High pH Stress Research Articles

Recent advances in protein precipitation-based methods for drug-target screening

Drug targets are biological macromolecules that bind drug molecules in vivo. Therefore, the system-wide identification of drug targets plays a vital role in fully understanding the mechanism of drug action, efficacy, and side effects. The unbiased screening of drug targets may accelerate the process of drug discovery and candidate screening. Mass spectrometry is a key tool for large-scale protein identification and accurate quantification owing to its high acquisition speed, resolution, and sensitivity. Mass spectrometry-based proteomics has been widely used for drug-target screening. It can systematically identify the protein-target landscape of a drug and elucidate drug-protein interactions. Commonly used drug-target characterization methods, such as labeling-based affinity enrichment, require the chemical derivatization of drug molecules, which is not only time-consuming but may also affect the affinity of the drug towards its targets. Furthermore, the spatial effects of the derivatization groups may block interactions between the drug and its targets. Considering the disadvantages of affinity-enrichment methods, strategies that do not require chemical derivatization have received widespread attention. Proteins may undergo denaturation, unfolding, and precipitation under different conditions such as high temperatures, extreme pH, denaturants, and mechanical stress. Binding to small-molecule drugs may alter the folding balance of target proteins. The conformational stability of target proteins can be stabilized by binding with drugs, and protein-drug complexes are more resistant than free proteins to the precipitation induced by different conditions. Based on this mechanism, various large-scale drug-target identification methods using protein precipitation have been developed by combining proteomics and mass spectrometry analysis, including thermal proteome profiling and solvent-, mechanical stress-, and pH-induced protein precipitation. These methods have been successfully applied to the characterization of small-molecule drug targets. In this review, we describe the protein precipitation-based methods used for the high-throughput discovery of drug targets and elucidation of the interactions between drugs and proteins in the past decade. We also summarize the characteristics of each method and discuss their application potential in drug-efficacy evaluation and drug discovery.

Physiological Adaptation of Fenneropenaeus chinensis in Response to Saline-Alkaline Stress Revealed by a Combined Proteomics and Metabolomics Method.

The rapid development of the mariculture industry has been hindered by limited coastal aquaculture space. To utilize the abundant inland saline-alkaline water, we studied the physiological effects of high carbonate alkalinity stress and high pH stress on Fenneropenaeus chinensis. The study employed quantitative proteomics by tandem mass tag (TMT) and non-targeted metabolomics analysis using a liquid chromatograph mass spectrometer (LC-MS) to understand the physiological and biochemical adaptive mechanisms of the hepatopancreas of F. chinensis in response to saline-alkaline stress at the molecular level. We designed two stress groups as follows: a high carbonate alkalinity (CA) group and a combined high carbonate alkalinity and high pH (CP) group. The study found that the protein and metabolic profiles of the two stress groups were changed, and the CP group, which was exposed to dual stresses, incurred more severe damage to the hepatopancreas compared to that of the CA group. After exposure to CA and CP, the hepatopancreas of F. chinensis showed significant alterations in 455 proteins and 50 metabolites, and 1988 proteins and 272 metabolites, respectively. In addition, F. chinensis upregulated the level of energy metabolism in the hepatopancreas to defend against osmotic imbalance caused by CA or CP stress, which was demonstrated by the significant upregulation of important proteins and metabolites in glycolysis, pyruvate metabolism, TCA cycle, and fatty acid oxidation. Additionally, pattern recognition receptors, the phenol oxidase system, and various immune-related metabolic enzymes and metabolites were also affected. The immune homeostasis of F. chinensis was affected by the alteration of the antioxidant system following exposure to CA or CP. These findings provide valuable information for F. chinensis saline-alkaline water cultivation practices.

Regulation of Root Exudation in Wheat Plants in Response to Alkali Stress.

