油菜素内酯和脱落酸调控葡萄果实花色苷合成的研究

【目的】为研究2,4-表油菜素内酯(2,4-epibrassinolide,EBR)和脱落酸(abscisic acid,ABA)处理对葡萄花色苷合成与可溶性固形物含量、果皮苯丙氨酸解氨酶(PAL)和类黄酮糖基转移酶(UFGT)活性以及果实内源ABA含量的影响,探索油菜素内酯调控葡萄果实成熟及花色苷合成的机理。【方法】以酿酒葡萄‘赤霞珠’(Cabernet Sauvignon)和‘烟73’(Yan 73)为试材,在葡萄转色前分别用0.1、0.4、0.8 mg.L-1EBR,1mg.L-1Brz(brassinazole,BR生物合成抑制剂)和200 mg.L-1ABA,均匀喷施于葡萄果实,在葡萄成熟过程中测定葡萄果皮花色苷含量及PAL和UFGT酶活性,同时测定果实ABA和可溶性固形物含量。【结果】在果实着色初期,‘赤霞珠’和‘烟73’葡萄果皮PAL和UFGT活性及果实内源ABA含量均逐渐升高,当果实接近成熟花色苷含量基本稳定时ABA含量开始降低。与对照相比,0.4mg.L-1EBR和200 mg.L-1ABA处理显著增加了果实内源ABA含量,提高了果皮PAL和UFGT活性,促进了果皮花色苷的合成和果实可溶性固形物的积累。0.1 mg.L-1和0.8 mg.L-1EBR处理总体增加了果实内源ABA含量,促进了花色苷的合成和可溶性固形物的积累,并提高了UFGT和PAL酶活性,但比0.4 mg.L-1EBR处理提高的幅度小,且差异显著。1 mg.L-1Brz处理使果实ABA的合成推迟,果实可溶性固形物含量、果皮PAL和UFGT活性以及花色苷含量均低于对照,但差异不显著。【结论】外源EBR和ABA处理促进了葡萄成熟和花色苷合成,在不同浓度EBR处理中,以0.4 mg.L-1处理效果较好;内源ABA可能参与了EBR对葡萄成熟和花色苷合成的调控。     

葡萄种质资源果形性状的分析

通过分析葡萄种质的果形性状以及不同类型的果形特点,为认识葡萄种质资源果粒形状特点和育种亲本选择提供一定依据。选取820份葡萄种质材料（286份的性状来自田间实地调查,534份来自《中国葡萄志》和《中国葡萄品种》记载）,对不同类型、种（species）和用途的葡萄果形特点、不同果形单果质量的分布、以及质量与其纵径和横径的相关性、不同年代选育不同果形的数量等进行分析。结果表明,倒卵圆形和长椭圆形的果粒较大,而圆形的果粒较小。欧亚种葡萄中长椭圆形和卵圆形的较多,欧美杂种葡萄中倒卵圆形的较多。鲜食葡萄较酿酒、制干和制汁葡萄果形更丰富且更狭长。黑色葡萄以圆形和近圆形为主,白色和红色葡萄中果形较为丰富。从育种进程来看,早期选育的葡萄果形以圆形为主,之后变为以圆形、近圆形和椭圆形为主,现代越来越多果形相继出现。圆形、卵圆形和倒卵圆形果粒的横、纵径与果粒质量拟合度较好。同一个葡萄品种果粒形状可能存在差异,其中椭圆形变化较为普遍。     

葡萄离体叶片和果实白藜芦醇及其糖苷合成对UV-C辐射诱导的响应

以‘北丰’和‘北全’葡萄离体叶片和果实为试材,研究了葡萄不同器官内白藜芦醇（Res）及其糖苷合成对紫外线C（UV-C）辐射诱导的响应,结果表明：不同器官反式白藜芦醇（trans-Res）合成积累对UV-C辐射诱导反应存在差异。叶片适宜的UV-C辐射剂量为1.8 kJ · m-2,而果皮为1.8 ~ 5.4 kJ · m-2,果实Res合成积累对UV-C辐射诱导的敏感性低于叶片。3.6 kJ · m-2 UV-C辐射果粒对种子中trans-Res含量没有显著影响,直接辐射完整种子或剖开种子切面也不能显著提高trans-Res含量。UV-C辐射处理的叶片和果皮中trans-Res含量随辐射后常温避光孵育时间的延迟表现出先升后降的趋势,而反式白藜芦醇苷（trans-PD）和顺式白藜芦醇苷（cis-PD）含量在12 ~ 48 h内迅速提高,之后维持在相对稳定状态。种子中trans-Res含量在整个孵育过程中与对照之间均不存在显著差异。     

