Scalable Synthesis of High-Quality Ultrathin Ferroelectric Magnesium Molybdenum Oxide.

The development of ultrathin, stable ferroelectric materials is crucial for advancing high-density, low-power electronic devices. Nonetheless, ultrathin ferroelectric materials are rare due to the critical size effect. Here, a novel ferroelectric material, magnesium molybdenum oxide (Mg2Mo3O8) is presented. High-quality ultrathin Mg2Mo3O8 crystals are synthesized using chemical vapor deposition (CVD). Ultrathin Mg2Mo3O8 has a wide bandgap (≈4.4 eV) and nonlinear optical response. Mg2Mo3O8 crystals of varying thicknesses exhibit out-of-plane ferroelectric properties at room temperature, with ferroelectricity retained even at a 2 nm thickness. The Mg2Mo3O8 exhibits a relatively large remanent polarization ranging from 33 to 52 µC cm- 2, which is tunable by changing its thickness. Notably, Mg2Mo3O8 possesses a high Curie temperature (>980 °C) across various thicknesses. Moreover, the as-grown Mg2Mo3O8 crystals display remarkable stability under harsh environments. This work introduces nolanites-type crystal into ultrathin ferroelectrics. The scalable synthesis of stable ultrathin ferroelectric Mg2Mo3O8 expands the scope of ferroelectric materials and may prosper applications of ferroelectrics.

Association between thyroid autoimmunity and bone mineral density in patients with type 2 diabetes mellitus: a cross-sectional study.

PURPOSE: The purpose of this study was to analyze the relationship between thyroid autoimmunity and bone mineral density (BMD) in patients with type 2 diabetes mellitus (T2DM), and to further explore the influence of thyroid autoimmunity on diabetic osteoporosis. METHODS: A total of 601 T2DM patients were included and divided into two groups according to thyroid autoantibodies, namely thyroid autoimmunity positive group (TPOAb+ or TGAb + ) and thyroid autoimmunity negative group (TPOAb- and TGAb-). Clinical data were collected and BMD was determined by dual-energy X-ray absorptiometry (DXA). SPSS26.0 software was used to data analysis. Model regression was used to analyze the influencing factors of BMD, and ROC curve was used to analyze the optimal cut-off point of thyroid peroxidase antibody (TPOAb) for screening osteoporosis. RESULTS: TPOAb and thyroglobulin antibody (TGAb) were negatively correlated with BMD and T-score (LS, FN and WB) (P < 0.01), and TGAb was negatively correlated with 25(OH)D (P < 0.05). Multiple linear regression analysis showed that TPOAb was an independent influence factor on LS, FN and WB BMD. ROC curve analysis showed that the optimal threshold of TPOAb for predicting osteoporosis was 12.35. CONCLUSIONS: In T2DM patients, TPOAb and TGAb levels are negatively correlated with LS, FN and WB BMD, and TPOAb is an independent influencing factor for diabetic osteoporosis, and TPOAb has a certain predictive value for the occurrence and development of diabetic osteoporosis clinically.

Association of serum trimethylamine N-oxide levels and bone mineral density in type 2 diabetes mellitus.

PURPOSE: The relationship between trimethylamine N-oxide (TMAO) and bone mineral density (BMD) in type 2 diabetes mellitus (T2DM) is unclear. We explore the relationship between TMAO levels and BMD in T2DM. METHODS: This is a cross-sectional study. 254 T2DM patients were enrolled and divided into three groups by TMAO tertiles, and the clinical data were collected. BMD was determined by dual-energy X-ray absorptiometry (DXA) and serum TMAO levels was determined by stable isotope dilution high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). RESULTS: Patients in the highest tertile of TMAO levels (TMAO > 6.72 µmol/L) showed relatively low BMD and a higher number of fracture history, osteoporosis (OP) than those in the lower tertiles. Spearman correlation analysis showed that serum TMAO was negatively correlated with BMD of whole body (WB), lumbar spine (LS) and femoral neck (FN), while TMAO was positive correlated with osteoporotic fracture (p < 0.05). Logistic regression models showed that TMAO was an independent influencing factor of fracture history after adjusting for confounders in TMAO > 6.72 µmol/L group. CONCLUSIONS: There is a significant linear correlation between TMAO levels and BMD in T2DM patients. Especially in TMAO > 6.72 µmol/L group, TMAO was negatively correlated with WB, LS, and FN BMD, and was positive correlated with osteoporotic fracture in T2DM patients. The findings suggest that elevated TMAO levels are associated with OP and osteoporotic fracture in T2DM patients.

