Anthocyanins and proanthocyanidins are considered to be essential secondary metabolites in grapes and are used to regulate metabolic processes, while miRNAs are involved in their synthesis of anthocyanins and proanthocyanidins to regulate metabolic processes. The present research work was carried out to investigate the underlying regulatory mechanism of target genes in the grape cultivars ‘Italia’ and ‘Benitaka’. miRNA and transnscriptomic sequencing technology were employed to characterize both the profiles of miRNAs and the transcripts of grape peels at 10 and 11 weeks post flowering (10 wpf and 11 wpf). The results revealed that the expression level of vvi-miR828a in ‘Italia’ at 10 and 11 wpf was significantly higher than that in ‘Benitaka’. miRNA-seq analysis predicted MYBPA1 to be the target gene of vvi-miR828a. In transcriptome analysis, the expression level of the VvMYBPA1 gene in ‘Benitaka’ was significantly higher than that in ‘Italia’; in addition, the TPM values (expression levels) of VvMYBPA1 and miR828a also showed an evident negative correlation. The determination of the proanthocyanidin (PA) content in ‘Italia’ and ‘Benitaka’ peels at 11 wpf demonstrated that the PA content of ‘Benitaka’ was significantly higher than that of ‘Italia’. The outcomes of RT-qRCR analysis exhibited that the expression levels of the VdPAL, VdCHS, VdCHI, VdDFR, VdMYB5b, VdANR, and VdMYBPA1 genes related anthocyanin and proanthocyanidin pathways were reduced, while the expression levels of all of the above genes were increased after the transient expression of the VvMYBPA1 vector into grape leaves. The results of the transient overexpression experiment of vvi-miR828a before the veraison period of strawberry fruits showed that vvi-miR828a can significantly slow down the coloration of strawberries. The vvi-miR828a negatively regulates the accumulation of proanthocyanidins in grape fruits by inhibiting the expression of VvMYBPA1.

为研究不同时期主梢修剪对"夏黑"葡萄二次结果的生长习性以及果实品质的影响,探索适宜湖南地区的二次结果技术,以9年生"夏黑"葡萄为试验材料,设置常规栽培和延后栽培处理,延后栽培是在1年生枝条去除一次花后,对"夏黑"葡萄在不同时期对主梢第6节位修剪促顶端冬芽萌发进行二次结果,观测物候期和生长习性,测定成熟期的果实品质.结果表明,"夏黑"葡萄在4月30日、5月30日、6月30日修剪促萌的二次果均能正常成熟,成功将"夏黑"成熟期延后至11月上旬.3个熟期的二次果在萌芽率、结果系数方面差异不大,均能整齐萌芽,但6月30日修剪的二次果在结果枝率方面显著低于其他两个处理,且主芽坏死严重,无法达到生产要求.3个熟期的二次果的果实品质虽略差于一次果,但二次果在果实硬度、果梗拉力、还原糖和糖酸比等方面优于一次果,且可滴定酸低于一次果,二次果的口感和风味较好,在一定程度上达到商品性要求.随着修剪时期往后推移果皮转色越快,着色越深.5月30日对"夏黑"葡萄主梢进行修剪生产的二次果综合性状最优,研究结果对湖南地区二次果的果粒增大、产量和品质提升提供了一定的理论指导.

