葡萄(Vitis vinifera L.)试管繁殖中芽增殖能力衰减的原因及其防治方法的研究

以20个葡萄品种试管苗作为实验材料,经四年、四十多代的继代培养反复对比实验发现:在葡萄试管繁殖中,连续恒温(25±1℃)条件下培养或只采用固体培养方法,不适期进行继代培养均会导致芽增殖能力的衰减,根据生物统计分析,当继代培养超过16～17代后,芽增殖能力即随着继代培养次数的增加而逐渐下降,采取变温培养、变换培养方法、适期继代培养或在增殖培养基中附加适量的椰子乳汁等方法均可防治芽增殖能力的哀减,保持高的芽增殖能力。

葡萄(Vitis Vinifera L.)X-射线能谱元素微区污染分析研究

对葡萄果实不同处理的细胞组织多位点进行X -射线能谱微区元素分析。结果 :葡萄不同处理的细胞组织中共发现Mg、Al、Si、P、S、Cl、K、Ca、Fe、Cu十种元素。且不同处理的组织细胞中各元素含量分布有差异 ,未清洗果实表皮处理的Al、Si、S、Ca、Cu含量均显著高于对照 。

VitisMod:一个葡萄基因共表达数据库

葡萄转录组已在各种组织、发育阶段、生物胁迫、非生物胁迫和其他条件下被测定。目前,仍没有简单实用的网络工具来探索这些宝贵的数据。本文从美国国立生物技术信息中心的基因表达数据库(NCBI GEO)下载1019个基因表达芯片数据,进行权重基因共表达网络分析(WGCNA)。鉴定到41个基因共表达模块。功能富集分析发现这些模块具有不同的功能,并与实验/表型相关。通过模块内连接度筛选枢纽基因,这些基因可能具有重要功能。通过关联推定(Guilt-by-association)原理对模块内功能未知的基因进行功能预测。最后,构建了免费的网络工具VitisMod,为葡萄的基因功能研究提供新资源,网址为:http://bioinformatics.fafu.edu.cn/grape。

The Establishment of an Agrobacterium-Mediated Transformation Platform for the Non-Embryogenic Calli of Vitis vinifera L.

Non-embryogenic calli （NEC） was inevitably and heavily produced when grape embryogenic calli （EC） was induced from explants or during the subculture of EC.A stable and highly efficient NEC transformation platform is required to further sort out and verify key genes which determine/switch the identity of NEC and EC.In this research,a vector pA5 containing a chitinase signal sequence fused to gfp （green fluorescent protein） and an HDEL motive was used to target and immobilize into Agrobacterium strain EHA105 to establish a transformation platform for Vitis vinifera L.cv.Chardonnay NEC.It was determined that NEC 10 d after subculture was the best target tissue;30 min for inoculation followed by 3 d co-cultivation with the addition of 200 μmol L-1 acetosyringone （AS） was optimized as protocol.The use of bacterial densities as 1.0 at OD600 did not result in serious tissue hypersensitive reaction and it had higher efficiency.Kanamycin at 200 mg L-1 was picked for positive expression selection.The stable transformation of NEC was proved by reverse transcription-polymerase chain reaction techniques （RT-PCR） and fluorescent microscopy after three sub-cultures of the selected cell line.Highly efficient genetic transformation protocol of grape NEC was achieved and some of the optimized parameters were different from that reported for EC.This transformation platform could facilitate the verification of candidate somatic embryogenesis （SE） decisive genes,and the successfully transformed NEC with certain genes can also be used as bioreactors for the production of functional products,as NEC not only proliferates fast,but also keeps in a rather stable condition.

