Search Results (16)

Shaker K⁺ channel proteins are responsible for potassium (K⁺) uptake and transport, playing a critical role in plant growth, development, and adaptation to K⁺ deficiency. Cassava, a key tropical root crop, is known for its characteristic of resilience to nutrient-poor soil and drought stress. However, the Shaker K⁺ channel gene family in cassava has not yet been characterized. In this study, 13 Shaker channel genes were identified from the near telomere-to-telomere (T2T) cassava genome using bioinformatics analysis. Phylogenetic relationships classified these genes into five distinct subfamilies, and all encoded proteins contained the conserved GYGD/GYGE motif typical of Shaker channels. Protein interaction network predictions revealed potential interactions among the Shaker family, as well as with the potassium transporter HAK5. Tissue-specific expression pattern analysis showed that MeGORK and MeAKT1.2 were expressed in all tissues. Furthermore, quantitative real-time PCR (qRT-PCR) analysis was conducted to examine the transcriptional levels of Shaker K⁺ channel gene family members in the roots and leaves of two cassava germplasms with different low-potassium tolerance after one month of low-potassium treatment. The results revealed that MeAKT1.2, MeAKT2.2, and MeKAT1 exhibited distinct expression patterns between the two germplasms, with higher expression levels observed in the potassium-tolerant germplasm. Therefore, these three genes may serve as important candidate genes for potassium stress tolerance in cassava. In summary, this study provides valuable insights into the characteristics and biological functions of the Shaker K⁺ channel gene family in cassava and identifies potential candidate genes for breeding or engineering potassium-efficient cassava cultivars. Full article

Shaker potassium channel proteins are a class of voltage-gated ion channels responsible for K⁺ uptake and translocation, playing a crucial role in plant growth and salt tolerance. In this study, bioinformatic analysis was performed to identify the members within the Shaker gene family. Moreover, the expression patterns of rice Shaker(OsShaker) K⁺ channel genes were analyzed in different tissues and salt treatment by RT–qPCR. The results revealed that there were eight OsShaker K⁺ channel genes distributed on chromosomes 1, 2, 5, 6 and 7 in rice, and their promoters contained a variety of cis-regulatory elements, including hormone-responsive, light-responsive, and stress-responsive elements, etc. Most of the OsShaker K⁺ channel genes were expressed in all tissues of rice, but at different levels in different tissues. In addition, the expression of OsShaker K⁺ channel genes differed in the timing, organization and intensity of response to salt and chilling stress. In conclusion, our findings provide a reference for the understanding of OsShaker K⁺ channel genes, as well as their potential functions in response to salt and chilling stress in rice. Full article

Stenotaphrum secundatum is an excellent shade-tolerant warm-season turfgrass. Its poor cold resistance severely limits its promotion and application in temperate regions. Mining cold resistance genes is highly important for the cultivation of cold-resistant Stenotaphrum secundatum. Although there have been many reports on the role of the Shaker potassium channel family under abiotic stress, such as drought and salt stress, there is still a lack of research on their role in cold resistance. In this study, the transcriptome database of Stenotaphrum secundatum was aligned with the whole genome of Setaria italica, and eight members of the Shaker potassium channel family in Stenotaphrum secundatum were identified and named SsKAT1.1, SsKAT1.2, SsKAT2.1, SsKAT2.2, SsAKT1.1, SsAKT2.1, SsAKT2.2, and SsKOR1. The KAT3-like gene, KOR2 homologous gene, and part of the AKT-type weakly inwardly rectifying channel have not been identified in the Stenotaphrum secundatum transcriptome database. A bioinformatics analysis revealed that the potassium channels of Stenotaphrum secundatum are highly conserved in terms of protein structure but have more homologous members in the same group than those of other species. Among the three species of Oryza sativa, Arabidopsis thaliana, and Setaria italica, the potassium channel of Stenotaphrum secundatum is more closely related to the potassium channel of Setaria italica, which is consistent with the taxonomic results of these species belonging to Paniceae. Subcellular location experiments demonstrate that SsKAT1.1 is a plasma membrane protein. The expression of SsKAT1.1 reversed the growth defect of the potassium absorption-deficient yeast strain R5421 under a low potassium supply, indicating that SsKAT1.1 is a functional potassium channel. The transformation of SsKAT1.1 into the cold-sensitive yeast strain INVSC1 increased the cold resistance of the yeast, indicating that SsKAT1.1 confers cold resistance. The transformation of SsKAT1.1 into the salt-sensitive yeast strain G19 increased the resistance of yeast to salt, indicating that SsKAT1.1 is involved in salt tolerance. These results suggest that the manipulation of SsKAT1.1 will improve the cold and salt stress resistance of Stenotaphrum secundatum. Full article

