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Proanthocyanidins (PAs) are important metabolites that enhance freezing tolerance of plants. Actinidia arguta, especially freezing-tolerant germplasms, accumulate abundant PAs in dormant shoots and thereby enhance freezing tolerance, but the underlying mechanism is unknown. In this study, we used two A. arguta with contrasting cold-resistant phenotypes, KL and RB, to explore the mechanisms in response to cold tolerance. We determined that a leucoanthocyanidin reductase gene (AaLAR1) was more highly expressed in freezing-tolerant KL than in freezing-sensitive RB. Moreover, overexpressing AaLAR1 in kiwifruit promoted PAs biosynthesis and enhanced cold tolerance. The AaLAR1 promoters of various A. arguta germplasms differ due to the presence of a 60-bp deletion in cold-tolerant genotypes that forms a functional binding site for MYC-type transcription factor. Yeast one-hybrid and two-hybrid, dual-luciferase reporter, bimolecular fluorescence complementation and coimmunoprecipitation assays indicated that the AaMYC2a binds to the MYC-core cis-element in the AaLAR1 promoter with the assistance of AaMYB5a, thereby promoting PAs accumulation in the shoots of cold-tolerant kiwifruit. We conclude that the variation in the AaLAR1 promoter and the AaMYC2a-AaMYB5a-AaLAR1 module shape freezing tolerance in A. arguta. The identification of a key structural variation in the AaLAR1 promoter offers a new target for resistance breeding of kiwifruit.
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The defense response of peach (Prunus persica) to insect attack involves changes in gene expression and metabolites. Piercing/sucking insects such as green peach aphid cause direct damage by obtaining phloem nutrients and indirect damage by spreading plant viruses. To investigate the response of peach trees to aphids, the leaf transcriptome and metabolome of two genotypes with different sensitivities to green peach aphid (GPA, Myzus persicae) were studied. The transcriptome analysis of infected peach leaves showed two different response patterns. The gene expression of aphid-susceptible peach plants infected by aphids was more similar to that of the control plants, while the gene expression of aphid-resistant peach plants infected by aphids showed strongly induced changes in gene expression compared with the response in the control plants. Furthermore, gene transcripts in defense-related pathways, including plant-pathogen interaction, MAPK signaling, and several metabolic pathways, were more strongly enriched upon aphid infestation. Untargeted secondary metabolite profiling confirmed that aphid treatment induced larger changes in aphid-resistant peaches than in aphid-susceptible peaches. Consistent with transcriptomic alterations, nine triterpenoids showed extremely significant GPA-induced accumulation in aphid-resistant peaches, whereas triterpenoid abundance remained predominantly unchanged or undetected in aphid- susceptible peaches. Furthermore, some types of transcription factors (including WRKYs, ERFs, NACs, etc.) were more strongly induced upon GPA infestation in aphid-resistant peaches but not in aphid-susceptible peaches. Aphid feeding-dependent transcriptome and metabolite profiles provide the foundation for understanding the molecular mechanisms underlying the response of peach to aphid infestation. These results suggested that accumulation of specialized triterpenoids and the corresponding pathway transcripts may play a key role in peach GPA resistance.
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BACKGROUND: Freezing injury, which is an important abiotic stress in horticultural crops, influences the growth and development and the production area of kiwifruit (Actinidia Lind1). Among Actinidia species, Actinidia arguta has excellent cold resistance, but knowledge relevant to molecular mechanisms is still limited. Understanding the mechanism underlying cold resistance in kiwifruit is important for breeding cold resistance. RESULTS: In our study, a population resulting from the cross of A. arguta 'Ruby-3' × 'Kuilv' male was generated for kiwifruit hardiness study, and 20 cold-tolerant and 20 cold-sensitive populations were selected from 492 populations according to their LT50. Then, we performed bulked segregant RNA-seq combined with single-molecule real-time sequencing to identify differentially expressed genes that provide cold hardiness. We found that the content of soluble sucrose and the activity of ß-amylase were higher in the cold-tolerant population than in the cold-sensitive population. Upon - 30 °C low-temperature treatment, 126 differentially expressed genes were identify; the expression of 59 genes was up-regulated and that of 67 genes was down-regulated between the tolerant and sensitive pools, respectively. KEGG pathway analysis showed that the DEGs were primarily related to starch and sucrose metabolism, amino sugar and nucleotide sugar metabolism. Ten major key enzyme-encoding genes and two regulatory genes were up-regulated in the tolerant pool, and regulatory genes of the CBF pathway were found to be differentially expressed. In particular, a 14-3-3 gene was down-regulated and an EBF gene was up-regulated. To validate the BSR-Seq results, 24 DEGs were assessed via qRT-PCR, and the results were consistent with those obtained by BSR-Seq. CONCLUSION: Our research provides valuable insights into the mechanism related to cold resistance in Actinidia and identified potential genes that are important for cold resistance in kiwifruit.
