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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.

Heat shock protein 70 (HSP70) is one kind of molecular chaperones which are widely found in organisms, and its members are highly conserved among each other, with important roles in plant growth and development. In this study, 56 HSP70 genes were identified from the apple genome database. Analysis of gene duplication events showed that tandem and segmental duplication events play an important role in promoting the amplification of the MdHSP70 gene family. Collinearity analysis showed that HSP70 family members of apple were more closely related to HSP70 family members of Arabidopsis, tomato and soybean. The promoter region of the apple HSP70 genes contains a large number of cis-acting elements in response to hormones and stress. Tissue-specific expression analysis showed that some of the genes were associated with various stages of the apple growth process. Codon preference analysis showed small differences between codon bases 1 and 3 in the apple HSP70 genome, and the codon base composition had a small effect on codon usage preference. The multiple expression patterns of the MdHSP70 gene suggested that MdHSP70 gene members play important roles in growth and development and in response to hormonal and abiotic stresses. The yeast two-hybrid (Y2H) demonstrated that MdHSP70-53 interacts with MdDVH24_032563. The qRT-PCR analysis showed that most MdHSP70 members' hormonal and abiotic stresses (MdHSP70-6, MdHSP70-26 and MdHSP70-45) appeared to be highly expressed. To further elucidate the function of MdHSP70 (6, 26, 45), we introduced them into tobacco to confirm subcellular locations and noted that these genes are located in the cytoplasm and cell membrane. This study serves as a theoretical basis for further studies of the MdHSP70 gene and helps to further investigate the functional characterization of MdHSP70 gene. [ABSTRACT FROM AUTHOR]

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]

Background: The lignification of branches can promote the accumulation of nutrients, increase plant survival and resistance to biotic and abiotic stresses. As an important carbon source for plants, glucose is also the carbon skeleton for lignin synthesis. Grapevine is a perennial cash crop, and highly lignified branches are essential to ensure the growth of the grapevine plant and the development of the fruit. Methods: Here, 'Red Globe' grape (Vitis vinifera L.) plantlets were selected as the material and cultured with different concentrations of glucose: 0 g/L, 20 g/L, 40 g/L and 60 g/L glucose (G0, G20, G40 and G60). Among them, G0 group as control. Lignin, anthocyanins and glucose contents, plant height and microstructure were measured at each glucose concentration after 40-, 50-, and 60-day treatments. Transcriptome and metabolome were used to analyze the difference in genes and metabolites after 50 days of growth. Results: After 50 days of cultivation, the lignin content in G40 group was the highest. And the xylem cells number also increased. To further, transcriptome and metabolome have identified a total of 3638 differentially expressed genes (DEGs) (including 245 TFs) and 510 differently accumulated metabolites (DAMs) in three comparison groups. In-depth joint analysis revealed that phenylpropanoid biosynthesis pathway significantly respond to exogenous glucose, and 37 DEGs were identified. Therefore, the phenylpropane biosynthesis pathway may be the key to exogenous glucose to increase lignin levels in grapes, with differential expression of genes being a top priority. These findings provide a new perspective for understanding the relationship between glucose and lignin in grape. [ABSTRACT FROM AUTHOR]

Background: Grape is a plant that is sensitive to low temperature and vulnerable to low-temperature damage. However, little is known about the roles of lncRNAs, miRNAs and mRNAs in regulating the hypothermia response mechanism in Vitis amurensis Rupr. Methods: In this study, the expression and regulatory network of low-temperature response genes were studied in the phloem of grape under different low-temperature stress. Results: Here, we performed analyses related to RNA-seq and miRNA-seq on grape phloem tissues from five periods of cold resistance campaigns. Three RNAs (lncRNAs, miRNAs and mRNAs) obtained by KEGG and GO analyses were used to identify starch and sucrose metabolism associated with cold resistance, and specific changes in BP, CC, and MF were identified in four comparisons. Venn diagrams, thermograms and pathway maps were used to analyze the differentially expressed genes (DEGs), and their specific gene expression during the cold exercise were obtained. The six DEGs finally selected were used for qRT-PCR to verify the RNA-seq data. In addition, we found that the regulatory networks of miRNAs and lncRNAs correspond to the six DEGs. This study will contribute to further experimental studies to elucidate the cold resistance mechanism of Vitis amurensis Rupr. Conclusions: The low-temperature response genes of grape are mainly enriched in the starch and sucrose metabolism, and they are regulated by miRNAs and lncRNAs. The conclusions will provide basic information for further understanding of the cold resistance mechanism of grape in the future. [ABSTRACT FROM AUTHOR]