Soil alkalization is an important environmental factor limiting crop production. Despite the importance of root secretion in the response of plants to alkali stress, the regulatory mechanism is unclear. In this study, we applied a widely targeted metabolomics approach using a local MS/MS data library constructed with authentic standards to identify and quantify root exudates of wheat under salt and alkali stresses. The regulatory mechanism of root secretion in alkali-stressed wheat plants was analyzed by determining transcriptional and metabolic responses. Our primary focus was alkali stress-induced secreted metabolites (AISMs) that showed a higher secretion rate in alkali-stressed plants than in control and salt-stressed plants. This secretion was mainly induced by high-pH stress. We discovered 55 AISMs containing -COOH groups, including 23 fatty acids, 4 amino acids, 1 amino acid derivative, 7 dipeptides, 5 organic acids, 9 phenolic acids, and 6 others. In the roots, we also discovered 29 metabolites with higher levels under alkali stress than under control and salt stress conditions, including 2 fatty acids, 3 amino acid derivatives, 1 dipeptide, 2 organic acids, and 11 phenolic acids. These alkali stress-induced accumulated carboxylic acids may support continuous root secretion during the response of wheat plants to alkali stress. In the roots, RNAseq analysis indicated that 5 6-phosphofructokinase (glycolysis rate-limiting enzyme) genes, 16 key fatty acid synthesis genes, and 122 phenolic acid synthesis genes have higher expression levels under alkali stress than under control and salt stress conditions. We propose that the secretion of multiple types of metabolites with a -COOH group is an important pH regulation strategy for alkali-stressed wheat plants. Enhanced glycolysis, fatty acid synthesis, and phenolic acid synthesis will provide more energy and substrates for root secretion during the response of wheat to alkali stress.

Functional Differentiation of the Duplicated Gene BrrCIPK9 in Turnip (Brassica rapa var. rapa).

Gene duplication is a key biological process in the evolutionary history of plants and an important driving force for the diversification of genomic and genetic systems. Interactions between the calcium sensor calcineurin B-like protein (CBL) and its target, CBL-interacting protein kinase (CIPK), play important roles in the plant's response to various environmental stresses. As a food crop with important economic and research value, turnip (Brassica rapa var. rapa) has been well adapted to the environment of the Tibetan Plateau and become a traditional crop in the region. The BrrCIPK9 gene in turnip has not been characterized. In this study, two duplicated genes, BrrCIPK9.1 and BrrCIPK9.2, were screened from the turnip genome. Based on the phylogenetic analysis, BrrCIPK9.1 and BrrCIPK9.2 were found located in different sub-branches on the phylogenetic tree. Real-time fluorescence quantitative PCR analyses revealed their differential expression levels between the leaves and roots and in response to various stress treatments. The differences in their interactions with BrrCBLs were also revealed by yeast two-hybrid analyses. The results indicate that BrrCIPK9.1 and BrrCIPK9.2 have undergone Asparagine-alanine-phenylalanine (NAF) site divergence during turnip evolution, which has resulted in functional differences between them. Furthermore, BrrCIPK9.1 responded to high-pH (pH 8.5) stress, while BrrCIPK9.2 retained its ancestral function (low K+), thus providing further evidence of their functional divergence. These functional divergence genes facilitate turnip's good adaptation to the extreme environment of the Tibetan Plateau. In summary, the results of this study reveal the characteristics of the duplicated BrrCIPK9 genes and provide a basis for further functional studies of BrrCBLs-BrrCIPKs in turnip.

Morphology, Development, and Pigment Production of Talaromyces marneffei are Diversely Modulated Under Physiologically Relevant Growth Conditions.

Talaromyces marneffei is an opportunistic pathogenic fungus that mainly affects HIV-positive individuals endemic to Southeast Asia and China. Increasing efforts have been made in the pathogenic mechanism and host interactions understanding of this pathogen in the last two decades; however, there are still no conclusions on how T. marneffei was transmitted from the donor bamboo rats to humans. A perception that the failure of fungus isolation from soil was attributed to the low salt tolerance of T. marneffei. Therefore, the effect of environmental fluctuations in fungal growth and development is fundamental for the characterization of its origin and fungal biology understanding. Herein, we characterized high osmolarity, pH, metal ions, nutrients, and oxidative stress have versatile effects on T. marneffei hyphal or yeast growth, conidia generation, and pigment production. Among these, high pH, low glucose amounts, and the inorganic nitrogen ammonium tartrate stimulated the red pigment production, whereas high osmolarity, high pH, and the inorganic nitrogen sodium nitrate could significantly accelerate the conidia generation. Specifically, zinc starvation repressed conidia generation and prevented the wrinkled yeast colony formation, indicating the function of zinc regulators in pathogenicity regulation. Since conidia are recognized as the infectious propagules, the effects characterization of different environmental factors in T. marneffei morphology in this work will not only expand the growth and pathogenic biology understanding of the fungus but also provide more clues for the T. marneffei infection transmission origin investigation.