Anthocyanin profi les and color properties of red wines made from Vitis davidii and Vitis vinifera grapes

Spine grape(Vitis davidii Foex.)is an important wild grape species native to China.Fifteen red spine grape clones and three red Vitis vinifera grape varieties were used to evaluate the differences in the anthocyanin profiles and color properties of wines made from V.davidii and V.vinifera grapes.Among spine wines,‘Junzi#2’wine had the highest total phenolic and total anthocyanin,‘Xiangzhenzhu’wine had the highest total flavonoids,and‘Junzi#1’wine had the highest total tannin.The anthocyanin compositions of all of the spine wines were dominated by Mv-3,5-diglucoside.The total individual anthocyanin contents in spine wines,except‘Gaoshan#5’,‘Junzi#5055’,‘Junzi#5061’,and‘Junzi#5044’,were signifi cantly higher than in V.vinifera wines.Most of the spine wines had a stronger red intensity and a brighter chroma with a bluer hue than V.vinifera wines.Correlation analysis revealed that the color properties were closely related to the anthocyanin composition.These results suggest that wines made from spine grapes may be useful for the wine industry for their color properties and high individual anthocyanin contents.

The potential role of grape (Vitis vinifera L.) in prevention of threatened abortion via immunomodulatory and anti-inflammatory abilities: a hypothesis

Threatened abortion is a common problem in early pregnancy. This early vaginal bleeding happens in about 25% of pregnant women. The medications including progesterone, uterine muscle relaxant or human chorionic gonadotropin have essential effects in developing pregnancy, but the clinical data are insufficient to prescribe them. In recent decades, medicinal herbs can help us to present new treatments. Grape (Vitis vinifera L.) can protect the fetus from the perspective of Persian medicine. So, we hypothesize about the salutary effects of grape in miscarriage prevention. We found five standard expected mechanisms of grape to prevent threatened abortion: immunomodulatory and anti-inflammatory, anti-oxidant, anti-contraction, hormonal and anti-stress activities. Grape reduces nitric oxide, prostaglandin E2, expression of nuclear factor κB and other pro-inflammatory cytokines like IL (Interleukin)-1β, IL-6, IL-8, and tumor necrosis factor-α. It also elevates anti-inflammatory mediators and expression of peroxisome proliferator-activated receptor-γ. Grape polyphenols have a crucial role in fetus protection with high antioxidant power and other functions such as prevention of stress-triggered abortion with proanthocyanidins, or hormonal effects and inhibition of uterine contractions with resveratrol. So according to these studies, grape probably has effects on the immune and endocrine factors involved in threatened miscarriage.

Somatic embryogenesis and histological analysis from zygotic embryos in Vitis vinifera L.‘Moldova’

We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape ＇Moldova＇ （Vitis vinifera U ＇Moldova＇）. Primary calli were initiated on Nitsch and Nitsch （NN） medium supplemented with 1.0 mg·L^-1 2,4-D and 0.5 mg·L^-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg·L^-1 6-BA and 2 mg·L^-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg·L^-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dynamic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic embryos, including typical epiderma, cotyledon primordium and vascular tissue.

Rooting Abilities in <i>Vitis vinifera</i>L.: Total Lipid and Fatty Acids Evolution in Perle Noir Cutting

Rooting ability in Vitis vinifera L. is the phenomena studied. The evolution of the total lipids contents and fatty acids amount of cuttings during the rooting were determined. Total lipid extraction was carried out by an extraction solvent consisting of a mixture of chloroform and methanol. Methylation of fatty acids was carried out by the boron trifluoride methanol complex. Their analysis is performed by gas chromatography. We have obtained a content of merithallus lipids equivalent with then in bibliography. In other hand 3 fatty acids were obtained: eicosapentaenoic acid, oleic acid and docosanoic acid. Among the studied of rooting abilities, we observed decreased levels of total lipids and in the essential fatty acids: eicosapentaenoic acid C20:5 n-3 and oleic acid C18:1 n-9. Minor fatty acids cuttings experimental have a variable evolution. Lipids and certain fatty acids may be markers of rooting in the vine.

