ATPase Gene Research Articles

Cupriavidus sp. strain Ni-2 resistant to high concentration of nickel and its genes responsible for the tolerance by genome comparison.

The widespread use of metals influenced many researchers to examine the relationship between heavy metal toxicity and bacterial resistance. In this study, we have inoculated heavy metal-contaminated soil from Janghang region of South Korea in the nickel-containing media (20mM Ni2+) for the enrichment. Among dozens of the colonies acquired from the several transfers and serial dilutions with the same concentrations of Ni, the strain Ni-2 was chosen for further studies. The isolates were identified for their phylogenetic affiliations using 16S rRNA gene analysis. The strain Ni-2 was close to Cupriavidus metallidurans and was found to be resistant to antibiotics of vancomycin, erythromycin, chloramphenicol, ampicillin, gentamicin, streptomycin, and kanamycin by disk diffusion method. Of the isolated strains, Ni-2 was sequenced for the whole genome, since the Ni-resistance seemed to be better than the other strains. From the genome sequence we have found that there was a total of 89 metal-resistance-related genes including 11 Ni-resistance genes, 41 heavy metal (As, Cd, Zn, Hg, Cu, and Co)-resistance genes, 22 cation-efflux genes, 4 metal pumping ATPase genes, and 11 metal transporter genes.

The effects of macrophages on cardiomyocyte calcium-handling function using in vitro culture models.

Following myocardial infarction (MI), myocardial inflammation plays a crucial role in the pathogenesis of MI injury and macrophages are among the key cells activated during the initial phases of the host response regulating the healing process. While macrophages have emerged as attractive effectors in tissue injury and repair, the contribution of macrophages on cardiac cell function and survival is not fully understood due to complexity of the in vivo inflammatory microenvironment. Understanding the key cells involved and how they communicate with one another is of paramount importance for the development of effective clinical treatments. Here, novel in vitro myocardial inflammation models were developed to examine how both direct and indirect interactions with polarized macrophage subsets present in the post‐MI microenvironment affect cardiomyocyte function. The indirect model using conditioned medium from polarized macrophage subsets allowed examination of the effects of macrophage‐derived factors on stem cell‐derived cardiomyocyte function for up to 3 days. The results from the indirect model demonstrated that pro‐inflammatory macrophage‐derived factors led to a significant downregulation of cardiac troponin T (cTnT) and sarcoplasmic/endoplasmic reticulum calcium ATPase (Serca2) gene expression. It also demonstrated that inhibition of macrophage‐secreted matricellular protein, osteopontin (OPN), led to a significant decrease in cardiomyocyte store‐operated calcium entry (SOCE). In the direct model, stem cell‐derived cardiomyocytes were co‐cultured with polarized macrophage subsets for up to 3 days. It was demonstrated that anti‐inflammatory macrophages significantly increased cardiomyocyte Ca2+ fractional release while macrophages independent of their subtypes led to significant downregulation of SOCE response in cardiomyocytes. This study describes simplified and controlled in vitro myocardial inflammation models, which allow examination of potential beneficial and deleterious effects of macrophages on cardiomyocytes and vise versa. This can lead to our improved understanding of the inflammatory microenvironment post‐MI, otherwise difficult to directly investigate in vivo or by using currently available in vitro models.

Expression Analysis of Genes Encoding NHX2 Antiporter and Subunit A of Vacuolar H+-ATPase Pump in Salt-Resistant and Salt-Sensitive Barley (Hordeum vulgare L.) Cultivars under Salt Stress

