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Abstract [Objective] The study aims to analyze the effects of melatonin (MT) on the growth, biochemical characteristics, and expression of cold-responsive genes in grapevine (Vitis vinifera L.) suspension cells under low temperature stress, to screen the appropriate MT concentrations and to investigate the mechanism of MT in enhancing cold tolerance of grapevine. [Methods] Using ‘Pinot Noir’ grapevine suspension cells as test materials, we investigated the effects of different concentrations of MT [0 (CK), 50 (T1), 100 (T2), 200 (T3), 300 μmol/L (T4)] on the growth of the suspension cells, the content of malondialdehyde (MDA), the content of proline (Pro), and the antioxidant enzyme activities, and changes in gene expression related to cold response to T2 treatment, under low temperature (4 ℃) stress, [Results] (1) Under low temperature stress, the dry weight of cells in each MT treatment showed a trend of first increase then decrease, which was finally stable or decreased, and was higher than CK at the end of treatment. (2) MDA and Pro contents of each treatment were firstly increased and then decreased with the time of treatment, which reached the peak at 24 h or 48 h. MDA content of each MT treatment was lower than that of CK, while the Pro content was higher than that of CK, with T2 treatment being the lowest or the highest. (3) The antioxidant enzyme activities of each MT treatment were higher than that of CK, with T2 treatment being the highest. With the prolongation of time, the activities of POD and CAT of each treatment firstly rose and then fell, while the activities of SOD and APX firstly fell and then rose, which reached the highest at 24 h or 6 h, respectively. (4) The relative expression of cold-responsive genes (VvCBFs, VvICE1, VvRD29B, and VvKIN-2) in T2-treated suspension cells were up-regulated compared with CK. [Conclusion] Under low temperature stress, the appropriate concentration (100 μmol/L) of MT increases the expression of antioxidant enzymes and cold-response genes, increases Pro content, decreases MDA content, and enhances low temperature tolerance of grapevine suspension cells.

Abstract [Objective] This study aims to identify and analyze the members of the tonoplast intrinsic protein (TIP) subfamily in apple, and to investigate the expression patterns under drought stress. It provides information for further research on utilization of drought resistance gene resources in apple. [Methods] The MdTIPs in apple genome was identified by bioinformatics methods. The physicochemical properties, gene structures, conserved motifs, cis-regulatory elements, and phylogenetic trees, etc. of the subfamily members were analyzed. The expression patterns of MdTIPs in different organs under drought stress were analyzed by qRT-PCR. [Results] A total of 13 MdTIP genes were identified in the apple genome, and most members were localized at the plasma membrane. Chromosome localization analysis suggested that all members were distributed on 10 chromosomes, with 1-3 members on each chromosome. Besides, the promoter regions of the genes contained response elements for hormonal and adversity stresses. qRTPCR showed that MdTIP members were up-regulated in roots except MdTIP1;1, of which MdTIP1;3 and MdTIP1;4 were up-regulated 5.27 times and 5.69 times, respectively, compared with the control, suggesting that these two genes might be critical in response to drought stress. [Conclusion] Identification of MdTIP subfamily members is provided in this study. 10 MdTIP members are differentially expressed in roots, stems, and leaves, and 12 members are highly expressed in roots.

The water extract contents, polyphenol compositions, total free amino acid contents, caffeine (CAF) contents, amino acid compositions and catechin compositions of spring and summer tea made from six varieties planted in Motuo, namely Tezao 213, Zhuyeqi, Zhongcha 302, Tieguanyin, Yinghong 9 and Fenghuang Dancong were determined and compared. The results indicated significant differences between spring and summer tea in all quality components except tea polyphenols. The average phenol/ammonia ratio of spring and summer tea was 4.60 and 7.99, respectively, indicating summer tea to be suitable for processing into green and black tea. Altogether 18 amino acids were detected in the six cultivars, and the contents of different groups of amino acid were in the decreasing order of umami amino acids > sweet amino acids > bitter amino acids > aromatic amino acids. The contents of umami and sweet amino acids in spring tea were significantly higher than those in summer tea. Correlation analysis revealed that the synthesis of various metabolites were highly correlated. Principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) showed that season had a more significant effect on tea metabolites than variety. A total of 16 compounds were identified as differential metabolites between spring and summer tea, including 7 amino acids, 8 catechins and CAF, which could be used as potential markers to distinguish spring and summer tea. This study provides a valuable reference for tea processing.