基于改进BiSeNet的葡萄黑麻疹病害程度分级预测

为了准确对葡萄(Vitis vinifera L.)黑麻疹病害程度进行分级预测,通过语义分割模型将叶片部分和病斑部分分割出来,以同一叶片上病斑面积与总叶面积的比值作为疾病严重程度分级的依据,对葡萄黑麻疹病害程度进行分级预测。精确标注了PlantVillage公开数据库中的419张葡萄疾病图像,细分为背景、叶片和病斑3个类别,并应用了数据增强技术增加样本多样性。以BiSeNet作为基准模型,引入GhostNet作为上下文路径的主干提取网络,不仅保持了较小的模型参数量,而且在精度上实现了明显提升,满足病害程度分级预测的需求。提出了累加空洞空间金字塔池化(CASPP)模块,用来替换BiSeNet模型中单一的上下文嵌入模块,以增强BiSeNet模型的多尺度上下文信息提取能力,提高了模型的分割精度。经过测试,本研究模型在测试集中的平均交并比为94.11%,在对葡萄黑麻疹病害程度进行分级预测时,准确率达98.21%,能够精确地对葡萄黑麻疹病害程度进行分级预测。

硫华菊混种葡萄对两种植物镉积累的影响

为降低果树镉积累,采用盆栽试验,将观赏型镉富集植物硫华菊(Cosmos sulphureus)和葡萄(Vitis vinifera L.)混种在镉污染的土壤上,研究了硫华菊混种葡萄对2种植物镉积累的影响。结果表明,硫华菊混种葡萄提高了硫华菊植株的生物量和镉含量,分别较硫华菊单种提高8.63%和22.40%;降低了葡萄植株的生物量和镉含量,分别较葡萄单种降低6.74%和34.84%。硫华菊混种葡萄提高了硫华菊植株的镉积累量,降低了葡萄植株的镉积累量,其中,混种的硫华菊植株镉积累量较其单种提高33.00%,而混种的葡萄植株镉积累量较其单种降低39.21%。此外,2种植物的整株生物量与整株镉含量、整株镉积累量、富集系数均呈负相关,整株镉含量与整株镉积累量、富集系数呈正相关,整株镉积累量与富集系数也呈正相关。因此,硫华菊混种葡萄能促进硫华菊对镉的积累、降低葡萄对镉的积累。

Effects of mesoclimate and microclimate variations mediated by high altitude and row orientation on sucrose metabolism and anthocyanin synthesis in grape berries

Climate change and extreme weather pose significant challenges to the traditional viticulture regions.Emerging high-altitude grape-producing regions with diverse orientations have shown great potential in coping with this challenge.Stable,high-quality wine grape production may be achieved by synchronizing the meso-and microclimate.To clarify the role of high altitude and row orientation in meso-and microclimate and the response of berries to it,we evaluated seven years(2012-2018)of climate data,two years of basic grape(Cabernet Sauvignon,Vitis vinifera L.)quality,and one-year microclimate from veraison to harvest.By comparing two locations(Sidon 2047 m,Sinon 2208 m)in Yunnan Province,China,we found that the average temperature has been stable at approximately 15℃ for seven years,with no extreme weather or,noticeable global warming.The light intensity(LI)in the north-south(NS)was more balanced than the east-west(EW)direction,and the east-west to the south(EW-S)canopy side was almost higher than the other sides.High LI was associated with high photosynthetically active radiation(PAR),ultraviolet(UV),and infrared(IR)light and vice versa.The north-south to the east(NS-E)and east-west to the north(EWN)sides were characterized by lower LI and higher UV and IR light,and higher total anthocyanin content.Most anthocyanin synthesis-related genes,for example,VvF3'H and VvF3'5'H,were highly expressed in NS-E from veraison to maturity.Perhaps UV and IR light induced their expression.This study provides new insights on the role of differently orientated rows in controlling grape quality due to varied light quality.The findings are globally significant,particularly in the context of climate change,and offer fresh insights into berry physiological responses and decision-making for the management of existing vineyards.

