VvWRKY13 enhances ABA biosynthesis in Vitis vinifera

Abscisic acid (ABA) plays critical roles in plant growth and development as well as in plants’ responses to abiotic stresses. We previously isolated VvWRKY13, a novel transcription factor, from Vitis vinifera (grapevine), and here we present evidence that VvWRKY13 may regulate ABA biosynthesis in plants. When VvWRKY13 was ectopically expressed in Arabidopsis, the transgenic lines showed delayed seed germination, smaller stomatal aperture size, and several other phenotypic changes, indicating elevated ABA levels in these plants. Sequence analysis of several genes that are involved in grapevine ABA synthetic pathway identified WRKY-specific binding elements (W-box or W-like box) in the promoter regions. Indeed, transient overexpression of VvWRKY13 in grapevine leaves significantly increased the transcript levels of ABA synthetic pathway genes. Taken together, we conclude that VvWRKY13 may promote ABA production by activating genes in the ABA synthetic pathway

VvBAP1 Is Involved in Cold Tolerance in Vitis vinifera L.

The majority of commercial grape cultivars originate from the European grape. While these cultivars have excellent organoleptic qualities, they suffer from a relatively poor tolerance to the cold experienced during winter, resulting in significant damage to grapevines. Thus, low temperature is one of the bottlenecks that restrict the further growth of the grape industry. Research on the mechanism of cold tolerance in grapes is therefore very important. BON association protein 1 (BAP1) is a recently discovered phospholipid-binding protein. In Arabidopsis, the expression of AtBAP1 can be regulated via low temperature; however, the function of BAP1 in the grapevine has not been reported. The VvBAP1 gene was cloned in our previous studies in grapes, and bioinformatics analysis showed that it harbors the conservative calcium-dependent C2 protein domain. However, little is known about its function and underlying mechanism. In this study, cold treatment was applied to the cold-resistant grape varieties ‘F-242’ and ‘Zuoyouhong’ as well as to the cold-sensitive grape varieties ‘Cabernet Sauvignon’ and ‘Chardonnay.’ The expression level of VvBAP1 in the cold-resistant varieties was significantly higher than in the cold-sensitive varieties, indicating that VvBAP1 could be associated with the cold response processes in the grapevine. Using the cold-resistant grape variety ‘F-242’ as material, with the 4°C and CaCl2 treatment, the relative expression of VvBAP1 was determined via qRT-PCR. Both low temperature and low-temperature signal Ca2+ induced VvBAP1 expression. In addition, the VvBAP1 gene was cloned and transferred into Arabidopsis to generate VvBAP1 overexpressing plants. Biochemical assays and gene expression analyses were conducted on plants subjected to low temperature treatments (4 and -8°C). The obtained results showed that the activities of superoxide dismutase and peroxidase in these transgenic plants were higher than those in wild type (WT) plants, and that cell membrane permeability and malondialdehyde content were both lower compared to WT plants. Furthermore, the content of soluble sugars and the expression levels of sugar-metabolizing related genes, such as BAM4-7, SS4, and G6PD5, were significantly higher than those of WT plants. Furthermore, the expression of low temperature response signal genes, including CBF1, CBF3, COR15a, COR6.6, COR27, and KIN1, were also enhanced. In summary, these results showed that VvBAP1 could strengthen the cold resistance in the grapevine through adjusting and controlling the sugar content and activating antioxidant enzyme activity

Transcriptomic Analysis Elaborates the Resistance Mechanism of Grapevine Rootstocks against Salt Stress