Multifunctional role of dietary copper to regulate stress-responsive gene for mitigation of multiple stresses in Pangasianodon hypophthalmus

It is an urgent needs to address climate change and pollution in aquatic systems using suitable mitigation measures to avoid the aquatic animals' extinction. The vulnerability and extinction of the aquatic animals in the current scenario must be addressed to enhance safe fish food production. Taking into consideration of such issues in fisheries and aquaculture, an experiment was designed to mitigate high temperature (T) and low pH stress, as well as arsenic (As) pollution in fish using copper (Cu) containing diets. In the present investigation, the Cu-containing diets graded with 0, 4, 8, and 12 mg kg-1 were prepared and fed to Pangasianodon hypophthalmus reared under As, low pH, and high-temperature stress. The gene expression was highly affected in terms of the primary, secondary, and tertiary stress response, whereas supplementation of Cu-containing diet mitigates the stress response. Oxidative stress genes such as catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) were significantly upregulated by stressors (As, As + T, and As + pH + T). Whereas, heat shock protein (HSP 70), inducible nitric oxide synthase (iNOS), metallothionine (MT), caspase 3a (Cas 3a), and cytochrome P450 (CYP 450) were highly upregulated by stressors, while dietary Cu at 8 mg kg-1 diet significantly downregulated these gene expressions. Indeed, the immunity-related genes viz. TNFα, Ig, TLR, and immune-related attributes viz. albumin, globulin, total protein, A:G ratio, blood glucose, NBT, and myeloperoxidase (MPO) were also improved with Cu-containing diets. Cu containing diets substantially improved neurotransmitter enzyme (AChE) and vitamin C (Vit C). DNA damage was also reduced with supplementation of Cu at 8 mg kg-1 diet. The growth index viz. final body weight gain (%), specific growth rate, protein efficiency ratio, food conversion ratio, relative feed intake, and daily growth index were noticeably enhanced by Cu diets (4 and 8 mg kg-1 diet). The growth-related genes expressions viz. growth hormone (GH), growth hormone regulator 1 (Ghr1), growth hormone regulator β (Ghrβ,) myostatin (MYST), and somatostatin (SMT) supported the growth enhancement with Cu at 8 mg kg-1 diet. The bioaccumulation of As was reduced with Cu-containing diets. The fish were infected with Aeromonas hydrophila at the end of the 105 days experimental trial. Cu at 8 mg kg-1 diet improved immunity, reduced the cumulative mortality, and enhanced the relative percentage survival of the fish. The results revealed that the innovative Cu diets could reduce the extinction of the fish against climate change and pollution era and produce the safest production that is safe to humans for consumption.

Comparison of stress tolerance of hybrid and selfed offspring of two populations of Litopenaeus vannamei