养分综合管理模式对葡萄产量及土壤养分含量的影响

为了研究生物有机肥和葡萄专用水溶肥在葡萄上的施用效果,连续2 a(2020年和2021年)采用小区试验,以常规施肥为对照(FP处理),分别设置了施用生物有机肥(SR处理)和施用生物有机肥+专用水溶肥(ISSM处理),研究其对葡萄产量和土壤养分含量的影响。结果表明,连续2 a试验ISSM处理产量均显著高于其他处理(P<0.05),分别为11985和10939 kg·hm^(-2),与FP比,ISSM处理增产效果最明显,增幅分别为2550和2528 kg·hm^(-2),增产率达到了27.03%和30.05%。在葡萄园地土壤养分含量方面,ISSM处理对提高土壤pH值、有效磷含量、速效钾含量、交换性钙镁含量效果显著,但对提高土壤全氮含量和有机质含量效果不明显。在土壤养分含量与葡萄产量相关性方面,2 a的葡萄产量与土壤速效钾含量和交换性钙含量存在显著或极显著差异。综合分析,生物有机肥配施专用水溶肥(ISSM处理)效果最佳。

温度对巨峰葡萄节间和带节茎段导水特性的影响

[目的]研究旨在明确巨峰葡萄枝条木质部导水能力与温度的量化关系,探索节对水分运输效率与安全性的影响。[方法]以巨峰葡萄1年生枝条为材料,在0.5~40℃温度范围,利用低压液流计法测试节间和带节茎段的导水率、比导率及导水率损失(PLC)随温度的变化,分析节与节间茎段导水特性的差异。[结果]巨峰葡萄茎段导水率和比导率均与温度呈极显著的线性关系。当温度从0.5℃升高至40℃,节间茎段的初始导水率、最大导水率、初始比导率、最大比导率分别升高154%、198%、157%和200%。节间茎段的导水率和比导率显著大于带节茎段,且差异随着温度升高呈增大趋势。节间茎段的PLC显著大于带节茎段,而带节茎段的导水率和比导率受温度的影响小于节间茎段。利用线性回归方程可对不同温度下的导水率和比导率进行预测。导水率和比导率与各温度下水的粘滞系数进行权衡后,各温度下的权衡导水率和权衡比导率没有明显差异。[结论]葡萄茎段导水率和比导率随着温度升高而增大,节对水分运输的效率和安全性有权衡作用。

木醋液对葡萄灰霉病菌的抑制作用

为挖掘更多葡萄(Vitis vinifera L.)灰霉病的生防资源,分析不同浓度木醋液对葡萄灰霉病菌菌丝生长、孢子量、孢子萌发的抑制作用以及对葡萄离体叶片灰霉病菌的防治效果。结果表明,木醋液浓度为0.500 0%,灰霉病菌菌株HM22-5菌丝生长和产孢受到明显抑制,菌丝生长抑制率为55.93%,分生孢子量显著降低,分生孢子萌发抑制率为97.08%;当木醋液浓度为1.000 0%时,葡萄灰霉病菌菌丝生长抑制率可达90.00%以上,分生孢子萌发抑制率为100.00%,对葡萄离体叶片的防效达77.32%;木醋液浓度为2.000 0%时,葡萄灰霉病菌菌丝生长抑制率为100.00%,对葡萄离体叶片的防效达89.69%。

葡萄及其制品中赭曲霉毒素A的污染现状及检测技术研究进展

赭曲霉毒素A(ochratoxin A,OTA)是由青霉属与曲霉属真菌产生的次级代谢产物之一,容易污染葡萄及其制品,是葡萄产业面临的重要问题之一。OTA具有急性、慢性、亚慢性毒性,对消费者健康存在潜在威胁。鉴于OTA的污染和毒性,世界各地对其制定了严苛的限量标准。因此,快速、高效、灵敏的OTA检测技术的开发与应用备受关注。本文综述了葡萄及其制品中OTA的污染现状,以及葡萄和葡萄制品生产过程中影响OTA污染的因素,总结近年来OTA传统检测技术的发展,重点介绍标记型与非标记型电化学适配体传感器的研究进展,讨论不同检测技术的优势和局限性,展望OTA检测技术的发展,以期为葡萄及其制品中OTA的研究提供理论基础。

The Influence of Photoperiod on the Regulation of Root and Callus Initiation of Perle Noir (<i>V. vinifera</i>L.): Expression of MADS-Box Gene

