Type 2C Research Articles

OsPP2C49, a Negative Regulatory Factor in the Abscisic Acid Signaling Pathway, Positively Regulates Grain Yield in Rice

Clade A type 2C protein phosphatases (PP2Cs) are crucial components of the abscisic acid (ABA) signaling pathway. Research on clade A PP2Cs has focused more on their roles related to ABA signaling and stress responses than on the molecular mechanisms mediating their effects on plant growth and grain yield. Rice (Oryza sativa L.) is an important food crop worldwide. We previously determined that OsPP2C49, which encodes a rice clade A PP2C family member, negatively controls rice responses to drought, salt, and high-temperature stresses. In this study, we investigated the regulatory effects of OsPP2C49 on ABA responses and rice grain yield. By analyzing potential interactions with core ABA components, including pyrabactin resistance 1 (PYR1)/PYR1-like (PYL)/regulatory component of the ABA receptor (RCAR) and stress-activated protein kinases (SAPKs), we confirmed that OsPP2C49 is involved in the ABA signaling pathway. OsPP2C49 overexpression led to decreased ABA sensitivity and increased rice grain yield; the opposite phenotypes were observed in the ospp2c49 knockout mutants. Therefore, OsPP2C49 negatively regulates ABA responses, but positively modulates rice grain yield. Furthermore, we found that OsPP2C49 can interact with and dephosphorylate five OsSAPKs in vitro. Unlike OsPP2C49, these OsSAPKs positively modulate ABA responsiveness, but negatively affect rice yield. These findings indicate that OsPP2C49 may partially regulate ABA responses and rice grain production by dephosphorylating OsSAPKs. This study preliminarily explored the molecular basis of the regulatory effects of OsPP2C49 on rice plant growth and grain yield.

A Positive Role for CaMEKK17 in Response to Drought Stress, Modulated by Clade A PP2Cs.

The abscisic acid (ABA) signaling pathway is essential for plant response to abiotic stresses and can be modulated positively or negatively by MAPKKK proteins. This study focuses on the functional characterization of CaMEKK17, a MAPKKK previously recognized for its rapid induction under drought stress. Functional analyses demonstrated that CaMEKK17 is an active serine/threonine kinase with a conserved catalytic domain that is crucial for its kinase activity. CaMEKK17 silencing in pepper plants resulted in reduced drought tolerance, characterized by increased transpirational water loss and impaired ABA-mediated stomatal closure. Conversely, CaMEKK17 overexpression in Arabidopsis increased kinase activity, enhancing ABA sensitivity and drought tolerance. Further investigation revealed that CaMEKK17 interacts with pepper group A type 2C protein phosphatases (PP2Cs), particularly CaAITP1 and CaAIPP1, which inhibit its kinase activity. Protein-protein interactions mediated inhibition by CaAITP1, whereas CaAIPP1 relied on its phosphatase activity. Double gene silencing of CaMEKK17 and CaAITP1 demonstrated that CaMEKK17 functions downstream of CaAITP1 in ABA-mediated drought tolerance. Taken together, our findings suggest that CaMEKK17 positively modulates drought tolerance in pepper plants but may be inhibited by PP2Cs.