Shaker potassium channels play a crucial role in potassium (K+) nutrition and stress resistance in plants. However, systematic research on Shaker K+ channels in Chinese cabbage [Brassica rapa var. chinensis (L.) Kitamura] remains scarce. This study identified 13 Shaker K+ channel members within the cabbage genome, which are unevenly distributed across eight chromosomes. Notably, the number of Shaker K+ channel members in Chinese cabbage exceeds that found in the model plants Arabidopsis (9) and rice (10). This discrepancy is attributed to a higher number of homologous proteins in Groups II and V of Chinese cabbage, with gene segmental duplication in these two subgroups being a significant factor contributing to the expansion of the Shaker K+ channel gene family. Interspecies collinearity analysis revealed that the whole genome and the Shaker K+ channel family of Chinese cabbage show greater similarity to those of Arabidopsis than to those of rice, indicating that Shaker K+ channels from the Brassicaceae family have a closer relationship than that from the Poaceae family. Given that gene expansion occurs in Group II, we investigated whether a functional difference exists between BrKAT1.1 and BrKAT1.2 using yeast assays and promoter analysis. The expression of two BrKAT1 genes in the potassium uptake-deficient yeast mutant R5421 can restore growth under low potassium conditions, indicating their role in potassium absorption. Truncation of the N-terminal 63 amino acids of BrKAT1.2 resulted in the loss of potassium absorption capability, suggesting that the N-terminus is essential for maintaining the potassium absorption function of BrKAT1.2. Furthermore, the expression of the two BrKAT1 genes in the salt-sensitive yeast G19 enhances yeast tolerance to salt stress. These results demonstrate that BrKAT1.1 and BrKAT1.2 exhibit similar abilities in potassium uptake and salt tolerance. The difference between BrKAT1.1 and BrKAT1.2 lay in their promoter regulatory elements, suggesting that differences in transcriptional regulation contributed to the functional differentiation of BrKAT1.1 and BrKAT1.2. These findings provide a foundation for understanding the evolution and functional mechanisms of the Shaker K+ channel family in Chinese cabbage and for improving potassium nutrition and salt tolerance in this species through the manipulation of BrKAT1. Full article

K⁺ channels of the Shaker family have been shown to play crucial roles in K⁺ uptake and transport. Cotton (Gossypium hirsutum) is an important cash crop. In this study, the 24 Shaker family genes were identified in cotton. Phylogenetic analysis suggests that they were assigned to five clusters. Additionally, their chromosomal location, conserved motifs and gene structure were analyzed. The promoter of cotton Shaker K⁺ channel genes comprises drought-, low-temperature-, phytohormone-response elements, etc. As indicated by qRT-PCR (quantitative real-time PCR), cotton Shaker K⁺ channel genes responded to low K⁺ and NaCl, and especially dehydration stress, at the transcript level. Moreover, one of the Shaker K⁺ channel genes, GhKAT1aD, was characterized. This gene is localized in the plasma membrane and is predicted to contain six transmembrane segments. It restored the growth of the yeast mutant strain defective in K⁺ uptake, and silencing GhKAT1a via VIGS (virus-induced gene silencing) resulted in more severe symptoms of K⁺ deficiency in cotton leaves as well as a lower net K⁺ uptake rate. The results of this study showed the overall picture of the cotton Shaker K⁺ channel family regarding bioinformatics as well as the function of one of its members, which provide clues for future investigations of cotton K⁺ transport and molecular insights for breeding K⁺-efficient cotton varieties. Full article