Correlation Between Insulin Resistance and Urinary Incontinence in Female Patients with Type 2 Diabetes Mellitus.

INTRODUCTION AND HYPOTHESIS: Patients with type 2 diabetes mellitus (T2DM) are at a high risk of developing urinary incontinence; however, its pathogenesis is unclear. The purpose of this study is to explore the relationship between insulin resistance and urinary incontinence and its severity in female patients with T2DM. METHODS: A total of 366 women with T2DM aged ≥18 years were enrolled in this study. Insulin resistance was evaluated by the homeostasis model insulin resistance (HOMA-IR) index and urinary incontinence was assessed by the International Consultation on Incontinence Questionnaire Short Form (ICIQ-SF). All subjects were divided into four groups according to HOMA-IR quartiles. Logistic regression analysis was performed to investigate the relationship between insulin resistance and urinary incontinence and its severity. RESULTS: Among the 366 patients, 186 (50.8%) had urinary incontinence. The prevalence of urinary incontinence increased significantly with HOMA-IR quartiles (p < 0.001). Adjusted logistic regression analysis showed that compared with HOMA-IR ≤ 1.76, 2.81 ≤ HOMA-IR ≤ 4.27 was associated with a significantly increased risk of moderate incontinence (OR = 2.197, 95% CI 1.031-4.683, p = 0.041), and HOMA-IR ≥ 4.28 was associated with a significantly increased risk of severe incontinence (OR = 5.699, 95% CI 1.685-19.276, p = 0.005). Binary logistic regression analysis showed that HOMA-IR was the independent risk factor for urinary incontinence (p < 0.001). CONCLUSIONS: Higher levels of insulin resistance are associated with urinary incontinence and its severity in female patients with type 2 diabetes mellitus.

Zika virus infection triggers caspase cleavage of STAT1.

IMPORTANCE: Zika virus (ZIKV) is a re-emerging flavivirus. Similar to other flaviviruses, ZIKV antagonizes the host interferon (IFN) signaling pathway to establish infection. Understanding the molecular mechanism by which ZIKV antagonizes IFN-induced antiviral signaling may lead to a new antiviral strategy by cracking the IFN antagonism. Flaviviruses have been reported to employ NS5-dependent and -independent mechanisms to block STAT2-mediated signaling, whereas whether flaviviruses target STAT1 remains controversial. Herein, we found that ZIKV infection triggered caspase-dependent cleavage of STAT1 at the aspartic acid 694 during late infection, whereas murine STAT1 (mSTAT1) was resistant to cleavage. Intriguingly, ectopically expressed cleavage-resistant human STAT1.D694A or complementation of cleavable mSTAT1.D695G exerted comparable anti-ZIKV activity with their counterparts, challenging the role of caspase-mediated STAT1 cleavage in the IFN antagonism in ZIKV-infected cells. These data may also imply a dominant role of the antagonism of STAT2 but not STAT1 in ZIKV-infected cells.

High-Resolution Genomic Profiling of a Genotype 3b Hepatitis C Virus from a Flare of an Occult Hepatitis Patient with Acute-on-Chronic Liver Failure.

Acute-on-chronic liver failure (ACLF) is defined as a syndrome of acutely decompensated cirrhosis in patients with chronic liver disease (CLD). Here we report an ACLF case caused by a flare of occult hepatitis C infection. This patient was infected with hepatitis C virus (HCV) more than a decade ago and hospitalized due to alcohol-associated CLD. Upon admission, the HCV RNA in the serum was negative and the anti-HCV antibody was positive, whereas the viral RNA in the plasma dramatically increased during hospitalization, which suggests an occult hepatitis C infection. Overlapped fragments encompassing the nearly whole HCV viral genome were amplified, cloned, and sequenced. Phylogenetic analysis indicated an HCV genotype 3b strain. Sanger sequencing to 10-fold coverage of the 9.4-kb nearly whole genome reveals high diversity of viral quasispecies, an indicator of chronic infection. Inherent resistance-associated substitutions (RASs) in the NS3 and NS5A but not in the NS5B regions were identified. The patient developed liver failure and accepted liver transplantation, followed by direct-acting antiviral (DAA) treatment. The hepatitis C was cured by the DAA treatment despite the existence of RASs. Thus, care should be taken for occult hepatitis C in patients with alcoholic cirrhosis. The analysis of viral genetic diversity may help to identify an occult hepatitis C virus infection and predict the efficacy of anti-viral treatment.

