Comparing two spatial variables with the probability of agreement.

Computing the agreement between 2 continuous sequences is of great interest in statistics when comparing 2 instruments or one instrument with a gold standard. The probability of agreement quantifies the similarity between 2 variables of interest, and it is useful for determining what constitutes a practically important difference. In this article, we introduce a generalization of the PA for the treatment of spatial variables. Our proposal makes the PA dependent on the spatial lag. We establish the conditions for which the PA decays as a function of the distance lag for isotropic stationary and nonstationary spatial processes. Estimation is addressed through a first-order approximation that guarantees the asymptotic normality of the sample version of the PA. The sensitivity of the PA with respect to the covariance parameters is studied for finite sample size. The new method is described and illustrated with real data involving autumnal changes in the green chromatic coordinate (Gcc), an index of "greenness" that captures the phenological stage of tree leaves, is associated with carbon flux from ecosystems, and is estimated from repeated images of forest canopies.

From growth promotion to intestinal inflammation alleviation: Unraveling the potential role of Lactobacillus rhamnosus GCC-3 in juvenile grass carp (Ctenopharyngodon idella).

Lactobacillus rhamnosus is a probiotic, which not only promotes the growth of animals, but also has anti-inflammatory effects. However, the mechanism by which Lactobacillus rhamnosus regulates intestinal immunity is not well comprehended. Hence, the study aimed to research how Lactobacillus rhamnosus affects the intestinal immunity using juvenile grass carp (Ctenopharyngodon idella) as a model. We selected 1800 juvenile grass carp for testing. They were divided into six treatments and fed with six gradients of Lactobacillus rhamnosus GCC-3 (0.0, 0.5, 1.0, 1.5, 2.0, 2.5 g/kg) for 70 days. Enteritis was subsequently induced with dextroside sodium sulfate. Results indicated that dietary Lactobacillus rhamnosus GCC-3 addition improved growth performance. Meanwhile, appropriate levels of Lactobacillus rhamnosus GCC-3 alleviated excessive inflammatory response by down-regulating the expression of TLR4 and NOD receptors, up-regulating the expression of TOR, and then down-regulating the expression of NF-κB. Additionally, appropriate Lactobacillus rhamnosus GCC-3 improved intestinal immunity by reducing pyroptosis triggered by NLRP3 inflammasome and mediated by GSDME. Furthermore, 16 S rRNA sequencing showing appropriate levels of Lactobacillus rhamnosus GCC-3 increased Lactobacillus and Bifidobacterium abundance and decreased Aeromonas abundance. These results suggest that Lactobacillus rhamnosus GCC-3 can alleviate intestinal inflammation through down-regulating NF-κB and up-regulating TOR signaling pathways, as well as by inhibiting pyroptosis.

The human SMAD9 (GCC) repeat links to natural selection and late-onset neurocognitive disorders.

INTRODUCTION: Whereas (GCC)-repeats are overrepresented in genic regions, and mutation hotspots, they are largely unexplored with regard to their link with natural selection. Across numerous primate species and tissues, SMAD9 (SMAD Family Member 9) reaches highest level of expression in the human brain. This gene contains a (GCC)-repeat in the interval between + 1 and + 60 of the transcription start site, which is in the high-ranking (GCC)-repeats with respect to length. METHODS: Here we sequenced this (GCC)-repeat in 396 Iranian individuals, consisting of late-onset neurocognitive disorder (NCD) (N = 181) and controls (N = 215). RESULTS: We detected two predominantly abundant alleles of 7 and 9 repeats, forming 96.2% of the allele pool. The (GCC)7/(GCC)9 ratio was in the reverse order in the NCD group versus controls (p = 0.005), resulting from excess of (GCC)7 in the NCD group (p = 0.003) and (GCC)9 in the controls (p = 0.01). Five genotypes, predominantly consisting of (GCC)7 and lacking (GCC)9 were detected in the NCD group only (p = 0.008). The patients harboring those genotypes received the diagnoses of Alzheimer's disease (AD) and vascular dementia (VD). Five genotypes consisting of (GCC)9 and lacking (GCC)7 were detected in the control group only (p = 0.002). The group-specific genotypes formed approximately 4% of the genotype pool in the human samples studied. CONCLUSION: We propose natural selection and a novel locus for late-onset AD and VD at the SMAD9 (GCC)-repeat in humans.