查耳酮异构酶在葡萄果实发育过程中的表达及亚细胞定位研究

以‘赤霞珠’葡萄果实为材料,利用制备好的葡萄查耳酮异构酶(CHI;EC 5.5.1.6)CHI多克隆抗体,采用RT-PCR、蛋白质印记杂交和胶体金免疫电镜技术对葡萄果实发育过程中CHI基因、蛋白表达及亚细胞定位进行了研究。结果表明:CHI基因和蛋白的表达随着果实发育有着明显的变化,在果实发育早期和转色期表达量较高,同果实中总类黄酮含量的积累变化一致;胶体金免疫定位显示在果实发育早期CHI主要分布于葡萄果皮细胞的细胞质中,在液泡及质体中也有少量分布;在转色期主要定位于细胞质、质体和细胞核中,其数量明显增多。在成熟果实中,主要存在于细胞质,少量存在于质体和液泡中。     

UV-C辐射对盆栽葡萄北丰叶片和邻近果穗果实白藜芦醇及其糖苷质量分数的影响

以1a生北丰(Vitis thunbergii×V.vinifera)葡萄盆栽结果树为试材,在果实始熟期采用紫外线C(Ultraviolet C,UV-C)辐射结合韧皮部环剥的方法,研究了UV-C辐射对葡萄叶片和邻近果穗果实白藜芦醇(Resveratrol,Res)及其糖苷质量分数的影响。结果表明,UV-C辐射诱导葡萄果实Res合成积累的敏感性低于叶片,辐射葡萄叶片对邻近果穗上果皮Res的影响远远小于UV-C直接辐射葡萄果穗,且辐射诱导葡萄果皮中Res质量分数与叶片中Res质量分数存在着一定的相关性。当叶片位于果穗下方时,叶片中反式白藜芦醇(trans-Resveratrol,trans-Res)、反式白藜芦醇苷(trans-Piceid,trans-PD)和顺式白藜芦醇苷(cis-Piceid,cis-PD)质量分数表现出环剥处理显著高于不环剥处理,然而果皮中的结果正好相反。当叶片位于果穗上方时,果皮中trans-Res、trans-PD和cis-PD质量分数表现出不环剥处理显著高于环剥处理,暗示葡萄Res可能存在韧皮部运输途径,且运输方向是双向的。此外,UV-C直接辐射葡萄果穗处理果皮trans-Res和cis-PD质量分数显著高于对照和UV-C辐射葡萄叶片处理。种子中仅检测到trans-Res,且仅环剥处理和辐射下方叶片不环剥处理显著高于对照。     

无核葡萄种质果粒遗传多样性及数量性状评价标准探讨

 对国家果树种质资源郑州葡萄圃内的94 份无核葡萄和40 份巨峰系葡萄的果粒颜色、形状、质量和大小进行了连续3 年的调查和统计分析。结果表明, 无核葡萄种质果粒质量和果粒大小遗传变异系数比巨峰系大得多, 无核葡萄种质的平均果粒质量主要在110～510 g 的范围内, 将近50 %无核葡萄种质的果粒为115～215 g , 为中等。因此目前国际及国内的现有描述评价标准都不适合评价无核葡萄种质的果粒质量及大小。根据本项研究结果提出了适合无核葡萄种质新的附加分级评价标准。     

葡萄品种资源裂果性状调查与分析

选取252 个鲜食葡萄品种（111 个欧美杂种,141 个欧亚种）,分别在转色期、成熟期、过熟期采集果实,通过清水浸泡试验,了解比较其裂果情况。结果表明,葡萄裂果症状有环裂、纵裂、纵环裂和龟裂等,龟裂只在欧亚种葡萄出现;欧亚种葡萄比欧美杂种葡萄裂果严重;成熟期裂果最严重;欧美杂种小果粒葡萄品种更容易裂果;果皮厚、果肉质地软的葡萄不易裂果。不同葡萄品种的裂果倾向性存在较大差异,通过研究清水浸泡后葡萄发生裂果的情况,对葡萄裂果有了初步认识,为葡萄品种选择及葡萄育种亲本选择提供一定理论支撑。     