Salinity is one of the most important factors causing limiting growth aspects. Salinity tolerance is a multigenic trait activating mechanisms such as high H+ pumping activity, like NHX2 that is a tonoplast localized Na+/H+ exchanger and actively transfers the excess sodium ions from the cytoplasm into the vacuole. Vacuolar (v-) H+-ATPase pump is also a tonoplast localized, providing the ATP energy required for tonoplast antiporters. Evaluation of the expression level of HvNHX2 and subunit A of v-H+-ATPase (VHA-A) genes using real-time RT-PCR, in root and shoot tissue of salt-sensitive Reihan and salt-tolerant Afzal barley (Hordeum vulgare L.) cultivars under different salinity concentrations indicated that increasing the concentrations or duration of NaCl stress would decrease the K+/Na ratio that is a signal for cascade responses to the stress but fluctuated expression level of studied genes in some time point. However, promoter analysis of studied genes showed that both have CAAT, TATA and W-box promoter motifs which involve pathogen, salt and ABA-responsive signaling pathways but differed in TC-rich repeats, A-box, TCA-element and GARE-motif that may be one of the reason of observed different.

Phytohormones enabled endophytic Penicillium funiculosum LHL06 protects Glycine max L. from synergistic toxicity of heavy metals by hormonal and stress-responsive proteins modulation.

This study investigates the stress-mitigating effects of endophytic Penicillium funiculosum LHL06 on soybean roots via modulation of physio-biochemical, molecular, and proteomic responses to combined heavy metal (Ni, Cu, Pb, Cr, and Al) toxicity. Preliminary screening revealed that LHL06 can tolerate and remediate combined heavy metal contamination in its media and upregulate gibberellins (GA1, GA3, GA4, GA7 and GA9) and indole-3-acetic acid (IAA) production. Inoculation of LHL06 resulted in marked reduction of metals uptake in roots and shoots by downregulating heavy metal ATPase genes (GmHMA13, GmHMA14, GmHMA19) and GmMATE1 compared to non-inoculated plants; in turn, this decreased abscisic acid and jasmonic acid levels. Moreover, triggering of free amino acid metabolism in LHL06-inoculated roots significantly upregulated expression of stress-related proteins (glutathione S-transferase L3, isoflavone reductase-like, chalcone isomerase A, NAD(P)H dehydrogenase (quinone), FQR1-like 1 isoform X2, and Peroxidase 3) to combat metals toxicity. Compared to non-inoculated-plants, LHL06-inoculated-plants exhibited higher antioxidant activity and transcript accumulation of glutathione S-transferase (GmGST8 and GmGST3), G6PDH, and GmSOD1[Cu-Zn], which decreased metal-induced reactive oxygen species. Therefore, LHL06-inoculation remediate combined metal contamination in soil, activate signaling network of stress-responsive hormones and antioxidant systems for promoting growth and tolerance, and reduce metal-accumulation, thereby making plants safer for consumption.

Effects of stocking density on stress response, innate immune parameters, and welfare of turbot (Scophthalmus maximus)

This experiment was conducted to investigate the growth performance, stress and immune responses, and welfare of juvenile turbot (Scophthalmus maximus) under three different densities (initial density 9.3, 13.6, and 19.1 kg m−2) for 120 days in a recirculating aquaculture system. Turbot were measured every 20 days to evaluate growth biometrically and sampled every month to measure biochemical parameters and mRNA levels of some stress-related genes. No significant differences were detected in the parameters and gene expression among density groups until the final sampling, except Fulton’s condition factor, lysozyme, immunoglobulin M, and complement C3. At the end of the experiment (final density 26.11, 38.22, and 52.25 kg m−2), turbot reared in the high-density (HD) group had lower body mass increase, specific growth rate, and Fulton’s condition factor, as well as higher feed conversion ratio and coefficient of variation for weight than those reared in the low-density (LD) group (P < 0.05). Fish in the HD group had higher serum cortisol, glucose, lactate, and cholesterol levels than fish in the other groups, whereas they had lower lysozyme, immunoglobulin M, and complement C3 and C4 contents (P < 0.05). Fish reared in the HD group also had higher serum chloride and osmolality levels and higher sodium-potassium adenosine triphosphatase (Na+,K+-ATPase) activity and higher Na+,K+-ATPase gene expression levels in gills compared to the other groups (P < 0.05). The mRNA levels of cytochrome P450 1A (CYP1A) and heat-shock proteins 70 and 90 (HSP70 and HSP90) were significantly upregulated, whereas glutathione S-transferase (GST) mRNA levels were significantly downregulated in the head kidney of fish in the HD group relative to fish in the other groups at the end of this trial (P < 0.05). These results indicated that overly high stocking (~50 kg m−2) density can negatively affect the growth performance, serum biochemical parameters, osmolality levels, stress-related gene expression, and overall welfare of turbot.

