scholarly journals The Lysine 299 Residue Endows the Multisubunit Mrp1 Antiporter with Dominant Roles in Na+Resistance and pH Homeostasis inCorynebacterium glutamicum

2018 ◽  
Vol 84 (10) ◽  
Author(s):  
Ning Xu ◽  
Yingying Zheng ◽  
Xiaochen Wang ◽  
Terry A. Krulwich ◽  
Yanhe Ma ◽  
...  
Keyword(s):  
Corynebacterium Glutamicum ◽  
Expression Patterns ◽  
Alkaline Stress ◽  
Alkaline Ph ◽  
Ph Homeostasis ◽  
Conserved Residues ◽  
Content Type ◽  
Ion Translocation ◽  
Alkaline Tolerance ◽  
Alkaline Conditions

ABSTRACTCorynebacterium glutamicumis generally regarded as a moderately salt- and alkali-tolerant industrial organism. However, relatively little is known about the molecular mechanisms underlying these specific adaptations. Here, we found that the Mrp1 antiporter played crucial roles in conferring both environmental Na+resistance and alkali tolerance whereas the Mrp2 antiporter was necessary in coping with high-KCl stress at alkaline pH. Furthermore, the Δmrp1Δmrp2double mutant showed the most-severe growth retardation and failed to grow under high-salt or alkaline conditions. Consistent with growth properties, the Na+/H+antiporters ofC. glutamicumwere differentially expressed in response to specific salt or alkaline stress, and an alkaline stimulus particularly induced transcript levels of the Mrp-type antiporters. When the major Mrp1 antiporter was overwhelmed,C. glutamicummight employ alternative coordinate strategies to regulate antiport activities. Site-directed mutagenesis demonstrated that several conserved residues were required for optimal Na+resistance, such as Mrp1A K299, Mrp1C I76, Mrp1A H230, and Mrp1D E136. Moreover, the chromosomal replacement of lysine 299 in the Mrp1A subunit resulted in a higher intracellular Na+level and a more alkaline intracellular pH value, thereby causing a remarkable growth attenuation. Homology modeling of the Mrp1 subcomplex suggested two possible ion translocation pathways, and lysine 299 might exert its effect by affecting the stability and flexibility of the cytoplasm-facing channel in the Mrp1A subunit. Overall, these findings will provide new clues to the understanding of salt-alkali adaptation duringC. glutamicumstress acclimatization.IMPORTANCEThe capacity to adapt to harsh environments is crucial for bacterial survival and product yields, including industrially usefulCorynebacterium glutamicum. AlthoughC. glutamicumexhibits a marked resistance to salt-alkaline stress, the possible mechanism for these adaptations is still unclear. Here, we present the physiological functions and expression patterns ofC. glutamicumputative Na+/H+antiporters and conserved residues of Mrp1 subunits, which respond to different salt and alkaline stresses. We found that the Mrp-type antiporters, particularly the Mrp1 antiporter, played a predominant role in maintaining intracellular nontoxic Na+levels and alkaline pH homeostasis. Loss of the major Mrp1 antiporter had a profound effect on gene expression of other antiporters under salt or alkaline conditions. The lysine 299 residue may play its essential roles in conferring salt and alkaline tolerance by affecting the ion translocation channel of the Mrp1A subunit. These findings will contribute to a better understanding of Na+/H+antiporters in sodium antiport and pH regulation.

scholarly journals Escherichia coli YqjA, a Member of the Conserved DedA/Tvp38 Membrane Protein Family, Is a Putative Osmosensing Transporter Required for Growth at Alkaline pH

2015 ◽  
Vol 197 (14) ◽  
pp. 2292-2300 ◽  
Author(s):  
Sujeet Kumar ◽  
William T. Doerrler
Keyword(s):  
Escherichia Coli ◽  
Membrane Protein ◽  
Amino Acid Identity ◽  
Protein Family ◽  
Cytoplasmic Ph ◽  
Alkaline Ph ◽  
Ph Homeostasis ◽  
Content Type ◽  
E Coli ◽  
Alkaline Conditions