葡萄果实发育后期脱落酸来源的研究

  以‘品丽珠’(Vitis vinifera L. cv. Cabernet-Franc) 葡萄为试材, 对发育后期果实中脱落酸(ABA) 的来源进行了研究。结果表明, 叶片合成的ABA 可能是果实中ABA 的重要来源, 而根部ABA 的合成对果实中ABA 含量贡献不大。种子中ABA 的合成可能不是果肉中ABA 的重要来源。果实具有独立合成和降解ABA 的能力。对果实内ABA 含量变化的代谢调控问题进行了讨论。     

Enzyme activity changes during anthocyanin synthesis in ‘Olympia’ grape berries

Anthocyanin synthesis began in the skin of ‘Olympia’ (Vitis labruscana Bailey) grape when the berry growth reached a late stage in the second sigmoid growth-curve. During berry development, shikimate NADP oxidoreductase (SORase, EC.1.1.1.25), phenylalanine ammonia-lyase (PAL, EC.4.3.1.5), and chalcone flavanone isomerase (CHFI, EC.5.5.1.6) were investigated in the skin. These enzyme activities were high during early development of the young berry, but rapidly decreased thereafter, and were activated again when rapid anthocyanin synthesis subsequently occurred. When PAL from the young berry was analyzed by isoelectricfocusing polyacrylamide gel electrophoresis (IEF-PAGE), it showed a single activity band with a pI value of approximately 5.4, whereas PAL from the ripe berry showed two activity bands with pIs of 5.4 and 6.5. Thus, SORase, PAL, and CHFI seem to be involved in anthocyanin accumulation simultaneously, and newly synthesized isozyme of PAL may play a key role in pigment formation in ‘Olympia’ skin. The function of these enzyme activities in young berry is also discussed in this text.     

几种配方肥对“巨峰”葡萄坐果率与果实品质的影响

以欧美杂种鲜食葡萄"巨峰"为试材,研究了配方肥花多乐、果多乐以及养多乐不同喷施浓度及喷施次数对葡萄坐果率以及果实品质的影响。结果表明:初花期喷施花多乐1次可提高"巨峰"葡萄坐果率;幼果期喷施果多乐600倍液2次或1 200倍液1次可显著增加葡萄单粒重、单穗重、果实横径以及果实含糖量;喷施花多乐、果多乐、养多乐均可提高"巨峰"果皮中花色素含量。     

养多乐对酿酒葡萄“赤霞珠”果实品质的影响

以欧亚种酿酒葡萄"赤霞珠"为试材,研究了养多乐(Sugar mover)不同喷施次数与喷施浓度对酿酒葡萄果实品质的影响。结果表明:随着养多乐喷施次数的增加,其果实还原糖含量增加。喷施1 200倍液养多乐"赤霞珠"果实还原糖最高。随着养多乐喷施次数的增加,"赤霞珠"果实总花色素含量增加,连续喷施4次600倍养多乐效果最佳。喷施养多乐对"赤霞珠"果实总酚含量影响不大。     

Application timing and concentration of abscisic acid affect the quality of ‘Redglobe’ grapes

Summary

The plant hormone abscisic acid (ABA), the concentration of which increases in grape berry skins at the onset of maturation (veraison), appears to be involved in the regulation of anthocyanin accumulation. Preliminary tests suggested that exogenous applications of ABA could improve grape berry colour, but its high cost precluded the development of commercial applications. Recently, a lower-cost ABA production method was developed, which led to the evaluation of different concentrations of ABA, applied at or around veraison, on the quality of ‘Redglobe’ grapes. In two of three years of tests, several ABA treatments enhanced the anthocyanin content of grape skins. ABA, applied at approx. 300 mg l^–1 at veraison, may be required to reliably increase pigmentation, and improve the colour of ‘Redglobe’ grapes. Although the total anthocyanin content was increased by ABA treatment, anthocyanin composition was not affected. Applications of ABA had few effects on fruit size or composition, although they did cause fruit softening, which is undesirable. A secondary objective of this study was to determine how the anthocyanin content of berry skins affected berry colour characteristics. Strong curvilinear relationships between anthocyanin content and lightness and hue showed that these colour characteristics were saturated by anthocyanin contents over 0.02 mg cm^–2 of skin. These data suggest that colour measurements may be needed to evaluate the effect of cultural practices on colour in table grapes.     

