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。     

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。     

Changes in sugar content and relative enzyme activity in grape berry in response to root restriction

Root restriction often depresses photosynthetic capacity and reduces shoot growth, but the photosynthate accumulation in fruit under restriction is higher than in control fruit. However, changes in metabolism responsible for these differences are unclear. To identify the metabolic mechanism by which root restriction affects sugar accumulation in fruit of ‘Kyoho’ grape (Vitis vinifera × V. labruscana), sugar metabolism and related enzyme activities in grape berries produced with and without root restriction were compared. Total sugar content of fruit produced under root restriction was higher than that of control fruit. Acid invertase (AI, EC 3.2.1.26) activity, which increases with berry development, was significantly higher in root-restricted berries than in control berries. Neutral invertase (NI, EC 3.2.1.26) activity showed a similar trend to AI, but the amount of NI activity was lower than AI in both treatments. Sucrose phosphate synthase (SPS, EC 2.4.1.14) and sucrose synthase (SS, EC 2.4.1.13) activity changed slightly with berry development, and there was no significant difference in SS and SPS activity between root restriction and control treatments. Therefore, AI appears to be the key enzyme induced by root restriction that explains the higher sugar content found in grape berry produced under root restriction.     

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.     

In vitro propagation of Symphytum species

Methods are described for the in vitro propagation of Symphytum × uplandicum Nyman from bud, root and stem explants. Highest shoot numbers were produced from root explants > 4 mm in diameter, cultured vertically with their distal cut surface on the medium. The most suitable medium for shoot production was Murashige and Skoog's (MS) with 0.3 mg l⁻¹ 6-benzylaminopurine (BAP). These shoots developed roots on MS medium without hormones and were successfully transplanted into pots. Subculturing shoots onto MS medium containing BAP, kinetin (K), 6-λ,λ-(dimethylallylamino)-purine or gibberellic acid (GA₃) at 0.5, 1.0, 5.0, 10.0 or 30.0 mg l⁻¹ failed to stimulate outgrowth of axillary buds in culture. In vitro propagation from root explants was also achieved with S. asperum Lepech., S. officinale L. and S. × uplandicum cultivars ‘Bocking 1’, ‘Bocking 2’, ‘Bocking 4’ and ‘Bocking 17’, but not with S. bohemicum, S. grandiflorum DC, S. tuberosum L. or S. × uplandicum ‘Bocking 7’ and ‘Variegatum’.     

红富士苹果采后1-MCP 处理对果皮色素及其相关酶活性的影响

 以‘红富士’苹果为试材,研究了采后1–甲基环丙烯（1-MCP）处理对冷藏期间果实硬度, 叶绿素、类胡萝素及花青苷含量变化的影响,并重点研究了花青苷含量变化与其合成相关酶（PAL、CHI、 DFR、UFGT）和降解相关酶PPO、POD 活性变化的关系,结果显示：1-MCP 处理能抑制叶绿素降解及 贮藏前期花青苷的积累,延缓类胡萝素的积累及花青苷的后期降解;贮藏期间花青苷的合成与CHI、DFR 活性不相关,与PAL、UFGT 活性正相关;贮藏期间花青苷降解相关酶PPO、POD 活性都呈现先上升后 下降的趋势,对照的PPO 活性始终高于1-MCP 处理,1-MCP 处理明显抑制贮藏前期果皮POD 活性的上 升及贮藏后期POD 活性的下降。     

Hardy framework influence (1969–1983) on yield, tree size and yield efficiency of ‘Wellspur’ and ‘Goldspur’ apple

Cumulative yield, trunk cross-sectional area, and yield efficiency were measured between 1969 and 1983 in an experimental planting (established in 1964 and top-worked in 1965) of ‘Wellspur’ and ‘Goldspur’ apple top-worked on frameworks of 19 cold-hardy cultivars (‘Alnarp 2’, ‘Antonovka’, ‘Beacon’, ‘Canada Baldwin’, ‘Delcon’, ‘Dr. Bill’, ‘Duchess’, ‘Haralson’, ‘Hawkeye Greening’, ‘Heyer 12’, ‘Heyer 20’, ‘J. Luke Seedling’, ‘McIntosh’, ‘Minnesota 447’, ‘Ottawa 271’, ‘Ottawa 292’, ‘Red Astrachan’, ‘Robusta 5’, ‘Yellow Transparent’) that were grafted on domestic seedling roots. There were significant differences in yield, trunk diameter and yield efficiency related to the interstock. ‘Duchess’ and ‘Heyer 12’ ranked highest in yield efficiency and lowest in tree size. ‘Red Astrachan’ ranked lowest in efficiency and largest in tree size. ‘McIntosh’ also tended to be low in efficiency. No severe winter temperatures occurred to test differential cold hardiness.     

