基于GRAPES全球分析系统的Hybrid-3DVAR混合同化研究

本文基于我国自主研发的GRAPES全球3DVAR同化系统,利用NCEP全球集合预报产品和time-lagged方法,针对膨胀系数、集合样本数和集合权重系数,开展了每日4次、连续一周的GRAPES全球Hybrid-3DVAR混合同化研究。结果表明:所有试验中,集合样本取60个、集合权重取0.5时,得到的混合同化分析和预报误差最小;在该混合同化系统中,在高层也考虑静态背景误差协方差和集合背景误差协方差的耦合,可避免混合同化方案分析场误差在150 hPa及以上过大,并超过3DVAR分析场误差的情况。

The Use of Secondary Grape Biomass in Beef Cattle Nutrition on Carcass Characteristics, Quality and Shelf Life of Meat

We determined whether the inclusion of 100 g/kg dry matter of grape pomace silage (GPS) and grape pomace bran (GPB) as substitutes for other traditional fiber sources in the diet of steers (Charolais x Nellore) would improve carcass characteristics, meat quality and composition, and shelf life. Twenty-four animals (248 ± 19.32 kg of initial body weight) were fed a high concentrate diet for 121 days. Carcass characteristics were measured, and the longissimus dorsi muscle was analyzed for fatty acid (FA) profile and composition. The meat was sliced and stored in air-permeable packages for 10 days. On each sampling day (d 1, 3, 7, and 10), oxidative stability, bacterial load, lipid and protein oxidation, and staining were analyzed. The experimental diets influenced the pH of cold carcasses only. The GPS group had a higher pH than the control. The GPS and GPB groups showed improved oxidant status (i.e., lower lipid peroxidation and concentrations of reactive oxygen species were in the meat of both groups than in control). On the first day of storage, the antioxidant enzyme glutathione S-transferase activity was more significant in the meat of the GPS and GPB groups than in the control. The bacterial loads in the meat were attenuated by GPS inclusion;there were lower total coliform counts and a trend toward lower counts for enterobacteria in the control group. The diets altered the FA profile of the meat;i.e., the GPB diet allowed for a more significant amount of the n-6 omegas in the meat, while the GPS diet showed a tendency for a more significant amount of n-6 and 9 omegas. Both diets (GPS and GPB) increased the amounts of long-chain FAs. The GPS diet decreased saturated FA levels. We conclude that the dietary treatments GPS and GPB are a promising alternative to maintain meat quality standards throughout in real-world retail conditions. These treatments gave rise to an improvement in the nutritional value of the meat due to the more significant amounts of FAs that improve human health.

Identification,characterization,and verification of miR399 target gene in grape

The microRNA miR399 plays an important role in phosphorus signal transduction pathways in plants.Previously,miR399 was shown to be closely associated with berry ripening in grape(Vitis vinifera).The objective of the present study was to elucidate the evolutionary characteristics of the miR399 gene family in grape and to verify the cleavage effect on the target genes.Grape miR399s were identified by miRNA sequencing and retrieval from the miRBase database.The mature sequences and precursor sequences were subjected to phylogenetic analysis to reconstruct evolutionary trees,as well as secondary structure analysis of the precursor sequence,and prediction of target genes.The cisacting elements in the miR399 promoter were predicted and the cleavage effect of grape miR399b on its target genes was verified.The grape miR399 family comprised nine precursor sequences and nine mature sequences.The precursor sequences formed a typical and stable stem—loop structure.The minimum folding free energy ranged from-55.70 kcal·mol^(-1)to-37.40 kcal·mol^(-1).Multiple sequence alignment revealed that the miR399 family was highly conserved.The grape miR399 family was phylogenetically closely related to peach,apple,and citrus miR399s.Grape miR399s were predicted to target inorganic phosphate transporter 1—3,phospholipase D delta-like,and beta-glucuronosyltransferase.The cleavage effect of grape miR399b on the target genes was verified by means of a dual-luciferase assay and 5’RLM-RACE.Histochemical GUS staining showed that the promoter activity of miR399b was promoted by GA3treatment.