Artificial Neuronal Devices Based on Emerging Materials: Neuronal Dynamics and Applications.

Artificial neuronal devices are critical building blocks of neuromorphic computing systems and currently the subject of intense research motivated by application needs from new computing technology and more realistic brain emulation. Researchers have proposed a range of device concepts that can mimic neuronal dynamics and functions. Although the switching physics and device structures of these artificial neurons are largely different, their behaviors can be described by several neuron models in a more unified manner. In this paper, the reports of artificial neuronal devices based on emerging volatile switching materials are reviewed from the perspective of the demonstrated neuron models, with a focus on the neuronal functions implemented in these devices and the exploitation of these functions for computational and sensing applications. Furthermore, the neuroscience inspirations and engineering methods to enrich the neuronal dynamics that remain to be implemented in artificial neuronal devices and networks toward realizing the full functionalities of biological neurons are discussed.

Circuit-Level Memory Technologies and Applications based on 2D Materials.

Memory technologies and applications implemented fully or partially using emerging 2D materials have attracted increasing interest in the research community in recent years. Their unique characteristics provide new possibilities for highly integrated circuits with superior performances and low power consumption, as well as special functionalities. Here, an overview of progress in 2D-material-based memory technologies and applications on the circuit level is presented. In the material growth and fabrication aspects, the advantages and disadvantages of various methods for producing large-scale 2D memory devices are discussed. Reports on 2D-material-based integrated memory circuits, from conventional dynamic random-access memory, static random-access memory, and flash memory arrays, to emerging memristive crossbar structures, all the way to 3D monolithic stacking architecture, are systematically reviewed. Comparisons between experimental implementations and theoretical estimations for different integration architectures are given in terms of the critical parameters in 2D memory devices. Attempts to use 2D memory arrays for in-memory computing applications, mostly on logic-in-memory and neuromorphic computing, are summarized here. Finally, challenges that impede the large-scale applications of 2D-material-based memory are reviewed, and perspectives on possible approaches toward a more reliable system-level fabrication are also given, hopefully shedding some light on future research.

Identifying A New Phylogeographic Population of the Blyth's Tragopan (Tragopan blythii) through Multi-locus Analyses.

Geographically and morphologically distinct populations within a species are frequently the focus of conservation efforts, especially when the populations are evolutionarily significant units. The Blyth's Tragopan (Tragopan blythii) is a globally-threatened species confined to South and Southeast Asia. During our field surveys in western Myanmar, we discovered a distinct group of individuals that differed in their appearance relative to all other populations. We further examined the differences in their DNA sequence using three nuclear introns and three mitochondrial genes through phylogenetic analytical methods. Our results showed the population from Mount Kennedy, Chin Hills formed reciprocal monophyletic groups with the nominate subspecies from Mount Saramati, Sagaing Division. Species delimitation analyses further confirmed this differentiation. Geographical isolation by the intervening lowlands found between high elevation habitats may have been the main cause of their differentiation. Hence, we propose that the Mount Kennedy population be viewed as a distinct evolutionarily significant unit and be given special priority for conservation.

NS5-independent Ablation of STAT2 by Zika virus to antagonize interferon signalling.

Flavivirus genus includes numerous arthropod-borne human pathogens that are clinically important. Flaviviruses are notorious for their ability to antagonize host interferon (IFN) induced anti-viral signalling. It has been documented that NS5s of flaviviruses mediate proteasome degradation of STAT2 to evade IFN signalling. Deciphering the molecular mechanism of the IFN antagonism by the viruses and reversing this antagonism may dictate anti-viral responses and provide novel antiviral approaches. In this report, by using Zika virus (ZIKV) as a model, we first demonstrated that ZIKV antagonized interferon signalling in an infectious dose-dependent manner; in other words, the virus antagonized interferon signalling at a high multiple of infection (MOI) and was sensitive to interferon signalling at a low MOI. Mechanistically, we found that ZIKV infection triggered degradation of ubiquitinated STAT2 and host short-lived proteins while didn't affect the proteasome activity per se. ZIKV infection resulted in suppression of host de novo protein synthesis. Overexpression of NS5 alone only marginally reduced STAT2 and had no effect on the host de novo protein synthesis. Ectopically expressed murine STAT2 that was resistant to NS5- and ZIKV-induced ablation exaggerated the IFN-induced anti-viral signalling. These data favour a new model of the innate immune evasion of ZIKV in which the viral infection triggers suppression of host de novo protein synthesis to accelerate the degradation of short-lived, ubiquitinated STAT2. As flaviviruses share a very conserved replication strategy, the mechanisms of IFN antagonism elucidated here might also be employed by other flaviviruses.