Na+-K+-ATPase and nka genes in spotted sea bass (Lateolabrax maculatus) and their involvement in salinity adaptation

Euryhaline teleosts can survive in a wide salinity range via alteration of the molecular mechanisms to maintain internal ionic and osmotic balance in osmoregulatory organs such as gill，kidney and intestine. Na+/K+-ATPase (NKA), plays a crucial role in sustaining intracellular homeostasis and is characterized by association of multiple isoforms of α- and β-subunits. To gain insight into the potential function of nka genes in salinity adaptation, 5 nkaα genes (nkaα1a, nkaα1b, nkaα2, nkaα3a, nkaα3b) and 7 nkaβ genes (nkaβ1a, nkaβ1b, nkaβ2a, nkaβ2b, nkaβ3a, nkaβ3b and nkaβ4) were identified from transcriptomic and genomic databases of Lateolabrax maculatus. The annotation and evolutionary footprint of these nka genes was revealed via the analysis of phylogenetic tree, gene synteny, copy numbers, exon-intron structures and motif compositions. The expressions of 12 nka genes in spotted sea bass was tested in ten tissues (kidney, gonad, stomach, intestine, gill, muscle, heart, spleen, liver and brain) and 6 genes (nkaα1a, nkaα1b, nkaα3a, nkaα3b, nkaβ1b and nkaβ2a) showed high expression in osmoregulatory organs. Furthermore, the responses of NKA and potential salinity-sensitive nka genes were examined under different salinity treatment (0 ppt, 12 ppt, 30 ppt, 45 ppt). Results showed that the enzyme activity of NKA was highest in gill and exhibited salinity dependent variation, with the highest activity identified in 45 ppt. Different nkaα/β-isoforms showed their diverse responses to salinity changes and the expression of nka genes including nkaα1a, nkaα3b, nkaβ1b in gill, nkaα3a in kidney and nkaβ2a in intestine were transcriptionally regulated by altered salinity. Notably, the expression patterns of nkaα1a and nkaβ1b in gill showed similar variation trend with NKA activity, suggesting that nkaα1a/β1b could be the major function isoforms involved in primary ion transport during salinity adaptation. Our results provided insights into the roles of nkas in osmotic regulation and a theoretical basis for future studies that focus on detailed molecular mechanisms in salinity adaptation of euryhaline teleosts.

Molecular insight into the expression of metal transporter genes in Chryseobacterium sp. PMSZPI isolated from uranium deposit.

Metal tolerant bacterium Chryseobacterium sp. PMSZPI previously isolated and characterized from uranium ore deposit was studied for elucidating the role of metal transporter genes belonging to the Cation Diffusion Facilitator (CDF), Root-Nodulation-Division (RND) and PIB-type ATPase family in cadmium and uranium tolerance. The bacterium showed tolerance towards cadmium (MIC~6mM) and uranium (MIC~2mM) and was found to harbor metal transporter genes belonging to CDF, RND and PIB-type ATPase family of proteins. Expression studies by real-time PCR showed an upregulation of czcA(RND), czcD(CDF) and cadA(PIB-type ATPase) genes in presence of cadmium or uranium. Higher expression of czcA and czcD was found when the bacterium was treated with cadmium and uranium respectively. This study provides significant insight into the molecular mechanism that plays a role in cadmium and uranium tolerance in bacteria.