We evaluated the effects of a new extract (70% acetonitrile, 2E0217022196DIPFARMTDA) of Lens culinaris Medik (Terre di Altamura SRL, Altamura BA) to prevent cytotoxic damage from cisplatin, staurosporine, irinotecan, doxorubicin, and the glucocorticoid dexamethasone. The acetonitrile–water extract (range 0.1–5 mg/mL) was obtained by extracting 10 g of lentil flour with 50 milliliters of the acetonitrile–water extraction mixture in a 70:30 ratio, first for 3 h and then overnight in a shaker at room temperature. The next day, the extract was filtered and passed through a Rotavapor to obtain only the aqueous component and eliminate that with acetonitrile, and then freeze-dried to finally have the powdered extract. In vitro experiments showed that the extract prevented the cytotoxic damage induced by cisplatin, irinotecan, and doxorubicin on HEK293 and SHSY5Y cell lines after 24–96 h. In murine osteoblasts after 24–72 h of incubation time, the extract was cytoprotective against all chemicals. The extract was effective against dexamethasone, leading to synergic cell proliferation in all cell types. In bone marrow cells, the extract is cytoprotective after 72 h against doxorubicin, staurosporine, and dexamethasone. Instead, on muscle fibers, the extract has a synergic effect with chemotherapeutics, increasing cytotoxicity induced by doxorubicin and staurosporine. LC-MS attested to the existence of several phenolic structures in the extract. The most abundant families of compounds were flavonoids (25.7%) and mellitic acid (18%). Thus, the development of this extract could be implemented in the area of research related to the chemoprevention of damage to renal, neuronal, bone marrow cells, and osteoblasts by chemotherapeutics; moreover, it could be used as a reinforcer of cytotoxic action of chemotherapeutics on muscle fibers. Full article

Potassium ion (K⁺) channels and transporters are key components of plant K⁺ absorption and transportation and play an important role in plant growth and development. This study revealed that K⁺ channels and transporters are involved in the salt tolerance molecular mechanism and metabolites of the halophyte representative plant Tamarix ramosissima (T. ramosissima) in response to NaCl stress, providing a theoretical basis for the mitigation of salt stress using halophytes. Through transcriptome sequencing and metabolite detection analysis of 0 h, 48 h and 168 h by applying exogenous K⁺ to the roots of T. ramosissima under NaCl stress, 15 high-quality Clean Data bases were obtained, Q20 reached more than 97%, Q30 reached more than 92%, and GC content reached 44.5%, which is in line with further bioinformatics analysis. Based on the Liquid chromatography–mass spectrometry (LC-MS) analysis, the roots of T. ramosissima were exposed to exogenous potassium for 48 h and 168 h under NaCl stress, and 1510 and 1124 metabolites were identified in positive and negative ion mode, respectively. Through orthogonal projections to latent structures discriminant analysis (OPLS-DA) model analysis, its metabolomic data have excellent predictability and stability. The results of this study showed that there were 37 differentially expressed genes (DEGs) annotated as Class 2 K⁺ channels (Shaker-like K⁺ channel and TPK channel) and Class 3 K⁺ transporters (HAK/KUP/KT, HKT and CPAs transporter families). Among them, 29 DEGs were annotated to the gene ontology (GO) database, and the most genes were involved in the GO Biological Process. In addition, the expression levels of Unigene0014342 in the HAK/KUP/KT transporter and Unigene0088276 and Unigene0103067 in the CPAs transporter both first decreased and then increased when treated with 200 mM NaCl for 48 h and 168 h. However, when treated with 200 mM NaCl + 10 mM KCl for 48 h and 168 h, a continuous upward trend was shown. Notably, the expression level of Unigene0016813 in CPAS transporter continued to increase when treated with 200 mM NaCl and 200 mM NaCl + 10 mM KCl for 48 h and 168 h. 3 DEGs, Unigene0088276, Unigene0016813 and Unigene0103067, were dominated by the positive regulation of their related metabolites, and this correlation was significant. The results showed that these DEGs increased the absorption of K⁺ and the ratio of K⁺/Na⁺ under NaCl stress at 48 h and 168 h after adding exogenous potassium and enhanced the salt tolerance of T. ramosissima. Notably, the expression level of Unigene0103067 in the CPAs transporter was consistently upregulated when 200 mM NaCl + 10 mM KCl was treated for 48 h and 168 h. The positive regulatory metabolites were always dominant, which better helped T. ramosissima resist salt stress. Unigene0103067 plays an important role in enhancing the salt tolerance of T. ramosissima and reducing the toxicity of NaCl in roots. Additionally, phylogenetic tree analysis showed that Unigene0103067 and Reaumuria trigyna had the closest genetic distance in the evolutionary relationship. Finally, 9 DEGs were randomly selected for quantitative real-time PCR (qRT-PCR) verification. Their expression trends were completely consistent with the transcriptome sequencing analysis results, proving that this study’s data are accurate and reliable. This study provides resources for revealing the molecular mechanism of NaCl stress tolerance in T. ramosissima and lays a theoretical foundation for cultivating new salt-tolerant varieties. Full article