A New High-Frequency Ultrasound Classification of De Quervain Tenosynovitis

PurposeThis study aimed to introduce a new high-frequency ultrasound classification of De Quervain tenosynovitis based on a large group of patients. Detailed characteristics of classification are also reported. MethodsFrom January 2014 to February 2024, patients diagnosed with De Quervain tenosynovitis were retrospectively reviewed. High-frequency ultrasound (7-14 MHz) scanning was performed to identify the anatomy of the extensor pollicis brevis and abductor pollicis longus tendons, presence of intertendinous septa, and pathologic changes. The affected wrist was compared to the contralateral wrist. Results453 patients were included: 65 male and 388 female; mean age, 46±27 years (range, 24–65 years). Symptom duration was 14±27 weeks. Disease types were type 0 (n= 5), type 1 (n=195), type 2a (n=72), type 2b (n=18), type 2c (n=50), type 3 (n=59), type 4 (n=45), and type 5 (n=9). Mean retinacular thickness was 2.1±0.5 mm in affected wrists and 0.4±0.1 mm in contralateral wrists (p<0.01). The intercompartmental septum was significantly thicker on the affected side (1.2±0.7 mm) than on the asymptomatic side (0.1±0.4 mm) (p<0.01). ConclusionsThis novel classification provides detailed ultrasonographic characteristics of De Quervain tenosynovitis, based on a large population of patients. It may help in selecting treatment and predicting outcome. However, relevance and therapeutic significance remain to be demonstrated. Level of EvidenceTherapeutic study, Level IV.

Transcription factor SlSTOP1 regulates Small Auxin-Up RNA Genes for tomato root elongation under aluminum stress.

Aluminum (Al) stress, a prevalent constraint in acidic soils, inhibits plant growth by inhibiting root elongation through restricted cell expansion. The molecular mechanisms of Al-induced root inhibition, however, are not fully understood. This study aimed to elucidate the role of Small Auxin-up RNAs (SlSAURs), which function downstream of the key Al stress-responsive transcription factor SENSITIVE TO PROTON RHIZOTOXICITY 1 (SlSTOP1) and its enhancer STOP1-INTERACTING ZINC-FINGER PROTEIN 1 (SlSZP1), in modulating root elongation under Al stress in tomato (Solanum lycopersicum). Our findings demonstrated that tomato lines with knocked out SlSAURs exhibited shorter root lengths when subjected to Al stress. Further investigation into the underlying mechanisms revealed that SlSAURs interact with Type 2C Protein Phosphatases (SlPP2Cs), specifically D-clade Type 2C Protein Phosphatases (SlPP2C.Ds). This interaction was pivotal as it suppresses the phosphatase activity, leading to the degradation of SlPP2C.D's inhibitory effect on plasma membrane H+-ATPase. Consequently, this promoted cell expansion and root elongation under Al stress. These findings increase our understanding of the molecular mechanisms by which Al ions modulate root elongation. The discovery of the SlSAUR-SlPP2C.D interaction and its impact on H+-ATPase activity also provides a perspective on the adaptive strategies employed by plants to cope with Al toxicity, which may lead to the development of tomato cultivars with enhanced Al stress tolerance, thereby improving crop productivity in acidic soils.

MANAGEMENT OF CHRONIC PELVIC DISCONTINUITY DURING REVISION HIP ARTHROPLASTY USING THE “ACETABULAR DISTRACTION TECHNIQUE”: CLINICAL AND RADIOLOGICAL OUTCOMES AT TWO TO TEN YEARS FROM A TWO-CENTRE STUDY

Chronic pelvic discontinuity (CPD) during revision hip arthroplasty is a challenging entity to address. The aim of this study was to evaluate the clinical and radiologic outcomes, and complications of the “acetabular distraction technique” for the management of CPD during revision hip arthroplasty.Patients with CPD, who underwent acetabular revision between 2014 and 2022 at two tertiary care centres, using an identical distraction technique, were evaluated. Demographic parameters, pre-operative acetabular bone loss, duration of follow-up, clinical and radiologic outcomes, and survivorship were evaluated.In all, 46 patients with a mean follow-up of 34.4 (SD=19.6, range: 24–120) months were available for evaluation. There were 25 (54.3%) male, and 21 (45.7%) female patients, with a mean age of 58.1 (SD=10.5, range: 40–81) years at the time of revision surgery. Based on the Paprosky classification of acetabular bone loss, 19 (41.3%), 12 (26.1%), and 15 (32.6%) patients had type 3b, 3a, and 2c defects. All patients were managed using the Trabecular Metal™ Acetabular Revision System; 16 patients required additional Trabecular Metal™ augments. The mean HHS improved from 50.1 (SD=7.6, range: 34.3 – 59.8) pre-operatively, to 86.6 (SD=4.2, range: 74.8 -91.8) at the last follow-up. Two patients (4.3 %) developed partial sciatic nerve palsy, two (4.3%) had posterior dislocation, and one (2.2%) required re-revision for aseptic loosening. Radiologically, 36 (78.3%) patients showed healing of the pelvic discontinuity. The Kaplan-Meier construct survivorship was 97.78% when using re-revision for aseptic acetabular loosening as an endpoint.The acetabular distraction technique has good clinical and radiologic outcomes in the management of CPD during revision hip arthroplasty.