Quasispecies dynamics of a hepatitis E virus 4 from the feces and liver biopsy of an acute hepatitis E patient during virus clearance.

The presence of a quasispecies in hepatitis E virus (HEV) infection has been documented, however, the implications of a quasispecies in HEV-host interaction are poorly understood. Here, we analyzed the whole genome sequences of a HEV 4d from the feces and liver biopsy of a patient during the icteric and convalescent phases in an acute hepatitis E infection. Viral RNAs were extracted, reversely transcribed and seven fragments encompassing the entire viral genome were amplified and cloned. By sequencing multiple colonies of each cloned viral genome amplicon with Sanger sequencing, we verified the existence of the HEV quasispecies or intra-host genetic variations within the fecal and liver biopsy samples. There were broader genetic variations in the HEV ORF1 region including the PCP, HPX, and RdRp regions during the convalescent phase whereas more genetic variations in the ORF2 P domain during the icteric phase. The quasispecies dynamics might reflect host immune pressure during viral clearance.

An infectious clone of enterovirus 71(EV71) that is capable of infecting neonatal immune competent mice without adaptive mutations.

Enterovirus 71 (EV71) is a major pathogen that causes hand, foot and mouth disease (HFMD), which is a life threatening disease in certain children. The pathogenesis of EV71-caused HFMD is poorly defined due to the lack of simple and robust animal models with severe phenotypes that recapitulate symptoms observed in humans. Here, we generated the infectious clone of a clinical isolate from a severe HFMD patient. Virus rescued from the cDNA clone was infectious in cell lines. When administrated intraperitoneally to neonatal ICR, BALB/c and C57 immune competent mice at a dosage of1.4 × 104 pfu per mouse, the virus caused weight loss, paralysis and death in the infected mice after 4-5 days of infection. In the infected mice, detectable viral replication was detected in various tissues such as heart, liver, brain, lung, kidney, small intestine, leg skeletal muscle and medulla oblongata. The histology of the infected mice included massive myolysis, glomerular atrophy, villous blunting in small intestine, widened alveolar septum, diminished alveolar spaces and lymphocytes infiltration into the lung. By using the UV-inactivated virus as a control, we elucidated that the virus first amplified in the leg skeletal muscle tissue and the muscle tissue served as a primary viral replication site. In summary, we generated a stable EV71 infectious clone that is capable of infecting neonatal immune competent mice without adaptive mutations and provide a simple, valuable animal model for the studies of EV71pathogenesis and therapy.

Strongly Surface State Carrier-Dependent Spin-Orbit Torque in Magnetic Topological Insulators.

The topological surface states (TSS) in topological insulators (TIs) can exert strong spin-orbit torque (SOT) on adjacent magnetization, offering great potential in implementing energy-efficient magnetic memory devices. However, there are large discrepancies among the reported spin Hall angle values in TIs, and its temperature dependence still remains elusive. Here, the spin Hall angle in a modulation-doped Cr-Bix Sb2- x Te3 (Cr-BST) film is quantitatively determined via both transport and optic approaches, where consistent results are obtained. A large spin Hall angle of ≈90 in the modulation-doped Cr-BST film is demonstrated at 2.5 K, and the spin Hall angle drastically decreases to 0.3-0.5 as the temperature increases. Moreover, by tuning the top TSS carrier concentration, a competition between the top and bottom TSS in contributing to SOT is observed. The above phenomena can account for the large discrepancies among the previously reported spin Hall angle values and reveal the unique role of TSS in generating SOT.

Dual role of the amphipathic helix of hepatitis C virus NS5A in the viral polyprotein cleavage and replicase assembly.