Utilization of plant allelopathic potential to control weed infestations provides an effective, cost-efficient, labor-free, and environmentally acceptable alternative to traditional chemical and mechanical methods. Conocarpus erectus, known as buttonwood, belongs to the Combretaceae family with high contents of phytochemicals and antioxidant activity. There have been no studies on the allelopathic potential of C. erectus. The present study (1) examined the allelopathic potential of C. erectus against selected weeds (Chenopodium murale and Amaranthus viridis) and crops (Solanum lycopersicum and Cucumis sativus) via investigating the growth inhibition ability of its aqueous extract, and (2) identified the potential allelochemicals found in this plant. Aqueous extracts were prepared from leaves, roots, and seeds of C. erectus by immersing the dried powder of the examined plant parts in sterile distilled water for 24 h on a shaker set to 180 rpm. The resulting filtrate was considered as 100% solution, and then dilutions were made to various concentrations (75%, 50%, and 25%). C. erectus leaves and seeds showed the highest rate of inhibition at all concentrations against Chenopodium murale and Amaranthus viridis grown in either Petri dishes or pots. Conversely, all the studied extracts did not show any toxic effects against tomato and cucumber plants grown in pots. In Petri dishes, a slight reduction in growth was observed. HPLC analysis of total phenolic contents in C. erectus methanolic extracts showed that leaves have the highest contents of gallic acid, caffeic acid, and ferulic acid (153.963, 69.135, and 39.801 ppm, respectively). The finding of the current study demonstrated that the part of the plant and the concentration of extraction have a significant effect on phytotoxicity. The positive results of this study might be used to develop environmentally-friendly herbicides for agricultural purposes. Full article

Potassium (K⁺) is one of the most important cations that plays a significant role in plants and constitutes up to 10% of plants’ dry weight. Plants exhibit complex systems of transporters and channels for the distribution of K⁺ from soil to numerous parts of plants. In this study, we have identified 39 genes encoding putative K⁺ transport-related genes in Vigna radiata. Chromosomal mapping of these genes indicated an uneven distribution across eight out of 11 chromosomes. Comparative phylogenetic analysis of different plant species, i.e., V. radiata, Glycine max, Cicer arietinum, Oryza sativa, and Arabidopsis thaliana, showed their strong conservation in different plant species. Evolutionary analysis of these genes suggests that gene duplication is a major route of expansion for this family in V. radiata. Comprehensive promoter analysis identified several abiotic stresses related to cis-elements in the promoter regions of these genes, suggesting their role in abiotic stress tolerance. Our additional analyses indicated that abiotic stresses adversely affected the chlorophyll concentration, carotenoids, catalase, total soluble protein concentration, and the activities of superoxide and peroxidase in V. radiata. It also disturbs the ionic balance by decreasing the uptake of K⁺ content and increasing the uptake of Na⁺. Expression analysis from high-throughput sequencing data and quantitative real-time PCR experiments revealed that several K⁺ transport genes were expressed in different tissues (seed, flower, and pod) and in abiotic stress-responsive manners. A highly significant variation of expression was observed for VrHKT (1.1 and 1.2), VrKAT (1 and 2) VrAKT1.1, VrAKT2, VrSKOR, VrKEA5, VrTPK3, and VrKUP/HAK/KT (4, 5, and 8.1) in response to drought, heat or salinity stress. It reflected their potential roles in plant growth, development, or stress adaptations. The present study gives an in-depth understanding of K⁺ transport system genes in V. radiata and will serve as a basis for a functional analysis of these genes. Full article