玫瑰香葡萄生长期酚类物质含量及抗氧化活性的变化

 以玫瑰香(Muscat Hamburg)葡萄为试材,通过测定不同生长发育期酚类物质含量和其清除自由基的能力,对酚类物质的动态变化及其与抗氧化活性的关系进行了研究。结果表明,叶片、茎、卷须的总酚,类黄酮和原花色素含量随果实的发育呈先下降后上升的趋势,在果实快速生长期达到最低点,之后有所增加。叶片酚类物质含量最高,茎中含量最低。果皮酚类物质含量的变化趋势与叶、茎、卷须中相同,在果实成熟期达最低点。果肉中酚类物质含量极低,且在整个生长期内无明显变化。各器官的抗氧化活性与酚类物质含量呈正相关。     

Dynamics of anthocyanin and flavonol profiles in the ‘Crimson Seedless’ grape berry skin during development and ripening

‘Crimson Seedless’ grapes (Vitis vinifera L.) do not develop adequate berry colour in different parts of the world including Australia and USA leading to serious economic losses to the growers. In the present study, various anthocyanins and flavonols were identified in the skin of the ‘Crimson Seedless’ grape berries using LC/PDA/ESI-MS and their changes in the berry skin during development and ripening of ‘Crimson Seedless’ grape berries were investigated during 2005–2006 and 2006–2007. Eleven anthocyanins and two flavonols were identified in the berry skin using LC/PDA/ESI-MS. Of the anthocyanins identified, four anthocyanins including cyanidin 3-O-(6″-O-acetyl)-glucoside, peonidin 3-O-(6″-O-acetyl)-glucoside, malvidin 3-O-(6″-O-acetyl)-glucoside and malvidin 3-O-(6″-O-coumaroyl)-glucoside were not reported earlier. During both the years, the concentration of the 3-O-glucosides of delphinidin, petunidin, peonidin, and malvidin as well as the acetyl and coumaroyl esters of the 3-O-glucosides of cyanidin, peonidin, and malvidin in the berry skin increased during berry development and ripening. During 2006–2007, the concentration of cyanidin 3-O-glucoside in the berry skin increased during the early stages of berry ripening and subsequently declined till harvest while in 2005–2006, the concentration increased during the initial phase of berry ripening and remained relatively stable thereafter till harvest. The concentration of total anthocyanins in the berry skin was higher during 2006–2007 as compared to 2005–2006. During both years, the concentration of quercetin 3-O-glucoside in the berry skin increased during berry development and ripening while the concentration of quercetin 3-O-glucuronide in the berry skin decreased during the same period. To the best of our knowledge, this is the first report on the evolution of different anthocyanins and flavonols in the ‘Crimson Seedless’ berry skin during berry development and ripening.     

根域限制对巨玫瑰葡萄果实外观、色素及内在品质的影响

针对巨玫瑰葡萄在浙江存在的着色差、品质下降等问题对其进行了根域限制栽培试验。结果表明根域限制栽培可以明显改善果实着色,增加果皮花青苷、类胡萝卜素、类黄酮和总酚质量分数,使着色后期叶绿素质量分数有所下降。与花青苷合成相关的PAL酶(苯丙氨酸解氨酶)活性也明显升高。根域限制提高了果肉中的葡萄糖、果糖和蔗糖3种可溶性糖的质量分数;虽然对果肉中的苹果酸影响不大,但是降低了酒石酸的质量分数。另外,根域限制还显著提高了果皮、果肉和种子中白藜芦醇的质量分数。因此,根域限制栽培不仅改善了巨玫瑰的果实外观,而且提高了果实的内在品质。     

晚熟鲜食葡萄新品种‘华葡翠玉’

‘华葡翠玉’葡萄是从‘红地球’‘玫瑰香’杂交后代中选出的晚熟新品种。果穗圆锥形,平均单穗质量862.4 g。果粒椭圆形,黄绿色,平均单粒质量11.3 g。果肉硬脆,汁液较多,味甜,有玫瑰香味。可溶性固形物含量18.7%,可滴定酸含量0.51%,鲜食品质好。每果粒含种子3 ~ 4粒。果实成熟后不脱粒,耐贮运。在辽宁兴城地区果实10月上、中旬充分成熟。结果稳定,早果性强。极丰产,产量37 500 kg • hm-2。适合设施栽培。     