ABSTRACTThe ability to persist and grow under alkaline conditions is an important characteristic of many bacteria. In order to survive at alkaline pH,Escherichia colimust maintain a stable cytoplasmic pH of about 7.6. Membrane cation/proton antiporters play a major role in alkaline pH homeostasis by catalyzing active inward proton transport. The DedA/Tvp38 family is a highly conserved membrane protein family of unknown function present in most sequenced genomes. YqjA and YghB are members of theE. coliDedA family with 62% amino acid identity and partially redundant functions. We have shown thatE. coliwith ΔyqjAand ΔyghBmutations cannot properly maintain the proton motive force (PMF) and is compromised in PMF-dependent drug efflux and other PMF-dependent functions. Furthermore, the functions of YqjA and YghB are dependent upon membrane-embedded acidic amino acids, a hallmark of several families of proton-dependent transporters. Here, we show that the ΔyqjAmutant (but not ΔyghB) cannot grow under alkaline conditions (ranging from pH 8.5 to 9.5), unlike the parentE. coli. Overexpression ofyqjArestores growth at alkaline pH, but only when more than ∼100 mM sodium or potassium is present in the growth medium. Increasing the osmotic pressure by the addition of sucrose enhances the ability of YqjA to support growth under alkaline conditions in the presence of low salt concentrations, consistent with YqjA functioning as an osmosensor. We suggest that YqjA possesses proton-dependent transport activity that is stimulated by osmolarity and that it plays a significant role in the survival ofE. coliat alkaline pH.IMPORTANCEThe ability to survive under alkaline conditions is important for many species of bacteria.Escherichia colican grow at pH 5.5 to 9.5 while maintaining a constant cytoplasmic pH of about 7.6. Under alkaline conditions, bacteria rely upon proton-dependent transporters to maintain a constant cytoplasmic pH. The DedA/Tvp38 protein family is a highly conserved but poorly characterized family of membrane proteins. Here, we show that the DedA/Tvp38 protein YqjA is critical forE. colito survive at pH 8.5 to 9.5. YqjA requires sodium and potassium for this function. At low cation concentrations, osmolytes, including sucrose, can facilitate rescue ofE. coligrowth by YqjA at high pH. These data are consistent with YqjA functioning as an osmosensing cation-dependent proton transporter.

scholarly journals Abscisic Acid Priming Creates Alkaline Tolerance in Alfalfa Seedlings (Medicago sativa L.)

Agriculture ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 608
Author(s):  
Tian-Jiao Wei ◽  
Ming-Ming Wang ◽  
Yang-Yang Jin ◽  
Guo-Hui Zhang ◽  
Miao Liu ◽  
...  
Keyword(s):  
Abscisic Acid ◽  
Molecular Mechanisms ◽  
Leaf Damage ◽  
Alkaline Stress ◽  
Acid Pretreatment ◽  
Solution Ph ◽  
Expression Of Genes ◽  
Alkaline Tolerance ◽  
Alkaline Conditions ◽  
Medicago Sativa L

Soil alkalization triggers ion toxicity and osmotic and alkaline (high pH) stresses in plants, damaging their growth and productivity. Therefore, we investigated whether priming with abscisic acid (ABA) increases the tolerance of alfalfa seedlings to alkaline stress, and then examined the underlying molecular mechanisms. Alfalfa seedlings were pretreated with ABA (10 μM) for 16 h and then subjected to alkaline stress using a 15 mM Na2CO3 solution (pH 10.87). Compared with the control, ABA pretreatment significantly alleviated leaf damage and improved the fresh weight, water content, and survival rate of alfalfa seedlings under alkaline conditions. Abscisic acid pretreatment reduced accumulation of reactive oxygen species (ROS), increased activities of the antioxidant enzymes superoxide dismutase (SOD) and peroxidase (POD), maintained higher ratios of K+/Na+, Ca2+/Na+, and Mg2+/Na+, and increased accumulation of proline. In addition, ABA upregulated the expression of genes involved in proline biosynthesis (P5CS) and the sequestration of Na+ in vacuoles (NHX1 and AVP) under alkaline conditions. Abscisic acid priming increased tolerance to alkaline stress by maintaining homeostasis of ROS and metal ions and upregulating osmoprotection and the expression of stress tolerance-related genes.