Potassium is the most important and abundant inorganic cation in plants and it can comprise up to 10% of a plant’s dry weight. Plants possess complex systems of transporters and channels for the transport of K⁺ from soil to numerous parts of plants. Cajanus cajan is cultivated in different regions of the world as an economical source of carbohydrates, fiber, proteins, and fodder for animals. In the current study, 39 K⁺ transport genes were identified in C. cajan, including 25 K⁺ transporters (17 carrier-like K⁺ transporters (KUP/HAK/KTs), 2 high-affinity potassium transporters (HKTs), and 6 K⁺ efflux transporters (KEAs) and 14 K⁺ channels (9 shakers and 5 tandem-pore K⁺ channels (TPKs). Chromosomal mapping indicated that these genes were randomly distributed among 10 chromosomes. A comparative phylogenetic analysis including protein sequences from Glycine max, Arabidopsis thaliana, Oryza sativa, Medicago truncatula Cicer arietinum, and C. cajan suggested vital conservation of K⁺ transport genes. Gene structure analysis showed that the intron/exon organization of K⁺ transporter and channel genes is highly conserved in a family-specific manner. In the promoter region, many cis-regulatory elements were identified related to abiotic stress, suggesting their role in abiotic stress response. Abiotic stresses (salt, heat, and drought) adversely affect chlorophyll, carotenoids contents, and total soluble proteins. Furthermore, the activities of catalase, superoxide, and peroxidase were altered in C. cajan leaves under applied stresses. Expression analysis (RNA-seq data and quantitative real-time PCR) revealed that several K⁺ transport genes were expressed in abiotic stress-responsive manners. The present study provides an in-depth understanding of K⁺ transport system genes in C. cajan and serves as a basis for further characterization of these genes. Full article

The Kunitz/BPTI peptide family includes unique representatives demonstrating various biological activities. Electrophysiological screening of peptides HCRG1 and HCRG2 from the sea anemone Heteractis crispa on six Kv1.x channel isoforms and insect Shaker IR channel expressed in Xenopus laevis oocytes revealed their potassium channels blocking activity. HCRG1 and HCRG2 appear to be the first Kunitz-type peptides from sea anemones blocking Kv1.3 with IC₅₀ of 40.7 and 29.7 nM, respectively. In addition, peptides mainly vary in binding affinity to the Kv1.2 channels. It was established that the single substitution, Ser5Leu, in the TRPV1 channel antagonist, HCRG21, induces weak blocking activity of Kv1.1, Kv1.2, and Kv1.3. Apparently, for the affinity and selectivity of Kunitz-fold toxins to Kv1.x isoforms, the number and distribution along their molecules of charged, hydrophobic, and polar uncharged residues, as well as the nature of the channel residue at position 379 (Tyr, Val or His) are important. Testing the compounds in a model of acute local inflammation induced by the introduction of carrageenan administration into mice paws revealed that HCRG1 at doses of 0.1–1 mg/kg reduced the volume of developing edema during 24 h, similar to the effect of the nonsteroidal anti-inflammatory drug, indomethacin, at a dose of 5 mg/kg. ELISA analysis of the animals blood showed that the peptide reduced the synthesis of TNF-α, a pro-inflammatory mediator playing a leading role in the development of edema in this model. Full article

The dynamic development of venomics in recent years has resulted in a significant increase in publicly available proteomic data. The information contained therein is often used for comparisons between different datasets and to draw biological conclusions therefrom. In this article, we aimed to show the possible differences that can arise, in the final results of the proteomic experiment, while using different research workflows. We applied two software solutions (PeptideShaker and MaxQuant) to process data from shotgun LC-MS/MS analysis of Naja ashei venom and collate it with the previous report concerning this species. We were able to provide new information regarding the protein composition of this venom but also present the qualitative and quantitative limitations of currently used proteomic methods. Moreover, we reported a rapid and straightforward technique for the separation of the fraction of proteins from the three-finger toxin family. Our results underline the necessary caution in the interpretation of data based on a comparative analysis of data derived from different studies. Full article