Alternative splicing of the PECTINESTERASE gene encoding a cell wall-degrading enzyme affects postharvest softening in grape

The firmness of table grape berries is a crucial quality parameter. Despite extensive research on postharvest fruit softening, its precise molecular mechanisms remain elusive. To enhance our comprehension of the underlying molecular factors, we initially identified differentially expressed genes(DEGs) by comparing the transcriptomes of folic acid(FA)-treated and water-treated(CK) berries at different time points. We then analyzed the sequences to detect alternatively spliced(AS) genes associated with postharvest softening. A total of 2,559 DEGs were identified and categorized into four subclusters based on their expression patterns, with subcluster-4 genes exhibiting higher expression in the CK group compared with the FA treatment group. There were 1,045 AS-associated genes specific to FA-treated berries and 1,042 in the CK-treated berries, respectively. Gene Ontology(GO) annotation indicated that the AS-associated genes in CK-treated berries were predominantly enriched in cell wall metabolic processes,particularly cell wall degradation processes. Through a comparison between treatment-associated AS genes and subcluster-4 DEGs, we identified eight genes, including Pectinesterase 2(VvPE2, Vitvi15g00704), which encodes a cell wall-degrading enzyme and was predicted to undergo an A3SS event. The reverse transcription polymerase chain reaction further confirmed the presence of a truncated transcript variant of VvPE2 in the FA-treated berries.Our study provides a comprehensive analysis of AS events in postharvest grape berries using transcriptome sequencing and underscores the pivotal role of VvPE2 during the postharvest storage of grape berries.

Genome-Wide Exploration of the Grape GLR Gene Family and Differential Responses of VvGLR3.1 and VvGLR3.2 to Low Temperature and Salt Stress

Grapes,one of the oldest tree species globally,are rich in vitamins.However,environmental conditions such as low temperature and soil salinization significantly affect grape yield and quality.The glutamate receptor(GLR)family,comprising highly conserved ligand-gated ion channels,regulates plant growth and development in response to stress.In this study,11 members of the VvGLR gene family in grapes were identified using whole-genome sequence analysis.Bioinformatic methods were employed to analyze the basic physical and chemical properties,phylogenetic trees,conserved domains,motifs,expression patterns,and evolutionary relationships.Phylogenetic and collinear analyses revealed that the VvGLRs were divided into three subgroups,showing the high conservation of the grape GLR family.These members exhibited 2 glutamate receptor binding regions(GABAb and GluR)and 3-4 transmembrane regions(M1,M2,M3,and M4).Real-time quantitative PCR analysis demonstrated the sensitivity of all VvGLRs to low temperature and salt stress.Subsequent localization studies in Nicotiana tabacum verified that VvGLR3.1 and VvGLR3.2 proteins were located on the cell membrane and cell nucleus.Additionally,yeast transformation experiments confirmed the functionality of VvGLR3.1 and VvGLR3.2 in response to low temperature and salt stress.Thesefindings highlight the significant role of the GLR family,a highly conserved group of ion channels,in enhancing grape stress resistance.This study offers new insights into the grape GLR gene family,providing fundamental knowledge for further functional analysis and breeding of stress-resistant grapevines.

基于Wink软件的微视频教学资源制作方法研究

在数字化教育的快速发展中,作为一种新兴的教学方式,微视频教学资源凭借其简洁、明了、易于消化的特点,受到广泛的关注和应用。作为一款功能全面的屏幕录制和视频编辑工具,Wink软件为教育工作者提供制作高质量教学视频的便利。文章首先介绍Wink软件的基本功能和优点,然后,详细描述使用Wink软件进行微视频教学资源制作的全过程,包括前期的教学内容规划、视频录制和后期编辑,最后,探讨Wink软件在未来教学资源制作中的潜在改进空间和应用方向。