Spatial Localization of a Transformer Robot Based on Ultrasonic Signal Wavelet Decomposition and PHAT-ß-γ Generalized Cross Correlation.

Because large oil-immersed transformers are enclosed by a metal shell, the on-site localization means it is difficult to achieve the accurate location of the patrol micro-robot inside a given transformer. To address this issue, a spatial ultrasonic localization method based on wavelet decomposition and PHAT-ß-γ generalized cross correlation is proposed in this paper. The method is carried out with a five-element stereo ultrasonic array for the location of a transformer patrol robot. Firstly, the localization signal is decomposed into wavelet coefficients of different scales, which would realize the adaptive decomposition of the frequency of the localization signal from low frequencies to high frequencies. Then, the wavelet coefficients are denoised and reconstructed by using the semi-soft threshold function. Second, a modified phase transform-beta-gamma (PHAT-ß-γ) method is used to calculate the exact time delay between different sensors by increasing the weights of the PHAT weighting function and introducing a correlation function. Finally, by using the proposed method, the accurate localization of the transformer patrol micro-robot is achieved with a five-element stereo ultrasonic array. The simulation and test results show that inside a transformer experimental oil tank (120 cm × 100 cm × 100 cm, L × W × H), the relative error of transformer patrol micro-robot spatial localization is within 4.1%, and the maximum localization error is less than 3 cm, which meets the requirement of engineering localization.

Shifts in sage-grouse arthropod food sources across grazing and environmental gradients in upland meadow communities.

Groundwater dependent systems are extremely important habitats for a wide variety of taxa in the Great Basin of North America. The impacts of grazing on these habitats cause shifts in resources and subsequent change in species composition. The Greater sage-grouse, a keystone species of Great Basin ecosystems, rear offspring in these areas during spring and summer months using forbs and arthropods. To examine the impact of grazing on arthropod abundance in these ecosystems, seven meadows, each made up of three unique vegetative communities, were grazed at three intensities across two years (2019-2020) and monitored for environmental variables and abundance of arthropods during peak sage-grouse utilization periods. Additionally, the relationship of field measurements and near-surface digital cameras (phenocams) was examined to better understand how remote sensing technologies can be used to monitor these insect abundance shifts on larger scales. Arthropod taxa abundance responded differently to grazing management and environmental variables. Coleoptera abundance during peak sage-grouse usage periods increased roughly 50% in some meadows with increased grazing intensity. For year-to-year environmental variability in precipitation, Lepidoptera abundance was 114% higher in the drier year, while Coleoptera was 39% lower. Near-surface cameras had varied success with predicting peak insect abundance levels. Lepidoptera and Coleoptera capture rates had strong correlations with phenological indices derived from phenocams, while Formicidae had much weaker relationships.

GCC Countries Strategic Options in a Global Transition to Zero-Net Emissions.

Using a multi-level perspective approach combined with top-down macroeconomic models, we analyze the situation of the GCC countries in the perspective of a global transition to zero-net emissions before the end of the century. Based on these analyses, we propose strategic and political options for these oil and gas exporting countries. We show that it would be unwise for GCC member states to adopt an obstructionist strategy in international climate negotiations. On the contrary, these countries could be proactive in developing international emissions trading market and exploiting negative emissions obtained from CO2 direct reduction technologies, in particular direct air capture with CO2 sequestration, and thus contribute to a global net-zero-emissions regime in which clean fossil fuels are still used.

Role of uroguanylin's signalling pathway in the development of ischaemic stroke.