To study the influence of photoperiod on roots differentiation in the Tunisian grapevine (Vitis vinifera L.) cultivar Perle noir, roots and callus initiation were analyzed under three different conditions of day length: long day (LD), short day (SD) and darkness (D). The photoperiod influenced the number of callus and roots per cuttings;it has a significant effect on the roots and callus initiation. Expression profile analysis of six MADS-box genes (VTM8, VSEP2, VAG12, VAG17-1, VAG17-2 and VSOC1.3) during root and callus development is in agreement with the above-mentioned observation. The expression of the MADS-box genes during root and callus development fluctuated in a tissue-dependent manner. These data suggest that all genes are expressed in roots under three photoperiods. Total darkness gives the number of the most important root per cutting compared to the other two conditions. This photoperiodic condition gave the most important expression of the studied genes VAG12, VAG17-2, VAG17-1, VTM8 and VSEP2 transcripts were not found in callus grown in the dark or in LD conditions, respectively. VSOC1.3 transcripts were not found in callus grown in the dark or in SD conditions, respectively. Transcript abundance of VTM8 and VSOC1 was highest in LD.

欧洲葡萄VvJAZ9基因互作蛋白的筛选与验证

【目的】探究葡萄JAZ家族基因特点及在低温胁迫下的表达情况,以期筛选出与低温胁迫相关的家族成员,并进一步筛选其互作蛋白,为后续葡萄抗寒机制研究提供理论依据。【方法】以拟南芥和葡萄基因组数据为基础,获得葡萄JAZ家族成员基因并进行同源克隆、染色体定位、理化性质以及蛋白家族聚类等分析,利用实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)技术分析VvJAZ家族成员基因在低温胁迫下的表达情况,应用酵母双杂交技术进行互作蛋白筛选和验证,并通过双分子荧光互补试验验证候选互作蛋白与VvJAZ9蛋白的互作关系。【结果】葡萄中有11个JAZ家族基因,随机分布于7条染色体上,都具有JAZ家族蛋白中特有且高度保守的TIFY和Jas结构域,均属于碱性不稳定亲水蛋白;聚类分析表明,葡萄JAZ家族蛋白可分为3个亚族,其中VvJAZ9与拟南芥AtJAZ1和AtJAZ2的亲缘关系最近,VvJAZ1与VvJAZ11、VvJAZ5与VvJAZ6同源关系最近。qRT-PCR结果表明,VvJAZ9是葡萄JAZ家族成员基因中受低温诱导表达最为明显的1个基因。通过酵母双杂交试验筛选获得26个可能与VvJAZ9存在互作关系的候选蛋白;通过q RT-PCR分析候选蛋白的相关基因在低温胁迫下的表达模式,结果表明HAP5A基因受低温诱导表达趋势与筛选出的VvJAZ9基因类似,推测HAP5A可能是JAZ9候选互作蛋白中响应低温最为显著的1个蛋白;通过同源克隆获得了VvHAP5A基因序列,全长636 bp,编码211个氨基酸,进一步分析发现该基因是NF-YC类型的转录因子,通过酵母双杂交和双分子荧光互补试验也证实了JAZ9与HAP5A蛋白全长有互作关系。【结论】鉴定并克隆到了葡萄的11个VvJAZs基因,VvJAZ9和VvHAP5A基因均响应葡萄低温诱导表达,并且二者还具有蛋白互作关系,可为进一步研究葡萄JAZ基因参与低温胁迫的机制提供参考。

壳聚糖对葡萄幼苗养分吸收的影响

为筛选出有利于葡萄(Vitis vinifera L.)幼苗养分吸收的壳聚糖浓度,为葡萄生产提供参考,试验以葡萄幼苗为材料,对其喷施不同浓度的壳聚糖溶液,研究壳聚糖对葡萄幼苗养分吸收的影响。结果表明,喷施壳聚糖溶液提高了葡萄幼苗整株氮、磷、钾含量,在壳聚糖浓度为4 g/L时达最大值,分别较对照增加25.53%、18.67%、12.63%。壳聚糖处理也提高了土壤碱解氮、速效磷、速效钾和有机质含量,其中,壳聚糖浓度4 g/L时,土壤有机质含量较对照增加42.58%。葡萄幼苗的中微量元素钠和钙含量随壳聚糖浓度的增大呈先降后升再降的变化趋势,而镁含量呈先升后降的变化趋势。就土壤酶活性而言,壳聚糖提高了土壤蔗糖酶、脲酶、过氧化氢酶和磷酸酶活性。4 g/L壳聚糖最有利于葡萄幼苗养分的吸收。