Characterization of a Novel Megalocytivirus Isolated from European Chub (Squalius cephalus).

A novel virus from moribund European chub (Squalius cephalus) was isolated on epithelioma papulosum cyprini (EPC) cells. Transmission electron microscopic examination revealed abundant non-enveloped, hexagonal virus particles in the cytoplasm of infected EPC cells consistent with an iridovirus. Illumina MiSeq sequence data enabled the assembly and annotation of the full genome (128,216 bp encoding 108 open reading frames) of the suspected iridovirus. Maximum Likelihood phylogenetic analyses based on 25 iridovirus core genes supported the European chub iridovirus (ECIV) as being the sister species to the recently-discovered scale drop disease virus (SDDV), which together form the most basal megalocytivirus clade. Genetic analyses of the ECIV major capsid protein and ATPase genes revealed the greatest nucleotide identity to members of the genus Megalocytivirus including SDDV. These data support ECIV as a novel member within the genus Megalocytivirus. Experimental challenge studies are needed to fulfill River’s postulates and determine whether ECIV induces the pathognomonic microscopic lesions (i.e., megalocytes with basophilic cytoplasmic inclusions) observed in megalocytivirus infections.

Etiology of Alternaria Leaf Spot of Cotton in Southern New Mexico.

Alternaria leaf spot (caused by Alternaria spp.) is one of the most common foliar diseases of cotton (Gossypium spp.) and occurs in most cotton-growing regions of the world. In surveys of commercial cotton fields, Alternaria leaf spot has increased in prevalence and incidence in southern New Mexico due to favorable environmental conditions in recent years. Incidence, severity, and etiology of leaf spot of cotton in southern New Mexico were determined. Fourteen cotton fields with plants exhibiting leaf spot symptoms were evaluated in October and November 2016, when plants were at late growth stage. Disease incidence was 100% in 13 of the fields, and averaged 70% in the 14th field. Average disease severity index for all fields ranged from 21.5 to 87.0. For identification of the causal agent, 14 isolates (one from each field) were characterized based on morphological features and PCR using universal primers ITS4/ITS5 and primers targeting the plasma membrane ATPase gene. Colonies of all 14 isolates were olive green with distinct white margins and relatively small spores when compared with reference isolates of large-spored species. All 14 isolates were identified as A. alternata. The fungus grew on potato dextrose agar from 5 to 35°C, and optimum growth occurred at temperatures between 20 and 30°C. Cotton plants inoculated with selected isolates of A. alternata displayed symptoms similar to those observed under field conditions. This is the first report of A. alternata as a causal agent of Alternaria leaf spot on cotton in southern New Mexico.

Defective Expression of Mitochondrial, Vacuolar H+-ATPase and Histone Genes in a C. elegans Model of SMA.

Spinal muscular atrophy (SMA) is a severe motor neuron degenerative disease caused by loss-of-function mutations in the survival motor neuron gene SMN1. It is widely posited that defective gene expression underlies SMA. However, the identities of these affected genes remain to be elucidated. By analyzing the transcriptome of a Caenorhabditis elegans SMA model at the pre-symptomatic stage, we found that the expression of numerous nuclear encoded mitochondrial genes and vacuolar H+-ATPase genes was significantly down-regulated, while that of histone genes was significantly up-regulated. We previously showed that the uaf-1 gene, encoding key splicing factor U2AF large subunit, could affect the behavior and lifespan of smn-1 mutants. Here, we found that smn-1 and uaf-1 interact to affect the recognition of 3′ and 5′ splice sites in a gene-specific manner. Altogether, our results suggest a functional interaction between smn-1 and uaf-1 in affecting RNA splicing and a potential effect of smn-1 on the expression of mitochondrial and histone genes.