Stroke is one of the leading causes of mortality and disability worldwide. By affecting bradykinin function, activation of guanylate cyclase (GC)-A has been shown to have a neuroprotective effect after ischaemic stroke, whereas the same has not been confirmed for GC-B; therefore, we aimed to determine the possible role of GC-C and its agonist, uroguanylin (UGN), in the development of stroke. In this study, middle cerebral artery occlusion (MCAO) was performed on wild-type (WT), GC-C KO and UGN KO mice. MR images were acquired before and 24 h after MCAO. On brain slices 48 h after MCAO, the Ca2+ response to UGN stimulation was recorded. Our results showed that the absence of GC-C in GC-C KO mice resulted in the development of smaller ischaemic lesions compared with WT littermates, which is an opposite effect compared with the effects of GC-A agonists on brain lesions. WT and UGN KO animals showed a stronger Ca2+ response upon UGN stimulation in astrocytes of the peri-ischaemic cerebral cortex compared with the same cortical region of the unaffected contralateral hemisphere. This stronger activation was not observed in GC-C KO animals, which may be the reason for smaller lesion development in GC-C KO mice. The reason why GC-C might affect Ca2+ signalling in peri-ischaemic astrocytes is that GC-C is expressed in these cells after MCAO, whereas under normoxic conditions, it is expressed mainly in cortical neurons. Stronger activation of the Ca2+ -dependent signalling pathway could lead to the stronger activation of the Na+ /H+ exchanger, tissue acidification and neuronal death.

The introduction of tobacco excise taxation in the Gulf Cooperation Council Countries: a step in the right direction of advancing public health.

BACKGROUND: The Gulf Cooperation Council (GCC) countries relied, until recently, solely on import duties for tobacco products. The agreement for the introduction of an excise and value added tax (VAT) in 2016 and 2017, respectively, in most GCC countries, was a major breakthrough for public health. There is, however, ample room for improvement. METHODS: The study examines the outcomes of tax reforms, for both public health and public finances, based on the World Health Organization (WHO) recommendations and best practices worldwide. Tax simulations were performed using the WHO TaXSiM model. The study is based on data from Saudi Arabia, the only GCC country for which sufficient data existed. RESULTS: We recommend a stepwise tax reform, which involves increasing the current ad valorem excise tax rate, phasing out import duties keeping total tax share constant and introducing a minimum excise, and finally switching to a revenue-neutral specific excise. Specific excises must be adjusted for inflation and income increases. If implemented, cigarette tax reform simulations show that the recommended reforms would lead to a higher than 50% increase in cigarette prices, 16% reduction in cigarette sales and almost 50% increase in total cigarette tax revenue. A significant number of cigarette-related deaths would be averted. CONCLUSIONS: The recommended tax reforms are expected to lead to significant improvements in both public health and tobacco tax revenues. Our results provide useful insights that are of relevance to the whole GGC region. The effectiveness of the reforms, however, requires a strong tax and customs administration, including the establishment of a good database to monitor and advance public health.

Optimization Algorithm for Delay Estimation Based on Singular Value Decomposition and Improved GCC-PHAT Weighting.

The accuracy of time delay estimation seriously affects the accuracy of sound source localization. In order to improve the accuracy of time delay estimation under the condition of low SNR, a delay estimation optimization algorithm based on singular value decomposition and improved GCC-PHAT weighting (GCC-PHAT-ργ weighting) is proposed. Firstly, the acoustic signal collected by the acoustic sensor array is subjected to singular value decomposition and noise reduction processing to improve the signal-to-noise ratio of the signal; then, the cross-correlation operation is performed, and the cross-correlation function is processed by the GCC-PHAT-ργ weighting method to obtain the cross-power spectrum; finally, the inverse transformation is performed to obtain the generalized correlation time domain function, and the peak detection is performed to obtain the delay difference. The experiment was carried out in a large outdoor pool, and the experimental data were processed to compare the time delay estimation performance of three methods: GCC-PHAT weighting, SVD-GCC-PHAT weighting (meaning: GCC-PHAT weighting based on singular value decomposition) and SVD-GCC-PHAT-ργ weighting (meaning: GCC-PHAT-ργ weighting based on singular value decomposition). The results show that the delay estimation optimization algorithm based on SVD-GCC-PHAT-ργ improves the delay estimation accuracy by at least 37.95% compared with the other two methods. The new optimization algorithm has good delay estimation performance.