scholarly journals Contribution of the Cell Wall Component Teichuronopeptide to pH Homeostasis and Alkaliphily in the Alkaliphile Bacillus lentus C-125

1999 ◽  
Vol 181 (21) ◽  
pp. 6600-6606 ◽  
Author(s):  
Rikizo Aono ◽  
Masahiro Ito ◽  
Takayoshi Machida
Keyword(s):  
Cell Wall ◽  
Amino Acid Residues ◽  
Alkaline Ph ◽  
Structural Components ◽  
Chromosomal Dna ◽  
Ph Homeostasis ◽  
Cell Wall Component ◽  
Bacillus Lentus ◽  
Defective Mutant ◽  
Alkaline Conditions

ABSTRACT A teichuronopeptide (TUP) is one of major structural components of the cell wall of the facultative alkaliphilic strain Bacillus lentus C-125. A mutant defective in TUP synthesis grows slowly at alkaline pH. An upper limit of pH for growth of the mutant was 10.4, while that of the parental strain C-125 was 10.8. GenetupA, directing synthesis of TUP, was cloned from C-125 chromosomal DNA. The primary translation product of this gene is likely a cytoplasmic protein (57.3 kDa) consisting of 489 amino acid residues. Introduction of the tupA gene into the TUP-defective mutant complemented the mutation responsible for the pleiotropic phenotypes of the mutant, leading to simultaneous disappearance of the defect in TUP synthesis, the diminished ability for cytoplasmic pH homeostasis, and the low tolerance for alkaline conditions. These results demonstrate that the acidic polymer TUP in the cell wall plays a role in pH homeostasis in this alkaliphile.

scholarly journals TheBeauveria bassianaGas3 β-Glucanosyltransferase Contributes to Fungal Adaptation to Extreme Alkaline Conditions

2018 ◽  
Vol 84 (15) ◽  
Author(s):  
Zhibing Luo ◽  
Tongbing Zhang ◽  
Pengfei Liu ◽  
Yuting Bai ◽  
Qiyan Chen ◽  
...  
Keyword(s):  
Cell Wall ◽  
Pathogenic Fungus ◽  
Growth Conditions ◽  
Alkaline Ph ◽  
Ph Responsive ◽  
Content Type ◽  
Cell Wall Remodeling ◽  
Alkaline Conditions ◽  
Increased Sensitivity ◽  
Ph Dependent