[Objective] The study aims to explore the effects of different foliar fertilizers on the photosyn thetic characteristics, fruit quality, and yield of greenhouse strawberries, and to screen suitable foliar fer tilizers. [Methods] The strawberry variety “Monterey” planted in the greenhouse of Moujiatai Strawberry in Xigu District of Lanzhou City, Gansu Province was used as test materials. Under sufficient fertilization, five types of foliar fertilizers, including classic 750×liquid (T1), Huapu 1 500×liquid (T2), and Runliang 750×liquid (T3), 500×Adela fruit (T4), and 1500×potassium dihydrogen phosphate (T5), were ap plied topdressing at four growth stages, with water as control. The photosynthetic parameters, fruit qual ity, biomass, and yield were measured. [Results] (1) After spraying different foliar fertilizers, the bio mass and chlorophyll content of strawberry plants were significantly increased, and the net photosynthetic rate and transpiration rate of leaves were significantly increased, while the intercellular carbon dioxide con centration was higher than that of CK. (2) For fruit quality, T2 and T3 treatments significantly improved the strawberry fruit shape index, while T1 and T5 treatments significantly increased fruit volume and weight. Among them, the average single fruit mass of T1 was increased by 35.04% compared with CK, while the transverse and longitudinal diameters of T5 fruits were increased by 49.39% and 40.64%, re spectively. (3) The quality of strawberry fruits was effectively improved by various foliar fertilizer treat ments, and T1 treatment was the best. The content of vitamin C, soluble solids, soluble proteins, reduc ing sugars, and soluble sugars were increased by 30.84%, 33.5%, 38.5%, 27.9%, and 36.2%, respec tively, compared with CK. The content of titratable acid was the lowest, and the sugar acid ratio was the highest, with significant difference compared with CK. (4) Spraying appropriate fertilizer significantly in creased plant biomass, max single fruit weight, and fruit yield; treatment T1 had the highest yield, an in crease of 13.65% compared with CK. [Conclusion] The application of appropriate foliar fertilizer effective ly improves the nutritional growth status of strawberries, enhances the quality of strawberry fruits, and significantly increases yield. The comprehensive effect is best when applying classic 750×liquid.

Objective To investigate the rehabilitation effects of vestibular rehabilitation in acute brainstem infarction patients with subjective visual vertical (SVV) tilting, and the correlation between the SVV, balance confidence, dizziness/vertigo and walking function. Methods All 45 acute brainstem infarction patients with SVV tilting who were hospitalized in Tianjin Huanhu Hospital from July 2022 to July 2023 were collected. Patients were divided into vestibular rehabilitation group (n = 25) and general rehabilitation group (n = 20), Bucket Test was used to test SVV tilting angle, Activities-Specific Balance Confidence Scale (ABC) was used to evaluate balance confidence, Visual Analog Scales (VAS) was used to evaluate dizziness/vertigo and Functional Ambulation Category Scale (FAC) was used to evaluate walking function before and after 2 weeks of treatment. Results The SVV tilting angle (F = 4.356, P = 0.043), ABC score (F = 4.389, P = 0.042), dizziness/vertigo VAS score (F = 4.138, P = 0.048) were significantly different between vestibular rehabilitation group and general rehabilitation group. After treatment, the SVV tilting angle (t = - 2.139, P = 0.038) and the dizziness/vertigo VAS score (t = - 2.952, P = 0.005) in vestibular rehabilitation group were lower than those in general rehabilitation group, and the ABC score was higher than that in general rehabilitation group (t = 2.920, P = 0.006). SVV tilting angle (F = 196.923, P = 0.000), ABC score (F = 89.050, P = 0.000), dizziness/vertigo VAS score (F = 81.803, P = 0.000), FAC grade (F = 72.866, P = 0.000) were statistically significant. The SVV tilting angle (t = 0.763, P = 0.000; t = 0.972, P = 0.000) and dizziness/vertigo VAS score (t = 8.815, P = 0.000; t = 5.107, P = 0.000) after treatment were lower than those before treatment of 2 groups, and the ABC score (t = 0.689, P = 0.001; t = 0.703, P = 0.001) and FAC grade (t = - 6.721, P = 0.000; t = - 5.772, P = 0.000) were higher than those before treatment of 2 groups. Correlation analysis showed a positive correlation between SVV tilting angle and dizziness/vertigo VAS score in acute brainstem infarction patients with SVV tilting (r = 0.627, P = 0.000). Conclusions Vestibular rehabilitation can effectively improve the SVV tilting, balance confidence and dizziness/vertigo symptoms of acute brainstem infarction patients with SVV tilting, and the SVV tilting angle is closely related to the degree of dizziness/vertigo.