Adsorption Effect of Phosphate Modified Grape Branch Biochar on Cd2

Two major problems facing agriculture at present are soil pollution and the disposal of solid wastes generated during plant growth. The method of preparing biochar from solid wastes produced by plants is a means of maximizing the use of resources to combat the problem of soil pollution. In this study, we did not choose straw in the traditional sense but the waste branches from grape pruning, which has higher lignin cellulose, as the raw material. The biochar derived from grape branches pyrolyzed at 300˚C for two hours was utilized as a raw material to prepare modified biochar with varying concentrations of phosphoric acid. The adsorption performance and mechanism of Cd2 were explored through experiments involving different concentrations, addition amounts, reaction times, kinetic analyses, and isothermal adsorption tests. The findings indicated that the optimal adsorption of Cd2 occurred with a 20% phosphoric acid concentration, achieving the highest adsorption rate of 84.62%. At a dosage of 10 g/L, the maximum adsorption capacity reached 7.02 mg/g. The adsorption kinetics and isothermal adsorption of Cd2 on biochar modified with 0.2% phosphoric acid (0.2 PB) closely followed the pseudo-first-order kinetics model (R2 > 0.98) and the Freundlich model (R2 > 0.97), respectively. This suggests that the adsorption process involves both physical and chemical mechanisms. SEM and FTIR analyses revealed that phosphoric acid modification primarily increased the biochar’s specific surface area and enhanced certain original functional groups. The adsorption process predominantly involved rapid ion diffusion and chemical adsorption, as confirmed by kinetic analysis and isothermal adsorption model analysis. In summary, the adsorption efficiency of 0.2 PB significantly improved, showing potential and feasibility for heavy metal remediation in soil. This supports the environmentally friendly concept of “treating waste with waste”.

Wink切口下显微镜精索静脉结扎术与腹腔镜下精索静脉高位结扎术疗效对比

目的:通过对比研究,分析精索静脉曲张患者采用Wink切口下显微镜精索静脉结扎术和腹腔镜精索静脉高位结扎术的治疗效果。方法:回顾性选取2020年7月-2021年12月在无锡市人民医院行精索静脉结扎术的50例患者的资料,根据治疗方法差异将其分为Ⅰ组和Ⅱ组,各25例。Ⅰ组采取Wink切口下显微镜精索静脉结扎术,Ⅱ组采取腹腔镜下精索静脉高位结扎术。记录两组患者手术时间、手术出血量及住院时间;分别于术前及术后6个月评估精液质量;记录术后并发症发生情况,术后6个月复发情况;评估患者治疗总满意度。结果:Ⅰ组手术时间长于Ⅱ组,差异有统计学意义(P<0.05),两组住院时间、手术出血量比较,差异均无统计学意义(P>0.05)。术后6个月,两组精液质量较术前均有所改善,且Ⅰ组精子密度、前向运动比例(proportion of sperm forward movement,PR)及精子存活率均高于Ⅱ组,差异均有统计学意义(P<0.05)。Ⅰ组并发症总发生率、术后6个月复发率均低于Ⅱ组,差异均有统计学意义(P<0.05)。Ⅰ组治疗总满意度高于Ⅱ组,差异有统计学意义(P<0.05)。结论:Wink切口下显微镜精索静脉结扎术用于精索静脉曲张治疗效果优于腹腔镜精索静脉高位结扎术,虽然显微镜下操作手术时间略长,但术后并发症少,复发率低,患者满意度高,切口更隐匿美观。

QTL mapping for berry shape based on a high-density genetic map constructed by whole-genome resequencing in grape

Grape berry shape is an important agricultural trait.Clarifying its genetic basis is significant for cultivating grape varieties that meet market demands.However,the current study by forward genetics has not achieved in-depth results.Here,a high-density map was constructed to identify quantitative trait loci(QTLs)for berry shape.A total of 358709 polymorphic SNPs were obtained using whole-genome resequencing(WGS)based on 208 F2 individuals derived from round grape‘E42-6’and oblong grape‘Rizamat’.The 1635.65 cM high-density map was divided into 19 linkage groups with an average distance of 0.37 cM.Using this map,three significant QTLs for fruit shape index(ShI:ratio of berry length to berry width)identified over three years were mapped onto LG4 and LG5,including one stable QTL on Chr5 with the genomic region of 0.47–1.94 Mb.Combining with gene annotation and expression patterns based on RNA-seq data from two contrasting F2 individuals with round and oblong berry(their average ShI was 1.89 and 1.10,respectively)at four developmental stages,four candidate genes were selected from the above QTLs.They were mainly involved in DNA replication,cell wall modification,and phytohormone biosynthesis.Further analysis of RNA-seq data revealed that several important phytohormone synthesis and metabolic pathways were enriched based on differentially expressed genes(DEGs),which was consistent with the results of QTL mapping for genes related to plant hormone biosynthesis in the F2 population.Furthermore,a comparison of plant hormone content showed that there were significant differences in IAA and tZ content between the two contrasting F2 individuals at different developmental stages.Our findings provide molecular insights into the genetic variation in grape berry shape.Stable QTLs and their tightly linked markers offer the possibility of marker-assisted selection to accelerate berry shape breeding.