Related type 2C protein phosphatases Pic3 and Pic12 negatively regulate immunity in tomato to Pseudomonas syringae.

Type 2C protein phosphatases (PP2Cs) constitute a large family in most plant species but relatively few of them have been implicated in immunity. To identify and characterize PP2C phosphatases that affect tomato (Solanum lycopersicum) immunity, we used CRISPR/Cas9 to generate loss-of-function mutations in 11 PP2C-encoding genes whose expression is altered in response to immune elicitors or pathogens. We report that two closely related PP2C phosphatases, Pic3 (PP2C immunity-associated candidate 3) and Pic12, are involved in regulating resistance to the bacterial pathogen Pseudomonas syringae pv. tomato (Pst). Loss-of-function mutations in Pic3 led to enhanced resistance to Pst in older but not younger leaves, whereas such mutations in Pic12 resulted in enhanced resistance in both older and younger leaves. Overexpression of Pic3 and Pic12 proteins in leaves of Nicotiana benthamiana inhibited resistance to Pst, and this effect was dependent on Pic3/12 phosphatase activity and an N-terminal palmitoylation motif associated with localization to the cell periphery. Pic3, but not Pic12, had a slight negative effect on flagellin-associated reactive oxygen species generation, although their involvement in the response to Pst appeared independent of flagellin. RNA-sequencing analysis of Rio Grande (RG)-PtoR wild-type plants and two independent RG-pic3 mutants revealed that the enhanced disease resistance in RG-pic3 older leaves is associated with increased transcript abundance of multiple defense related genes. RG-pic3/RG-pic12 double mutant plants exhibited stronger disease resistance than RG-pic3 or RG-pic12 single mutants. Together, our results reveal that Pic3 and Pic12 negatively regulate tomato immunity in an additive manner through flagellin-independent pathways.

PP2C phosphatase Pic14 negatively regulates tomato Pto/Prf-triggered immunity by inhibiting MAPK activation.

Type 2C protein phosphatases (PP2Cs) are emerging as important regulators of plant immune responses, although little is known about how they might impact nucleotide-binding, leucine-rich repeat (NLR)-triggered immunity (NTI). We discovered that expression of the PP2C immunity-associated candidate 14 gene (Pic14) is induced upon activation of the Pto/Prf-mediated NTI response in tomato. Pto/Prf recognizes the effector AvrPto translocated into plant cells by the pathogen Pseudomonas syringae pv. tomato (Pst) and activate a MAPK cascade and other responses which together confer resistance to bacterial speck disease. Pic14 encodes a PP2C with an N-terminal kinase-interacting motif (KIM) and a C-terminal phosphatase domain. Upon inoculation with Pst-AvrPto, Pto/Prf-expressing tomato plants with loss-of-function mutations in Pic14 developed less speck disease, specifically in older leaves, compared to wild-type plants. Transient expression of Pic14 in leaves of Nicotiana benthamiana and tomato inhibited cell death typically induced by Pto/Prf and the MAPK cascade members M3Kα and Mkk2. The cell death-suppressing activity of Pic14 was dependent on the KIM and the catalytic phosphatase domain. Pic14 inhibited M3Kα- and Mkk2-mediated activation of immunity-associated MAPKs and Pic14 was shown to be an active phosphatase that physically interacts with and dephosphorylates Mkk2 in a KIM-dependent manner. Together, our results reveal Pic14 as an important negative regulator of Pto/Prf-triggered immunity by interacting with and dephosphorylating Mkk2.