In recent years, as the marine environment has been deteriorated, the aquaculture water environment has also been negatively affected to varying degrees. Negative environmental factors make extremely damaging to organisms, resulting in stress-induced diseases and high mortality rates of cultured shrimp. Therefore, stress resistance breeding of Litopenaeus vannamei and evaluating the stress tolerance of the breeding population are urgently needed now. Litopenaeus vannamei collected from Thailand (T) and the United States (M) were used as parents, while four progeny populations of TT, MM, TM, and MT were constructed by diallel cross, with a total of 20 families. The tolerance of young shrimp to high ammonia-N, high pH, and low salt was compared through a 96-h acute toxicity test; the heterosis of each mating combination was analyzed; and the tolerance of parents and offspring was evaluated. Here we show that under 96 h of high ammonia-N, high pH, and low salt stress, the mortality rates of each family were 19.52%–92.22%, 23.33%–92.22%, and 19.33%–80.00%, respectively. There were significant differences in the tolerance of different families to ammonia-N, pH, and salinity stress (P < 0.05). The population with a female parent from the United States had stronger tolerance to ammonia-N, pH, and low salt stress than the population with a female parent from Thailand. The population with a male parent from Thailand had weaker tolerance to pH and low salt stress than the population with a male parent from the United States, but it was superior to the population with a male parent from the United States under ammonia-N stress. The heterosis rates of the hybrid population TM in acute high ammonia-N, high pH and low salinity were 81.67%, 44.58% and −10.13%, respectively; However, the heterosis rates of the MT population were 14.89%, 38.89%, and −8.96%, respectively. The overall resistance of the four populations showed MM > MT > TT > TM. The population TM had obvious heterosis in high ammonia-N and high pH tolerance traits, and the family MM7 had a strong low salt tolerance, so it can be considered a candidate family for subsequent breeding work. Moreover, variability in stress resistance and heterosis of stress resistance of different populations obtained by family selection was discussed in this paper. The experimental results provide a basis for screening new strains of vannamei shrimp with strong stress resistance through family breeding.

Benefits of biological nitrification inhibition of Leymus chinensis under alkaline stress: the regulatory function of ammonium-N exceeds its nutritional function.

The production of root exudates with biological nitrification inhibition (BNI) effects is a strategy adopted by ammonium-N () tolerant plant species that occur in N-limited environments. Most knowledge on BNI comes from plant species that occur in acidic soils. Here, combining field sampling and laboratory culture, we assessed the BNI-capacity of Leymus chinensis, a dominant grass species in alkaline grasslands in eastern Asia, and explored why L. chinensis has BNI ability. The results showed that L. chinensis has strong BNI-capacity. At a concentration of 1 mg mL-1, L. chinensis' root exudates inhibited nitrification in soils influenced by Puccinellia tenuiflora by 72.44%, while DCD only inhibited it by 68.29%. The nitrification potential of the soil of L. chinensis community was only 53% of the P. tenuiflora or 41% of the Suaeda salsa community. We also showed that the supply of driven by L. chinensis' BNI can meet its requirements . In addition, can enhance plant adaptation to alkaline stress by regulating pH, and in turn, the uptake of nitrate-N (). We further demonstrated that the regulatory function of is greater than its nutritional function in alkaline environment. The results offer novel insights into how L. chinensis adapts to high pH and nutrient deficiency stress by secreting BNIs, and reveal, for the first time, differences in the functional roles of and in growth and adaptation under alkaline conditions in a grass species.

Identification of key immune and stress related genes and pathways by comparative analysis of the gene expression profile under multiple environmental stressors in pacific white shrimp (Litopenaeus vannamei)

Water salinity, pH, and nitrite concentration are considered environmental factors affecting the growth rate, survival, health, and physiological conditions of aquatic animals. The identification of key genes that are involved in the response to environmental stressors is essential for controlling stress in aquatic animals and sustainable aquaculture. In this study, RNA sequencing was performed to identify the differentially expressed genes (DEGs) and biological pathways that are involved in the response of the hepatopancreas to environmental stressors, including low salinity stress, nitrite stress, low pH stress, and high pH stress. The DEGs were enriched in biological pathways related to immune response, energy metabolism, oxidative stress response, hemostasis, and enzymatic activity of the hepatopancreas. In addition to the identification of DEGs related to each stressor, some DEGs were found to be expressed among all groups. The most important overlapping DEGs under multiple stressors were juvenile hormone esterase-like protein 2 (JHE-like), myosin light chain, C-type lectin 2, myosin-9-like, anti-lipopolysaccharide factor 1 (ALF-1), peroxisomal acyl-coenzyme An oxidase 1-like (ACX1), hepatic lectin-like, venom phosphodiesterase 2-like, hemolymph clottable protein-like (CP), cathepsin L, and Ras-like protein 2. The results of the present study provide additional information regarding the transcriptional response of the hepatopancreas to low salinity, nitrite, low pH, and high pH stress. Moreover, the discovery of several overlapping DEGs among different stressors provided a better understanding of the molecular function of the hepatopancreas.

