Identification of Some Anion Transporter Genes in the Halophyte Suaeda altissima (L.) Pall. and Their Expression under Nitrate Deficiency and Salinity

Shuvalov, Alexey; Yurchenko, A. A.; Nedelyaeva, O.I.; Myasoedov, N. A.; Karpichev, Igor V.; Халилова, Л. А.; Попова, Л. Г.; Balnokin, Yu. V.

doi:10.1134/s1021443721050174

Cited by 7 publications

(15 citation statements)

References 28 publications

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“…Coding nucleotide sequences of SaCLCd , SaCLCf , and SaCLCg , genes from the halophyte S. altissima , were determined based on the putative similarity of these genes to homologous genes from the halophyte S. fruticosa . As a result of in silico searches of sequences related to the CLC family in the de novo assembled transcriptome of S. fruticosa [ 51 , 56 ], the contigs containing the partial coding regions of three sequences homologous to the A. thaliana CLC genes were designated by us as SfCLCd , SfCLCf , and SfCLCg. The contigs from S. fruticosa served as a base for identification of the full-size coding sequences of the target S. altissima genes by a rapid amplification of 3´- and 5´-cDNA ends.…”

Section: Resultsmentioning

confidence: 99%

“…First, we performed an in silico search for the sequences homologous to the AtCLCd, AtCLCf , and AtCLCg genes in the de novo assembled by us transcriptomes of Suaeda fruticosa (L.) Forssk, which is a closely related species of S. altissima [ 51 , 56 ]. To do this, the contigs of the assembled transcriptomes were translated into amino acid sequences and search for the sequences related to the CLC family proteins was accomplished in the obtained arrays.…”

Section: Methodsmentioning

confidence: 99%

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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

Попова

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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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

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

Попова

Volkov

et al. 2022

Plants

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“…In the current work, cDNAs of two members of the CLC family chloride channels, SaCLCa2 and SaCLCc2 , from euhalophyte S. altissima , have been cloned in addition to SaCLCa1 , SaCLCc1 , SaCLCd , SaCLCf , and SaCLCg , cloned previously [ 31 , 32 , 33 ]. Coding sequences of SaCLCa2 and most parts of SaCLCc2 are similar to the CDSs of SfCLCa2 and SfCLCc2 , respectively, identified earlier in silico in the de novo assembled transcriptome of S. fruticosa , a closely related species to S. altissima [ 31 , 35 ]. However, the nucleotide sequences of SaCLCc2 cDNA at the 5´-end and amino acid sequence at the N-end of the encoded protein, respectively, are longer than those in the S. fruticosa homolog.…”

Section: Discussionmentioning

confidence: 60%

“…Taking into account the data obtained earlier [ 31 , 32 , 33 ], altogether 7 representatives of CLC family in the halophyte S. altissima ( Table 1 )—which corresponds to 7 full size or partial CDSs found in silico in transcriptomes of a close related species S. fruticosa [ 31 , 35 ]—are identified by us. Two of the identified proteins, SaCLCa1 and SaCLCa2, are likely to be NO 3 − /H + antiporters, while the other proteins are likely to be Cl − /H + antiporters (SaCLCc1, SaCLCc2, SaCLCd) or chloride channels (SaCLCf, SaCLCg).…”

Section: Discussionmentioning

confidence: 87%

“…Coding nucleotide sequences (CDSs) of SaCLCa2 and SaCLCc2 , the genes from the halophyte S. altissima , were determined based on the putative similarity of these genes to homologous genes from the halophyte S. fruticosa . As a result of in silico search of sequences related to the CLC family in the de novo assembled transcriptome of S. fruticosa [ 31 , 35 ], the contigs containing the coding regions of two sequences homologous to the A. thaliana CLC genes were found and designated by us as SfCLCa2 and SfCLCc2 . The contigs from S. fruticosa served as a base for identification of full-size coding sequences of the target S. altissima genes by a rapid amplification of 3’- and 5´-cDNA ends.…”