[目的]研究不同葡萄品种叶片组织结构特征及其光谱响应差异,揭示葡萄叶片光谱反射率差异的主要影响因素,为提高葡萄叶片营养光谱诊断精度提供参考.[方法]在河北廊坊葡萄园,采集夏黑、意大利、红宝石、秋黑4个葡萄品种的叶片,用Fieldspec FR2500光谱仪测定叶片光谱数据,常规化学方法测定叶片含氮量,通过扫描电镜(SU8010)冷冻传输技术观察和测量叶片组织结构,进行不同葡萄品种的叶片光谱与叶片组织结构的相关分析.[结果]葡萄叶片气孔主要分布在叶片反面,是正反面光谱反射率产生差异的主要原因,在可见光波段范围内,叶片反面光谱反射率皆高于叶片正面光谱反射率,在近红外波段范围,叶片正面光谱反射率普遍高于叶片反面光谱反射率;不同葡萄品种叶片表面气孔数量和分布不同,栅栏组织细胞厚度以及海绵组织厚度存在差异,在叶片含氮量相近条件下,不同品种的光谱反射率曲线有差异,主要是叶片组织结构和形态差异所致;叶片正面光谱的红边参数λred(在660～770 nm波长范围内,当光谱反射率的一阶微分值达最大时所对应的波长)与不同品种叶片厚度相关性均达到显著或极显著水平,光谱红边参数与其它叶片结构参数也有较强相关性,其中栅栏组织和海绵组织的厚度在不同品种的光谱响应中是不可忽略的因素.[结论]明确了叶片正反面表皮细胞形态、叶片内部结构差异及其与光谱特征的相关性,不同葡萄品种叶片厚度变化均可以用红边参数λred来反映,但若考虑品种因素,还可以针对每个品种选择相关性更好的参数,这为后期葡萄叶片营养光谱诊断模型的建立和优化提供了依据:为提高叶片营养光谱诊断模型的精度,在利用光谱技术进行叶片营养诊断时需要考虑叶片结构因素的影响.

郴州市葡萄种植主要集中在苏仙区和宜章县;主栽品种为巨峰、无核白、阳光玫瑰,其中巨峰葡萄种植最为普遍;栽培模式仍处露地栽培向避雨栽培的转变期;不同葡萄园因主栽品种、栽培模式的不同而产生了极大的效益差异,进而影响了果农对园区的期许目标.分析了郴州市葡萄产业现状和优劣势,根据郴州市葡萄产业发展的潜力,提出了切实可行的对策建议.

探索葡萄冬芽主芽坏死的规律和分析影响主芽坏死的可能原因,利用解剖观测的方式对湖南长沙和山东济南种植的“红地球”冬芽主芽坏死进行对比;在葡萄萌芽期、摘心期、盛花期、第一次膨大期、浆果生长期、浆果成熟期、采果期和休眠期,对湖南长沙种植的“美人指”‘香妃”‘红罗莎里奥”“爱神玫瑰”“白罗莎里奥”“甲斐乙女”“红地球”7个欧亚种品种和“摩尔多瓦”“金手指“阳光玫瑰”“夏黑无核”4个欧美种品种的冬芽主芽坏死情况解剖观测和统计分析.结果表明:湖南长沙和山东济南种植的“红地球”葡萄冬芽均存在主芽坏死现象,且湖南长沙更严重.湖南长沙11个葡萄品种均出现了主芽坏死现象,且品种间差异有显著性(p＜0.05),欧亚种的坏死程度普遍比欧美种严重.主芽坏死一经形成不可逆转,主芽坏死发生高峰期为第一次膨大期和浆果成熟期.结果枝主芽坏死率较营养枝高,且各品种结果枝主芽坏死率较高的节位均集中在第7～8节位.分析认为,品种基因型、品种属性、树体营养不足、较高的赤霉素含量均可能是影响葡萄冬芽主芽坏死的主要因素.

利用SSR分子标记技术,对刺葡萄与玫瑰香、金手指等葡萄品种种间杂交的64株后代进行真实性鉴定,筛选出在双亲间有特异性位点的SSR引物.结果表明,64株杂交后代中有61个单株具有父母本特异性条带,结合田间形态学观察,确定为双亲杂交后代.因此,SSR标记可以有效地对葡萄属种间杂交后代进行真实性鉴定,可以作为刺葡萄种质创新的有效辅助手段.

以腺枝葡萄新类型为试材,采用田间调查与果实品质分析等方法,研究了腺枝葡萄新类型的物候期及相关农艺性状.结果表明:腺枝葡萄新类型果实成熟期比刺葡萄推迟1个月左右,果皮紫黑色,果肉及其汁液均为淡紫红色,甜酸适口,有淡淡青草香味,可溶性固形物含量达21.0％.该类型栽培性状良好,花芽分化能力强,抗病性强,农艺性状均优于腺枝葡萄,为晚熟酿酒葡萄新类型,且实施高效无公害化栽培技术有利于在湖南推广种植.

