provide a mechanistic understanding of the interactions between a beneficial bacterial strain and plants under alkaline stress.

Examination of the relationship of carbonate alkalinity stress and ammonia metabolism disorder-mediated apoptosis in the Chinese mitten crab, Eriocheir sinensis: Potential involvement of the ROS/MAPK signaling pathway

The current global status of freshwater resources and deterioration of water quality in general have led to limitations on the scope of freshwater aquaculture. The development of technology that would enable aquaculture under saline-alkaline conditions would be valuable. Therefore, the exploration is needed to understand the damage mechanism of saline-alkaline water, particularly the impact of alkalinity, on fish and shellfish. In this study, Chinese mitten crabs (Eriocheir sinensis) were subjected to different concentrations of carbonate alkalinity (0.00, 4.38, 8.75, 17.50, and 35.00 mmol/L) for 96 h. The survival rates in the 17.50 and 35.00 mmol/L alkalinity groups showed a significant decrease compared to the control group and the groups exposed to lower carbonate alkalinity concentrations (4.38 and 8.75 mmol/L). The exposure to carbonate alkalinity had a significant inhibitory effect on the ammonia excretion rate and led to an increase of ammonia content in hemolymph. However, in contrast to the low carbonate alkalinity exposure groups (4.38 and 8.75 mmol/L), the high carbonate alkalinity exposure group (35.00 mmol/L) exhibited a significant reduction in the concentrations of ammonia and urea nitrogen in hemolymph. Moreover, high carbonate alkalinity exposure was found to induce oxidative stress, as indicated by increased levels of reactive oxygen species (ROS) and malondialdehyde. Additionally, there were notable alterations observed in the activities of antioxidant enzymes such as superoxide dismutase, catalase, and glutathione. Flow cytometry analysis and ultrastructural observations revealed that exposure to carbonate alkalinity can induce apoptosis. In the 8.75, 17.50, and 35.00 mmol/L alkalinity groups, the activation of p38 and JNK pathways was observed, along with an increase in the Bax/Bcl-2 ratio and the expression of caspase 3. In summary, the findings of this study suggested that exposure to carbonate alkalinity can disrupt ammonia metabolism, leading to apoptosis through the activation of the ROS/MAPK signaling pathway. This ultimately results in a decrease in the survival rate of crabs. These results provide valuable insights into the toxicological effects of high carbonate alkalinity exposure and shed light on the tolerance mechanisms of high carbonate alkalinity exposure in crabs.

GhIMP10D, an inositol monophosphates family gene, enhances ascorbic acid and antioxidant enzyme activities to confer alkaline tolerance in Gossypium hirsutum L.

BackgroundInositol monophosphates (IMP) are key enzymes in the ascorbic acid (AsA) synthesis pathways, which play vital roles in regulating plant growth and development and stresses tolerance. To date, no comprehensive analysis of the expression profile of IMP genes and their functions under abiotic stress in cotton has been reported.ResultsIn this study, the genetic characteristics, phylogenetic evolution, cis-acting elements and expression patterns of IMP gene family in cotton were systematically analyzed. A total of 28, 27, 13 and 13 IMP genes were identified in Gossypium hirsutum (G. hirsutum), Gossypium barbadense (G. barbadense), Gossypium arboreum (G. arboreum), and Gossypium raimondii (G. raimondii), respectively. Phylogenetic analysis showed that IMP family genes could cluster into 3 clades. Structure analysis of genes showed that GhIMP genes from the same subgroup had similar genetic structure and exon number. And most GhIMP family members contained hormone-related elements (abscisic acid response element, MeJA response element, gibberellin response element) and stress-related elements (low temperature response element, defense and stress response element, wound response element). After exogenous application of abscisic acid (ABA), some GhIMP genes containing ABA response elements positively responded to alkaline stress, indicating that ABA response elements played an important role in response to alkaline stress. qRT-PCR showed that most of GhIMP genes responded positively to alkaline stress, and GhIMP10D significantly upregulated under alkaline stress, with the highest up-regulated expression level. Virus-induced gene silencing (VIGS) experiment showed that compared with 156 plants, MDA content of pYL156:GhIMP10D plants increased significantly, while POD, SOD, chlorophyII and AsA content decreased significantly.ConclusionsThis study provides a thorough overview of the IMP gene family and presents a new perspective on the evolution of this gene family. In particular, some IMP genes may be involved in alkaline stress tolerance regulation, and GhIMP10D showed high expression levels in leaves, stems and roots under alkaline stress, and preliminary functional verification of GhIMP10D gene suggested that it may regulate tolerance to alkaline stress by regulating the activity of antioxidant enzymes and the content of AsA. This study contributes to the subsequent broader discussion of the structure and alkaline resistance of IMP genes in cotton.