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Actinidia , Actinidia/genética , Respuesta al Choque por Frío/genética , Frutas , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Masculino , FitomejoramientoRESUMEN
BACKGROUND: Kiwifruit (Actinidia Lindl.) is considered an important fruit species worldwide. Due to its temperate origin, this species is highly vulnerable to freezing injury while under low-temperature stress. To obtain further knowledge of the mechanism underlying freezing tolerance, we carried out a hybrid transcriptome analysis of two A. arguta (Actinidi arguta) genotypes, KL and RB, whose freezing tolerance is high and low, respectively. Both genotypes were subjected to - 25 °C for 0 h, 1 h, and 4 h. RESULTS: SMRT (single-molecule real-time) RNA-seq data were assembled using the de novo method, producing 24,306 unigenes with an N50 value of 1834 bp. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of DEGs showed that they were involved in the 'starch and sucrose metabolism', the 'mitogen-activated protein kinase (MAPK) signaling pathway', the 'phosphatidylinositol signaling system', the 'inositol phosphate metabolism', and the 'plant hormone signal transduction'. In particular, for 'starch and sucrose metabolism', we identified 3 key genes involved in cellulose degradation, trehalose synthesis, and starch degradation processes. Moreover, the activities of beta-GC (beta-glucosidase), TPS (trehalose-6-phosphate synthase), and BAM (beta-amylase), encoded by the abovementioned 3 key genes, were enhanced by cold stress. Three transcription factors (TFs) belonging to the AP2/ERF, bHLH (basic helix-loop-helix), and MYB families were involved in the low-temperature response. Furthermore, weighted gene coexpression network analysis (WGCNA) indicated that beta-GC, TPS5, and BAM3.1 were the key genes involved in the cold response and were highly coexpressed together with the CBF3, MYC2, and MYB44 genes. CONCLUSIONS: Cold stress led various changes in kiwifruit, the 'phosphatidylinositol signaling system', 'inositol phosphate metabolism', 'MAPK signaling pathway', 'plant hormone signal transduction', and 'starch and sucrose metabolism' processes were significantly affected by low temperature. Moreover, starch and sucrose metabolism may be the key pathway for tolerant kiwifruit to resist low temperature damages. These results increase our understanding of the complex mechanisms involved in the freezing tolerance of kiwifruit under cold stress and reveal a series of candidate genes for use in breeding new cultivars with enhanced freezing tolerance.
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Aclimatación/genética , Actinidia/genética , Actinidia/fisiología , Congelación , Regulación de la Expresión Génica de las Plantas , Frutas/genética , Frutas/fisiología , Perfilación de la Expresión Génica , Redes Reguladoras de Genes , Sistema de Señalización de MAP Quinasas , Anotación de Secuencia Molecular , Fosfatidilinositoles/metabolismo , Fitomejoramiento , Reguladores del Crecimiento de las Plantas/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Almidón/metabolismo , Sacarosa/metabolismoRESUMEN
PURPOSE: To compare the relative clinical efficacy of preoperative computed tomography (CT)-guided methylene-blue (MB) and coil localization for lung nodules (LNs). MATERIAL AND METHODS: Between January 2013 and December 2018, a total of 89 patients with LNs underwent CT-guided MB or coil localization and subsequent video-assisted thoracoscopic surgery (VATS)-guided wedge resection in our hospital. We compared the technical success of localization and wedge resection between two groups. RESULTS: In MB group, 47 LNs in 39 patients were localized, with successful localization and wedge resection rates of 97.9% and 97.9%, respectively. In the coil group, 64 LNs in 50 patients were localized, with successful localization and wedge resection rates of 96.9% and 96.9%, respectively. There were no significant differences in technical success rates of localization and wedge resection between the two groups (p = 1.000 and 1.000). The coil group sustained a longer duration between localization and VATS relative to the MB group (14.4 h vs. 1.6 h, p = .001). CONCLUSION: Both MB and coil localization were safe and effective techniques to establish a high success rate of VATS-guided wedge resection for LNs. Relative to MB localization, coil localization might be compatible with a longer delay between localization and VATS.