Detection of canine parvovirus type 2c (CPV-2c) in Palestine.

The objective of the present study was to report, for the first time, the presence of canine parvovirus type 2c (CPV-2c) in domesticated dogs with acute gastroenteritis and to characterize the antigenic variants circulating in Palestine. A veterinary clinical-based epidemiological study was carried out between December 2022 and April 2023. Fifty fecal samples were collected from dogs with gastroenteritis and screened for CPV-2 infection by polymerase chain reaction. The distribution of positive cases according to various epidemiological factors was studied. Partial sequencing of the viral protein 2 (VP2) gene was performed for the analysis of CPV-2 variants. Most of the investigated samples (60%; n = 50) during the study period were found positive for CPV-2 infection. There was no difference in the distribution of positive cases of CPV-2 infection based on age group, gender, location, and vaccination status. The analysis of nucleotide and amino acid sequences from amplified products, as well as phylogenetic analysis, revealed the presence of CPV-2c clustered with Asian CPV-2c variants. In summary, this study represents the initial genetic analysis of CPV-2 present in Palestinian dogs with gastroenteritis and provides evidence that confirms the existence of the CPV-2c variants. To determine the prevailing CPV-2 variant associated with the infection, it is crucial to conduct further sequence analysis using large populations of both domestic and wild canines.

SlPP2C2 interacts with FZY/SAUR and regulates tomato development via signaling crosstalk of ABA and auxin.

Abscisic acid (ABA) signaling interacts frequently with auxin signaling when it regulates plant development, affecting multiple physiological processes; however, to the best of our knowledge, their interaction during tomato development has not yet been reported. Here, we found that type 2C protein phosphatase (SlPP2C2) interacts with both flavin monooxygenase FZY, an indole-3-acetic acid (IAA) biosynthetic enzyme, and small auxin upregulated RNA (SAUR) of an IAA signaling protein and regulates their activity, thereby affecting the expression of IAA-responsive genes. The expression level of SlPP2C2 was increased by exogenous ABA, IAA, NaCl, or dehydration treatment of fruits, leaves, and seeds, and it decreased in imbibed seeds. Manipulating SlPP2C2 with overexpression, RNA interference, and CRISPR/Cas9-mediated genome editing resulted in pleiotropic changes, such as morphological changes in leaves, stem trichomes, floral organs and fruits, accompanied by alterations in IAA and ABA levels. Furthermore, the RNA-seq analysis indicated that SlPP2C2 regulates the expression of auxin-/IAA-responsive genes in different tissues of tomato. The results demonstrate that SlPP2C2-mediated ABA signaling regulates the development of both vegetative and reproductive organs via interaction with FZY/SAUR, which integrates the cross-talk of ABA and auxin signals during development and affects the expressions of development-related genes in tomato.

Evaluation of Ultrasound Screening Method and Prevalence for Developmental Hip Dysplasia In The Central Anatolia

Objectives:This study aimed to assess the frequency and subsequent results of developmental dysplasia of the hip (DDH) in newborns in Central Anatolia who were admitted to Konya City Hospital and evaluate the efficiency of the existing screening and treatment methods in detecting and managing DDH in infants. Methods: Total of 10,650 infants were evaluated who had Graf USG screening in Konya City Hospital. İnfants born at or after 38 weeks of gestation were included, while premature infants were excluded. Additionally, we included infants who underwent USG screening between 30 and 90 days after birth, and whose hip development physiology was adequately visualized and assessed during the USG examination. Results: The study examined 8,695 term infants (52.5% male, 47.5% female) who underwent USG screening between 2020 and 2022. Participants mean gestational age at the time of the first USG examination was 7.94±2.07 weeks. Graf type 1 was higher in males (%97-96,5) and graf type 2 was higher in females (%7,2-7,8). Radiologists prefer to re-examine after 1 month for type 2a(%84,49-82,02) but orthopaedic consultation for type Type-2B, 2C, and type-3. The majority of infants (93.6%) underwent only one USG screening. No patients who continued regular USG control and treatment required surgical treatment requiring osteotomy. Conclusion: USG is an early diagnostic method for developmental hip dysplasia (DDH), which allows for simple treatment options and prevention of complications. It is a simple, inexpensive, and non-invasive method. Our study supports that regular USG screenings in infants eliminate the need for surgical procedures requiring osteotomy.