The influence of stress factors on selected phenotypic and genotypic features of Listeria monocytogenes – a pilot study

BackgroundListeria monocytogenes are Gram-positive rods, widespread in the environment due to their wide tolerance to changing conditions. The apilot study aimed to assess the impact of six various stresses (heat, cold, osmotic, acid, alkali, frozen) on phenotypic features: MIC of antibiotics (penicillin, ampicillin, meropenem, erythromycin, co-trimoxazole; gradient stripes), motility, ability to form a biofilm (crystal violet method) and growth rate (OD and quantitative method), expression level of sigB (stress induced regulator of genes), agrA, agrB (associated with biofilm formation) and lmo2230, lmo0596 (acid and alkali stress) (qPCR) for three strains of L. monocytogenes.ResultsApplied stress conditions contributed to changes in phenotypic features and expression levels of sigB, agrA, agrB, lmo2230 and lmo0596. Stress exposure increased MIC value for penicillin (ATCC 19111 - alkaline stress), ampicillin (472CC - osmotic, acid, alkaline stress), meropenem (strains: 55 C - acid, alkaline, o smotic, frozen stress; 472CC - acid, alkaline stress), erythromycin (strains: 55 C - acid stress; 472CC - acid, alkaline, osmotic stress; ATCC 19111 - osmotic, acid, alkaline, frozen stress), co-trimoxazole (strains: 55 C - acid stress; ATCC 19111 - osmotic, acid, alkaline stress). These changes, however, did not affect antibiotic susceptibility. The strain 472CC (a moderate biofilm former) increased biofilm production after exposure to all stress factors except heat and acid. The ATCC 19111 (a weak producer) formed moderate biofilm under all studied conditions except cold and frozen stress, respectively. The strain 55 C became a strong biofilm producer after exposure to cold and produced a weak biofilm in response to frozen stress. Three tested strains had lower growth rate (compared to the no stress variant) after exposure to heat stress. It has been found that the sigB transcript level increased under alkaline (472CC) stress and the agrB expression increased under cold, osmotic (55 C, 472CC), alkali and frozen (472CC) stress. In contrast, sigB transcript level decreased in response to acid and frozen stress (55 C), lmo2230 transcript level after exposure to acid and alkali stress (ATCC 19111), and lmo0596 transcript level after exposure to acid stress (ATCC 19111).ConclusionsEnvironmental stress changes the ability to form a biofilm and the MIC values of antibiotics and affect the level of expression of selected genes, which may increase the survival and virulence of L. monocytogenes. Further research on a large L. monocytogenes population is needed to assess the molecular mechanism responsible for the correlation of antibiotic resistance, biofilm formation and resistance to stress factors.