Bud-notching in pear varieties with weak-branches enhances branch development, hormone distribution, and germination, promoting healthier growth and improving early yield. To examine the regulatory mechanisms of endogenous hormones on lateral bud germination in Pyrus spp. (cv. 'Huangguan') (Pyrus bretschneideri Rehd.), juvenile buds were collected from 2-year-old pear trees. Then, a comprehensive study, including assessments of endogenous hormones, germination and branching rates, RNA-seq analysis, and gene function analysis in these lateral buds was conducted. The results showed that there was no significant difference in germination rate between the control and bud-notching pear trees, but the long branch rate was significantly increased in bud-notching pear trees compared to the control (p < 0.05). After bud-notching, there was a remarkable increase in IAA and BR levels in the pruned section of shoots, specifically by 141% and 93%, respectively. However, the content of ABA in the lateral buds after bud-notching was not significantly different from the control. Based on RNA-seq analysis, a notable proportion of the differentially expressed genes (DEGs) were linked to the plant hormone signal transduction pathway. Notably, the brassinosteroid signaling pathway seemed to have the closest connection with the branching ability of pear with the related genes encoding BRI1 and CYCD3, which showed significant differences between lateral buds. Finally, the heterologous expression of PyCYCD3 has a positive regulatory effect on the increased Arabidopsis growth and branching numbers. Therefore, the PyCYCD3 was identified as an up-regulated gene that is induced via brassinosteroid (BR) and could act as a conduit, transforming bud-notching cues into proliferative signals, thereby governing lateral branching mechanisms in pear trees. [ABSTRACT FROM AUTHOR]

In order to solve the problem of water waste and environmental pollution in arid and semiarid areas, this study aimed to investigate the effects of different water and nitrogen treatments on the growth of apple leaves, photosynthetic physiology, nutrient uptake and nitrogen use efficiency. A two-factor experimental design was used in this experiment. The first factor was three levels of irrigation (40–50%, 50–65%, 65–80% RWC, relative water content) and the second factor was four levels of nitrogen application (0, 150, 300, 600 kg/ha), which were a completely randomized combination. The results showed that irrigation and nitrogen application had significant effects on the growth, photosynthetic rate, chlorophyl and mineral element contents of apple tree leaves, but excessive input of water and nitrogen would restrain the growth of apple tree leaves. The Ndff% of different organs for apple trees were evaluated by 15N isotope tracer technology, and the highest Ndff% value was found in leaves, ranging from 1.01–1.23‰. Next were roots with 0.29–0.43‰, and the lowest Ndff% was found in fruits with 0.03–0.08‰. The calculation results of 15N utilization in apple trees showed that nitrogen use efficiency (NUE) under the medium-water low nitrogen (W1N1) treatment had the highest value with 43.33%. In conclusion, considering the growth physiology and nitrogen utilization of apple trees, it is recommended that a water and fertilizer coupling combination scheme for apple trees in arid and semi-arid areas of the Loess Plateau should be that of a medium-water and medium-nitrogen mode (W1N2, 50–65% RWC, 300 kg/ha). [ABSTRACT FROM AUTHOR]

SNARE proteins (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) play a key role in mediating a variety of plant biological processes. Currently, the function of the SNARE gene family in phytohormonal and abiotic stress treatments in grapevine is currently unknown, making it worthwhile to characterize and analyze the function and expression of this family in grapevine. In the present study, 52 VvSNARE genes were identified and predominantly distributed on 18 chromosomes. Secondary structures showed that the VvSNARE genes family irregular random coils and α-helices. The promoter regions of the VvSNARE genes were enriched for light-, abiotic-stress-, and hormone-responsive elements. Intraspecific collinearity analysis identified 10 pairs collinear genes within the VvSNARE family and unveiled a greater number of collinear genes between grapevine and apple, as well as Arabidopsis thaliana, but less associations with Oryza sativa. Quantitative real-time PCR (qRT-PCR) analyses showed that the VvSNARE genes have response to treatments with ABA, NaCl, PEG, and 4 °C. Notably, VvSNARE2, VvSNARE14, VvSNARE15, and VvSNARE17 showed up-regulation in response to ABA treatment. VvSNARE2, VvSNARE15, VvSNARE18, VvSNARE19, VvSNARE20, VvSNARE24, VvSNARE25, and VvSNARE29 exhibited significant up-regulation when exposed to NaCl treatment. The PEG treatment led to significant down-regulation of VvSNARE1, VvSNARE8, VvSNARE23, VvSNARE25, VvSNARE26, VvSNARE31, and VvSNARE49 gene expression. The expression levels of VvSNARE37, VvSNARE44, and VvSNARE46 were significantly enhanced after exposure to 4 °C treatment. Furthermore, subcellular localization assays certified that VvSNARE37, VvSNARE44, and VvSNARE46 were specifically localized at the cell membrane. Overall, this study showed the critical role of the VvSNARE genes family in the abiotic stress response of grapevines, thereby providing novel candidate genes such as VvSNARE37, VvSNARE44, and VvSNARE46 for further exploration in grapevine stress tolerance research. [ABSTRACT FROM AUTHOR]