Investigating the role of export product diversification for renewable, and non-renewable energy consumption in GCC (gulf cooperation council) countries: does the Kuznets hypothesis exist?

This study explores the effects of renewable and nonrenewable energy demand on export product diversification, economic growth, natural resources, human capital, and trade in GCC (Gulf Cooperation Council) countries using data of six countries from 1990 to 2019. The empirical analysis integrates the panel unit root tests (IPS and CIPS), panel quantile regression, and fully modified OLS models. The empirical results confirm that there exists a significant negative relationship between renewable energy and export diversification; signifying that diversification of products will reduce renewable energy. Similarly, when compared to the square of export product diversification, it shows a positive and significant correlation. The empirical findings highlighted the presence of Kuznets's hypothesis between export product diversification, renewable, and non-renewable energy consumption. Furthermore, the findings suggest that natural resources and economic growth may increase overall energy consumption in GCC countries. It implies an important policy suggestion that encouraging export diversification will reduce GCC countries' reliance on oil to meet energy demand.

Impact of global health crisis and oil price shocks on stock markets in the GCC.

This study examines the impact of global COVID-19 cases and oil price shocks on the stock markets in the GCC. Using the Kalman filter to generate the unexpected oil price shocks, we find that, with the exception of Oman, the GCC markets responded to positive and negative oil price shocks before and during the pandemic, with impacts of higher magnitude since March 11, 2020. We also find that the spread of global COVID-19 cases had in itself no meaningful impact on the GCC stock markets.

EgJUB1 and EgERF113 transcription factors as potential master regulators of defense response in Elaeis guineensis against the hemibiotrophic Ganoderma boninense.

BACKGROUND: Hemibiotrophic pathogen such as the fungal pathogen Ganoderma boninense that is destructive to oil palm, manipulates host defense mechanism by strategically switching from biotrophic to necrotrophic phase. Our previous study revealed two distinguishable expression profiles of oil palm genes that formed the basis in deducing biotrophic phase at early interaction which switched to necrotrophic phase at a later stage of infection. RESULTS: The present report is a continuing study from our previous published transcriptomic profiling of oil palm seedlings against G. boninense. We focused on identifying differentially expressed genes (DEGs) encoding transcription factors (TFs) from the same RNA-seq data; resulting in 106 upregulated and 108 downregulated TFs being identified. The DEGs are involved in four established defense-related pathways responsible for cell wall modification, reactive oxygen species (ROS)-mediated signaling, programmed cell death (PCD) and plant innate immunity. We discovered upregulation of JUNGBRUNNEN 1 (EgJUB1) during the fungal biotrophic phase while Ethylene Responsive Factor 113 (EgERF113) demonstrated prominent upregulation when the palm switches to defense against necrotrophic phase. EgJUB1 was shown to have a binding activity to a 19 bp palindromic SNBE1 element, WNNYBTNNNNNNNAMGNHW found in the promoter region of co-expressing EgHSFC-2b. Further in silico analysis of promoter regions revealed co-expression of EgJUB1 with TFs containing SNBE1 element with single nucleotide change at either the 5th or 18th position. Meanwhile, EgERF113 binds to both GCC and DRE/CRT elements promoting plasticity in upregulating the downstream defense-related genes. Both TFs were proven to be nuclear-localized based on subcellular localization experiment using onion epidermal cells. CONCLUSION: Our findings demonstrated unprecedented transcriptional reprogramming of specific TFs potentially to enable regulation of a specific set of genes during different infection phases of this hemibiotrophic fungal pathogen. The results propose the intricacy of oil palm defense response in orchestrating EgJUB1 during biotrophic and EgERF113 during the subsequent transition to the necrotrophic phase. Binding of EgJUB1 to SNBE motif instead of NACBS while EgERF113 to GCC-box and DRE/CRT motifs is unconventional and not normally associated with pathogen infection. Identification of these phase-specific oil palm TFs is important in designing strategies to tackle or attenuate the progress of infection.

Partial protection of Sinopharm vaccine against SARS COV2 during recent outbreak in Bahrain.