ABSTRACTFungal β-1,3-glucanosyltransferases are cell wall-remodeling enzymes implicated in stress response, cell wall integrity, and virulence, with most fungal genomes containing multiple members. The insect-pathogenic fungusBeauveria bassianadisplays robust growth over a wide pH range (pH 4 to 10). A random insertion mutant library screening for increased sensitivity to alkaline (pH 10) growth conditions resulted in the identification and mapping of a mutant to a β-1,3-glucanosyltransferase gene (Bbgas3).Bbgas3expression was pH dependent and regulated by the PacC transcription factor, which activates genes in response to neutral/alkaline growth conditions. Targeted gene knockout ofBbgas3resulted in reduced growth under alkaline conditions, with only minor effects of increased sensitivity to cell wall stress (Congo red and calcofluor white) and no significant effects on fungal sensitivity to oxidative or osmotic stress. The cell walls of ΔBbgas3aerial conidia were thinner than those of the wild-type and complemented strains in response to alkaline conditions, and β-1,3-glucan antibody and lectin staining revealed alterations in cell surface carbohydrate epitopes. The ΔBbgas3mutant displayed alterations in cell wall chitin and carbohydrate content in response to alkaline pH. Insect bioassays revealed impaired virulence for the ΔBbgas3mutant depending upon the pH of the media on which the conidia were grown and harvested. Unexpectedly, a decreased median lethal time to kill (LT50, i.e., increased virulence) was seen for the mutant using intrahemocoel injection assays using conidia grown at acidic pH (5.6). These data show that BbGas3 acts as a pH-responsive cell wall-remodeling enzyme involved in resistance to extreme pH (>9).IMPORTANCELittle is known about adaptations required for growth at high (>9) pH. Here, we show that a specific fungal membrane-remodeling β-1,3-glucanosyltransferase gene (Bbgas3) regulated by the pH-responsive PacC transcription factor forms a critical aspect of the ability of the insect-pathogenic fungusBeauveria bassianato grow at extreme pH. The loss ofBbgas3resulted in a unique decreased ability to grow at high pH, with little to no effects seen with respect to other stress conditions, i.e., cell wall integrity and osmotic and oxidative stress. However, pH-dependent alternations in cell wall properties and virulence were noted for the ΔBbgas3 mutant. These data provide a mechanistic insight into the importance of the specific cell wall structure required to stabilize the cell at high pH and link it to the PacC/Pal/Rim pH-sensing and regulatory system.

scholarly journals Anaerobic Growth of Corynebacterium glutamicum via Mixed-Acid Fermentation

2015 ◽  
Vol 81 (21) ◽  
pp. 7496-7508 ◽  
Author(s):  
Andrea Michel ◽  
Abigail Koch-Koerfges ◽  
Karin Krumbach ◽  
Melanie Brocker ◽  
Michael Bott
Keyword(s):  
Amino Acids ◽  
Corynebacterium Glutamicum ◽  
Tricarboxylic Acid Cycle ◽  
Model Organism ◽  
Anaerobic Growth ◽  
Efficient Production ◽  
Ph Homeostasis ◽  
Anaerobic Conditions ◽  
Acid Fermentation ◽  
Content Type

ABSTRACTCorynebacterium glutamicum, a model organism in microbial biotechnology, is known to metabolize glucose under oxygen-deprived conditions tol-lactate, succinate, and acetate without significant growth. This property is exploited for efficient production of lactate and succinate. Our detailed analysis revealed that marginal growth takes place under anaerobic conditions with glucose, fructose, sucrose, or ribose as a carbon and energy source but not with gluconate, pyruvate, lactate, propionate, or acetate. Supplementation of glucose minimal medium with tryptone strongly enhanced growth up to a final optical density at 600 nm (OD600) of 12, whereas tryptone alone did not allow growth. Amino acids with a high ATP demand for biosynthesis and amino acids of the glutamate family were particularly important for growth stimulation, indicating ATP limitation and a restricted carbon flux into the oxidative tricarboxylic acid cycle toward 2-oxoglutarate. Anaerobic cultivation in a bioreactor with constant nitrogen flushing disclosed that CO2is required to achieve maximal growth and that the pH tolerance is reduced compared to that under aerobic conditions, reflecting a decreased capability for pH homeostasis. Continued growth under anaerobic conditions indicated the absence of an oxygen-requiring reaction that is essential for biomass formation. The results provide an improved understanding of the physiology ofC. glutamicumunder anaerobic conditions.

scholarly journals Pho85 Kinase, a Cyclin-Dependent Kinase, Regulates Nuclear Accumulation of the Rim101 Transcription Factor in the Stress Response of Saccharomyces cerevisiae