Genome Sequences of Canine Parvovirus Type 2c Prevalent in Western Mexico.

Canine parvovirus type 2 (CPV-2) is one of the main etiologies of viral gastroenteritis in dogs across the globe. This disease is mainly characterized by the presence of diarrhea, abdominal pain, vomiting, anorexia, and dehydration. This virus is responsible for high mortality and morbidity rates in unvaccinated dogs and those younger than three months. The monitoring of viral variants in our region has demonstrated that in the last seven years, variant CPV-2c has been circulating exclusively, which is unusual if we consider that in the rest of the world, at least two variants co-circulate among dog populations. To the best of our knowledge, no studies in Mexico have reported genomic sequences of CPV-2,which are relevant for population comparisons at the genetic level. Therefore, the present study aimed to sequence genomes associated with CPV-2c. To meet this objective, rectal swab samples were collected from dogs with suspected CPV-2 infection. Five positive cases diagnosed by lateral flow testing and polymerase chain reactionwere selected for viral genome sequencing. Comparative analyses illustrated that the obtained genome sequences were > 99% homologous to those reported for CPV-2 in the GenBank. On the other hand, 52 nucleotide mutations were identified in thevp1/vp2gene, out of which three impacted amino acid transition (T226S, F267Y, and A440T). Phylogenetic analysis of thevp1/vp2gene demonstrated that the five sequences clustered in a clade called "III", pertaining to sequences from USA and Uruguay. To our knowledge, this was the first report of genomic sequences associated with CPV-2 in Mexico, which is of great relevance for the epidemiological-molecular understanding and evolution of the virus.

Functional screening of the Arabidopsis 2C protein phosphatases family identifies PP2C15 as a negative regulator of plant immunity by targeting BRI1-associated receptor kinase 1.

Genetic engineering using negative regulators of plant immunity has the potential to provide a huge impetus in agricultural biotechnology to achieve a higher degree of disease resistance without reducing yield. Type 2C protein phosphatases (PP2Cs) represent the largest group of protein phosphatases in plants, with a high potential for negative regulatory functions by blocking the transmission of defence signals through dephosphorylation. Here, we established a PP2C functional protoplast screen using pFRK1::luciferase as a reporter and found that 14 of 56 PP2Cs significantly inhibited the immune response induced by flg22. To verify the reliability of the system, a previously reported MAPK3/4/6-interacting protein phosphatase, PP2C5, was used; it was confirmed to be a negative regulator of PAMP-triggered immunity (PTI). We further identified PP2C15 as an interacting partner of BRI1-associated receptor kinase 1 (BAK1), which is the most well-known co-receptor of plasma membrane-localized pattern recognition receptors (PRRs), and a central component of PTI. PP2C15 dephosphorylates BAK1 and negatively regulates BAK1-mediated PTI responses such as MAPK3/4/6 activation, defence gene expression, reactive oxygen species bursts, stomatal immunity, callose deposition, and pathogen resistance. Although plant growth and 1000-seed weight of pp2c15 mutants were reduced compared to those of wild-type plants, pp2c5 mutants did not show any adverse effects. Thus, our findings strengthen the understanding of the mechanism by which PP2C family members negatively regulate plant immunity at multiple levels and indicate a possible approach to enhance plant resistance by eliminating specific PP2Cs without affecting plant growth and yield.