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Neoplasias Pulmonares , Azul de Metileno , Humanos , Pulmón , Neoplasias Pulmonares/diagnóstico por imagen , Neoplasias Pulmonares/cirugía , Neumonectomía , Estudios Retrospectivos , Tomografía Computarizada por Rayos XRESUMEN
OBJECTIVE: The aim of this study was to evaluate the changes of the iodine value quantified on the Couinaud segments measured in port-venous phase using the iodine-mixed technique of contrast-enhanced dual-source dual-energy computed tomography (CT) scanning in different Child-Pugh stages of hepatitis B-induced liver cirrhosis. METHODS: Patients clinically diagnosed with hepatitis B-induced cirrhosis were prospectively engaged in our study. Each patient underwent multiphase iodine agent contrast-enhanced dual-source dual-energy CT scanning, and then the iodine-mixed imaging of port-venous phase was postprocessed. Iodine concentration was obtained for each segment based on the Couinaud segments. The volume of each segment and the total of the liver were measured and calculated using the postprocessing software of volume. All the cirrhosis patients were grouped into 3 subgroups based on the Child-Pugh stage method. Patients without cirrhosis were engaged for the control group. The iodine concentration, volume, and iodine storage among groups were analyzed by SPSS version 19.0. Single energy was used for the nonenhanced phase scanning, which was used for the radiation dosage comparison with dual-energy CT scanning. RESULTS: Two hundred three patients were ultimately enrolled in our study, including 148 patients with cirrhosis (Child A, 69; Child B, 51; Child C, 28) and 55 patients without cirrhosis as control subjects. The total volume and iodine storage of cirrhosis group were smaller than those of the control group (P < 0.001). Compared with the control group, the iodine concentration in each segment decreased with progression of cirrhosis. The volume, iodine concentration, and iodine storage of the right hepatic lobe and left medial segment decreased with cirrhosis severity (P < 0.001). There was no significant difference in the volume of right hepatic lobe between Child C group and Child B group, whereas the iodine storage of Child C group was lower than that of Child B group (P < 0.05). The volume and iodine storage of left lateral segment increased with the progression of liver cirrhosis in the Child A and Child B groups (P < 0.05), whereas there was no statistical difference between the Child B and Child C groups, and the iodine storage in the Child C group was lower than that of the Child B group (P < 0.05). The radiation dose of dual-energy scanning was lower than that of single-energy scanning (P < 0.001). The iodine concentration 1.512 mg/mL on the left medial segment reached the most optimal evaluation on cirrhosis, with a sensitivity of 100%, specificity of 0.722, and area under the curve of 0.914. CONCLUSIONS: Iodine concentration in portal phase measurement can evaluate and reflect the severity of cirrhosis. Iodine content segmental quantification can analyze the changes of the liver storage with a progression of cirrhosis. Dual-energy scanning reduced the radiation damage in patients and is valuable for a further study and clinical application.