根区施用硅肥对‘玫瑰香’葡萄果实品质的影响

为调控果实采后的保鲜品质,在采前对9年生倾斜独龙干型展穗期的‘玫瑰香’树体在根区施用了不同剂量的硅肥,并在果实成熟期采用离子迁移谱(gas chromatography-ion mobility spectroscopy,GC-IMS)等方法测定了果实的挥发性组分及常规品质指标。结果表明,不同硅肥施用量(250、500、800 g·株^(-1))下‘玫瑰香’果实的几种常规品质指标(总糖、总酸、维生素C、可溶性固形物)含量分别为16.47%、17.97%、16.87%;0.325%、0.225%、0.300%;12.703、18.870、19.487 mg·100g^(-1);23.82%、22.59%、30.18%。硅肥施用量为800 g·株^(-1)时,总糖与总酸相较CK无显著变化(P>0.05),但维生素C和可溶性固形物大幅增加(P<0.05)。对果实挥发性物质分析发现,一些脂类物质的峰体积与硅肥施用量增加呈正相关,在800 g·株^(-1)时达到峰值,乙酸乙酯与丙酸乙酯的峰体积相较于CK变幅分别为70.89%、368.49%。表明增施硅肥达到一定剂量时对定向改变‘玫瑰香’果实品质有促进作用,为进一步探究这些物质对采后保鲜的影响奠定了理论依据。

露地栽培‘大青葡萄’架面不同位置果实品质指标分析

为了探究露地栽培‘大青葡萄’(Vitis vinifera L.Daqing)架面及单穗不同位置果实理化指标及挥发性物质的特点,并建立其果实质量评价体系,在果实成熟期,通过常规指标检测、全质构分析及气相离子迁移谱(GC-IMS)技术,测定了宁夏‘大青葡萄’架面及单穗不同位置果实的相关品质指标。结果表明:架面后部果实总糖和Vc含量较中部和前部果实分别显著(P<0.05)提高了8.92%、10.38%和22.29%、62.00%;总酸含量最高的是前部果实,较中部和后部分别高出11.81%、17.21%。单穗肩部果实在可溶性固形物、总糖、Vc含量上均显著(P<0.05)的高于尖部,分别高出5.62%、6.10%、13.02%;总酸含量最高的是尖部果实,较肩部高出9.46%。质构指标分析发现,架面后部果实在单粒重和硬度上较中部、前部果实分别高出14.84%、19.12%和4.44%、5.36%,存在显著差异(P<0.05);单穗肩部果实在单粒重和硬度上均高于尖部,分别高出6.02%、3.22%。通过气相离子迁移谱技术对挥发性物质分析发现,架面前部果实表现出青草、麦芽香气特点;中部果实体现脂肪香气;后部果实具备水果、青草香气特征;在一些酯类和萜烯类物方面,后部果实与中部和前部位置果实存在显著差异(P<0.05)。单穗尖部果实青草和奶油香气表现突出,肩部果实则是果香和坚果香味较为明显;肩部果实一些脂类物质和萜烯类物质(β-蒎烯、α-水芹烯、乙酸乙酯等)的积累量显著(P<0.05)高于尖部的果实,分别高出27.68%、50.88%、785.43%。结合常规品质指标、质构指标以及挥发性物质特征,‘大青葡萄’果实在架面不同空间位置的品质高低综合排序为后部>中部>前部;单穗不同部位的果实为肩部优于尖部。露地大棚架栽培‘大青葡萄’存在架面不同空间位置上的果实品质与挥发性物质上的差异,并且单穗果实同样表现出不同部位的品质区别。研究结果为宁夏露地大棚架栽培‘大青葡萄’建立了果实质量评价体系,以指导采后保鲜与供应不同目标市场。