2010 ◽  
Vol 9 (6) ◽  
pp. 943-951 ◽  
Author(s):  
Masafumi Nishizawa ◽  
Mirai Tanigawa ◽  
Michio Hayashi ◽  
Tatsuya Maeda ◽  
Yoshiaki Yazaki ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Transcription Factor ◽  
Cellular Response ◽  
Proteolytic Processing ◽  
Yeast Cells ◽  
Nuclear Accumulation ◽  
Alkaline Stress ◽  
Content Type ◽  
Its Gene ◽  
Alkaline Conditions

ABSTRACT The budding yeast Saccharomyces cerevisiae alters its gene expression profile in response to changing environmental conditions. The Pho85 kinase, one of the yeast cyclin-dependent kinases (CDK), is known to play an important role in the cellular response to alterations in parameters such as nutrient levels and salinity. Several genes whose expression is regulated, either directly or indirectly, by the Rim101 transcription factor become constitutively activated when Pho85 function is absent,. Because Rim101 is responsible for adaptation to alkaline conditions, this observation suggests an interaction between Pho85 and Rim101 in the response to alkaline stress. We have found that Pho85 affects neither RIM101 transcription, the proteolytic processing that is required for Rim101 activation, nor Rim101 stability. Rather, Pho85 regulates the nuclear accumulation of active Rim101, possibly via phosphorylation. Additionally, we report that Pho85 and the transcription factor Pho4 are necessary for adaptation to alkaline conditions and that PTK2 activation by Pho4 is involved in this process. These findings illustrate novel roles for the regulators of the PHO system when yeast cells cope with various environmental stresses potentially threatening their survival.

scholarly journals The alkaliphilic side of Staphylococcus aureus

2019 ◽  
Author(s):  
Manisha Vaish ◽  
Amyeo Jereen ◽  
Amall Ali ◽  
Terry Ann Krulwich
Keyword(s):  
Staphylococcus Aureus ◽  
Salt Tolerance ◽  
Significant Role ◽  
Catalytic Properties ◽  
Stress Conditions ◽  
Infection Model ◽  
Alkaline Stress ◽  
Alkaline Ph ◽  
Ph Homeostasis ◽  
Synergistic Relationship

AbstractThe genome of Staphylococcus aureus has eight structurally distinct cation/proton antiporters (CPA) that play significant roles in maintaining cytoplasmic pH and ions in extreme conditions. These antiporters enable S. aureus to persist under conditions that are favorable to the bacterium but unfavorable to animal host including humans. In this study, we report physiological roles and catalytic properties of NhaC (NhaC1, NhaC2 and NhaC3), CPA1 (CPA1-1 and CPA1-2) and CPA2 family antiporters and how these antiporters crosstalk with Mnh1, a CPA3 family antiporter, recently shown to play important roles in virulence and pH tolerance. Catalytic properties of antiporters were determined by Na+/H+ and K+/H+ antiport assays using everted membrane vesicles of a CPA-deficient E. coli KNabC host. NhaC and CPA1 candidates exhibited Na+/H+ and K+/H+ antiporter activity in the pH range between pH 7 to 9.5 but did not show significant role in halotolerance and osmotolerance alone. Interestingly, NhaC3 exhibited significant antiporter activity at alkaline pH and play major roles in pH and salt tolerance. CPA2 neither exhibited Na+or K+/H+ exchange nor showed any active role in pH and salt tolerance. Double deletion of mnhA1 with nhaC1, nhaC3, cpa1-1 or cpa1-2 respectively, made S. aureus severely sensitive at pH 7.5 under stress conditions indicating synergistic relationship of Mnh1 with these antiporters. The functional loss study of these antiporters in in-vivo mouse infection model, nhaC3 deletion showed significant loss of S. aureus virulence. Altogether, the current study indicates NhaC3 as a potential target against S. aureus virulence under extreme pH and salt conditions.ImportanceIn this study, we established catalytic properties and physiological roles of S. aureus NhaC, CPA1 and CPA2 family antiporters and their importance under salt and alkaline stress conditions. Except CPA2, all five antiporters of both families were active for Na+/H+ and K+/H+ exchange. CPA1-1 showed significant role in pH homeostasis at pH 7.5 whereas CPA1-2 and NhaCs were major contributors to halotolerance and osmotolerance at alkaline pH. The severity of growth deficit in double knockouts of mnhA1 with each of nhaC1, nhaC2, nhaC3, cpa1-1 or cpa1-2 establishes their synergistic relationship in regulating pH and salt homeostasis. Deletion of cpa1-1, cpa1-2 and nhaC1, nhaC2, and nhaC3 were assessed in mice model and NhaC3 was shown to play a major role in S. aureus virulence.