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Hepatitis B/complicaciones , Hepatitis B/diagnóstico por imagen , Cirrosis Hepática/diagnóstico por imagen , Cirrosis Hepática/virología , Imagen Radiográfica por Emisión de Doble Fotón/métodos , Tomografía Computarizada por Rayos X/métodos , Adulto , Anciano , Medios de Contraste , Femenino , Humanos , Masculino , Persona de Mediana Edad , Vena Porta/diagnóstico por imagen , Estudios Prospectivos , Dosis de Radiación , Interpretación de Imagen Radiográfica Asistida por Computador/métodos , Sensibilidad y Especificidad , Ácidos TriyodobenzoicosRESUMEN
BACKGROUND The aim of this study was to investigate whether the anisotropy parameters are helpful in the detection and discrimination of breast cancers, and to determine its value in predicting the risk of cancers. MATERIAL AND METHODS There were 56 patients with 56 lesions (34 malignant, 22 benign) included in the study. DTI was performed in every patient and apparent diffusion coefficient (ADC), fractional anisotropy (FA), and eigenvalues E1, E2, and E3 were measured in every lesion and the normal breast tissue. RESULTS ADC, FA, and eigenvalues of E1, E2, E3, and E1-E3 in breast cancers were all significantly lower than in normal tissue (P<0.001 for all) with mean reduction of (32 ± 17)%, (24 ± 13)%, (33 ± 19)%, (32 ± 17)%, (31 ± 18)%, and (37 ± 20)% for ADC, FA, E1, E2, E3, and E1-E3, respectively. These parameters were also statistically lower in cancers than in benign lesions (P<0.01 for all), except FA (P>0.05). ADC, E1, E2, and E3 were very similar in discriminating breast cancers and benign lesions, with area under the curve (AUC) 0.885-0.898, sensitivity 73.5-85.3%, and specificity 90.9-100%. CONCLUSIONS ADC, E1, E2, E3, and E1-E3 are much lower in breast cancers than in normal tissue and benign lesions. The reduction of ADC, E1, E2, E3, and E1-E3 of a mass in the breast is highly associated with the risk of breast cancer, but the FA has no utility in breast cancer risk prediction.
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Neoplasias de la Mama/diagnóstico , Neoplasias de la Mama/patología , Imagen de Difusión Tensora/métodos , Adulto , Anciano , Anisotropía , Mama/patología , Medios de Contraste , Diagnóstico Diferencial , Femenino , Humanos , Persona de Mediana Edad , Probabilidad , Curva ROC , Factores de RiesgoRESUMEN
CONTEXT: In this paper, we systematically studied the effects of non-metallic element (B, C, N, O, F) doping and biaxial stretching on the photoelectric properties of ZrS2/ZrSe2 heterostructures by using the first-principles calculation method based on density functional theory. The results show that the p-type doping is realized by B, C, and N atom doping, and the n-type doping is realized by O and F atom doping. The doping of B and C atoms produces impurity energy levels in the band gap, which affects the conductivity of the heterostructure. The band gap of N and O atom-doped heterostructures increases under tensile strain, but it is still a direct band gap. The analysis of the optical properties of the heterostructures shows that the doping of non-metallic atoms can adjust the optical absorption rate and reflectivity of the heterostructures. Under the action of tensile strain, the optical properties of the doped heterostructures have changed significantly in the low-energy region. This article provides a theoretical basis for the future application of ZrS2/ZrSe2 heterostructures. METHOD: This paper uses the first-principles calculation method based on density functional theory. The PBE exchange-correlation functional based on generalized gradient approximation (GGA) is selected for the specific calculation, and the crystal structure is geometrically optimized by the ultrasoft pseudopotential method. It is verified that when the cutoff energy of the ZrS2/ZrSe2 heterostructure is 500 eV, the K-point grid is selected to be 10 × 10 × 2 with the lowest energy, so the cutoff energy is selected to be 500 eV. The K-point grid is selected to be 10 × 10 × 2. The convergence limits for structural optimization are as follows: the maximum force between atoms is 0.01 eV/Å, the convergence threshold of the maximum energy change is set to 10-9 eV/atom, and the convergence threshold of the maximum displacement is 0.001 Å. In order to avoid the influence of atomic periodic motion between different atomic layers, a vacuum layer of 20 Å is added in the vertical direction. Considering the interaction of vdW between the interfaces, the DFT-D2 method is used to verify. The optical properties were calculated by the random phase approximation method, and the K-point grid was selected as 12 × 12 × 2.