Differential expression patterns of sodium potassium ATPase alpha and beta subunit isoforms in mouse brain during postnatal development

The sodium potassium ATPase (Na+/K+ ATPase) is essential for the maintenance of a low intracellular Na+ and a high intracellular K+ concentration. Loss of function of the Na+/K+ ATPase due to mutations in Na+/K+ ATPase genes, anoxic conditions, depletion of ATP or inhibition of the Na+/K+ ATPase function using cardiac glycosides such as digitalis, causes a depolarization of the resting membrane potential. While in non-excitable cells, the uptake of glucose and amino acids is decreased if the function of the Na+/K+ ATPase is compromised, in excitable cells the symptoms range from local hyper-excitability to inactivating depolarization. Although several studies have demonstrated the differential expression of the various Na+/K+ ATPase alpha and beta isoforms in the brain tissue of rodents, their expression profile during development has yet to be thoroughly investigated. An immunohistochemical analysis of postnatal day 19 mouse brain showed ubiquitous expression of Na+/K+ ATPase isoforms α1, β1 and β2 in both neurons and glial cells, whereas α2 was expressed mostly in glial cells and the α3 and β3 isoforms were expressed in neurons. Furthermore, we examined potential changes in the relative expression of the different Na+/K+ ATPase isoforms in different brain areas of postnatal day 6 and in adult 9 months old animals using immunoblot analysis. Our results show a significant up-regulation of the α1 isoform in cortex, hippocampus and cerebellum, whereas, the α2 isoform was significantly up-regulated in midbrain. The β3 isoform showed a significant up-regulation in all brain areas investigated. The up-regulation of the α3 isoform matched that of the β2 isoform which were both significantly up-regulated in cortex, hippocampus and midbrain, suggesting that the increased maturation of the neuronal network is accompanied by an increase in expression of α3/β2 complexes in these brain structures.

Calcium acetate-induced reduction of cadmium accumulation in Oryza sativa: Expression of auto-inhibited calcium-ATPase and cadmium transporters.

Calcium (Ca) signalling has an essential role in regulating plant responses to various abiotic stresses. This study applied Ca in various forms (Ca acetate and CaCl2 ) and concentrations to reduce cadmium (Cd) concentration in rice and propose a possible mechanism through which Ca acts to control the Cd concentration in rice. The results showed that supplementation of Cd-contaminated soil with Ca acetate reduced the Cd concentration in rice after exposure for 7days in both hydroponic and soil conditions. The possible involvement of the auto-inhibited Ca2+ -ATPase gene (ACA) might act to control the primary signal of the Cd stress response. The messages from ACA3 and ACA13 tended to up-regulate the low-affinity cation transporter (OsLCT1) and down-regulate Cd uptake and the Cd translocation transporter, including the genes, natural resistance-associated macrophage protein 5 (Nramp5) and Zn/Cd-transporting ATPase 2 (HMA2), which resulted in a reduction in the Cd concentration in rice. After cultivation for 120days, the application of Ca acetate into Cd-contaminated soil inhibited Cd uptake of rice. Increasing the Ca acetate concentration in the soil lowered the Cd concentration in rice shoots and grains. Moreover, Ca acetate maintained rice productivity and quality whereas both aspects decreased under Cd stress.

Na+ /K+ -ATPase gene duplications in clitellate annelids are associated with freshwater colonization.

