Chloride Channels and Transporters of the CLC Family in Plants

Nedelyaeva, O.I.; Shuvalov, Alexey; Balnokin, Yu. V.

doi:10.1134/s1021443720050106

Cited by 20 publications

(19 citation statements)

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“…The precisely regulated influx and efflux of Cl – and Na + ions across the cell and organelle membranes are essential for plant salt tolerance ( Arzani and Ashraf, 2016 ). This action is performed by three membrane-embedded proteinous structures known as ion channels, ion pumps, and ion transporters ( Teakle and Tyermam, 2010 ; Hedrich, 2012 ; Nedelyaeva et al, 2020 ). What do our transcriptome data suggest about how salt-tolerant wild barley performs the role of regulating the expression of transcripts capable to act in simultaneous multiple ion transporters and channels?…”

Section: Discussionmentioning

confidence: 99%

Physiological and Transcriptome Indicators of Salt Tolerance in Wild and Cultivated Barley

et al. 2022

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Barley is used as a model cereal to decipher salt tolerance mechanisms due to its simpler genome than wheat and enhanced salt tolerance compared to rice and wheat. In the present study, RNA-Seq based transcriptomic profiles were compared between salt-tolerant wild (Hordeum spontaneum, genotype no. 395) genotype and salt-sensitive cultivated (H. vulgare, ‘Mona’ cultivar) subjected to salt stress (300 mM NaCl) and control (0 mM NaCl) conditions. Plant growth and physiological attributes were also evaluated in a separate experiment as a comparison. Wild barley was significantly less impacted by salt stress than cultivated barley in growth and physiology and hence was more stress-responsive functionally. A total of 6,048 differentially expressed genes (DEGs) including 3,025 up-regulated and 3,023 down-regulated DEGs were detected in the wild genotype in salt stress conditions. The transcripts of salt-stress-related genes were profoundly lower in the salt-sensitive than the tolerant barley having a total of 2,610 DEGs (580 up- and 2,030 down-regulated). GO enrichment analysis showed that the DEGs were mainly enriched in biological processes associated with stress defenses (e.g., cellular component, signaling network, ion transporter, regulatory proteins, reactive oxygen species (ROS) scavenging, hormone biosynthesis, osmotic homeostasis). Comparison of the candidate genes in the two genotypes showed that the tolerant genotype contains higher functional and effective salt-tolerance related genes with a higher level of transcripts than the sensitive one. In conclusion, the tolerant genotype consistently exhibited better tolerance to salt stress in physiological and functional attributes than did the sensitive one. These differences provide a comprehensive understanding of the evolved salt-tolerance mechanism in wild barley. The shared mechanisms between these two sub-species revealed at each functional level will provide more reliable insights into the basic mechanisms of salt tolerance in barley species.

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Section: Discussionmentioning

confidence: 99%

Physiological and Transcriptome Indicators of Salt Tolerance in Wild and Cultivated Barley

et al. 2022

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“…In glycophytes, anion channels and transporters from the CLC (chloride channel) family play key roles in anionic homeostasis, salinity tolerance, and nitrogen nutrition [ 32 , 33 , 34 ]. CLC proteins are found in representatives of all kingdoms [ 35 , 36 , 37 , 38 ].…”

Section: Introductionmentioning

confidence: 99%

“…Seven genes of the CLC family have been cloned from A. thaliana , namely AtCLCa–e. While the functions and physiological roles of their products have been extensively investigated [ 32 , 33 , 34 , 46 ], the halophyte orthologs of CLC family proteins remain barely studied. The molecular cloning and functional characterization of proteins from halophytes are important for elucidating the mechanisms underlying plant salt tolerance and improving crop resistance to soil salinity by genetic manipulations [ 47 , 48 , 49 , 50 ].…”

Section: Introductionmentioning

confidence: 99%

Molecular Cloning and Characterization of SaCLCd, SaCLCf, and SaCLCg, Novel Proteins of the Chloride Channel Family (CLC) from the Halophyte Suaeda altissima (L.) Pall