Voltage-gated potassium (K_V) channels regulate diverse physiological processes and are an important target for developing novel therapeutic approaches. Sea anemone (Cnidaria, Anthozoa) venoms comprise a highly complex mixture of peptide toxins with diverse and selective pharmacology on K_V channels. From the nematocysts of the sea anemone Actinia bermudensis, a peptide that we named AbeTx1 was purified and functionally characterized on 12 different subtypes of K_V channels (K_V1.1–K_V1.6; K_V2.1; K_V3.1; K_V4.2; K_V4.3; K_V11.1; and, Shaker IR), and three voltage-gated sodium channel isoforms (Na_V1.2, Na_V1.4, and BgNa_V). AbeTx1 was selective for Shaker-related K⁺ channels and is capable of inhibiting K⁺ currents, not only by blocking the K⁺ current of K_V1.2 subtype, but by altering the energetics of activation of K_V1.1 and K_V1.6. Moreover, experiments using six synthetic alanine point-mutated analogs further showed that a ring of basic amino acids acts as a multipoint interaction for the binding of the toxin to the channel. The AbeTx1 primary sequence is composed of 17 amino acids with a high proportion of lysines and arginines, including two disulfide bridges (Cys1–Cys4 and Cys2–Cys3), and it is devoid of aromatic or aliphatic amino acids. Secondary structure analysis reveals that AbeTx1 has a highly flexible, random-coil-like conformation, but with a tendency of structuring in the beta sheet. Its overall structure is similar to open-ended cyclic peptides found on the scorpion κ-KTx toxins family, cone snail venoms, and antimicrobial peptides. Full article

In Brazil, Tityus serrulatus (Ts) is the species responsible for most of the scorpion related accidents. Among the Ts toxins, the neurotoxins with action on potassium channels (α-KTx) present high interest, due to their effect in the envenoming process and the ion channel specificity they display. The α-KTx toxins family is the most relevant because its toxins can be used as therapeutic tools for specific target cells. The improved isolation method provided toxins with high resolution, obtaining pure Ts6 and Ts7 in two chromatographic steps. The effects of Ts6 and Ts7 toxins were evaluated in 14 different types of potassium channels using the voltage-clamp technique with two-microelectrodes. Ts6 toxin shows high affinity for Kv1.2, Kv1.3 and Shaker IR, blocking these channels in low concentrations. Moreover, Ts6 blocks the Kv1.3 channel in picomolar concentrations with an IC₅₀ of 0.55 nM and therefore could be of valuable assistance to further designing immunosuppressive therapeutics. Ts7 toxin blocks multiple subtypes channels, showing low selectivity among the channels analyzed. This work also stands out in its attempt to elucidate the residues important for interacting with each channel and, in the near future, to model a desired drug. Full article

Background. Alveolar hypoxia is an important condition related to many disorders such as chronic pulmonary hypertension, pulmonary vasoconstriction, and pulmonary vascular remodeling. The aim of present study was to disclose the biological response and the potential transcriptome networks regulating the hypoxia response in the lungs.
Materials and Methods. In this study, the microarray dataset GSE11341 was used to construct a regulatory network and identify the potential genes related to alveolar hypoxia. In addition, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) term enrichment analyses were also performed.
Results. Hypoxia inducible factor 1 alpha (HIF-1α), peroxisome proliferator-activated receptor gamma (PPARγ), and nuclear factor of kappa light polypeptide gene enhancer in B cells (NF-кB) were to be the hub nodes in the transcriptome network. HIF-1α may regulate potassium voltagegated channel, shaker-related subfamily, member (5KCNA5), solute carrier family 2 (facilitated glucose transporter), member (1SLC2A1), and heme oxygenase (decycling) 1 (HMOX1) expression through the regulation of membrane potential, glucose metabolism, and anti-inflammation pathways. HMOX-1 mediates signaling pathways that relate to NF-кB. CCND1 (cyclin D1) expression could be regulated by PPARγ and HIF-1α via the cell cycle pathway. In addition, new transcriptional factors and target genes, such as phosphofructokinase (PFKL, liver), aldolase A (ALDOA, fructose-bisphosphate), and trefoil factor 3 (intestinal) (TFF3), were also identified.
Conclusions. Transcriptome network analysis is a helpful method for the identification of the candidate genes in alveolar hypoxia. The KEGG pathway and GO term analysis are beneficial in the prediction of the underlying molecular mechanism of these identified genes in alveolar hypoxia. Full article