Major habitat transitions, such as those from marine to freshwater habitats or from aquatic to terrestrial habitats, have occurred infrequently in animal evolution and may represent a barrier to diversification. Identifying genomic events associated with these transitions can help us better understand mechanisms that allow animals to cross these barriers and diversify in new habitats. Study of the Capitella telata and Helobdella robusta genomes allows examination of one such habitat transition (marine to freshwater) in Annelida. Initial examination of these genomes indicated that the freshwater leech H.robusta contains many more copies (12) of the sodium-potassium pump alpha-subunit (Na+ /K+ -ATPase) gene than does the marine polychaete C.telata (2). The sodium-potassium pump plays a key role in maintenance of cellular ionic balance and osmoregulation, and Na+ /K+ -ATPase duplications may have helped annelids invade and diversify in freshwater habitats. To assess whether the timing of Na+ /K+ -ATPase duplications coincided with the marine-to-freshwater transition in Clitellata, we used transcriptomic data from 18 annelid taxa, along with the two genomes, to infer a species phylogeny and identified Na+ /K+ -ATPase gene transcripts in order to infer the timing of gene duplication events using tree-based methods. The inferred timing of Na+ /K+ -ATPase duplication events is consistent with the timing of the initial marine-to-freshwater transition early in the history of clitellate annelids, supporting the hypothesis that gene duplications may have played a role in the annelid diversification into freshwater habitats.

MicroRNA Sequencing Revealed Citrus Adaptation to Long-Term Boron Toxicity through Modulation of Root Development by miR319 and miR171.

Boron (B) toxicity in Citrus is a common physiological disorder leading to reductions in both productivity and quality. Studies on how Citrus roots evade B toxicity may provide new insight into plant tolerance to B toxicity. Here, using Illumina sequencing, differentially expressed microRNAs (miRNAs) were identified in B toxicity-treated Citrus sinensis (tolerant) and C. grandis (intolerant) roots. The results showed that 37 miRNAs in C. grandis and 11 miRNAs in C. sinensis were differentially expressed when exposed to B toxicity. Among them, miR319, miR171, and miR396g-5p were confirmed via 5′-RACE and qRT-PCR to target a myeloblastosis (MYB) transcription factor gene, a SCARECROW-like protein gene, and a cation transporting ATPase gene, respectively. Maintenance of SCARECROW expression in B treated Citrus roots might fulfill stem cell maintenance, quiescent center, and endodermis specification, thus allowing regular root elongation under B-toxic stress. Down-regulation of MYB due to up-regulation of miR319 in B toxicity-treated C. grandis roots might decrease the number of root tips, thereby dramatically changing root system architecture. Our findings suggested that miR319 and miR171 play a pivotal role in Citrus adaptation to long-term B toxicity by targeting MYB and SCARECROW, respectively, both of which are responsible for root growth and development.

Retinal abiotrophy in mitochondrial pathology: NARP sindrome (a clinical case)

Purpose: to present a clinical case of retinal abiotrophy in mitochondrial pathology (NARP syndrome) caused by the mutation m.8993T&gt;G in the ATPase gene type 6 in order to improve the diagnosis of hereditary abiotrophies. Material and methods. The results of a clinical molecular genetic examination of the patient’s family, undertaken in order to clarify the diagnosis and determine the genetic risk, are presented. The family was found to have an isolated pathology of the eye. Results. DNA studies by MLPA method and the analysis of clinical data in the family revealed a hereditary syndromic pathology which caused changes in the eyes. The inheritance type was found to be maternal. Conclusion. NARP syndrome is a syndrome with the maternal type of inheritance in which retinal abiotrophy is primarily associated with the mutation m.8993T&gt;G mtDNA and can be considered as the main diagnostic feature among other clinical manifestations. The case demonstrates the difficulties of diagnosing hereditary syndromes accompanied by eye pathology.

Arbuscular mycorrhiza augments cadmium tolerance in soybean by altering accumulation and partitioning of nutrient elements, and related gene expression