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CONTEXT: This paper systematically analyzes the phonon dispersion curves of single-layer ZrS2, ZrSe2, and ZrS2/ZrSe2 heterostructures under different strains. The phonon spectra and thermal parameters of the three structures were obtained based on the density functional perturbation theory method. The upper limits of strain that different monolayers and heterojunctions can withstand were studied. The monolayers ZrSe2 and ZrS2 can withstand up to 8% biaxial tensile strain, and the ZrS2/ZrSe2 heterojunction can withstand up to 6% biaxial tensile strain. In addition, the van der Waals force of the heterojunction may cause phonon tearing in the vertical direction. The application of biaxial tensile strain can adjust the thermal properties of the system to a large extent, which is similar to the strain effect in the pristine case. When the temperature rises, the entropy enthalpy of the three structures also gradually increases, the free energy gradually decreases, and the heat capacity of the system gradually increases until it tends to be stable. Taking single-layer ZrS2 as an example, we analyzed the change curve of thermal properties of single-layer ZrS2 under tensile strain. The results show that with the gradual increase of strain, the crystal's entropy, enthalpy, and free energy change differently. In addition, the heat capacity increases slowly under high temperatures. When all systems reach the limit strain, the phonon spectrum appears to have an imaginary frequency, and the thermal properties decrease significantly. METHODS: This paper uses the first-principle calculation method based on density functional theory, and the PBE exchange-correlation function based on generalized gradient approximation (GGA) is selected for a specific calculation. The density functional perturbation theory (DFPT) calculates the full kinetic matrix. Because the lattice constants of ZrS2 and ZrSe2 are similar and have similar periodicity, the corresponding unit cells are used for structural optimization and property calculation. The Brillouin zone is integrated using the K points generated by the Monkhorst-pack method. For single-layers ZrS2 and ZrSe2, 8 × 8 × 1 K-point grid is selected, and for ZrS2/ZrSe2 heterojunction, 8 × 8 × 2 K-point grid is selected. A vacuum layer of 30 Å was added in the vertical direction to avoid interlayer interaction. The non-conservative pseudopotential method is used to optimize the structure, and the optimization convergence is set as follows: the cutoff energy is set to 700 eV, the convergence threshold of the maximum force between atoms is 0.01 eV/Å, the convergence threshold of the maximum energy change is set to 1 × 10-9 eV, and the convergence threshold of the maximum displacement is 0.001 Å.
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CONTEXT: Two-dimensional semiconductor materials have received much attention in recent years due to their wide variety of applications in the field of nano-optoelectronic devices. In this project, we applied stresses ranging from -6 to +6% to the ZrSe2/HfSe2 heterostructure and systematically investigated its electrical and optical properties. It is discovered that stress can effectively modulate the forbidden bandwidth of the ZrSe2/HfSe2 heterojunction; whereas, under compressive stress, the forbidden bandwidth of the material decreases further until the bandgap is zero, leading to the material's transformation from semiconductor to metal. The forbidden band gap of the ZrSe2/HfSe2 heterojunction increases with increasing horizontal biaxial tensile strain. We discovered that the light absorption performance of this heterostructure is significantly better than that of its similar monomolecular layer and that its light absorption intensity can reach an order of magnitude of 104. Under compressive and tensile stresses, the ZrSe2/HfSe2 heterojunctions exhibit different degrees of red or blue shift. The results indicate that constructing ZrSe2/HfSe2 heterojunctions and applying horizontal biaxial stresses to them can significantly modulate the optoelectronic properties of the materials. ZrSe2/HfSe2 heterojunction is a new type of high-performance photogenerated carrier transport device with a wide range of applications. METHODS: The calculations in this study are carried out the first principles approach of density functional theory, as implemented in the CASTEP module of Materials-Studio2019. The researchers used an ultrasoft reaction potential to calculate the interactions between the ion core and the electrons and applied the Perdew-Burke-Ernzerhof (PBE) and the generalized gradient approximation (GGA) to perform the calculations. The Monkhorst-Pack technique was employed to create the k-point samples utilized for integration on the Brillouin zone, and the k-point grid was uniformly 6 × 6 × 1. In addition, in order to avoid interactions between the atomic layers affecting the properties and stability of the material, such interactions were prevented by adding a 30 Å vacuum layer. Using a plane-wave energy cutoff of 500 eV and the convergence accuracy of the iterative process was set to 1 × 10-5 eV to ensure the accuracy of the computational results, and in addition. The maximum stress in the lattice was limited to less than 0.05 GPa or the interaction force between neighboring atoms was lower than 0.03 eV/Å. For the calculation of the properties of the optical properties, a k-point grid of 18 × 18 × 1 is used for optimization, and the polarization direction of the material is not taken into account, considering that the material is isotropic. This study proposes to apply the Tkatchenko-Scheffler (TS) dispersion correction method in DFT-D to appropriately represent the interlayer van der Waals interaction forces to solve inaccuracies in the computation of van der Waals interactions via density functional theory.