Assembling a viral replicase on host intracellular membranes is a common strategy for viral replication of almost all of the positive-strand RNA viruses. Understanding how the key modules of the replicase are involved in the replicase assembly may provide insights into the pathway of the replicase assembly. Herein, by using HCV as a model, we dissect the roles of the amphipathic helix (AH) of NS5A, a key repilcase component, in the viral replicase assembly. The results show that the AH is dispensable for membrane anchoring of NS5A. Instead, AH plays a dual role in the viral replicase assembly: positions a replicase module properly for efficient polyprotein processing and participates in protein-protein interactions within the replicase. This property of AH may serve as an attractive direct anti-viral target.

Growth and Thermo-driven Crystalline Phase Transition of Metastable Monolayer 1T'-WSe2 Thin Film.

Two-dimensional (2D) transition metal dichalcogenides MX2 (M = Mo, W, X = S, Se, Te) attracts enormous research interests in recent years. Its 2H phase possesses an indirect to direct bandgap transition in 2D limit, and thus shows great application potentials in optoelectronic devices. The 1T' crystalline phase transition can drive the monolayer MX2 to be a 2D topological insulator. Here we realized the molecular beam epitaxial (MBE) growth of both the 1T' and 2H phase monolayer WSe2 on bilayer graphene (BLG) substrate. The crystalline structures of these two phases were characterized using scanning tunneling microscopy. The monolayer 1T'-WSe2 was found to be metastable, and can transform into 2H phase under post-annealing procedure. The phase transition temperature of 1T'-WSe2 grown on BLG is lower than that of 1T' phase grown on 2H-WSe2 layers. This thermo-driven crystalline phase transition makes the monolayer WSe2 to be an ideal platform for the controlling of topological phase transitions in 2D materials family.

Hepatitis C virus NS5A inhibitor daclatasvir allosterically impairs NS4B-involved protein-protein interactions within the viral replicase and disrupts the replicase quaternary structure in a replicase assembly surrogate system.

Daclatasvir (DCV) is a highly potent direct-acting antiviral that targets the non-structural protein 5A (NS5A) of hepatitis C virus (HCV) and has been used with great clinical success. Previous studies have demonstrated its impact on viral replication complex assembly. However, the precise mechanisms by which DCV impairs the replication complex assembly remains elusive. In this study, by using HCV subgenomic replicons and a viral replicase assembly surrogate system in which the HCV NS3-5B polyprotein is expressed to mimic the viral replicase assembly, we assessed the impact of DCV on the aggregation and tertiary structure of NS5A, the protein-protein interactions within the viral replicase and the quaternary structure of the viral replicase. We found that DCV did not affect aggregation and tertiary structure of NS5A. DCV induced a quaternary structural change of the viral replicase, as evidenced by selective increase of NS4B's sensitivity to proteinase K digestion. Mechanically, DCV impaired the NS4B-involved protein-protein interactions within the viral replicase. These phenotypes were consistent with the phenotypes of several reported NS4B mutants that abolish the viral replicase assembly. The DCV-resistant mutant Y93H was refractory to the DCV-induced reduction of the NS4B-involved protein interactions and the quaternary structural change of the viral replicase. In addition, Y93H reduced NS4B-involved protein-protein interactions within the viral replicase and attenuated viral replication. We propose that DCV may induce a positional change of NS5A, which allosterically affects protein interactions within the replicase components and disrupts replicase assembly.

Replicase-mediated shielding of the poliovirus replicative double-stranded RNA to avoid recognition by MDA5.

Replication of the positive-strand RNA viruses generates double-stranded RNAs (dsRNAs) that are recognized by host pattern recognition receptors (PRRs) to trigger innate immune responses. Formation of the viral replication complex (RC) has been thought to shield dsRNA from being recognized by innate sensors. To elucidate the RC-mediated evasion of innate recognition, we selected poliovirus (PV) as a model. We first found that RNAs generated during PV replication were potent interferon (IFN) inducers upon transfection, while there was no obvious IFN production detected in PV-replicating cells. PV replication did not interfere with IFN production when IFN agonists were synchronously introduced with the replicating PV RNAs, and in PV-infected cells, IFN agonist-induced IFN production was only moderately impaired but not completely abolished. When PV-infected cells were in situ permeabilized by digitonin, viral dsRNAs were readily detected by an anti-dsRNA antibody and were resistant to RNase III digestion. When digitonin-permeabilized cells were further solubilized by 1 % triton X-100, the dsRNAs of PV became sensitive to RNase III digestion. A co-localization study showed that PV dsRNA did not co-localize with MDA5 in virally infected cells. Given that the PV replication complex is protruding single-membrane and tubular in form, viral replicative dsRNAs are probably shielded by the replication complex or the viral replicase to avoid being accessed by RNase III and MDA5. We propose that the replication complex- or replicase-mediated shielding of dsRNA may act as a means for innate evasion.