A Preliminary study was conducted on the botany characteristics, phenology period, bear fruit traits and disease resistance of the new type of variation of Summer Black Seedless grape. The results indicated that the new type of variation of summer black seedless grape hybrids for Europe and American. The ripening period is about 20 days earlier than that of the Summer Black Seedless grape. The peel is purple, the flesh and its juice are red, sweet and sour taste, and strawberry flavor is thicker, soluble solids Content of 19.8%~21.0%. In conclusion, this is a new type of special early fresh grapes with excellent agronomic traits. This paper summarized the efficient and pollution-free cultivation techniques of the new type of variation of Summer Black Seedless grape

[Objective] Vitis davidii Fo(e)x.is a special wild resource in China,which belongs to the east Asian population and is mainly distributed in Hunan,Jiangxi and Fujian provinces.It is typically characterized with the rawhide sting on the new tip,petiole and veins and strong resistance to humidity,high temperature and diseases.The fruits are rich in nutrition with abundant flavonoids (mainly anthocyanins),resveratrol,superoxide dismutase,vitamin C and other active ingredients,which is believed functionally beneficial to the health of human.Traditional research on the diversity of the V.davidii Fo(e)x.was mainly based on morphological features and biological characteristics with low accuracy.Therefore,ISSR marker system was established and employed to analyze the diversity of the species for providing the basis for protection and utilization.[Methods]52 samples of V.davidii Fo(e)x.(41 from Hunan province,1 from Fujian province,10 from Jiangxi province) and 8 varieties or accessions of the other species of Vitis were used as the materials.ISSR primers were selected at Canada Columbia University for PCR amplification.The genomic DNA was extracted by modified CATB method.Firstly,the healthy leaves were swabbed with 75％ alcohol and freezing liquid nitrogen and were stored in a refrigerator at-70 ℃.Then the dried leaves were treated by CATB-free buffer (0.2 mol· L-1 Tris-HCl,pH 8.0;50 mmol· L-1 EDTA,pH 8.0;0.25 mol· L-1 NaCl) and ice bath for 30 min;1 μL extract DNA (pH from 8.0 to 10.0) with 1.5 μL bromophenol blue solution (2.5 g· L-1 bromophenol blue,2.5 g· L-1 xylene cyanol,400 g· L-1 saccharose) of each sample was taken for electrophoresis (0.8％ agarose gel) with TE buffer(10 mmol· L-1 Tris-HCl,1 mmol· L-1 EDTA,pH 8.0).The concentration of Mg2+,dNTPs,primer,DNA template and the number of amplification cycles suitable for ISSR-PCR amplification reaction of V.davidii Fo(e)x were optimized using the multifactor and multilevel experiments.The PCR reaction cycling profile was modified as 94 ℃ for 5min,94 ℃ for 30 s,54-58 ℃ annealing for 1 min,72 ℃ for 5 min followed by 35 cycles,and a final extension at 72 ℃ for 7 min.SPSS 17.0 software program was employed for clustering analysis of the V.davidii Fo(e)x.The clustering figure was also established.[Results] The optimum ISSR-PCR reaction system was as follows:in the total reaction system with amount of 20.0 μL,there were 4 μL 10×PCR Buffer,1 UT Tag enzyme,2.5 mmol· L-1 Mg2+,0.25 mmol· L-1 dNTPs,0.6 μmol· L-1 primer,30 ng DNA template,respectively.In optimization of ISSR-PCR reaction system,13 primers from 100 arbitrary primers were selected.116 polymorphism bands among 139 amplified bands were obtained with high quality,purification and stability.The rate of polymorphism was 83.45％.The primers UBC807 and UBC834 had got the highest number of polymorphic loci (14 loci).According to genetic similarity coefficient clustering analysis,the 8 materials of the control were outside the V.davidii Fo(e)x species,The genetic distance between the V.pseudoreticulata W.T.Wang and V.davidii Fo(e)x was near.All the 52 samples of V.davidii Fo(e)x could be divided into three subgroups.[Conclusion] Vitis germplasm resources had abundant genetic diversity.ISSR markers could effectively reveal the genetic diversity and genetic relationship of the V.davidii Fo(e)x.The results would be beneficial to the protection and and utilization of the species.