GlPRMT5 inhibits GlPP2C1 via symmetric dimethylation and regulates the biosynthesis of secondary metabolites in Ganoderma lucidum

PRMT5, a type II arginine methyltransferase, is involved in transcriptional regulation, RNA processing and other biological processes and signal transduction. Secondary metabolites are vital pharmacological compounds in Ganoderma lucidum, and their content is an important indicator for evaluating the quality of G. lucidum. Here, we found that GlPRMT5 negatively regulates the biosynthesis of secondary metabolites. In further in-depth research, GlPP2C1 (a type 2C protein phosphatase) was identified out as an interacting protein of GlPRMT5 by immunoprecipitation-mass spectrometry (IP-MS). Further mass spectrometry detection revealed that GlPRMT5 symmetrically dimethylates the arginine 99 (R99) and arginine 493 (R493) residues of GlPP2C1 to weaken its activity. The symmetrical dimethylation modification of the R99 residue is the key to affecting GlPP2C1 activity. Symmetrical demethylation-modified GlPP2C1 does not affect the interaction with GlPRMT5. In addition, silencing GlPP2C1 clearly reduced GA content, indicating that GlPP2C1 positively regulates the biosynthesis of secondary metabolites in G. lucidum. In summary, this study reveals the molecular mechanism by which GlPRMT5 regulates secondary metabolites, and these studies provide further insights into the target proteins of GlPRMT5 and symmetric dimethylation sites. Furthermore, these studies provide a basis for the mutual regulation between different epigenetic modifications.

Functional evaluation of various modalities of management in floating knee injuries at a tertiary care centre in central India

Background: The injuries involving femur and tibia fractures due to high velocity are known as floating knee injuries. These fractures may involve the shaft, metaphysis or the articular surface. There are many complications associated with these injuries. This study evaluates the functional, clinical and radiological outcomes of floating knee injuries. Methods: A Prospective and interventional study was performed at MGMMC and MYH Hospital, Indore. We included 30 patients in our study. Femur fractures are managed by intramedullary nailing or distal femur plating. Tibia fractures are managed by Intramedullary nailing or tibia plating. Patient were called for regular follow up for a minimum of 6 months. Functional and clinical evaluation done by Karlstorm and olerud scoring system and radiological outcome by union on x rays were done. Results: Out of 30 patients 28 (93.33%) male and 2 (6.66%) female. According to Fraser classification, 17 (56.66%) type 1, 4 (13.33%) type 2A, 4 (13.33%) type2B, 5 (16.66%) type 2C injuries. A majority of the injuries occurred due to road traffic accidents involving right limb 21 (70%) more then left 9 (30%). Knee stiffness occurred in 8, infection in femur 3, infection in tibia 2, malunion of femur 2, limb length discrepancy in 2 patients. Outcome was excellent in 5 (16.66%), Good in 10 (33.33%), Acceptable in 9 (30%) and poor in 6 (20%). Conclusions: Fraser type 1 fracture has excellent results and Fraser type 2C has poor results. Closed fracture has better outcome compared to compound Fractures. A better functional outcome can be determined on the basis of implant choice based on Fraser Classification, level of injury, open or closed injury, simple or compound type of fracture.

Genome-Wide Identification, Expression and Interaction Analyses of PP2C Family Genes in Chenopodium quinoa.