scholarly journals Vibrio cholerae OmpR Contributes to Virulence Repression and Fitness at Alkaline pH

2020 ◽  
Vol 88 (6) ◽  
Author(s):  
D. E. Kunkle ◽  
X. R. Bina ◽  
J. E. Bina
Keyword(s):  
Vibrio Cholerae ◽  
Marine Ecosystem ◽  
Acid Tolerance ◽  
Alkaline Ph ◽  
Adaptive Responses ◽  
Gram Negative ◽  
Content Type ◽  
Regulatory Systems ◽  
Contaminated Food ◽  
Alkaline Conditions

ABSTRACT Vibrio cholerae is a Gram-negative human pathogen and the causative agent of the life-threatening disease cholera. V. cholerae is a natural inhabitant of marine environments and enters humans through the consumption of contaminated food or water. The ability to transition between aquatic ecosystems and the human host is paramount to the pathogenic success of V. cholerae. The transition between these two disparate environments requires the expression of adaptive responses, and such responses are most often regulated by two-component regulatory systems such as the EnvZ/OmpR system, which responds to osmolarity and acidic pH in many Gram-negative bacteria. Previous work in our laboratory indicated that V. cholerae OmpR functioned as a virulence regulator through repression of the LysR-family transcriptional regulator aphB; however, the role of OmpR in V. cholerae biology outside virulence regulation remained unknown. In this work, we sought to further investigate the function of OmpR in V. cholerae biology by defining the OmpR regulon through RNA sequencing. This led to the discovery that V. cholerae ompR was induced at alkaline pH to repress genes involved in acid tolerance and virulence factor production. In addition, OmpR was required for V. cholerae fitness during growth under alkaline conditions. These findings indicate that V. cholerae OmpR has evolved the ability to respond to novel signals during pathogenesis, which may play a role in the regulation of adaptive responses to aid in the transition between the human gastrointestinal tract and the marine ecosystem.

scholarly journals Characterization of alkaline stress tolerance mechanisms in Lotus forage species modulated by Pantoea eucalypti

2020 ◽  
Author(s):  
Maria Paula Campestre ◽  
Nazareno Luis Castagno ◽  
Cristian Javier Antonelli ◽  
Vanina Giselle Maguire ◽  
Francisco Jose Escaray ◽  
...  
Keyword(s):  
Considerable Increase ◽  
Physiological Responses ◽  
Dry Mass ◽  
Alkaline Treatment ◽  
Alkaline Stress ◽  
Tolerance Mechanisms ◽  
Alkaline Tolerance ◽  
Forage Species ◽  
Alkaline Conditions