Converting waste to resource: Utilisation of elemental S and consortia of halo–alkaliphilic S–oxidisers for preparation of acidified compost from organic wastes

Utilising the acidulated manures is a reliable option for plant nutrition and managing alkalinity stress. This study characterizes the common wastes and their acidulation potential through consortia of microbes like Priestia aryabhattai, Stutzerimonas stutzeri and Pseudomonas chloritidismutans and elemental sulphur (S°). The poultry manure (PM) and pressmud (Pmud) were rich in N, P, and phenol with a narrow C/N ratio. The water–soluble total N (TNsol), dissolved organic N (DON), NO3−–N was greater in cattle manure. Acidulation of manure declined pH by 0.5–2.5 units and increased titratable acidity (TA) and equivalent alkalinity neutralisation potential (EANP) by 5034.8 and 2,805,732.3 me kg−1, respectively. The incubation for 28 days was optimum for acidulation. The water–soluble SO42−, Mg2+, and DOC were the key variables explaining variability in EANP. The rapid acidulation of manure using microbial consortia and S° could be a pragmatic approach for recycling and reusing the waste.

Genome-Wide Characterization of Soybean Hexokinase Genes Reveals a Positive Role of GmHXK15 in Alkali Stress Response.

Hexokinase (HXK) proteins catalyze hexose phosphorylation and are important for the sensing and signaling of sugar. In order to determine the roles played by HXKs in soybean growth and stress responsiveness, seventeen HXK genes (GmHXK1-17) were isolated and analyzed. The phylogenic analysis and subcellular location prediction showed that GmHXKs were clearly classified into type A (GmHXK1-4) and type B (GmHXK5-17). There were similar protein structures and conserved regions in GmHXKs to the HXKs of other plants. An expression analysis of the GmHXK genes in soybean organs or tissues demonstrated that GmHXK3 and GmHXK12, 15, and 16 were the dominant HXKs in all the examined tissues. In addition, salt, osmotic, and alkaline stress treatments dramatically increased the activity and transcripts of GmHXKs. There is the possibility that a type-B isoform (GmHXK15) plays a crucial role in soybean adaptation to alkali, as the expression levels of this isoform correlate well with the HXK enzyme activity. Based on an enzyme assay performed on recombinant plant HXK15 proteins expressed in Escherichia coli, we found that GmHXK15 had functional HXK activities. A further analysis indicated that GmHXK15 specifically targeted the mitochondria, and the overexpression of the GmHXK15 gene could significantly enhance the resistance of transgenic soybean to alkali stress. The present findings will serve as a basis for a further analysis of the function of the GmHXK gene family.

Ochrobactrum sp. NBRISH6 Inoculation Enhances Zea mays Productivity, Mitigating Soil Alkalinity and Plant Immune Response.

Intensifying sodic land characterized by high alkaline pH is an incipient environmental hazard-limiting agricultural potential. In this study, we investigated the effects of plant growth-promoting bacteria Ochrobactrum sp. strain NBRISH6 on the growth and physiology of maize (Zea mays L.) grown under alkaline stress at two soil pH levels. Additionally, we also studied the effects of NBRISH6 on soil fertility parameters. A greenhouse experiment was designed using two live soils (pH 8.2 and 10.2) in earthen pots using maize as a host. Results revealed a significant increase in plant growth and a decrease in defense enzymes in both soil types due to NBRISH6 inoculation as compared to non-treated control. Furthermore, activities of all soil enzymes along with bacterial diversity increased in NBRISH6 treatment under normal as well as stressed conditions. In addition, field evaluation of NBRISH6 inoculation using maize was carried out under normal and alkaline conditions, which resulted in significant enhancement of all vegetative parameters as compared to respective controls. Therefore, the study suggested that Ochrobactrum sp. NBRISH6 can be used to develop a bioinoculant formulation to ameliorate abiotic stresses and enhanced crop productivity.

Characterization of structure and antioxidant activity of exopolysaccharides from endophytic Lysinibacillus sphaericus Ya6 under acid-base stress