BACKGROUND: In order to impart immunity against SARS COV 2 in the community, the oil rich countries of the Gulf Cooperation Council (GCC) provided citizens and expatriates with free vaccination. Different types of vaccination brands were utilized for this purpose. The purpose of this study is to determine the efficacy of the different types of vaccinations used. METHODS: This is an observational analytical case study of one Bahraini family who were vaccinated with 1st, 2nd or no dose. RESULTS: Out of 22 double dose recipients of SARS COV2 vaccine, 20 were infected. Those 20 were vaccinated against SARS COV 2 using Sinopharm, the rest (2) were in direct contact with the source but were vaccinated against SARS COV 2 using other type of vaccine. Out of 26 single dose recipients of Sinopharm vaccine, 23 were infected. The other three were not in direct contact with the infected source. Social gathering has been the main source of transmission. The infection has been mild with headache, chest pain. From 20 cases with double dose vaccinations only one had a lung infection and needed hospitalization. Out of 23 cases with single dose vaccinations 10 were hospitalized due to lung infections. All family members who were not vaccinated were infected, three were hospitalized one of which was deceased due to diabetes mellitus complications. CONLCUSION: Sinopharm provides partial protection against SARS COV 2 infection. That might be due to lack of its potential to detect recent variations in the protein structure of spike(S) protein of virus.

Cerebral trauma-induced dyschromatopsia in the left hemifield: case presentation.

BACKGROUND: Acquired color anomalies caused by cerebral trauma are classified as either achromatopsias or dyschromatopsias (Zeki, Brain 113:1721-1777, 1990). The three main brain regions stimulated by color are V1, the lingual gyrus, which was designated as human V4 (hV4), and the fusiform gyrus, designated as V4α. (Zeki, Brain 113:1721-1777, 1990). An acquired cerebral color anomaly is often accompanied by visual field loss (hemi- and quadrantanopia), facial agnosia, prosopagnosia, visual agnosia, and anosognosia depending on the underlying pathology (Bartels and Zeki, Eur J Neurosci 12:172-193, 2000), (Meadows, Brain 97:615-632, 1974), (Pearman et al., Ann Neurol 5:253-261, 1979). The purpose of this study was to determine the characteristics of a patient who developed dyschromatopsia following a traumatic injury to her brain. CASE PRESENTATION: The patient was a 24-year-old woman who had a contusion to her right anterior temporal lobe. After the injury, she noticed color distortion and that blue objects appeared green in the left half of the visual field. Although conventional color vision tests did not detect any color vision abnormalities, short wavelength automated perimetry (SWAP) showed a decrease in sensitivity consistent with a left hemi-dyschromatopsia. Magnetic resonance imaging (MRI) detected abnormalities in the right fusiform gyrus, a part of the anterior temporal lobe. At follow-up 14 months later, subjective symptoms had disappeared, but the SWAP abnormalities persisted and a thinning of the sectorial ganglion cell complex (GCC) was detected. CONCLUSION: The results indicate that although the subjective symptoms resolved early, a reduced sensitivity of SWAP remained and the optical coherence tomography (OCT) showed GCC thinning. We conclude that local abnormalities in the anterior section of fusiform gyrus can cause mild cerebral dyschromatopsia without other symptoms. These findings indicate that it is important to listen to the symptoms of the patient and perform appropriate tests including the SWAP and OCT at the early stage to objectively prove the presence of acquired cerebral color anomaly.

Co-movement between oil price, CO2 emission,renewable energy and energy equities: Evidence from GCC countries.

Using the data from GCC countries, this paper analyses the co-movement between oil price, EU carbon allowance prices, global clean energy index and equity index from three GCC counties, namely, Kuwait, Saudi Arabia and the United Arab Emirates. Almost no previous research has investigated the dynamic interrelations in the conventional energy markets, like those of the GCC countries, against the dramatic growth in clean energy production and the new emissions trading schemes. Employing three different multivariate GARCH models, we document the existence of volatility spillover effects and co-movement among global clean energy production, crude oil price, CO2 emission price and each of the three GCC energy stock markets. Furthermore, we found that the conditional variances of all return series are influenced by the shocks coming from the markets themselves. Volatilities in all the markets under consideration are highly persistent, and the long-run persistent volatilities are more pronounced especially for oil and CO2 emission prices. The forecasting exercise demonstrates the superior performance of the multivariate diagonal-BEKK GARCH models.