GRAPES对流尺度集合预报模式中不同尺度初始扰动能量的演变特征

对流尺度数值预报对初始场的微小扰动非常敏感,且初始扰动的演变具有模式依赖、环流依赖和尺度依赖特征,如何构建合理的初始扰动场是国内外对流尺度集合预报领域尚未解决的难点问题和研究前沿。本文基于中国气象局3 km水平分辨率的GRAPES(Global/Regional Assimilation and Prediction Enhanced System)对流尺度模式,利用其同化分析系统的背景误差和一个二维随机型,构建大、中、小三个尺度的初始扰动场,并选取中国夏季一次典型的多区域强降水天气个例,开展对流尺度集合预报试验,对比分析了大、中、小尺度初始扰动能量的时空演变和谱分解特征,以期为构建适用于GRAPES对流尺度集合预报的初始扰动场提供依据。研究结果表明,在GRAPES 3 km对流尺度模式中:(1)大、中、小尺度初始扰动总能量的增长过程具有明显差异。大尺度初始扰动总能量随着模式积分呈增长趋势,尤以对流层中高层的持续增长为甚;而中、小尺度初始扰动总能量随着模式积分以日变化为主,表现为下午至傍晚(夜晚至清晨),扰动总能量显著增加(减小),且扰动总能量小尺度分量的日变化占主导,这可能是由于太阳辐射引起地表加热,使得白天的对流活动比夜晚活跃,且对流直接影响了扰动总能量小尺度分量的变化。此外,大、中、小尺度初始扰动总能量增长均以扰动动能增长为主,扰动位能在对流层低层的增长不可忽略。(2)大、中、小尺度初始扰动总能量增长具有环流依赖特征。对北支气流控制的中高纬天气区,在斜压不稳定较强的低槽区,大尺度初始扰动总能量增长突出,而在槽后西北气流区,大、中、小尺度初始扰动总能量均不增长;对南北气流交汇区,仍以大尺度初始扰动总能量增长最为明显;对南海夏季风影响区,大、中、小尺度初始扰动总能量发展均较弱,扰动位能增长与区域降水大值率演变有较好的对应关系。(3)大、中、小尺度初始扰动总能量的谱分析结果显示,不同积分时段扰动总能量的多尺度串级特征有差异。积分前3 h主要为扰动总能量的大尺度分量向小尺度分量降尺度串级,积分6 h后则为中、小尺度分量的升尺度串级。上述研究结果表明,在天气系统复杂、动力不稳定时空分布不均匀的区域(如中国区域)发展对流尺度集合预报时,有必要针对不同的不稳定天气区,构建具有尺度依赖和环流依赖的初始扰动结构。

Development and application of KASP marker for high throughput detection of the seedless trait in grapevine

Molecular marker-assisted selection(MAS) can significantly accelerate and improve the efficiency of the breeding process in seedless grape cultivars. In this study, we developed the KASP_VviAGL11 and VviAGL11_410 markers based on a single nucleotide polymorphism(SNP) site(Chr18: 26889437(A/C)) of the VviAGL11 gene, and compared them with previously reported SSR markers p3_VvAGL11 and 5U_VviAGL11 by testing 101 cultivars and 81 F1 hybrid progenies. The results showed that both of the proposed markers obtained 100% accuracy rates in detecting allele A, which was closely associated with the seedless trait in grapes, while p3_VvAGL11 and 5U_VviAGL11 had lower accuracy rates due to their tendency to produce false positives. After careful evaluation of the technical advantages and disadvantages associated with these markers, we concluded that KASP_VviAGL11 was superior in terms of simplicity,cost-effectiveness, efficiency, and accuracy. Thus, we optimized the process of molecular MAS for seedless grapes,focusing on the KASP_VviAGL11 marker as a central component, to provide key technical support for the development of new seedless grape cultivars.