Plant protein phosphatase 2Cs (PP2Cs) function as inhibitors in protein kinase cascades involved in various processes and are crucial participants in both plant development and signaling pathways activated by abiotic stress. In this study, a genome-wide study was conducted on the CqPP2C gene family. A total of putative 117 CqPP2C genes were identified. Comprehensive analyses of physicochemical properties, chromosome localization and subcellular localization were conducted. According to phylogenetic analysis, CqPP2Cs were divided into 13 subfamilies. CqPP2Cs in the same subfamily had similar gene structures, and conserved motifs and all the CqPP2C proteins had the type 2C phosphatase domains. The expansion of CqPP2Cs through gene duplication was primarily driven by segmental duplication, and all duplicated CqPP2Cs underwent evolutionary changes guided by purifying selection. The expression of CqPP2Cs in various tissues under different abiotic stresses was analyzed using RNA-seq data. The findings indicated that CqPP2C genes played a role in regulating both the developmental processes and stress responses of quinoa. Real-time quantitative reverse transcription PCR (qRT-PCR) analysis of six CqPP2C genes in subfamily A revealed that they were up-regulated or down-regulated under salt and drought treatments. Furthermore, the results of yeast two-hybrid assays revealed that subfamily A CqPP2Cs interacted not only with subclass III CqSnRK2s but also with subclass II CqSnRK2s. Subfamily A CqPP2Cs could interact with CqSnRK2s in different combinations and intensities in a variety of biological processes and biological threats. Overall, our results will be useful for understanding the functions of CqPP2C in regulating ABA signals and responding to abiotic stress.

MdMYB44-like positively regulates salt and drought tolerance via the MdPYL8-MdPP2CA module in apple.

Abscisic acid (ABA) is involved in salt and drought stress responses, but the underlying molecular mechanism remains unclear. Here, we demonstrated that the overexpression of MdMYB44-like, an R2R3-MYB transcription factor, significantly increases the salt and drought tolerance of transgenic apples and Arabidopsis. MdMYB44-like inhibits the transcription of MdPP2CA, which encodes a type 2C protein phosphatase that acts as a negative regulator in the ABA response, thereby enhancing ABA signaling-mediated salt and drought tolerance. Furthermore, we found that MdMYB44-like and MdPYL8, an ABA receptor, form a protein complex that further enhances the transcriptional inhibition of the MdPP2CA promoter by MdMYB44-like. Significantly, we discovered that MdPP2CA can interfere with the physical association between MdMYB44-like and MdPYL8 in the presence of ABA, partially blocking the inhibitory effect of the MdMYB44-like-MdPYL8 complex on the MdPP2CA promoter. Thus, MdMYB44-like, MdPYL8, and MdPP2CA form a regulatory loop that tightly modulates ABA signaling homeostasis under salt and drought stress. Our data reveal that MdMYB44-like precisely modulates ABA-mediated salt and drought tolerance in apples through the MdPYL8-MdPP2CA module.

Calcium regulation of the Arabidopsis Na+/K+ transporter HKT1;1 improves seed germination under salt stress.

Calcium is known to improve seed-germination rates under salt stress. We investigated the involvement of calcium ions (Ca2+) in regulating HIGH-AFFINITY K+ TRANSPORTER 1 (HKT1; 1), which encodes a Na+/K+ transporter, and its post-translational regulator TYPE 2C PROTEIN PHOSPHATASE 49 (PP2C49), in germinating Arabidopsis (Arabidopsis thaliana) seedlings. Germination rates of hkt1 mutant seeds under salt stress remained unchanged by CaCl2 treatment in wild-type Arabidopsis, whereas pp2c49 mutant seeds displayed improved salt-stress tolerance in the absence of CaCl2 supplementation. Analysis of HKT1;1 and PP2C49 promoter activity revealed that CaCl2 treatment results in radicle-focused expression of HKT1;1 and reduction of the native radicle-exclusive expression of PP2C49. Ion-content analysis indicated that CaCl2 treatment improves K+ retention in germinating wild-type seedlings under salt stress, but not in hkt1 seedlings. Transgenic seedlings designed to exclusively express HKT1;1 in the radicle during germination displayed higher germination rates under salt stress than the wild type in the absence of CaCl2 treatment. Transcriptome analysis of germinating seedlings treated with CaCl2, NaCl, or both revealed 118 upregulated and 94 downregulated genes as responsive to the combined treatment. Bioinformatics analysis of the upstream sequences of CaCl2-NaCl-treatment-responsive upregulated genes revealed the abscisic acid response element CACGTGTC, a potential CaM-binding transcription activator-binding motif, as most prominent. Our findings suggest a key role for Ca2+ in mediating salt-stress responses during germination by regulating genes that function to maintain Na+ and K+ homeostasis, which is vital for seed germination under salt stress.