In this study, the structure and antioxidant activity of exopolysaccharides (EPS) produced by Lysinibacillus sphaericus Ya6 under different acid-base stresses were investigated. The structure of EPS was analyzed by UV-visible and mid-IR, and the monosaccharide composition and the molecular weight were determined by High-performance liquid chromatography (HPLC) and high-performance gel permeation chromatography respectively. The findings indicated that EPS produced by Lysinibacillus sphaericus Ya6 was a homogenous polysaccharide with a molecular weight of 79.25 kDa that was composed of mannose and glucose. The molecular weight and monosaccharide composition of polysaccharides changed under acid-base stress conditions. With the addition of the acid-base stress level, the logarithmic growth period of Lysinibacillus sphaericus Ya6 was prolonged for a short while, but later growth rates and EPS production under stress conditions were all higher than under the control. In vitro determinations found that acid-base stress boosted the antioxidant activity of EPS. Particularly under conditions of alkaline stress, DPPH radical scavenging capacity and superoxide anion scavenging of EPS were higher than those of the control, and their capacities to scavenge DPPH was increased from 39.30% (the control) to 95.65% at pH 10.0, which was close to ascorbic acid's (95.77%) as a positive control.

GsPKS24, a calcineurin B-like protein-interacting protein kinase gene from Glycine soja, positively regulates tolerance to pH stress and ABA signal transduction.

SOS2-like protein kinases (PKS/CIPK) family genes are known to be involved in various abiotic stresses in plants. Even though, its functions have been well characterized under salt and drought stresses. The roles of PKS genes associated with alkaline stress response are not fully established yet. In this study, we identified 56 PKS family genes which could be mainly classified into three groups in wild soybean (Glycine soja). PKS family genes transcript profiles revealed different expression patterns under alkali stress. Furthermore, we confirmed the regulatory roles of GsPKS24 in response to NaHCO3, pH and ABA treatments. Overexpression of GsPKS24 enhanced plant tolerance to pH stress in Arabidopsis and soybean hairy roots but conferred suppressed pH tolerance in Arabidopsis atpks mutant. Additionally, Overexpression of GsPKS24 decreased the ABA sensitivity compared to Arabidopsis atpks mutant which displayed more sensitivity towards ABA. Moreover, upregulated expression of stress responsive and ABA signal-related genes were detected in GsPKS24 overexpression lines. In conclusion, we identified the wild soybean PKS family genes, and explored the roles of GsPKS24 in positive response to pH stress tolerance, and in alleviation of ABA sensitivity.

Combined analysis of mRNA and miRNA reveals the mechanism of pacific white shrimp (Litopenaeus vannamei) under acute alkalinity stress.

The pacific white shrimp (Litopenaeus vannamei) is now a more common aquaculture species in saline-alkali waters, while alkalinity stress is considered to be one of the stressors for shrimp. Thus, an understanding of the molecular response to alkalinity stress is critical for advancing the sustainability of culture in pacific white shrimp. In this study, we aimed to explore the response mechanism to acute high-alkaline stress by RNA-seq at low-alkaline (50 mg/L) and high-alkaline (350 mg/L). We identified 215 differentially expressed mRNAs (DEGs) and 35 differentially expressed miRNAs (DEMs), of which 180 DEGs and 28 DEMs were up-regulated, 35 DEGs and 7 DEMs were down-regulated, respectively. The DEGs were enriched in several pathways, including carbohydrate digestion and absorption, pancreatic secretion, starch and sucrose metabolism, antigen processing and presentation and glutathione metabolism. The DEMs involved in lysosome and ion transport related pathways were significantly up-regulated. We also achieved 42 DEGs, which were targeted by DEMs. miRNA-mRNA regulatory network was constructed by integrated analysis of miRNA-mRNA data. We detected several genes and miRNAs which were identified as candidate regulators of alkalinity stress, and expression patterns of key genes related to alkalinity stress in pacific white shrimp. Among these genes, the expression levels of most key genes enriched in ion regulation, digestion and immunity were increased, and the expression levels of genes enriched in metabolism were down-regulated. This research indicated that the homeostatic regulation and digestion changed significantly under acute alkaline stress, and the variations from metabolic and immunity can cope with the osmotic shock of alkalinity stress in pacific white shrimp. This study provides key clues for exploring the molecular mechanism of pacific white shrimp under acute alkalinity stress, and also gives scientific basis for the optimisation of saline-alkaline aquaculture technology.