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CONTEXT: Based on the first principles under the framework of density functional theory, it calculates the effect of vacancy defects in single Zr and single Se atoms and the replacement of Se atoms in ZrSe2 with O, Se, and Te atoms on the optoelectronic properties of monolayer ZrSe2, including geometry, energy band structure, electronic density of states, and optical properties. The doping of the three non-metallic atoms was n-type doping for the O and S atoms and p-type doping for the Te atom. Defects in the Zr atoms and O-atom doping significantly affect the peak reflectance and absorption coefficient of the ZrSe2 system. METHODS: All Density Functional Theory calculations were carried out using the CASTEP module in the Materials-Studio (MS) software. The generalized gradient approximation plane-wave pseudopotential method and the Perdew-Burke-Ernzerfhof (PBE) generalized function were used for structural optimization and total energy calculation of the defect and doping systems. After convergence tests, the plane wave truncation energy was set to 500 eV, and the Brillouin zone K-point grid was set to 4 × 4 × 1. The atomic energy convergence criterion is 1.0 × 10-6 eV/atom, the interatomic interaction force convergence criterion is 0.02 eV/Å, the maximum atomic displacement convergence criterion is 0.001 Å, and the internal crystal stress convergence criterion is 0.05 GPa. In order to avoid the influence of the interaction forces between the layers, a vacuum layer of 15 Å is placed in the Z-axis direction.
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The Actinidia (kiwifruit) is an emerging fruit plant that is severely affected by salt stress in northern China. Plants have evolved several signaling network mechanisms to cope with the detrimental effects of salt stress. To date, no reported work is available on metabolic and molecular mechanisms involved in kiwifruit salt tolerance. Therefore, the present study aims to decipher intricate adaptive responses of two contrasting salt tolerance kiwifruit species Actinidia valvata [ZMH (an important genotype), hereafter referred to as R] and Actinidia deliciosa ['Hayward' (an important green-fleshed cultivar), hereafter referred to as H] under 0.4% (w/w) salt stress for time courses of 0, 12, 24, and 72 hours (hereafter refered to as h) by combined transcriptome and metabolome analysis. Data revealed that kiwifruit displayed specific enrichment of differentially expressed genes (DEGs) under salt stress. Interestingly, roots of R plants showed a differential expression pattern for up-regulated genes. The KEGG pathway analysis revealed the enrichment of DEGs related to plant hormone signal transduction, glycine metabolism, serine and threonine metabolism, glutathione metabolism, and pyruvate metabolism in the roots of R under salt stress. The WGCNA resulted in the identification of five candidate genes related to glycine betaine (GB), pyruvate, total soluble sugars (TSS), and glutathione biosynthesis in kiwifruit. An integrated study of transcriptome and metabolome identified several genes encoding metabolites involved in pyruvate metabolism. Furthermore, several genes encoding transcription factors were mainly induced in R under salt stress. Functional validation results for overexpression of a candidate gene betaine aldehyde dehydrogenase (AvBADH, R_transcript_80484) from R showed significantly improved salt tolerance in Arabidopsis thaliana (hereafter referred to as At) and Actinidia chinensis ['Hongyang' (an important red-fleshed cultivar), hereafter referred to as Ac] transgenic plants than in WT plants. All in all, salt stress tolerance in kiwifruit roots is an intricate regulatory mechanism that consists of several genes encoding specific metabolites.
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Beta-amylase (BAM) plays an important role in plant resistance to cold stress. However, the specific role of the BAM gene in freezing tolerance is poorly understood. In this study, we demonstrated that a cold-responsive gene module was involved in the freezing tolerance of kiwifruit. In this module, the expression of AaBAM3.1, which encodes a functional protein, was induced by cold stress. AaBAM3.1-overexpressing kiwifruit lines showed increased freezing tolerance, and the heterologous overexpression of AaBAM3.1 in Arabidopsis thaliana resulted in a similar phenotype. The results of promoter GUS activity and cis-element analyses predicted AaCBF4 to be an upstream transcription factor that could regulate AaBAM3.1 expression. Further investigation of protein-DNA interactions by using yeast one-hybrid, GUS coexpression, and dual luciferase reporter assays confirmed that AaCBF4 directly regulated AaBAM3.1 expression. In addition, the expression of both AaBAM3.1 and AaCBF4 in kiwifruit responded positively to cold stress. Hence, we conclude that the AaCBF-AaBAM module is involved in the positive regulation of the freezing tolerance of kiwifruit.