Reversible phosphorylation of a lectin-receptor-like kinase controls xylem immunity

Pattern-recognition receptors (PRRs) mediate basal resistance to most phytopathogens. However, plant responses can be cell type specific, and the mechanisms governing xylem immunity remain largely unknown. We show that the lectin-receptor-like kinase LORE contributes to xylem basal resistance in Arabidopsis upon infection with Ralstonia solanacearum, a destructive plant pathogen that colonizes the xylem to cause bacterial wilt. Following R.solanacearum infection, LORE is activated by phosphorylation at residue S761, initiating a phosphorelay that activates reactive oxygen species production and cell wall lignification. To prevent prolonged activation of immune signaling, LORE recruits and phosphorylates type 2C protein phosphatase LOPP, which dephosphorylates LORE and attenuates LORE-mediated xylem immunity to maintain immune homeostasis. A LOPP knockout confers resistance against bacterial wilt disease in Arabidopsis and tomatoes without impacting plant growth. Thus, our study reveals a regulatory mechanism in xylem immunity involving the reversible phosphorylation of receptor-like kinases.

Congenital myasthenic syndrome type 2C in a neonate: Redefining the phenotype of CHRNB1‐related myasthenic syndromes

AbstractObjectiveWe present a neonate with generalized weakness due to autosomal recessive congenital myasthenic syndrome type 2C (CMS2C) resulting from a compound heterozygous mutation in the CHRNB1 gene.Patient descriptionOur patient was determined by multiple methodologies to have a diagnosis of CMS2C (OMIM #616314). Whole‐genome sequencing revealed two distinct variants in the CHRNB1 gene (OMIM *100710): a maternally inherited 2 kb pathogenic microdeletion on chromosome 17p13.1 and a paternally inherited intronic deletion (c.1218‐9_1218‐7) that was reported by the laboratory as a variant of unknown significance.ConclusionsCMS2C is a rare autosomal recessive genetic condition associated with early‐onset muscle weakness. Our patient had a paternally inherited deletion in CHRNB1 (c.1218‐9_1218‐7) that was initially described as a variant of unknown significance. We suggest this finding is “likely pathogenic,” as this aberration has not been commonly described. He also had a partial deletion of CHRNB1 in the maternally inherited allele, which provides further evidence that partial gene deletions may be a more common molecular mechanism than previously known for this condition. The combination of the clinical presentation and electrophysiologic data allowed us to understand the molecular findings and ultimately diagnose CMS2C in our patient.

Inflammatory Factor IL1α Induces Aberrant Astrocyte Proliferation in Spinal Cord Injury Through the Grin2c/Ca2+/CaMK2b Pathway

Spinal cord injury (SCI) is one of the most devastating traumas, and the aberrant proliferation of astrocytes usually causes neurological deficits. However, the mechanism underlying astrocyte over-proliferation after SCI is unclear. Grin2c (glutamate ionotropic receptor type 2c) plays an essential role in cell proliferation. Our bioinformatic analysis indicated that Grin2c and Ca2+ transport functions were inhibited in astrocytes after SCI. Suppression of Grin2c stimulated astrocyte proliferation by inhibiting the Ca2+/calmodulin-dependent protein kinase 2b (CaMK2b) pathway in vitro. By screening different inflammatory factors, interleukin 1α (IL1α) was further found to inhibit Grin2c/Ca2+/CaMK2b and enhance astrocyte proliferation in an oxidative damage model. Blockade of IL1α using neutralizing antibody resulted in increased Grin2c expression and the inhibition of astrocyte proliferation post-SCI. Overall, this study suggests that IL1α promotes astrocyte proliferation by suppressing the Grin2c/Ca2+/CaMK2b pathway after SCI, revealing a novel pathological mechanism of astrocyte proliferation, and may provide potential targets for SCI repair.