In Silico Prediction, Molecular Docking and Dynamics Studies of Steroidal Alkaloids of Holarrhena pubescens Wall. ex G. Don to Guanylyl Cyclase C: Implications in Designing of Novel Antidiarrheal Therapeutic Strategies.

The binding of heat stable enterotoxin (STa) secreted by enterotoxigenic Escherichia coli (ETEC) to the extracellular domain of guanylyl cyclase c (ECDGC-C) causes activation of a signaling cascade, which ultimately results in watery diarrhea. We carried out this study with the objective of finding ligands that would interfere with the binding of STa on ECDGC-C. With this view in mind, we tested the biological activity of a alkaloid rich fraction of Holarrhena pubescens against ETEC under in vitro conditions. Since this fraction showed significant antibacterial activity against ETEC, we decided to test the screen binding affinity of nine compounds of steroidal alkaloid type from Holarrhena pubescens against extracellular domain (ECD) by molecular docking and identified three compounds with significant binding energy. Molecular dynamics simulations were performed for all the three lead compounds to establish the stability of their interaction with the target protein. Pharmacokinetics and toxicity profiling of these leads demonstrated that they possessed good drug-like properties. Furthermore, the ability of these leads to inhibit the binding of STa to ECD was evaluated. This was first done by identifying amino acid residues of ECDGC-C binding to STa by protein-protein docking. The results were matched with our molecular docking results. We report here that holadysenterine, one of the lead compounds that showed a strong affinity for the amino acid residues on ECDGC-C, also binds to STa. This suggests that holadysenterine has the potential to inhibit binding of STa on ECD and can be considered for future study, involving its validation through in vitro assays and animal model studies.

Optical coherence tomography indices in the diagnosis and discrimination of stages of primary open-angle glaucoma in an African population.

PURPOSE: The objective of this study was to determine the structure-function association of the optical coherence tomography (OCT) indices (retinal nerve fibre layer (RNFL), ganglion cell complex (GCC) and optic nerve head (ONH) parameters) with the visual field sensitivity and their diagnostic ability at different stages of primary open-angle glaucoma (POAG) among a population West-African descent. METHODS: The study was a clinic-based prospective study which employed purposive sampling in the recruitment of clinically diagnosed POAG and non-glaucoma patients. OCT and visual field test (VFT) results were collated. Classification of the POAG cases was done using the Hodapp-Parrish-Anderson criteria. ROC was used for assessing the diagnostic ability of OCT indices. Pearson's correlation was used to assess the structure-function association. P value < 0.05 was considered statistically significant. CONCLUSIONS: Of the 497 subjects (497 eyes), 301 (60.6%) were males and 196 (39.4%) were females. The average, superior and inferior RNFL and GCC showed a strong positive correlation with the visual field sensitivity, but the average RNFL had the highest correlation (r = 0.673, p < 0.001). The parameters of the ONH showed a moderate-to-weak correlation with the visual field sensitivity. However, there was no significant correlation between structure and function in early POAG (p > 0.05). Superior GCC showed the highest diagnostic ability for POAG (AUC = 0.655; p < 0.001) comparable to the RNFL. Macula (GCC) and (RNFL) have a comparable ability to diagnose POAG at all the stages of POAG and can be used complimentarily for glaucoma assessment and diagnosis.

Pharmacy practice in hospital settings in GCC countries: Prescribing and transcribing.