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Cold stress poses a serious treat to cultivated kiwifruit since this plant generally has a weak ability to tolerate freezing tolerance temperatures. Surprisingly, however, the underlying mechanism of kiwifruit's freezing tolerance remains largely unexplored and unknown, especially regarding the key pathways involved in conferring this key tolerance trait. Here, we studied the metabolome and transcriptome profiles of the freezing-tolerant genotype KL (Actinidia arguta) and freezing-sensitive genotype RB (A. arguta), to identify the main pathways and important metabolites related to their freezing tolerance. A total of 565 metabolites were detected by a wide-targeting metabolomics method. Under (-25°C) cold stress, KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway annotations showed that the flavonoid metabolic pathways were specifically upregulated in KL, which increased its ability to scavenge for reactive oxygen species (ROS). The transcriptome changes identified in KL were accompanied by the specific upregulation of a codeinone reductase gene, a chalcone isomerase gene, and an anthocyanin 5-aromatic acyltransferase gene. Nucleotides metabolism and phenolic acids metabolism pathways were specifically upregulated in RB, which indicated that RB had a higher energy metabolism and weaker dormancy ability. Since the LPCs (LysoPC), LPEs (LysoPE) and free fatty acids were accumulated simultaneously in both genotypes, these could serve as biomarkers of cold-induced frost damages. These key metabolism components evidently participated in the regulation of freezing tolerance of both kiwifruit genotypes. In conclusion, the results of this study demonstrated the inherent differences in the composition and activity of metabolites between KL and RB under cold stress conditions.
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Low temperature causes injuries to plants during winter, thereby it affects kiwi fruit quality and yield. However, the changes in metabolites and gene expression during cold acclimation (CA) and deacclimation (DA) in kiwi fruit remain largely unknown. In this study, freezing tolerance, carbohydrate metabolism, and ß-amylase gene expression in two Actinidia arguta cv. "CJ-1" and "RB-3" were detected from CA to DA stages. In all acclimation stages, the "CJ-1" was hardier than "RB-3" and possessed lower semi-lethal temperature (LT50). Furthermore, "CJ-1" had a more rapid acclimation speed than "RB-3". Changes of starch, ß-amylase, and soluble sugars were associated with freezing tolerance in both cultivars. Starch contents continued to follow a declining trend, while soluble sugars contents continuously accumulated in both cultivars during CA stages (from October to January). To investigate the possible molecular mechanism underlying cold response in A. arguta, in total, 16 AcBAMs genes for ß-amylase were identified in the kiwi fruit genome. We carried out localization of chromosome, gene structure, the conserved motif, and the analysis of events in the duplication of genes from AcBAMs. Finally, a strong candidate gene named AaBAM3 from AcBAMs was cloned in Actinidia arguta (A. arguta), The real-time qPCR showed that AaBAM3 gene expression in seasonal changes was consistent with changes of soluble sugars. These results reveal that AaBAM3 may enhance the freezing tolerance of A. arguta through increasing soluble sugar content.
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Among the genus Actinidia, Actinidia arguta possesses the strongest cold resistance and produces fresh fruit with an intense flavor. To investigate genomic variation that may contribute to variation in phenotypic traits, we performed whole-genome re-sequencing of four A. arguta genotypes originating from different regions in China and identified the polymorphisms using InDel markers. In total, 4,710,650, 4,787,750, 4,646,026, and 4,590,616 SNPs and 1,481,002, 1,534,198, 1,471,304, and 1,425,393 InDels were detected in the 'Ruby-3', 'Yongfeng male', 'Kuilv male', and 'Hongbei male' genomes, respectively, compared with the reference genome sequence of cv 'Hongyang'. A subset of 120 InDels were selected for re-sequencing validation. Additionally, genes related to non-synonymous SNPs and InDels in coding domain sequences were screened for functional analysis. The analysis of GO and KEGG showed that genes involved in cellular responses to water deprivation, sucrose transport, decreased oxygen levels and plant hormone signal transduction were significantly enriched in A. arguta. The results of this study provide insight into the genomic variation of kiwifruit and can inform future research on molecular breeding to improve cold resistance in kiwifruit.