Identification of UGT85A glycosyltransferases associated with volatile conjugation in grapevine(Vitis vinifera × Vitis labrusca)

Glycosylated volatiles are considered as hidden aroma precursors that affect fruit flavor and are involved in plant defense response. Several uridine-diphosphate(UDP)-glycosyltransferase(UGT) members related to the glycosylation of volatiles have been identified from grapevine berries. Although grapevine leaves produced much higher levels of glycosylated volatiles than berries, UGTs responsible for the production of glycosylated volatiles in leaves have not yet been identified. Here, eight novel UGT genes were cloned, among these four members that belong to UGT85A family(named UGT85A24, UGT85A25, UGT85A26, and UGT85A27) were localized in the cytoplasm and their recombinant proteins showed activity toward volatiles detected in grapevine, with geraniol as the preferred substrate in vitro. Two UGT genes with 98.1% homology differ in R/S-linalool selection. The UGT85A26-catalyzed substrate showed enrichment in S-linalool, whereas UGT85A27 showed a strong preference for R-linalool. UGT85A24 and UGT85A25 were expressed mainly in berries, and UGT85A26 and UGT85A27 were expressed predominantly in grapevine mature leaves and correlated with the highest content of glycosylated volatiles in the mature leaves. Methyl jasmonate(MeJA) induced the expression of UGT85A26 and UGT85A27 as well as the production of linalyl-β-glucoside, citronellyl-β-glucoside and phenethyl-β-glucoside. Transient overexpression of UGT85A26 and UGT85A27 in tobacco leaves led to the accumulation of linalyl-β-glucoside and an increase in citronellyl-β-glucoside in vivo. These results indicate that the expression of UGT85A26 and UGT85A27 is a major regulator affecting the content of glycosylated volatiles in grapevine.

Mining candidate genes of grape berry cracking based on high density genetic map

Fruit cracking is a phenomenon in which the peel cracks during grape berry development,which seriously affects the yield and quality of the fruit.However,there are few studies on the mining of candidate genes related to berry cracking.In order to better understand the genetic basis of berry cracking,we used the results of previous quantitative trait locus(QTL)mapping,combined with field surveys of berry-cracking types and the berry-cracking rate,to mine candidate berry-cracking genes.The results showed that three identical QTL loci were detected in two years(2019 and 2020);and three candidate genes were annotated in the QTL interval.In mature berries,the expressions of the candidate genes were more abundant in the cracking-susceptible parent(‘Crimson Seedless’)than in the cracking-resistant parent(‘Muscat Hamburg’).Grape berry cracking is a complex trait controlled by multiple genes,mainly including genes encoding cellulose synthase–like protein H1,glucan endo-1,3-beta-glucosidase 12,and brassinosteroid insensitive 1-associated receptor kinase 1.The high expression of the candidate berry-cracking genes may promote the occurrence of berry cracking.This study helps elucidate the genetic mechanism of grape berry cracking.

Intellectual Property Protection and High-quality Development of Grape Industrial Clusters in Xinjiang

This paper introduced the grape industry in Xinjiang and its agricultural intellectual property resources,and analyzed six major problems such as the ineffective leading role of intellectual property rights in high-quality development.Finally,it proposed eight strategies to build a national-level regional fine-grain breeding base for grapes and develop“agricultural chips”in the context of the strategy of strengthening the country with intellectual property and innovation-driven development strategy.

Winery by‑products as a feed source with functional properties:dose–response effect of grape pomace,grape seed meal,and grape seed extract on rumen microbial community and their fermentation activity in RUSITEC

Background Grape and winery by-products have nutritional values for cattle and also contain functional compounds like phenols,which not only bind to protein but can also directly affect microbiota and their function in the rumen.We characterized the nutritional and functional effects of grape seed meal and grape pomace as well as an effective dosage of grape phenols on ruminal microbiota and fermentation characteristics using a rumen simulation technique.Results Six diets(each n=8)were compared including a control diet(CON,no by-product),a positive control diet(EXT,CON+3.7%grape seed extract on a dry matter(DM)basis),two diets with grape seed meal at 5%(GS-low)and 10%(GS-high),and two diets with grape pomace:at 10%(GP-low)and 20%(GP-high),on a DM basis.The inclusion of the by-product supplied total phenols at 3.4%,0.7%,1.4%,1.3%,and 2.7%of diet DM for EXT,GS-low,GS-high,GP-low,and GP-high,respectively.Diets were tested in four experimental runs.All treatments decreased ammonia concentrations and the disappearances of DM and OM(P<0.05)compared to CON.EXT and GP-high lowered butyrate and odd-and branch-chain short-chain fatty acids while increased acetate compared to CON(P<0.05).Treatments did not affect methane formation.EXT decreased the abundance of many bacterial genera including those belonging to the core microbiota.GP-high and EXT consistently decreased Olsenella and Anaerotipes while increased Ruminobacter abundances.Conclusion The data suggest that the inclusion of winery by-products or grape seed extract could be an option for reducing excessive ammonia production.Exposure to grape phenols at a high dosage in an extract form can alter the rumen microbial community.This,however,does not necessarily alter the effect of grape phenols on the microbial community function compared to feeding high levels of winery by-products.This suggests the dominant role of dosage over the form or source of the grape phenols in affecting ruminal microbial activity.In conclusion,supplementing grape phenols at about 3%of diet DM is an effective dosage tolerable to ruminal microbiota.