Red globe grape was used as material, the process of flower bud differentiation was observed with the paraffin section method. The results indicated that the period for the main flower bud differentiation of winter buds of red globe grape can be divided into four stages: the Nondifferentiation (before June 3rd), Inflorescence perimordium differentiation (June 3rd to July 8th), Spikelet perimordium differentiation (July 8th to August 12th) and the period of dormancy.

Potassium is essential for vine growth,enzyme activation,photosynthesis,osmotic regulation,yield and quality.Potassium is one of the most abundant cations in grapes,the potassium ion concentration in cytoplasm can reach 100-200 mmol·L-1,even up to 500 mmol·L-1 in apoplast.Potassium as a highly mobile element,can easily transfer from old tissue to a young area,and primarily concentrate in the center of the active part,such as the growing point,buds,berries and so on.Grape berries are a strong sink for K,and the potassium content has been rising in the whole growth period.Plant nutrient analysis results show that:more K is accumulated particularly during ripening.In the mature period,the potassium ion can account for about 70％ of the total cation content and 66％ of the total potassium content of aboveground organs.Among berry tissues,K concentration per unit fresh weight is higher in the skin than in the fleshy pericarp.The K concentration in the seeds is lower than in the skin but slightly higher than in the flesh.K concentration in the skin is 1.7-6.9 times higher than that in the pulp and 1.6-4.3 times higher than in the seeds.The concentration in the skin can reach 50％ of the whole berry potassium concentration.Potassium transportation is done by ion channels.There are low affinity K+ absorption systems and high affinity K+ absorption systems in higher plants on the absorption of potassium,the former done in external high K+ concentration and the latter in low K+ concentration.In grapes,potassium ion channels belong to the Shaker gene families,and VvKUP 1 and VvKUP 2 are two genes encoding potassium transport carriers and are found in larger expressions before veraison and lower expressions after veraison.The expression of these carriers is restricted to the skin,the tissue in the berry with the highest K concentrations.Researches on the other types of K+ transport mechanisms in different berry tissues may better help in understanding the mechanism of berry K accumulation.Potassium movement occurs in both xylem and phloem.In grape berries,the xylem seems to be a minor route of K entry because xylem flow into the berry may be low due to the low transpiration rate of the berry.Potassium is the major cation in the leaf xylem sap and shoot xylem sap.While K concentration differs significantly with leaf age,leaf xylem sap flow is highest in the middle leaves and lowest in the old and young leaves.At the beginning of grape growth,potassium reflux occurs in the plant through the xylem,and remobilization of K from other organs to the berries can be reabsorbed into the phloem by apoplast.Between the veraison and mature period,the K+ concentration being gradually reduced in the different organs shows that potassium is being retransported into the berries.Many factors may affect K accumulation in berries including soil,climate,variety,phytohormones and cultural practice.K deficiency may be induced in saline conditions where Na+ is the predominant cation.Gravel soil is better for absorption of the potassium than sand and clay;high temperature,and shading treatment promotes the potassium accumulation in fruit;varieties differ on the potassium absorption efficiency,while higher efficiency is found in ‘ Cabernet Sauvignon’ than in ‘ Cabernet Franc’.Cultivation measures affect the potassium distribution and accumulation in berries.Canopy management is a technique that results in the altered position and density of leaves,shoots,and fruit to achieve a specific desired arrangement.Reasonable leaf canopy management helps to decrease excessive potassium accumulation in the pulp,and reduces the pH of the wine.Crop load affects berry K accumulation by changing the balance of the source/sink for K.It may also have indirect effects on berry K accumulation through changes in the degree of shading in the canopy,availability of assimilates and rates of berry growth.The effects of the K addition on berry K accumulation are variable,with more K being accumulated when more fertilizers are applied.Results from other studies indicate that berry,juice,or wine K is not a positive correlation to potassic fertilizer supplement.There are data suggesting that increased irrigation tends to increase berry K accumulation.Berry and wine K concentrations were higher for irrigated treatments than for non-irrigated treatments.Irrigation enhances the dissolution of K from clay particles and its movement in the soil solution,which facilitates its supply to roots and provides higher uptake.Potassium accumulation in berries is a very complicated process.Comprehension of berry K accumulation requires an understanding of the function of K in the berries,how berry K requirements are determined,and how K is translocated into and out of the berry,while the available information is still unknown and limited.This review summarizes the potassium nutrition from the distributions,functions of potassium,potassium characteristics and uptake in grape,emphasis on the berry accumulation and relative impact factors,to better provide the basis for regulating nutritional balance in grape berries and producing scientifically.