New Insight into Plant Saline-Alkali Tolerance Mechanisms and Application to Breeding.

Saline-alkali stress is a widespread adversity that severely affects plant growth and productivity. Saline-alkaline soils are characterized by high salt content and high pH values, which simultaneously cause combined damage from osmotic stress, ionic toxicity, high pH and HCO3-/CO32- stress. In recent years, many determinants of salt tolerance have been identified and their regulatory mechanisms are fairly well understood. However, the mechanism by which plants respond to comprehensive saline-alkali stress remains largely unknown. This review summarizes recent advances in the physiological, biochemical and molecular mechanisms of plants tolerance to salinity or salt- alkali stress. Focused on the progress made in elucidating the regulation mechanisms adopted by plants in response to saline-alkali stress and present some new views on the understanding of plants in the face of comprehensive stress. Plants generally promote saline-alkali tolerance by maintaining pH and Na+ homeostasis, while the plants responding to HCO3-/CO32- stress are not exactly the same as high pH stress. We proposed that pH-tolerant or sensitive plants have evolved distinct mechanisms to adapt to saline-alkaline stress. Finally, we highlight the areas that require further research to reveal the new components of saline-alkali tolerance in plants and present the current and potential application of key determinants in breed improvement and molecular breeding.

Comprehensive resistance evaluation of 15 blueberry cultivars under high soil pH stress based on growth phenotype and physiological traits.

High soil pH is one of the main abiotic factors that negatively affects blueberry growth and cultivation. However, no comprehensive evaluation of the high soil pH tolerance of different blueberry cultivars has been conducted. Herein, 16 phenotypic and physiological indices of 15 blueberry cultivars were measured through pot experiments, and the high-pH soil tolerance coefficient (HSTC) was calculated based on these indices to comprehensively evaluate the high-soil-pH tolerance of plants. The results demonstrated that high soil pH stress inhibited blueberry 77.growth, and MDA, soluble sugar (SS), and soluble protein (SP) levels increased in leaves. Moreover, in all cultivars, CAT activity in the antioxidant system was enhanced, whereas SOD activity was reduced, and the relative expression levels of the antioxidant enzyme genes SOD and CAT showed similar changes. In addition, the leaf chlorophyll relative content (SPAD), net photosynthetic rate (Pn), transpiration rate (E), and stomatal conductance (Gs) decreased, while changes in the intercellular CO2 concentration (Ci) were noted in different cultivars. Finally, according to the comprehensive evaluation value D obtained from the combination of principal component analysis (PCA) and membership function (MF), the 15 blueberry cultivars can be divided into 4 categories: high soil pH-tolerant type ['Briteblue' (highest D value 0.815)], intermediate tolerance type ('Zhaixuan 9', 'Zhaixuan 7', 'Emerald', 'Primadonna', 'Powderblue' and 'Chandler'), low high soil pH-tolerant type ('Brightwell', 'Gardenblue', 'Plolific' and 'Sharpblue') and high soil pH-sensitive type ['Legacy', 'Bluegold', 'Baldwin' and 'Anna' (lowest D value 0.166)]. Stepwise linear regression analysis revealed that plant height, SS, E, leaf length, Ci, SOD, and SPAD could be used to predict and evaluate the high soil pH tolerance of blueberry cultivars.

Morphological, physiological, and transcriptional responses of the freshwater diatom Fragilaria crotonensis to elevated pH conditions.