Ampelographic and Genetic Diversity Assessment of Some Local Grape Genotypes under Egyptian Conditions

Grape (Vitis vinifera L.) is assumed as one of the most valuable and popular fruit crops all over the world. In this study, ten of local grape germplasm were characterized considering phenotypic diversity and genetic features under Egyptian conditions. Selected local grapes;Baltim Eswid, Edkawy, Matrouh Eswid, Bez El-Naka, Bez El-Anza, Romy Abiad, Romy Ahmer, Gharibi, Fayoumi and Banaty, were spread geographically among seven national governorates. A total of 58 attributes were characterized grape germplasm for the distinctness of vine parts. For molecular characterization, 9 nuclear SSR loci were analysed. Data revealed a broad sense of similarity at the level of studied morphological attributes, where the highest similarity (0.93) was between Romy Ahmer and Romy Abiad. A total of 24 alleles, ranging from 124 - 253 bp in size, were detected at the nine tested loci with an average of 2.67 alleles per locus. The amplification products in all of the 9 SSRs loci showed polymorphism among the 10 grapevine cultivars. The genetic relatedness among most of the cultivars was in accordance with their identification based on ampelographic properties. Data of phenotypic and molecular analysis revealed high variability of Vitis germplasm in Egypt and contributed substantially to our awareness of valuable local grape genetic resources which are on the verge of extinction.

Identification of key gene networks controlling monoterpene biosynthesis during grape ripening by integrating transcriptome and metabolite profiling

Monoterpenes are typical aroma components of muscat grape cultivars,providing pleasant floral and fruity aromas to grapes and wines.However,the molecular mechanism of monoterpene biosynthesis between muscat and non-muscat grape remains unclear.Here,the muscat grape cultivar‘Jumeigui’and the non-muscat grape cultivar‘Kyoho’were chosen as plant materials for a comprehensive transcriptome and metabolite analysis.The gas chromatography-mass spectrometry(GC-MS)analysis demonstrated that a total of 27 and 23 monoterpene compounds were identified and quantified in the‘Jumeigui’and‘Kyoho’grape,respectively.‘Jumeigui’grape accumulated significantly higher concentrations of monoterpenes than‘Kyoho’grape.Furthermore,geraniol,linalool,geranic acid,and β-citronellol might be important odorants contributing to the floral character of the‘Jumeigui’grape due to the high levels odor activity values(OAVs).Transcriptome analysis demonstrated that the expression profiles of VvDXS,VvGGPPS.SSU1,Vv TPS-b/g showed a positive correlation with monoterpene accumulation in grapes.In addition,the expression patterns of the genes involved in jasmonic acid(JA)synthesis and signal were also positively correlated with monoterpene accumulation.All these results will help guide the functional verification of candidate genes related to monoterpene biosynthesis,as well as identify the master transcriptional and hormonal regulators of this pathway in grapes.