本文以水晶刺葡萄叶片和新梢为试验材料,采用超声波辅助提取白藜芦醇,高效液相色谱(HPLC)法检测其含量,并分别对烘干温度、破碎度对其白藜芦醇含量的影响进行研究.结果表明,水晶刺葡萄样品前处理在50~100℃范围内最佳烘干温度为90℃,在30~100目范围内最佳粉碎度为40目,在此处理下,水晶刺葡萄叶片白藜芦醇含量达到6.22±0.34μg/g,水晶刺葡萄新梢白藜芦醇含量达到11.23±0.29μg/g.

本文以腺枝葡萄、刺葡萄、毛葡萄、夕阳红、金星无核、红宝石无核为试材,运用LI-6400便携式光合作用测定系统测定了5个葡萄种类品种的净光合速率、气孔导度、胞间二氧化碳浓度、蒸腾速率等,结果表明:腺枝葡萄的净光合速率比普通栽培品种夕阳红要高,且不表现"午休现象".腺枝葡萄在光合性能方面比葡萄栽培品种具有更优越的表现,可以作为高光合育种材料.

以南方特色刺葡萄资源湘珍珠和紫秋为试验材料,结合现有刺葡萄的3种主要栽培架式:篱棚一体架、水平棚架和双十字V形架,对不同品种、不同栽培模式下果实的内在品质、外观品质及糖酸类物质代谢进行分析.结果表明:水平棚架栽培的刺葡萄无论是在产量、穗重和穗长等外观品质,还是在总糖、总酸、花青素含量等内在品质方面都显著优于篱棚一体架和双十字V形架.在刺葡萄成熟后不同时间采收发现,糖类物质代谢呈上升趋势,其中水平棚架中糖含量最高,积累速度较快;在3种架式中,酸类物质代谢速度明显低于糖类物质,酒石酸的含量与增长速度无显著差异,苹果酸含量则呈下降趋势.

The downy mildew resistance from 20 cultivars of table grapes and seveal progenies ofVitis davidii Fo?x were investigated by the method which the indoor leaf discs were inoculatedin vitro. The identification results showed that Golden Queen, Xiazhihong, Ruby Seedless, Giant Roses, Xiangfei characterized by higher resistance. Specially, the Golden Queen showed the highest resistance. Several other species of susceptible varieties, Otilia Seedless showed the highest susceptible. The results would provide certain reference to improve the downy mildew resistance of the progeny ofVitis davidiiFo?x.

25 samples ofVitis (21 from Hunan province, 1 from Fujian province, 3 from Jiangxi province) were used as materal, the characteristics of leaf, flower, fruit, and seed as index, to analyze the 16 classification traits by the variation and principal components analysis of taxonomic traits. The results showed that three groups of theVitis, Lixian–4 and Jishou–1, Lixian–5 and Jishou–3, Zhijiang60Co and Wangcheng, had the same foliar morphological codes of three lobed margin or entire, while the otherVitis had different foliar morphological codes. According to the ratio of polar and equatorial axis and ovary morphology, the ovary of male flower had a shape of degraded and irregularly, while the ovaries of bisexual and female flowers had four categories in morphology, namely narrow conical, narrower conical, wide conical and wider conical, which the wide conical was the majority. TheVitis had big difference in cluster weight and berry weight, the berries were round, the pericarp colors were green and pink, purple and dark purple. The seed of the Vitis was long oval with no horizontal furrow. Acorrding to the variation analysis of taxonomic traits, the variation range of 16 classification traits was 5.03% to 28.52%, the average variation coefficient was 12.46%, which berry weight was the greatest (28.57%), while the seed length was the smallest (5.03%). According to principal components analysis, the morphological characters of fruit grain was the first, the second and third was the shape of the leaf and the fourth was the morphological characters of seed. The total contribution of the four ingredients was 83.168%.