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Actinidia/genética , Genoma de Planta , ADN de Plantas/genética , Frutas/genética , Regulación de la Expresión Génica de las Plantas , Mutación INDEL , Proteínas de Plantas/genética , Polimorfismo de Nucleótido Simple , Secuenciación Completa del GenomaRESUMEN
PURPOSE: To determine the correlation between the parameters of MR diffusion kurtosis imaging (MR-DKI) and mismatch repair gene expression (MMR) for rectal carcinomas. MATERIALS AND METHODS: Data from 80 patients with rectal carcinoma were analyzed in this prospective study. High-resolution T2WI and DKI (b = 0, 800 and 1600 s/mm2, respectively) were performed. Mean kurtosis (MK) and mean diffusivity (MD) from DKI were measured. MMR-positive expression and HER-2 expression were classified into two groups. For comparison between different grades, the Mann-Whitney U test, receiver operating characteristic curve, and Spearman's correlation analysis were used for statistical analyses. RESULTS: The MK values in identifying positive MMR expressions (MLH1, MSH2, and MSH6) were more reliable than the MD values (rs value: 0.772 vs. 0.448, 0.733 vs. 0.499, and 0.828 vs. 0.633 respectively, P<0.01). Receiver operating curve analysis showed that the performances of the MK values were better than those of the MD values (z = 2.835, 2.000, and 2.827, respectively, P<0.05), while the performances of the MK and MD-MK values were not statistically significant (z = 0.808, 1.557, and 0.596, respectively, P>0.05). Similarly, MK values were better than MD values in identifying HER2 expression (z = 2.795, P<0.05). CONCLUSIONS: MK derived from DKI demonstrated a greater correlation than MD with MMR expression. It also showed better performance in differentiating between high- and low-grade positive MMR expression and HER2 expression. Thus, DKI may be valuable for the prognoses and evaluation of non-invasive therapies.
Asunto(s)
Reparación de la Incompatibilidad de ADN/genética , Imagen de Difusión por Resonancia Magnética , Regulación Neoplásica de la Expresión Génica , Neoplasias del Recto/diagnóstico , Neoplasias del Recto/genética , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Receptor ErbB-2/metabolismo , Neoplasias del Recto/metabolismoRESUMEN
OBJECTIVE: To investigate the apparent diffusion coefficient (ADC) and fractional anisotropy (FA) measured by diffusion tensor imaging (DTI), tissue cellularity and their relationship in breast malignant/benign lesions. METHODS: 88 patients with 88 breast lesions who underwent DTI and dynamic contrast-enhanced MR scanning between November 2013 and December 2014 were retrospectively analyzed. The diagnosis was confirmed pathologically. ADC and FA values as well as histopathological cellularity of different pathological types of lesions were analyzed and compared statistically. The Pearson's correlation between cellularity and ADC and FA was calculated. RESULTS: There were 59 cases of breast cancer and 29 cases of benign lesions included in the study. ADC values of breast cancers were statistically lower than that of benign lesions (p < 0.001). FA and cellularity were higher in cancers than in benign lesions with statistical significance (p < 0.05 and p < 0.001, respectively). The mean FA values in the patients with invasive ductal carcinoma (IDC) were higher than that in the patients with ductal carcinoma in situ (DCIS) without statistical difference (p > 0.05). The ADC and the cellularity in the IDC of grade III were statistically lower (p < 0.05) and higher (p < 0.05) than that in the DCIS and IDC of grade I-II, respectively. ADC was negatively correlated to cellularity (r = -0.8319, p < 0.001) and FA was positively correlated to cellularity (r = 0.4231, p < 0.001). CONCLUSION: ADC and FA values were statistically different between benign and malignant breast lesions and were significantly correlated to tissue cellularity. ADC and FA may help to discriminate malignant from benign breast lesions and to predict cellularity. ADC is helpful in the prediction of the grade of breast cancer. ADVANCES IN KNOWLEDGE: ADC and FA values were statistically different between benign and malignant breast lesions and were significantly correlated to tissue cellularity.