AbstractThis study was designed to elucidate the physiological responses of three Lotus forage accessions to alkaline stress and the influence of the inoculation of a Pantoea eucalypti endophyte strain on its mitigation. One-month-old diploid accessions of Lotus corniculatus (Lc) and Lotus tenuis (Lt), and the interspecific hybrid LtxLc obtained from these parental accessions, were exposed to alkaline stress (pH 8.2) by the addition of NaHCO3 10 mM to the nutrient solution for 2 weeks. The results indicated that Lt and the LtxLc hybrid are alkaline-tolerant compared to Lc, based on the observation that their dry mass is not reduced under stress, and symptoms of chlorosis do not appear on leaf blades, in contrast to observations of the Lc accession subjected to identical growth and stress conditions. In Lc and LtxLc accessions, the Fe2+ concentration decreased in the aerial part under stress and increased in the roots. Interveinal chlorosis observed in the youngest leaves of Lc during alkaline treatment was accompanied with a higher reduction of Fe2+ levels in shoots and a higher increment of Fe2+ in roots, compared to the other accession. Plant inoculation also tended to acidify the medium under alkalinity, contributing to Fe accumulation in the roots. Moreover, the inoculation caused a considerable increase in Fe2+ content in shoots in all three Lotus forage species under alkaline treatment.Fv/Fm and PIABS were only reduced in Lc under alkaline treatment. Inoculation reverted this effect and improved the ABS/RC and DIo/RC ratios in all three accessions. In addition, under alkaline conditions, Lc dissipated more energy than control plants. Expression of the metal-transporting gene NRAMP1 increased in the inoculated Lc accession under stress, while remaining unmodified in Lt and LtxLc hybrid.Altogether, the results obtained make clear the importance of inoculation with P. eucalypti, which contributed significantly to the mitigation of alkaline stress. Thus, all the results provide useful information for improving alkaline tolerance traits in Lotus forage species and their interspecific hybrids.

scholarly journals Physiological and transcriptomic analyses reveal novel insights into the cultivar-specific response to alkaline stress in alfalfa (Medicago sativa L.)

2021 ◽  
Author(s):  
Tian-Jiao Wei ◽  
Guang Li ◽  
Ming-Ming Wang ◽  
Yang-Yang Jin ◽  
Guo-Hui Zhang ◽  
...  
Keyword(s):  
Medicago Sativa ◽  
Chlorophyll Content ◽  
Soluble Sugar ◽  
Alkaline Treatment ◽  
Redox Status ◽  
Mapk Signaling ◽  
Specific Response ◽  
Alkaline Stress ◽  
Alkaline Tolerance ◽  
Alkaline Conditions

Abstract Key message Candidate pathways for alkaline tolerance in alfalfa seedlings were identified; these included those for homeostasis of ions and redox status, biosynthesis of phenylpropanoids, flavonoids, and amino acids, and MAPK signaling.Abstract Soil alkalization severely limits plant growth and development; however, the mechanisms of alkaline response remain largely unknown. In this study, we performed physiological and transcriptomic analyses using two alfalfa cultivars (Medicago sativa L.) with different sensitivities to alkaline conditions. The chlorophyll content and shoot fresh weight drastically declined in the alkaline-sensitive cultivar Algonquin (AG) following alkaline treatment (0-25 mM Na2CO3 solution), while the alkaline-tolerant cultivar Gongnong NO.1 (GN) maintained relatively stable growth and chlorophyll content. Physiological analysis revealed that compared with AG, GN had higher contents of Ca2+ and Mg2+; the ratios of Ca2+ and Mg2+ to Na+, proline and soluble sugar, and enzyme activities of peroxidase (POD) and catalase (CAT) decreased under the alkaline conditions. Further, transcriptomic analysis identified three categories of alkaline-responsive differentially expressed genes (DEGs) between the two cultivars: 48 genes commonly induced in both the cultivars (CAR), 574 genes from the tolerant cultivar (TAR), and 493 genes from the sensitive cultivar (SAR). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that CAR genes were mostly involved in phenylpropanoid biosynthesis, lipid metabolism, and DNA replication and repair; TAR genes were significantly enriched in metabolic pathways, biosynthesis of secondary metabolites, MAPK signaling pathway, and flavonoid and amino acid biosynthesis; the SAR genes were specifically enriched in vitamin B6 metabolism. Taken together, the results identified candidate pathways associated with genetic variation in response to alkaline stress, providing novel insights into the mechanisms underlying alkaline tolerance in alfalfa.

Sign in / Sign up

Export Citation Format

Share Document