UsingV. davidii Fo?x including Jiangxi Junzi Vally, Huaihua Bai and Zhejiang Bai as scion,V. Vinifera including Red Globe, Heifeng, Guibao and Beta as rootstock, to determine and compare with each grafting combination's affinity, related morphological indexes and physiological indexes, and observeing in resistance of leaves and fruits. The results indicated thatV. davidii Fo?x Zhejiang Bai/Beta was better than other grafting combination; about 40~65 days after flowering, total chlorophyll content up to the maximum value, after that rendering a downward trend; Chlorophyll a/b value rendered the tendency of first declined and then increased; with the growth and development of the leaves, carotenoids' content rendered downward trend, reaching the lowest value at about 60 d after flowering, then it raised, again it rendered downward trend; the incidence and disease index of downy mildew and anthrachose of grape were continued to decline throughout the entire disease cycle trends, using Beta and Hutai 8 as rookstock, the incidence and disease index of two kinds of disease were lower.

To screen V. davidii Foex varieties with high photosynthetic efficiency,16 samples were grafted onto six-year-old red globe grape trees as the base stock of SO4,the red globe plants as the control,the net pho?tosynthetic rates of different varieties of top grafting V. davidii Foex were determined.The leaf chlorophyll content and net photo synthetic rates of V. davidii Foex varieties were closely related,and the chlorophyll b content had the most significant effects on the net photosynthetic rate of V. davidii Foex varieties.The intercellular CO2 con?centration and transpiration rate were the major fectors in the level of net photosynthetic rate in V. davidii Foex varieties.From the comprehensive assessment,two V. davidii Foёx germplasm resources were screened.V. davidii Foёx shuijing was a type of high photosynthesis efficiency,high light utilization and water use efficiency.V. Davi?dii Foёx Zhijiang Co60 was a type of low consumption.V. clone Foёx Liuyang♂ was a type of small blade,early bloom,and mesh surface.

In the experiment, 16 samples of Vitis davidii(12 from Hunan province, 3 from Jiangxi provience and 1 from Fujian provience ) and Vitis pseudoreticulata W.T. (as CK1) and Vitis adenoclada Hand. Mazz (as CK2) were used as materials to study the phenological period and agronomic trait and cutting rooting of Vitis. The results showed that the phenological period of all samples were earlier than CK1 and CK2, except the sample from Fujian, which the coloring stage was later than CK1. The samples from Hunan had the earliest in germinaion, shoot-growing, flower bud period, maturity period and defoliation period, while the stage of physiological drop and fruit softening had little difference among the samples. All samples had similar agronomic trait, such as, the heart and simple shallow crack or entire edge, the irregular tendrils, conical inflorescence, tender tip with brownish yellow, green or light purple, new tip with brown. One of the samples from Jiangxi (Jiangxi–9) had the best cutting rooting, which had 38.67% of cutting’s survival rate, 5.9 roots and 4.57 mm of root thickness.

为培养园艺专业人才的综合素质和就业竞争力,培养符合社会需求的园艺专业高技术人才,校地合作是高校人才培养模式的一种有效教育形式,是高校人才培养的重要途径.本论文分析了园艺专业人才校地合作培养模式的重要性、校企合作人才培养面临的问题,及提出了园艺专业人才三种培养模式:新农村发展研究院建设背景下校企合作人才培养模式、"项目驱动"式人才培养、"引厂入校"人才培养模式.