In this study, we obtained and cloned VvSnRK2.7 by screening transcriptomic data to investigate the function of the grape sucrose non-fermenting kinase 2 (SnRK2) gene under stress conditions. A yeast two-hybrid (Y2H) assay was used to further screen for interaction proteins of VvSnRK2.7. Ultimately, VvSnRK2.7 was heterologously expressed in Arabidopsis thaliana, and the relative conductivity, MDA content, antioxidant enzyme activity, and sugar content of the transgenic plants were determined under drought treatment. In addition, the expression levels of VvSnRK2.7 in Arabidopsis were analyzed. The results showed that the VvSnRK2.7-EGFP fusion protein was mainly located in the cell membrane and nucleus of tobacco leaves. In addition, the VvSnRK2.7 protein had an interactive relationship with the VvbZIP protein during the Y2H assay. The expression levels of VvSnRK2.7 and the antioxidant enzyme activities and sugar contents of the transgenic lines were higher than those of the wild type under drought treatment. Moreover, the relative conductivity and MDA content were lower than those of the wild type. The results indicate that VvSnRK2.7 may activate the enzyme activity of the antioxidant enzyme system, maintain normal cellular physiological metabolism, stabilize the berry sugar metabolism pathway under drought stress, and promote sugar accumulation to improve plant resistance. [ABSTRACT FROM AUTHOR]

Cold stress adversely impacts grape growth, development, and yield. Therefore, improving the cold tolerance of grape is an urgent task of grape breeding. The Jasmonic acid (JA) pathway responsive gene JAZ plays a key role in plant response to cold stress. However, the role of JAZ in response to low temperatures in grape is unclear. In this study, VvJAZ13 was cloned from the 'Pinot Noir' (Vitis vinefera cv. 'Pinot Noir') grape, and the potential interacting protein of VvJAZ13 was screened by yeast two-hybrid (Y2H). The function of VvJAZ13 under low temperature stress was verified by genetic transformation. Subcellular localization showed that the gene was mainly expressed in cytoplasm and the nucleus. Y2H indicated that VvF-box, VvTIFY5A, VvTIFY9, Vvbch1, and VvAGD13 may be potential interacting proteins of VvJAZ13. The results of transient transformation of grape leaves showed that VvJAZ13 improved photosynthetic capacity and reduced cell damage by increasing maximum photosynthetic efficiency of photosystem II (Fv/Fm), reducing relative electrolyte leakage (REL) and malondialdehyde (MDA), and increasing proline content in overexpressed lines (OEs), which played an active role in cold resistance. Through the overexpression of VvJAZ13 in Arabidopsis thaliana and grape calli, the results showed that compared with wild type (WT), transgenic lines had higher antioxidant enzyme activity and proline content, lower REL, MDA, and hydrogen peroxide (H2O2) content, and an improved ability of scavenging reactive oxygen species. In addition, the expression levels of CBF1-2 and ICE1 genes related to cold response were up-regulated in transgenic lines. To sum up, VvJAZ13 is actively involved in the cold tolerance of Arabidopsis and grape, and has the potential to be a candidate gene for improving plant cold tolerance. [ABSTRACT FROM AUTHOR]

Mucoepidermoid carcinoma is a rare entity of lung malignancy that is subclassified into high-grade or low-grade types according to its histological features. High-grade mucoepidermoid carcinoma is a more aggressive form of malignancy, with a tendency towards lymph node involvement and distant metastasis. Cancer-related microangiopathic hemolytic anemia as a less common situation of paraneoplastic syndrome may be encountered with metastatic malignancy, but has not been reported previously in mucoepidermoid carcinoma of the lung. Herein, we report a 78-year-old male patient who presented with hemoptysis for one day. Laboratory tests showed microangiopathic hemolytic anemia and thrombocytopenia. A chest X-ray demonstrated consolidation in the left lung field. Chest computed tomography revealed a mass in the left upper lobe, and a subsequent bronchoscopic biopsy was performed. The histopathological results indicated a high-grade mucoepidermoid carcinoma. Magnetic resonance imaging of the brain demonstrated leptomeningeal carcinomatosis. The patient refused systemic chemotherapy, and palliative radiation therapy only was conducted for local disease control. The patient has performed well for 12 months to date since diagnosis of the tumor.

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