Nedelyaeva

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Volkov

et al. 2022

Plants

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Coding sequences of the CLC family genes SaCLCd, SaCLCf, and SaCLCg, the putative orthologs of Arabidopsis thaliana AtCLCd, AtCLCf, and AtCLCg genes, were cloned from the euhalophyte Suaeda altissima (L.) Pall. The key conserved motifs and glutamates inherent in proteins of the CLC family were identified in SaCLCd, SaCLCf, and SaCLCg amino acid sequences. SaCLCd and SaCLCg were characterized by higher homology to eukaryotic (human) CLCs, while SaCLCf was closer to prokaryotic CLCs. Ion specificities of the SaCLC proteins were studied in complementation assays by heterologous expression of the SaCLC genes in the Saccharomyces cerevisiae GEF1 disrupted strain Δgef1. GEF1 encoded the only CLC family protein, the Cl− transporter Gef1p, in undisrupted strains of this organism. Expression of SaCLCd in Δgef1 cells restored their ability to grow on selective media. The complementation test and the presence of both the “gating” and “proton” conservative glutamates in SaCLCd amino acid sequence and serine specific for Cl− in its selectivity filter suggest that this protein operates as a Cl−/H+ antiporter. By contrast, expression of SaCLCf and SaCLCg did not complement the growth defect phenotype of Δgef1 cells. The selectivity filters of SaCLCf and SaCLCg also contained serine. However, SaCLCf included only the “gating” glutamate, while SaCLCg contained the “proton” glutamate, suggesting that SaCLCf and SaCLCg proteins act as Cl− channels. The SaCLCd, SaCLCf, and SaCLCg genes were shown to be expressed in the roots and leaves of S. altissima. In response to addition of NaCl to the growth medium, the relative transcript abundances of all three genes of S. altissima increased in the leaves but did not change significantly in the roots. The increase in expression of SaCLCd, SaCLCf, and SaCLCg in the leaves in response to increasing salinity was in line with Cl− accumulation in the leaf cells, indicating the possible participation of SaCLCd, SaCLCf, and SaCLCg proteins in Cl− sequestration in cell organelles. Generally, these results suggest the involvement of SaCLC proteins in the response of S. altissima plants to increasing salinity and possible participation in mechanisms underlying salt tolerance.

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“…At present, the function and classification of Arabidopsis CLC family genes have been extensively and deeply studied. Specifically, members of the AtCLC gene family have been divided into two subclasses (I and II) and 7 subfamilies (-a, -b, -c, -d, -e, -f, and -g) ( Nedelyaeva et al, 2020 ). Subclass Ⅰ is mainly composed of AtCLC-a/-b/-c/-d/-g subfamily, while subclass II is composed of AtCLC-e/-f subfamily ( Nedelyaeva et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

“…Specifically, members of the AtCLC gene family have been divided into two subclasses (I and II) and 7 subfamilies (-a, -b, -c, -d, -e, -f, and -g) ( Nedelyaeva et al, 2020 ). Subclass Ⅰ is mainly composed of AtCLC-a/-b/-c/-d/-g subfamily, while subclass II is composed of AtCLC-e/-f subfamily ( Nedelyaeva et al, 2020 ). In addition, it was found that subclass I contained the patterns GxGIPE (I), GKxGPxxH (II), and PxxGxLF (III).…”

Section: Introductionmentioning

confidence: 99%

Genome-Wide Identification and Functional Characterization of the Chloride Channel TaCLC Gene Family in Wheat (Triticum aestivum L.)

et al. 2022

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In plants, chloride channels (CLC) are involved in a series of specific functions, such as regulation of nutrient transport and stress tolerance. Members of the wheat Triticum aestivum L. CLC (TaCLC) gene family have been proposed to encode anion channels/transporters that may be related to nitrogen transportation. To better understand their roles, TaCLC family was screened and 23 TaCLC gene sequences were identified using a Hidden Markov Model in conjunction with wheat genome database. Gene structure, chromosome location, conserved motif, and expression pattern of the resulting family members were then analyzed. Phylogenetic analysis showed that the TaCLC family can be divided into two subclasses (I and II) and seven clusters (-a, -c1, -c2, -e, -f1, -f2, and -g2). Using a wheat RNA-seq database, the expression pattern of TaCLC family members was determined to be an inducible expression type. In addition, seven genes from seven different clusters were selected for quantitative real-time PCR (qRT-PCR) analysis under low nitrogen stress or salt stress conditions, respectively. The results indicated that the gene expression levels of this family were up-regulated under low nitrogen stress and salt stress, except the genes of TaCLC-c2 cluster which were from subfamily -c. The yeast complementary experiments illustrated that TaCLC-a-6AS-1, TaCLC-c1-3AS, and TaCLC-e-3AL all had anion transport functions for NO3− or Cl−, and compensated the hypersensitivity of yeast GEF1 mutant strain YJR040w (Δgef1) in restoring anion-sensitive phenotype. This study establishes a theoretical foundation for further functional characterization of TaCLC genes and provides an initial reference for better understanding nitrate nitrogen transportation in wheat.

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Chloride Channels and Transporters of the CLC Family in Plants

Cited by 20 publications

References 79 publications

Physiological and Transcriptome Indicators of Salt Tolerance in Wild and Cultivated Barley

Physiological and Transcriptome Indicators of Salt Tolerance in Wild and Cultivated Barley

Molecular Cloning and Characterization of SaCLCd, SaCLCf, and SaCLCg, Novel Proteins of the Chloride Channel Family (CLC) from the Halophyte Suaeda altissima (L.) Pall

Genome-Wide Identification and Functional Characterization of the Chloride Channel TaCLC Gene Family in Wheat (Triticum aestivum L.)

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