Arbuscular mycorrhizal (AM) fungi can protect plants against cadmium (Cd) stress, and are the most prominent symbiotic fungi for contribution to phytoremediation. However, the tolerance mechanism for AM symbiosis on Cd toxicity still remains unclear, especially the related molecular mechanisms. In this study, different Cd treatments were applied to two soybean genotypes with different Cd tolerance in the presence or absence of AM fungal inoculation. The results showed that Cd addition obviously decreased AM colonization. AM symbiosis significantly increased plant dry weight, root growth, and P acquisition in Cd-tolerant HX3 genotype at Cd addition treatments. The effectiveness was associated with a concomitant increased expression of the AM inducible phosphate (Pi) transporter genes GmPT8, GmPT9, GmPT10, and upregulated expression of P-type heavy metal ATPase gene GmHMA19. Additionally, AM fungal inoculation effectively impacted the partitioning of Mg, Cu and Zn, including increased Mg, and decreased Cu and Zn relative concentrations in shoots of Cd tolerant HX3. Taken together, these results suggest that AM symbiosis can alleviate Cd toxicity in soybean through enhanced P nutrition, up-regulated expression of AM inducible GmPTs and GmHMA19, as well as, the alteration of the partitioning of essential nutrient elements.

P-type transport ATPases in Leishmania and Trypanosoma.

P-type ATPases are critical to the maintenance and regulation of cellular ion homeostasis and membrane lipid asymmetry due to their ability to move ions and phospholipids against a concentration gradient by utilizing the energy of ATP hydrolysis. P-type ATPases are particularly relevant in human pathogenic trypanosomatids which are exposed to abrupt and dramatic changes in their external environment during their life cycles. This review describes the complete inventory of ion-motive, P-type ATPase genes in the human pathogenic Trypanosomatidae; eight Leishmania species (L. aethiopica, L. braziliensis, L. donovani, L. infantum, L. major, L. mexicana, L. panamensis, L. tropica), Trypanosoma cruzi and three Trypanosoma brucei subspecies (Trypanosoma brucei brucei TREU927, Trypanosoma brucei Lister strain 427, Trypanosoma brucei gambiense DAL972). The P-type ATPase complement in these trypanosomatids includes the P1B (metal pumps), P2A (SERCA, sarcoplasmic-endoplasmic reticulum calcium ATPases), P2B (PMCA, plasma membrane calcium ATPases), P2D (Na+ pumps), P3A (H+ pumps), P4 (aminophospholipid translocators), and P5B (no assigned specificity) subfamilies. These subfamilies represent the P-type ATPase transport functions necessary for survival in the Trypanosomatidae as P-type ATPases for each of these seven subfamilies are found in all Leishmania and Trypanosoma species included in this analysis. These P-type ATPase subfamilies are correlated with current molecular and biochemical knowledge of their function in trypanosomatid growth, adaptation, infectivity, and survival.

Improving outcomes for patients with distal renal tubular acidosis: recent advances and challenges ahead.

Primary distal renal tubular acidosis (dRTA) is a rare genetic disorder caused by impaired distal acidification due to a failure of type A intercalated cells (A-ICs) in the collecting tubule. dRTA is characterized by persistent hyperchloremia, a normal plasma anion gap, and the inability to maximally lower urinary pH in the presence of systemic metabolic acidosis. Common clinical features of dRTA include vomiting, failure to thrive, polyuria, hypercalciuria, hypocitraturia, nephrocalcinosis, nephrolithiasis, growth delay, and rickets. Mutations in genes encoding three distinct transport proteins in A-ICs have been identified as causes of dRTA, including the B1/ATP6V1B1 and a4/ATP6V0A4 subunits of the vacuolar-type H+-ATPase (H+-ATPase) and the chloride–bicarbonate exchanger AE1/SLC4A1. Homozygous or compound heterozygous mutations in ATP6V1B1 and ATP6V0A4 lead to autosomal recessive (AR) dRTA. dRTA caused by SLC4A1 mutations can occur with either autosomal dominant or AR transmission. Red blood cell abnormalities have been associated with AR dRTA due to SLC4A1 mutations, including hereditary spherocytosis, Southeast Asia ovalocytosis, and others. Some patients with dRTA exhibit atypical clinical features, including transient and reversible proximal tubular dysfunction and hyperammonemia. Incomplete dRTA presents with inadequate urinary acidification, but without spontaneous metabolic acidosis and recurrent urinary stones. Heterozygous mutations in the AE1 or H+-ATPase genes have recently been reported in patients with incomplete dRTA. Early and sufficient doses of alkali treatment are needed for patients with dRTA. Normalized serum bicarbonate, urinary calcium excretion, urinary low-molecular-weight protein levels, and growth rate are good markers of adherence to and/or efficacy of treatment. The prognosis of dRTA is generally good in patients with appropriate treatment. However, recent studies showed an increased frequency of chronic kidney disease (CKD) in patients with dRTA during long-term follow-up. The precise pathogenic mechanisms of CKD in patients with dRTA are unknown.