Harmful algal blooms (HABs) caused by the toxin-producing cyanobacteria Microcystis spp., can increase water column pH. While the effect(s) of these basified conditions on the bloom formers are a high research priority, how these pH shifts affect other biota remains understudied. Recently, it was shown these high pH levels decrease growth and Si deposition rates in the freshwater diatom Fragilaria crotonensis and natural Lake Erie (Canada-US) diatom populations. However, the physiological mechanisms and transcriptional responses of diatoms associated with these observations remain to be documented. Here, we examined F. crotonensis with a set of morphological, physiological, and transcriptomic tools to identify cellular responses to high pH. We suggest 2 potential mechanisms that may contribute to morphological and physiological pH effects observed in F. crotonensis. Moreover, we identified a significant upregulation of mobile genetic elements in the F. crotonensis genome which appear to be an extreme transcriptional response to this abiotic stress to enhance cellular evolution rates-a process we have termed "genomic roulette." We discuss the ecological and biogeochemical effects high pH conditions impose on fresh waters and suggest a means by which freshwater diatoms such as F. crotonensis may evade high pH stress to survive in a "basified" future.

Brassinolide improves the tolerance of Malus hupehensis to alkaline stress.

Malus hupehensis is one of the most widely used apple rootstocks in china but is severely damaged by alkaline soil. Alkaline stress can cause more serious harmful effects on apple plants than salt stress because it also induces high pH stress except for ion toxicity, osmotic stress, and oxidative damage. Brassinolide (BL) plays important roles in plant responses to salt stress. However, its role and function mechanism in apple plants in response to alkaline stress has never been reported. This study showed that applying exogenous 0.2 mg/L BL significantly enhanced the resistance of M. hupehensis seedlings to alkaline stress. The main functional mechanisms were also explored. First, exogenous BL could decrease the rhizosphere pH and promote Ca2+ and Mg2+ absorption by regulating malic acid and citric acid contents and increasing H+ excretion. Second, exogenous BL could alleviate ion toxicity caused by alkaline stress through enhancing Na+ efflux and inhibiting K+ expel and vacuole compartmentalization. Last, exogenous BL could balance osmotic stress by accumulating proline and reduce oxidative damage through increasing the activities of antioxidant enzymes and antioxidants contents. This study provides an important theoretical basis for further analyzing the mechanism of exogenous BL in improving alkaline tolerance of apple plants.

Heterologous overexpression of HIOMT alleviates alkaline stress in apple plants by increasing melatonin concentration

Melatonin is an indole derivative of tryptophan that has an important function in alleviating abiotic stress. Previous studies have reported that exogenous melatonin increases the tolerance of plants to saline-alkali stress in a concentration-dependent manner, but the role of endogenous melatonin metabolism in the response of apple (Malus domestica) plants to alkali stress is unclear. In this study, the human melatonin biosynthesis-related enzyme N-acetyl-5-hydroxytryptamine methyltransferase gene (HIOMT) transformed in ‘GL-3’ apple was used to study the regulation of HIOMT in alkaline stress of apple plants. The heterologous overexpression of HIOMT under alkaline stress increased the concentration of endogenous melatonin in apple leaves, alleviated the stress-induced growth inhibition of the plants, enhanced root development, leaf photosynthesis, and antioxidant capacity, and stabilized cell membrane permeability and ionic balance. In addition, the transgenic lines also increased the concentrations of organic acids, such as oxalic acid, malic acid, malonic acid, citric acid, and succinic acid, which alleviated the high pH stress. These results provide evidence for the critical role of heterologous overexpression of HIOMT in the alkaline stress tolerance of apple plants.

Highly sensitive and specific responses of shrimp gill cells to high pH stress based on single cell RNA-seq analysis.

High pH is one of the main stressors affecting the shrimp survival, growth, and physiology in aquaculture ponds, but the cellular and molecular mechanism responsible for high pH stress has not been elucidated in shrimp. In this study, the shrimp acid-base disturbance and gill cell alterations were significantly observed and then single cell RNA-sequencing (scRNA-seq) was performed to study the sensitive and specific responses of gill cells to high pH stress. Three main gill cell types, including pillar cells, hemocytes and septal cells were identified. By comparative scRNA-seq analysis between control and pH group, the pillar cell was regarded as the target cell type in response to high pH stress with the down-regulation of ammonia excretion and H+ transport related genes and up-regulation of immune related genes. Notedly, high pH resulted in the emergence of a new immune cell subcluster in pillar cells, with immune activation and stress defense states. Pseudotime analysis also showed that the pillar cells could transform into the functionally inhibited ion cell subclusters and functionally activated immune cell subclusters after high pH stress. Further, the regulatory network of pillar cell population was predicted by WGCNA and two transcription factors were identified. In conclusion, these results provide key insights into the shrimp gill cell-type-specific mechanisms underlying high pH stress response at a single-cell resolution.