Section: Resultsmentioning

confidence: 99%

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Chloride Channel Family in the Euhalophyte Suaeda altissima (L.) Pall: Cloning of Novel Members SaCLCa2 and SaCLCc2, General Characterization of the Family

Nedelyaeva¹,

Попова²,

Khramov³

et al. 2023

IJMS

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CLC family genes, comprising anion channels and anion/H+ antiporters, are widely represented in nearly all prokaryotes and eukaryotes. CLC proteins carry out a plethora of functions at the cellular level. Here the coding sequences of the SaCLCa2 and SaCLCc2 genes, homologous to Arabidopsis thaliana CLCa and CLCc, were cloned from the euhalophyte Suaeda altissima (L.) Pall. Both the genes cloned belong to the CLC family as supported by the presence of the key conserved motifs and glutamates inherent for CLC proteins. SaCLCa2 and SaCLCc2 were heterologously expressed in Saccharomyces cerevisiae GEF1 disrupted strain, Δgef1, where GEF1 encodes the only CLC family protein, the Cl– transporter Gef1p, in undisrupted strains of yeast. The Δgef1 strain is characterized by inability to grow on YPD yeast medium containing Mn2+ ions. Expression of SaCLCa2 in Δgef1 cells growing on this medium did not rescue the growth defect phenotype of the mutant. However, a partial growth restoration occurred when the Δgef1 strain was transformed by SaCLCa2(C544T), the gene encoding protein in which proline, specific for nitrate, was replaced with serine, specific for chloride, in the selectivity filter. Unlike SaCLCa2, expression of SaCLCc2 in Δgef1 resulted in a partial growth restoration under these conditions. Analysis of SaCLCa2 and SaCLCc2 expression in the euhalophyte Suaeda altissima (L.) Pall by quantitative real-time PCR (qRT-PCR) under different growth conditions demonstrated stimulation of SaCLCa2 expression by nitrate and stimulation of SaCLCc2 expression by chloride. The results of yeast complementation assay, the presence of both the “gating” and “proton” glutamates in aa sequences of both the proteins, as well results of the gene expression in euhalophyte Suaeda altissima (L.) Pall suggest that SaCLCa2 and SaCLCc2 function as anion/H+ antiporters with nitrate and chloride specificities, respectively. The general bioinformatic overview of seven CLC genes cloned from euhalophyte Suaeda altissima is given, together with results on their expression in roots and leaves under different levels of salinity.

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Adaptive and viable characteristics of microclonal seedlings of hyperhylophyte of Suaeda arcuate under in vitro conditions

Khalbekova

2024

E3S Web of Conf.

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This article presents data on the introduction of in vitro culture and microclonal propagation of plants identified in the group of hyperhalophytes belonging to the Suaeda arcuata Bunge. This study was carried out to optimize the composition of nutrient media for the main stages of reproduction in vitro, as well as studies on the rooting and adaptation of regenerants for species of the Suaeda arcuata from axillary or apical buds, but more often from stem segments with a node. In this work, hormones of the cytokinin and auxin series, or a combination of them, were added to the nutrient environment for growth activation. The cultivation of regenerates on the environment 1/2 MS + 1 mg/l 6-BAP + 0.3 mg/l IAA + 2,4-D showed the best effect on the growth of regenerants, created the possibility of obtaining the maximum amount of biomass. Our study indicated that Suaeda arcuata is a highly salt-tolerant species that survived at 300 mM NaCl most conducive to plant growth, where the quantity of Pro and Met were bigger than the control.

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Identification of Some Anion Transporter Genes in the Halophyte Suaeda altissima (L.) Pall. and Their Expression under Nitrate Deficiency and Salinity

Cited by 7 publications

References 28 publications

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

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

Chloride Channel Family in the Euhalophyte Suaeda altissima (L.) Pall: Cloning of Novel Members SaCLCa2 and SaCLCc2, General Characterization of the Family

Adaptive and viable characteristics of microclonal seedlings of hyperhylophyte of Suaeda arcuate under in vitro conditions

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