PURPOSE: To outline hospital pharmacy practices across the Gulf Cooperation Councils (GCC) countries' hospitals. METHODS: A modified survey questionnaire was prepared from the original 2019 American Society of Health-System Pharmacist (ASHP) survey questions. Survey details were discussed with some pharmacy directors for clarity and relevance. A list of hospitals were obtained from the Ministry of Health of each of the targeted GCC countries. A secure invitation link containing a survey questionnaire was sent to the participants directly. RESULTS: Sixty four hospitals responded to this survey. The overall response rate was 52%. About 47% of the surveyed hospitals considered their drug formularies as closed, and strict. Additionally, only 44% of hospitals compare the effectiveness of products, when taking formulary decisions for drug inclusion. Forty-four percent of hospitals have computerized prescriber order entry (CPOE / EHR) system functionality for formulary system management. At about 39.1% hospitals, pharmacists have the responsibility for managing medication therapies, majority were engaged in providing anticoagulation therapies. About 61% of hospital pharmacies in GCC countries receive medication orders electronically, through CPOE/EHR. Majority (66%) of the hospitals in GCC countries have an active Antimicrobial Stewardship Program (ASP) while only 40% of pharmacists have a key role in providing clinical support. About 57.8% of hospital pharmacy directors reported that pharmacists do not provide ambulatory care clinical pharmacy services in their hospitals. CONCLUSION: In GCC countries' hospitals, there are major areas for improvement to patient care of which pharmacists are uniquely qualified as the medication experts to have the most meaningful outcomes in all of the domains of safe medication use, medication therapy management, antimicrobial stewardship program and participation in outpatient clinics.

The ethylene response factor VaERF092 from Amur grape regulates the transcription factor VaWRKY33, improving cold tolerance.

Cold stress is a major limiting factor in grape (Vitis) productivity. In this study, we characterized a cold-responsive ethylene response factor (ERF) transcription factor, VaERF092, from Amur grape (Vitis amurensis). VaERF092 expression was induced by both low temperatures and the ethylene precursor 1-aminocyclopropane-1-carboxylate (ACC), but was suppressed by treatment with the ethylene inhibitor aminoethoxyvinylglycine (AVG) under cold conditions. Ectopic expression of VaERF092 in Arabidopsis thaliana enhanced cold tolerance. Co-expression network analysis of V. vinifera genes indicated that WRKY33 might be a downstream target of VaERF092. This hypothesis was supported by the fact that VaWRKY33 was expressed temporally after VaERF092 expression and could also be induced by cold and ACC, and inhibited by AVG. Yeast one-hybrid, transient ß-glucuronidase (GUS) and dual-luciferase reporter assays provided evidence for an interaction between VaERF092 and a GCC-box element in the VaWRKY33 promoter. In addition, heterologous overexpression of VaWRKY33 in A. thaliana resulted in enhanced cold tolerance. VaERF092- and VaWRKY33 overexpressing grape calli showed lower low-temperature exothermic values than the empty vector (EV) calli, indicating enhanced tolerance to cold. Together, these results indicated that VaERF092 regulates VaWRKY33 through binding to its promoter GCC-box, leading to enhanced cold stress tolerance.

Structural insights into Arabidopsis ethylene response factor 96 with an extended N-terminal binding to GCC box.

The phytohormone ethylene is widely involved in many developmental processes and is a crucial regulator of defense responses against biotic and abiotic stresses in plants. Ethylene-responsive element binding protein, a member of the APETALA2/ethylene response factor (AP2/ERF) superfamily, is a transcription factor that regulates stress-responsive genes by recognizing a specific cis-acting element of target DNA. A previous study showed only the NMR structure of the AP2/ERF domain of AtERF100 in complex with a GCC box DNA motif. In this report, we determined the crystal structure of AtERF96 in complex with a GCC box at atomic resolution. We analyzed the binding residues of the conserved AP2/ERF domain in the DNA recognition sequence. In addition to the AP2/ERF domain, an N-terminal α-helix of AtERF96 participates in DNA interaction in the flanking region. We also demonstrated the structure of AtERF96 EDLL motif, a unique conserved motif in the group IX of AP2/ERF family, might involve in the transactivation of defense-related genes. Our study establishes the structural basis of the AtERF96 transcription factor in complex with the GCC box, as well as the DNA binding mechanisms of the N-terminal α-helix and AP2/ERF domain.