Grapes are subject to a wide range of climatic conditions during their life cycle, but the use of rootstocks can effectively ameliorate the effects of abiotic stress. However, the tolerance mechanism of different grape rootstock varieties varies under various stresses, and systematic research on this aspect is limited. On the basis of previous research, transcriptome sequencing was performed on three tolerant grape rootstock varieties (3309C, 520A, 1103P) and three intolerant grape rootstock varieties (5BB, 101–14, Beta). In total, 56,478,468 clean reads were obtained. One hundred and ten genes only existed in all combinations during P1 with a downregulated trend, and 178 genes existed only in P1 of tolerant grape rootstock varieties. Salt treatment firstly affected the photosynthesis of leaves, and tolerant varieties weakened or even eliminated this effect through their own mechanisms in the later stage. Tolerant varieties mobilized a large number of MFs during the P2 stage, such as hydrolase activity, carboxypeptidase activity, and dioxygenase activity. Carbon metabolism was significantly enriched in P1, while circadian rhythm and flavonoid biosynthesis were only enriched in tolerant varieties. In the intolerant varieties, photosynthesis-related pathways were always the most significantly enriched. There were large differences in the gene expression of the main signal pathways related to salt stress in different varieties. Salt stress affected the expression of genes related to plant abiotic stress, biotic stress, transcription factors, hormones, and secondary metabolism. Tolerant varieties mobilized more bHLH, WRKY, and MYB transcription factors to respond to salt stress than intolerant varieties. In the tolerant rootstocks, SOS was co-expressed. Among these, SOS1 and SOS2 were upregulated, and the SOS3 and SOS5 components were downregulated. The genes of heat shock proteins and the phenylalanine pathway were upregulated in the tolerant varieties. These findings outline a tolerance mechanism model for rootstocks for coping with osmotic stress, providing important information for improving the resistance of grapes under global climate change

Effect of BMI on cumulative live birth rates in patients that completed IVF treatment: a retrospective cohort study of 16,126 patients

Although several studies have reported that high maternal BMI could influence the cumulative live birth rate (CLBR) in fresh embryo transfer cycles, the association of BMI with CLBR remains unclear in patients that completed IVF treatment. In this study, we examined the association of maternal BMI with CLBR, including repetitive one oocyte pick-up (OPU) and all fresh and frozen embryo transfer until live birth or embryos were run out. A total of 16,126 patients’ data were included in the analysis and were divided into four groups based on BMI. We found that patients’ characteristics, embryo parameters, and pregnancy outcomes differed among different BMI groups. Multivariate logistic regression showed that being underweight was associated with a higher possibility of having live birth than the reference group (OR (95% CI) 1.40 (1.22–1.59), P < 0.001), whereas being overweight and obese were associated with a lower possibility of having live birth than the reference group ((OR (95% CI) 0.81 (0.74–0.90), P < 0.001) and (OR (95% CI) 0.68 (0.55–0.85), P < 0.001)). After adjustment for confounding factors, the reference group was associated with a higher possibility of having live birth, with a significant difference found between the obese and reference groups (OR (95% CI) 0.55 (0.43–0.70), P < 0.001). An association was found between CLBR and BMI, indicating that an increase in BMI results in a decline in CLBR. Moreover, the CLBR of patients with different characteristics differed in the various BMI groups. Taken together, our data show that maternal BMI has a significant impact on CLBR

A Single Aptamer-Dependent Sandwich-Type Biosensor for the Colorimetric Detection of Cancer Cells via Direct Coordinately Binding of Bare Bimetallic Metal&ndash;Organic Framework-Based Nanozymes

A typical colorimetric sandwich-type sensor relies on dual antibodies/aptamers to specifically visualize the targets. The requirement of dual antibodies/aptamers and low signal intensity inevitably increases the design difficulty and compromises the sensing sensitivity. In this work, a novel sandwich-type aptasensor was developed using single aptamer-functionalized magnetic nanoparticles as a specific recognition unit to target cancer cells and a bimetallic metal&ndash;organic frameworks (MOFs)-based nanozymes as a colorimetric signal amplification unit. The well-defined crystalline structure of UIO-66 MOFs enabled the introduction of Fe/Zr bimetal nodes, which possessed integrated properties of the peroxidase-like nanozyme activity and direct coordinately binding to the cell surface. Such a novel construction strategy of sandwich-type aptasensors achieved simple, sensitive, and specific detection of the target cancer cells, which will inspire the development of biosensors