Gene mutation associated with esl mediates shifts on fungal community composition in rhizosphere soil of rice at grain-filling stage

Generally, plant roots shape the rhizosphere fungal community but how individual plant genes involved in senescence affect this shaping is less studied. We used an early senescence leaf (esl) mutant rice and compared it with its isogenic wild type variety to evaluate the effect of the vacuolar H+-ATPase (VHA-A1) gene mutation on the rhizosphere fungal community structure and composition using a metagenomic pyrosequencing approach. The most predominate fungal phyla identified for both isogenic lines belonged to Ascomycota, Basidiomycota and Glomeromycota, where Ascomycota were more prevalent in the esl mutant than the wild type variety. Real-time quantitative PCR analysis confirmed a significant rise in the richness of Cladosporium cladosporioides in esl mutant rice than the wild type variety. Correlation analysis revealed four most abundant genera identified for the esl mutant and their close association with yield and biomass decline, lipid peroxidation, lower root vitality, chlorophyll degradation and limited VHA activity. Higher K+ efflux, H+ and a lower Ca2+ influx was also observed in the esl mutant which could be the reason for abnormal functioning of mutant plants. These results illustrate that besides the well-known effect of senescence on plant physiology and yield decline, it can further shape the rhizosphere fungal community.

Insights into regulatory roles of MAPK-cascaded pathways in multiple stress responses and life cycles of insect and nematode mycopathogens.

Fungal entomopathogenicity may have evolved at least 200 million years later than carnivorism of nematophagous fungi on Earth. This mini-review focuses on the composition and regulatory roles of mitogen-activated protein kinase (MAPK) cascades, which act as stress-responsive signaling pathways. Unveiled by genomic comparison, three MAPK cascades of these mycopathogens consist of singular MAPKs (Fus3/Hog1/Slt2), MAPK kinases (Ste7/Pbs2/Mkk1), and MAPK kinase kinases (Ste11/Ssk2/Bck1). All cascaded components characterized in fungal entomopathogens play conserved and special roles in regulating multiple stress responses and phenotypes associated with biological control potential. Fus3-cascaded components are indispensable for fungal growth on oligotrophic substrata and virulence, and mediate cell tolerance to Na+/K+ toxicity, which is often misinterpreted as hyperosmotic effect but readily clarified by transcriptional changes of Na+/K+ ATPase genes and/or cell responses to osmotic polyols. Hog1-cascaded components regulate osmotolerance positively and phenylpyrrole-type fungicide resistance negatively, and also play differential roles in cell growth, conidiation, virulence, and responses to other stress cues. Ste11 has no stress-responsive role in the Beauveria Hog1 cascade despite an essential role in branched yeast Hog1 cascade. Slt2-cascaded components are required for mediation of cell wall integrity and repair of cell wall damage. A crosstalk between Hog1 and Slt2 cascades ensures fungal osmotolerance inside or outside insect. In nematode-trapping fungi, Slt2 is indispensable for cell wall integrity, conidiation, and mycelial trap formation, suggesting that the Slt2 cascade could have evolved along a distinct trajectory required for fungal carnivorism and dispersal/survival in nematode habitats. Altogether, the MAPK cascades are major parts of signaling network that regulate fungal adaptation to insects and nematodes and their habitats.