Improved maceration techniques to study the fruit vascular anatomy of grape

The visualization and evaluation of fruit vascular bundles in grape are indispensable for understanding the development and growth of the fruit. The vascular bundles in grape are as thin as human hair, and strongly adhere to flesh cells, making it difficult to isolate individual intact vessel elements. Currently there is little information about methods to characterize grape vascular tissue. In this study, we describe an easy and effective technique to visualize the xylem cell structure of the ‘Red Globe' grape(Vitis L.). The intact vascular bundles of the grape were isolated through enzymatic degradation of flesh cells by hydrolases which were effective in removing flesh cells from vascular bundles. This enabled the illustration of the fine surface structure of vessel elements and their arrangement in the vascular bundles of the grape. This modified method to separate the vascular bundles into individual vessel elements was more effective than former methods of manually shaking to isolate individual vessels. Clear images of xylem vessel arrangement and structural characteristics of individual vessel element were acquired by light microscopy(BX51, Olympus, Tokyo, Japan), transmission electron microscopy(Tecnai 12, Philips, Netherlands) and scanning electron microscopy(GeminiSEM 300, Carl Zeiss, Germany). In addition, the 3D structure of vessel elements was observed using confocal laser scanning microscopy(Zeiss: model: LSM-880, Oberkochen, Germany). The imaging techniques for visualizing and analyzing the structure of xylem vessels in grape described in this study are an improvement of past methods. An effective method to isolate grape vascular bundles from flesh cells was also developed, which enables the imaging of the fine surface structure of vessel elements and their arrangement in grape vascular bundles. By adjusting the process of maceration of vascular bundles, an easy and effective method was developed to gently disrupt the plant tissue and isolate individual vessel elements. These improved techniques are suitable to observe the intact morphology of vascular bundles in the grape, improve experimental efficiency and accelerate new discoveries on the anatomical structure of the grape.

Quality assurance of postharvest grapes against Botrytis cinerea by terbinafine

Worldwide,fruit is an indispensable treasure house of nutrition for human beings,occupying a vital position of human diet.Postharvest fruit storage requires efficient antifungal agents to control Botrytis cinerea,which is a vital postharvest disease affecting fruit and leading to enormous losses.However,with the enormous abuse of existing antifungal drugs,the problem of drug-resistant fungi is imminent,making the controlling diseases caused by pathogenic fungi even more challenging.Drug repurposing is an efficient alternative method,we evaluated a well-known antifungal chemical,terbinafine,against the agricultural pathogen,B.cinerea in vitro,as a result,terbinafine showed strong anti-fungal activity.Furthermore,the in vivo antifungal activity of terbinafine was evaluated,the results showed that terbinafine could reduce the decay area on grapes.Terbinafine could disrupt the cell membrane integrity,increase cell membrane permeability,and eventual cell death of B.cinerea.In addition,terbinafine reduced decay incidence,and weight loss and maintained the soluble solids,titratable acidity,ascorbic acid,total phenolic,and malondialdehyde content during the storage period of grapes.Overall,terbinafine could be an antifungal preservative for postharvest table grapes fresh-keeping.

VvAGAMOUS Affect Development of Four Different Grape Species Ovary

Grape pistil has an important influence on fruit size and quality.However,there were few studies on grape ovary,and the development process of the ovary is still unclear.Therefore,in this paper,four different grape varieties with different lengths of small inflorescences,namely‘Musct Hambourg’grape(Vitis vinifera),‘Concord’grape(Vitis labrusca),‘ShanPuTao’grape(Vitis amurensis)and‘GongNiang2Hao’grape(Vitis amurensis×Vitis vinifera)were used as test materials.Four varieties ovary were significant differences by means of stereomicroscope,paraffin section.The expression of ovary determining gene VvAGAMOUS(VvAG)and its development related genes VvCRABS CLAW(VvCRC)and VvAGAMOUS-LIKE 11(VvAGL11)with similar functions during the development of different grape varieties were preliminarily explored using fluorescence quantitative test.The relationship between VvAG and VvCRC,VvAG and VvAGL11 were analyzed using Y1H assay.Our results showed that there were obvious abdominal sutures on the surface of expect for‘Musct Hambourg’grape,and existing poly carpels.The ovary development of‘ShanPuTao’and‘GongNiang2Hao’grape was completed when the inflorescence length was less than 1 cm,while the‘Concord’and‘Musct Hambourg’grape were fully developed when the length of inflorescence was 3–4 and 4–5 cm,respectively.VvAG and VvCRC began to express in large quantities after the formation of stamen primordia,while VvAGL11 during the forming of ovule primordia.Therefore,VvAG and VvCRC mainly regulated the development of stamens and carpels and also promote the development of ovules,while VvAGL11 major regulated the development of ovules.The promoters of VvCRC and VvAGL11 were bound by VvAG.This study provides an important theoretical basis for further research on the molecular mechanism of grape ovary development.