Magnetic Bead-Sensing-Platform-Based Chemiluminescence Resonance Energy Transfer and Its Immunoassay Application

A competitive immunoassay based on chemiluminescence resonance energy transfer (CRET) on the magnetic beads (MBs) is developed for the detection of human immunoglobulin G (IgG). In this protocol, carboxyl-modified MBs were conjugated with horseradish peroxidase (HRP)-labeled goat antihuman IgG (HRP-anti-IgG) and incubated with a limited amount of fluorescein isothiocyanate (FITC)-labeled human IgG to immobilize the antibody–antigen immune complex on the surface of the MBs, which was further incubated with the target analyte (human IgG) for competitive immunoreaction and separated magnetically to remove the supernatant. The chemiluminescence (CL) buffer (containing luminol and H₂O₂) was then added, and the CRET from donor luminol to acceptor FITC in the immunocomplex on the surface of MBs occured immediately. The present protocol was evaluated for the competitive immunoassay of human IgG, and a linear relationship between CL intensity ratio (<i><i>R</i> = I</i>₄₂₅/<i>I</i>₅₂₅) and human IgG concentration in the range of 0.2–4.0 nM was obtained with a correlation coefficient of 0.9965. The regression equation was expressed as <i>R</i> = 1.9871<i>C</i> + 2.4616, and a detection limit of 2.9 × 10^–11 M was obtained. The present method was successfully applied for the detection of IgG in human serum. The results indicate that the present protocol is quite promising for the application of CRET in immunoassays. It could also be developed for detection of other antigen–antibody immune complexes by using the corresponding antigens and respective antibodies

Image_1_VvBAP1 Is Involved in Cold Tolerance in Vitis vinifera L..pdf

<p>The majority of commercial grape cultivars originate from the European grape. While these cultivars have excellent organoleptic qualities, they suffer from a relatively poor tolerance to the cold experienced during winter, resulting in significant damage to grapevines. Thus, low temperature is one of the bottlenecks that restrict the further growth of the grape industry. Research on the mechanism of cold tolerance in grapes is therefore very important. BON association protein 1 (BAP1) is a recently discovered phospholipid-binding protein. In Arabidopsis, the expression of AtBAP1 can be regulated via low temperature; however, the function of BAP1 in the grapevine has not been reported. The VvBAP1 gene was cloned in our previous studies in grapes, and bioinformatics analysis showed that it harbors the conservative calcium-dependent C2 protein domain. However, little is known about its function and underlying mechanism. In this study, cold treatment was applied to the cold-resistant grape varieties ‘F-242’ and ‘Zuoyouhong’ as well as to the cold-sensitive grape varieties ‘Cabernet Sauvignon’ and ‘Chardonnay.’ The expression level of VvBAP1 in the cold-resistant varieties was significantly higher than in the cold-sensitive varieties, indicating that VvBAP1 could be associated with the cold response processes in the grapevine. Using the cold-resistant grape variety ‘F-242’ as material, with the 4°C and CaCl₂ treatment, the relative expression of VvBAP1 was determined via qRT-PCR. Both low temperature and low-temperature signal Ca²⁺ induced VvBAP1 expression. In addition, the VvBAP1 gene was cloned and transferred into Arabidopsis to generate VvBAP1 overexpressing plants. Biochemical assays and gene expression analyses were conducted on plants subjected to low temperature treatments (4 and -8°C). The obtained results showed that the activities of superoxide dismutase and peroxidase in these transgenic plants were higher than those in wild type (WT) plants, and that cell membrane permeability and malondialdehyde content were both lower compared to WT plants. Furthermore, the content of soluble sugars and the expression levels of sugar-metabolizing related genes, such as BAM4-7, SS4, and G6PD5, were significantly higher than those of WT plants. Furthermore, the expression of low temperature response signal genes, including CBF1, CBF3, COR15a, COR6.6, COR27, and KIN1, were also enhanced. In summary, these results showed that VvBAP1 could strengthen the cold resistance in the grapevine through adjusting and controlling the sugar content and activating antioxidant enzyme activity.</p