The role of Acpbs2 in the asexual sporulation, stress responseand carbon metabolism of Aspergillus cristatus.

Aspergillus cristatus is the dominant fungus during the fermentation of Fuzhuan brick tea, hypotonic conditions only induced its sexual development to produce ascospores, while hypertonic conditions only induced its asexual development to produce conidia, indicating that osmotic stress can regulate spore production in A. cristatus. However, the underlying regulatory mechanism is unclear. In this study, the roles of Acpbs2, which is homologous to pbs2 from Saccharomyces cerevisiae, in sporulation, stress responses, the color of colonies, and carbon metabolism were explored in A. cristatus. Deletion mutants of Acpbs2 were obtained by homologous recombination. The time required to produce conidia was delayed, and the number of conidia produced was significantly reduced in hypertonic media in ΔAcpbs2 by phenotypic observations, indicating that Acpbs2 plays a positive role in asexual development. Stress sensitivity tests showed that the order of the sensitivity of ΔAcpbs2 to different osmotic regulators was 3 M NaCl > 3 M sucrose > 3 M sorbitol. Moreover, the deletion mutants were sensitive to high oxidative stress. The growth of the Acpbs2 deletion mutant was inhibited under alkaline-pH stress, indicating that Acpbs2 is involved in high pH stress tolerance. Additionally, compared with the wild type, the colony color of the Acpbs2 deletion mutant became lighter. All the above developmental defects were reversed by the reintroduction of the Acpbs2 gene in ΔAcpbs2. Transcriptome data showed that Acpbs2 regulated the expression of several genes related to conidial development, osmotic stress, oxidative stress, and carbon metabolism. More importantly, the interaction between Acpbs2 and its downstream gene Achog1 was verified by yeast two-hybrid assays. We speculated that this interaction might regulate the osmotic stress response, the oxidative stress response, and asexual sporulation in A. cristatus, which will be one of the focuses of our future research.

Biogeographical Patterns and Assembly of Bacterial Communities in Saline Soils of Northeast China

Increasing salinity undermines soil fertility and imposes great threats to soil ecosystem productivity and ecological sustainability. Microbes with the ability to adapt to environmental adversity have gained increasing attention for maintenance and restoration of the salt-affected soil ecosystem structure and functioning; however, the characterization of microbial communities in saline–sodic soils remains limited. This study characterized the bacterial community composition and diversity in saline–sodic soils along a latitude gradient across Northeast China, aiming to reveal the mechanism of physicochemical and geographic characteristics shaping the soil bacterial communities. Our results showed that the bacterial community composition and diversity were significantly impacted by soil pH, electrical conductivity, Na+, K+, Cl−, and CO32−. Significant differences in bacterial diversity were revealed along the latitude gradient, and the soil factors accounted for 58.58% of the total variations in bacterial community composition. Proteobacteria, Actinobacteria, Gemmatimonadetes, Chloroflexi, and Bacteroidetes were dominant across all samples. Actinobacteria and Gemmatimonadetes were significantly enriched in high soil sodicity and salinity, while Acidobacteria and Proteobacteria were suppressed by high pH and salt stress in the saline–sodic soils. Increase in soil pH and salinity significantly decreased bacterial species richness and diversity. Community composition analysis indicated that bacterial taxonomic groups (e.g., Bacillus, Egicoccus, Truepera, Halomonas, and Nitrolancea) that may adapt well to high salinity were greatly enriched in the examined soils. The findings collectively evidenced that bacterial community composition and diversity in a broad biographic scale were determined by niche-based environmental characteristics and biotic interactions. The profiling of the soil bacterial communities along the latitude gradient will also provide a basis for a better understanding of the salt-affected soil ecosystem functioning and restoration of these soil ecosystems.