Affinity Purification of the Hepatitis C Virus Replicase Identifies Valosin-Containing Protein, a Member of the ATPases Associated with Diverse Cellular Activities Family, as an Active Virus Replication Modulator.

Like almost all of the positive-strand RNA viruses, hepatitis C virus (HCV) induces host intracellular membrane modification to form the membrane-bound viral replication complex (RC), within which viral replicases amplify the viral RNA genome. Despite accumulated information about how HCV co-opts host factors for viral replication, our knowledge of the molecular mechanisms by which viral proteins hijack host factors for replicase assembly has only begun to emerge. Purification of the viral replicase and identification of the replicase-associated host factors to dissect their roles in RC biogenesis will shed light on the molecular mechanisms of RC assembly. To purify the viral replicase in the context of genuine viral replication, we developed an HCV subgenomic replicon system in which two different affinity tags were simultaneously inserted in frame into HCV NS5A and NS5B. After solubilizing the replicon cells, we purified the viral replicase by two-step affinity purification and identified the associated host factors by mass spectrometry. We identified valosin-containing protein (VCP), a member of the ATPases associated with diverse cellular activities (AAA+ATPase) family, as an active viral replication modulator whose ATPase activity is required for viral replication. A transient replication assay indicated that VCP is involved mainly in viral genome amplification. VCP associated with viral replicase and colocalized with a viral RC marker. Further, in an HCV replicase formation surrogate system, abolishing VCP function resulted in aberrant distribution of HCV NS5A. We propose that HCV may co-opt a host AAA+ATPase for its replicase assembly. IMPORTANCE: Almost all of the positive-strand RNA viruses share a replication strategy in which viral proteins modify host membranes to form the membrane-associated viral replicase. Viruses hijack host factors to facilitate this energy-unfavorable process. Understanding of this fundamental process is hampered by the challenges of purifying the replicase because of the technical difficulties involved. In this study, we developed an HCV subgenomic replicon system in which two different affinity tags were simultaneously inserted in frame into two replicase components. Using this dual-affinity-tagged replicon system, we purified the viral replicase and identified valosin-containing protein (VCP) AAA+ATPase as a pivotal viral replicase-associated host factor that is required for viral genome replication. Abolishing VCP function resulted in aberrant viral protein distribution. We propose that HCV hijacks a host AAA+ATPase for its replicase assembly. Understanding the molecular mechanism of VCP regulates viral replicase assembly may lead to novel antiviral strategies targeting the most conserved viral replication step.

VdCrz1 is involved in microsclerotia formation and required for full virulence in Verticillium dahliae.

Calcium signaling plays crucial roles in ion stress tolerance, sporulation and pathogenicity in fungi. Although the signaling pathway mediated by calcineurin and the calcineurin-responsive zinc finger transcription factor Crz1 is well characterized in other fungi, this pathway is not well characterized in the phytopathogenic fungus, Verticillium dahliae. To better understand the role of this calcineurin-dependent transcription factor in V. dahliae, an ortholog of CRZ1, VdCrz1, was identified and characterized functionally. Transcriptional analysis of VdCrz1 and GFP expression driven by the VdCrz1 promoter indicated that VdCrz1 was involved in microsclerotia development. After targeted deletion of VdCrz1, microsclerotia formation and melanin accumulation were impaired. Furthermore, the ΔVdCrz1 mutants were hypersensitive to high concentrations of Ca(2+) and cell wall-perturbing agents, such as sodium dodecyl sulfate. The addition of Mg(2+) to the medium restores the microsclerotia formation in ΔVdCrz1 mutants. The ΔVdCrz1 mutants exhibited delayed Verticillium wilt symptoms on smoke tree. These results suggest that VdCrz1 plays important roles in Ca(2+) signaling, cell wall integrity, microsclerotia development and full virulence in V. dahliae.