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。     

Variations in contents of browning substrates and activities of some related enzymes during litchi fruit development

Pericarp browning is a major factor to limit storage and transportation of postharvest litchi fruit. Experiments were conducted firstly to determine variations in contents of browning substrate (−)-epicatechin in pericarp tissues of litchi fruit during development. Changes in the activities of polyphenoloxidase (PPO), peroxidase (POD) and phenylalanine ammonialyase (PAL) in association with the synthesis or oxidation of (−)-epicatechin were also examined. (−)-Epicatechin content tended to decrease during fruit development. PAL activity had a high level at the initial stage of fruit development, then decreased rapidly, and finally increased slightly, which was in agreement with the variation in (−)-epicatechin content. The higher contents of total phenolics and (−)-epicatechin of fruit of ‘Nuomici’ than ‘Feizixiao’ were also related to the higher PAL activity. PPO had a similar change to PAL in activity, but POD activity slowly decreased at the early fruit development and then increased markedly. It is suggested that (−)-epicatechin content might be largely regulated by PAL activity. This study could help to understand the variations in the content of the browning substrate and the activities of some substrate-related enzymes of litchi fruit during development and then control better pericarp browning of harvested litchi fruit by the application of appropriate preharvest treatments.     

不同颜色果袋对李果实着色及花色素苷合成的影响因素分析

以大石早生李为试材,研究了不同颜色的果袋对李(Prunus salicina Lindl.)果实外观色泽、花色素苷含量的影响,并探讨了果实中糖酸含量、PAL(苯丙氨酸氨解酶)和UFGT(类黄酮葡糖苷转移酶)酶活性与花色素苷合成的关系。结果表明,果袋的透光率与果皮外观色泽有密切关系,透光率越高,a*越大,果皮颜色越红。白色果袋与对照、红袋与黄袋果实外观无显著差异,白袋与红袋,黑袋(完全不透光)与黄袋,红袋与黄袋处理之间的果实外观色泽存在显著差异。果实成熟时果皮中花色素苷含量与纸袋的透光率呈正比,波长580nm以上橙、红光的透光率与果皮中花色素苷含量相关系数r>0.99。在不同颜色果袋影响果实成熟时果皮的糖酸积累方面:不同颜色果袋对果肉中糖酸含量无显著影响,果皮的糖含量与花色素苷含量亦无显著相关性,但成熟时果皮中酸含量与花色素苷含量呈显著正相关(r=0.9815*)。不同颜色果袋影响PAL、UFGT酶活性,果实成熟时不同果袋PAL酶活性与花色素苷含量相关性显著(r=0.9613*),UFGT酶活性与花色素苷含量相关性显著(r=0.9365*)。     

毛欧杂种葡萄‘NW196’花色苷组成与相关基因表达分析

以不同发育期毛欧杂种葡萄‘NW196’为试验材料,利用HPLC–MS检测其果皮中花色苷的组成及含量,Real-time PCR检测花色苷合成相关基因表达水平,并与欧亚种‘赤霞珠’比较。结果表明,成熟后期‘NW196’果皮中花色苷总量显著高于‘赤霞珠’;采收期‘NW196’葡萄果皮中共检测到23种花色苷,成熟后期单糖苷所占比例高于双糖苷,其中含量最高的是花翠素–3–O–葡萄糖苷,3’5’–羟基取代花色苷比例高于3’–羟基取代花色苷;与欧亚种‘赤霞珠’相比,‘NW196’果皮中酰化和甲基化花色苷比例较低。结论:毛欧杂种‘NW196’花色苷组成与欧亚种‘赤霞珠’差异明显,这与其品种特性有关,并受到类黄酮代谢路径相关基因的调控;此外,花色素双糖苷主要在转色初期合成。     

Growth and yield response of ‘Correll’ peach trees to fumigation with DBCP

A study was conducted to evaluate the effect of the nematicidal soil-fumigant DBCP (1,2-dibromo-3-chloropropane) on a peach (Prunus persica L. Batsch.) orchard planted on a site with high potential for “short life”. ‘Correll’ peach trees on ‘Lovell’ rootstock were planted on a Ruston fine sandy loam soil (Typic Paleudult, fine loamy, siliceous, thermic) on which 2 generations of peach trees had been grown. Increased tree growth and yield occurred when DBCP was applied twice (pre-plant and 1 year post-planting). After 4 years, there were no differences in the nematode populations or foliar nutrient content as a result of fumigation with DBCP.     