Strigolactone alleviates the salinity-alkalinity stress of Malus hupehensis seedlings.

Salinity-alkalinity stress can remarkably affect the growth and yield of apple. Strigolactone (SL) is a class of carotenoid-derived compounds that functions in stress tolerance. However, the effects and mechanism of exogenous SL on the salinity-alkalinity tolerance of apple seedlings remain unclear. Here, we assessed the effect of SL on the salinity-alkalinity stress response of Malus hupehensis seedlings. Results showed that treatment with 100 μM exogenous SL analog (GR24) could effectively alleviate salinity-alkalinity stress with higher chlorophyll content and photosynthetic rate than the apple seedlings without GR24 treatment. The mechanism was also explored: First, exogenous GR24 regulated the expression of Na+/K+ transporter genes and decreased the ratio of Na+/K+ in the cytoplasm to maintain ion homeostasis. Second, exogenous GR24 increased the enzyme activities of superoxide, peroxidase and catalase, thereby eliminating reactive oxygen species production. Third, exogenous GR24 alleviated the high pH stress by regulating the expression of H+-ATPase genes and inducing the production of organic acid. Last, exogenous GR24 application increased endogenous acetic acid, abscisic acid, zeatin riboside, and GA3 contents for co-responding to salinity-alkalinity stress indirectly. This study will provide important theoretical basis for analyzing the mechanism of exogenous GR24 in improving salinity-alkalinity tolerance of apple.

Effect of TMCP rolling schedules on the microstructure and performance of X70 steel

Four laboratory rolling schedules were devised and produced from a thick-wall API X70 steel plate. The microstructure and crystallographic texture were characterized followed by an evaluation of the performance of the rolled steels by investigating the mechanical properties and SCC susceptibility. High pH Stress Corrosion Cracking (SCC) is a damaging form of environmental corrosion that can cause rapid failure of buried gas transmission pipelines. It is a key question to ascertain if the manufacturing procedure has any bearing on the resistance of line pipe steels to SCC. Finish rolling above the recrystallization lower temperature, RLT (Tnr), resulted in the formation of large, equiaxed grains of upper bainite and granular bainite. Despite resulting in the highest Yield Strength (YS) and Ultimate Tensile Strength (UTS), the steel manufactured through this process had the highest SCC susceptibility. Finish rolling in the two-phase region increased the polygonal ferrite formation and deteriorated mechanical properties of the rolled steels. Finish rolling above the Ar3, combined with severe rough rolling, produced the lowest SCC susceptibility, with a primarily granular bainite microstructure and a high intensity of {332}<113> texture. Rolling strategies that balanced strength and ductility were shown to positively affect SCC performance.

Calcium Carbonate Can Be Used to Manage Soilless Substrate pH for Blueberry Production

Blueberry (Vacciniumcorymbosum interspecific hybrids) production in soilless substrates is becoming increasingly popular. Soilless substrates have low pH buffering capacity. Blueberry plants preferentially take up ammonium, which acidifies the rhizosphere. Consequently, soilless substrates where blueberry plants are grown exhibit a tendency to get acidified over time. Agricultural lime (CaCO3) is commonly used to raise soil and substrate pH in other crops, but it is rarely used in blueberry cultivation. We hypothesized that substrate amendment with low rates of agricultural lime increases substrate pH buffering capacity and provides nutritional cations that can benefit blueberry plants. We tested this hypothesis in a greenhouse experiment with ‘Emerald’ southern highbush blueberry plants grown in rhizoboxes filled with a 3:1 mix of coconut coir and perlite. We found that substrate amendment with CaCO3 did not cause high pH stress. This amendment maintained substrate pH between 5.5 and 6.5 and provided Ca and Mg for plant uptake. When blueberry plants were grown in CaCO3-amended substrate and fertigated with low pH nutrient solution (pH 4.5), they exhibited greater biomass accumulation than plants grown in unamended substrates. These results suggest that low rates of CaCO3 could be useful for blueberry cultivation in soilless substrates.