不同土壤条件对酿酒葡萄生理及果实品质的影响

【目的】为了探索不同土壤条件对酿酒葡萄生理及果实品质的影响,【方法】以‘蛇龙珠’酿酒葡萄为试材,研究了宁夏贺兰山东麓地区,黄河淤土、含石的沙壤土和风沙土3种土壤对葡萄糖分卸载的影响,测定了不同土壤栽培条件下葡萄果实的单果质量、可溶性固形物、可滴定酸、可溶性总糖、色素、单宁及葡萄叶片的光合速率。【结果】结果表明,在葡萄开始着色至葡萄完全成熟期间,在含石的沙壤土栽培条件下葡萄可溶性总糖含量(22.4%)比黄河淤土栽培条件下和风沙土栽培条件下分别高7%、9%;葡萄色素(5.14 mg.g-1)和叶片光合速率也最高,黄河淤土栽培条件下葡萄的含酸量(8.28 g.L-1)和单粒质量(2.32 g)较高,然而,风沙土条件下,单宁(籽单宁1.15 mg.g-1,皮单宁0.30 mg.g-1)的含量最高。【结论】土壤条件对酿酒葡萄果实品质影响较为显著,含石的沙壤土条件下葡萄品质最佳。     

金雀异黄素和环鸟苷酸调控离体葡萄果实花青苷积累

以离体‘巨峰’葡萄为材料,观察到金雀异黄素（GNT）对果皮花青苷积累的促进效应,而且证明GNT的诱导过程（约10 ~ 12 h）不需要光照,但其后的花青苷积累却依赖于光照。如果在GNT预处理后6 h内用环鸟苷酸（cGMP）处理,可以明显抑制GNT的促进效应,但在GNT预处理后12 h再用cGMP处理,则抑制效应消失。半定量RT-PCR检测表明,光和GNT诱导葡萄果皮花青苷合成相关基因PAL、CHS和UFGT表达量上调,而对LDOX表达量无明显影响;cGMP处理抑制了GNT诱导的PAL和CHS等基因表达的促进作用,说明GNT和cGMP相互拮抗,共同调控葡萄果皮花青苷积累。     

SO2对葡萄采后呼吸强度及内源激素的影响

采用气相色谱法和改进的高效液相色谱法研究发现：‘无核白’葡萄果粒为非跃变型呼吸类型，其乙烯释放量很低，但释放潜力很高，切伤后乙烯释放速率迅速增高；果梗和穗梗为跃变型呼吸类型，有很高的呼吸高峰和乙烯释放高峰。适量的ＳＯ２保鲜处理对葡萄的呼吸强度、脱落酸（ＡＢＡ）含量及乙烯释放具有明显抑制作用，对生长素（ＩＡＡ）、赤霉素（ＧＡ）有促进作用，既可抑制葡萄生理衰老，又可减轻葡萄的落粒，延长葡萄采后贮藏寿命     

Effect of bearing position on phenolics profiles in the skins of four cultivars of grapevine (Vitis vinifera L.)

Summary

The fan-shaped trellis training system, resulting in grapevines (Vitis vinifera L.) with more than two trunks, has been widely used in northern China. The fruiting zone of grapevines trained to the fan-shaped trellis system is distributed from the top to the bottom of the canopy. The phenolics profiles of the skins of ‘Cabernet Sauvignon’, ‘Merlot’, ‘Chardonnay’, and ‘Italian Riesling’ grape berries were analysed to measure the effect of differences in bearing position on the phenolic compound and anthocyanin compositions of grape berries.The results showed that the upper bearing position significantly increased the concentrations of most phenolic compounds in grape berry skins. Among the non-anthocyanin and anthocyanin phenolic compounds identified in this study, the upper bearing position promoted the accumulation of delphinidin-, cyanidin-, and petunidin-glucosides in the skin of the two red grape cultivars. However, lowering the bearing position was associated with greater methylation of anthocyanins, and the flow of photosynthate towards the biosynthesis of kaempferol. These results were correlated with micrometeorological alterations in the canopy at the different bearing positions. The findings of this study suggest that higher bearing positions can increase the accumulation of phenolic compounds in grape berry skins in four different grapevine cultivars.