外源NO和H2O2诱导不同颜色马铃薯花青素积累

以不同块茎颜色马铃薯品种陇薯8号、陇薯7号、LC310-2和山东彩肉为材料,研究外源NO、H2O2、NO+H2O2处理对淀粉积累期马铃薯块茎花青素合成及相关酶活性的影响。结果表明：经外源NO和H2O2处理后,4个基因型马铃薯薯皮和薯肉花青素含量均升高,PAL、PPO和CHI活性增强|诱导效果为0.1 mmol·L-1 SNP（NO的供体）+0.25 mmol·L-1 H2O2＞0.25 mmol·L-1 H2O2＞0.1 mmol·L-1 SNP＞清水（CK）,说明NO和H2O2处理对马铃薯花青素合成具有一定的促进作用,且表现为加性效应。     

Effects of temperature and plant growth regulators on anthocyanin synthesis and phenylalanine ammonia-lyase activity in chicory (Cichorium intybus L.)

Summary

This study addresses the effects of air temperature and plant growth regulators on anthocyanin synthesis, sugar content and phenylalanine ammonia-lyase (PAL) activity in chicory (Cichorium intybus L.). Anthocyanin in chicory was synthesised at the highest level under 15°/10°C (day/night) temperatures, followed by 20°/15°C, and 25°/20°C; while synthesis was inhibited > 90% at 30°/25°C, resulting in an almost green colour. Sugar contents paralleled anthocyanin development under the same temperatures. The plant growth regulators, abscisic acid (ABA), ethephon and uniconazole all stimulated anthocyanin synthesis, with uniconazole treatment showing the greatest effect. Gibberellic acid (GA₃) inhibited anthocyanin development, while GA₃ in combination with uniconazole alleviated this inhibition.

PAL activity was higher at 15°/10°C or 20°/15°C (day/night) temperatures when plants were treated with ABA, ethephon or GA₃, than at 25°/20°C and 30°/25°C (day/night) temperatures. These results suggest that, under lower temperatures, plant growth regulators may play an important role in anthocyanin synthesis and PAL activity in chicory.     

腐胺对藤稔葡萄果皮色素及相关酶的影响

通过对藤稔葡萄喷施不同浓度腐胺(Put),研究了腐胺对葡萄果皮色素及相关酶的影响.结果表明:不同浓度的Put都起到了延缓叶绿素降解的作用,影响花色素苷合成和色泽表现.Put影响葡萄果皮中花色素苷合成途径中苯丙氨酸氨解酶(PAL)、多酚氧化酶(PPO)、过氧化物酶(POD)3个酶活性.     

紫皮大蒜鳞茎外皮花青苷生物合成影响因素和相关酶的研究

 以紫皮大蒜品种‘G075’为试材,研究了鳞茎发育过程中鳞茎外皮花青苷的积累规律及与其生物合成相关的苯丙氨酸解氨酶（PAL）、查儿酮异构酶（CHI）和二氢黄酮醇还原酶（DFR）活性的关系,分析了不同设施栽培方式、基质pH值、基质氮素水平和磷素水平对花青苷生物合成的影响。研究结果表明：随着大蒜鳞茎的发育,鳞茎外皮花青苷含量逐步提高,CHI活性与鳞茎外皮花青苷积累关系密切,其活性变化与花青苷积累趋势吻合;露地栽培温度相对较低的紫皮大蒜花青苷积累高于保护地温度相对较高的栽培大蒜,基质pH 6.5和1/2氮素水平（7.5 mmol · L^-1）时对花青苷合成有利,花青苷积累随磷素水平提高呈现先升后降的变化趋势。     

Sweet-potato productivity as affected by recurrent use of vines as planting-material

Yield performances of apical shoot cuttings of sweet potato (Ipomoea batatas) obtained from crops raised 2 or 4 times recurrently through vines, were compared with those obtained from tubers. The recurrent use of vines as planting-material did not show any marked variation in market-grade tuber yield, but it significantly decreased vine yield and increased weevil infestation in tubers. Cultivars ‘Cross-4’, ‘RS-5’ differed significantly in performance     

紫外光对‘北陆’越橘转色果花青苷积累、关键酶活性及其基因表达的影响

以3年生‘北陆’越橘为试材,对发育过程中果实花青苷含量及合成酶活性变化进行研究,并进一步利用3种紫外光（UV-A、UV-B、UV-C）分别照射转色期越橘果实,测定了果实中花青苷等酚类物质的含量、生物合成酶活性及其基因表达,从底物、酶活性、基因转录水平阐明了不同紫外光对花青苷生物合成途径的影响。3种紫外光都明显诱导了果实花青苷的积累,特别是UV-C照射后果实花青苷含量提高了2.36倍。UV照射显著诱导果实PAL、UFGT酶活性升高,VcPAL、VcUFGT转录增强,但同时也抑制了DFR酶活性及基因表达,其中PAL、UFGT酶活性与花青苷的积累极显著正相关（r = 0.807**,r = 0.894**）,而DFR酶则相反（r =–0.854**）。结果表明,UV照射能诱导丙苯氨酸途径中响应紫外处理的一些关键基因的转录激活（如VcPAL,VcUFGT）或抑制（如VcDFR）,并影响相应酶活性的变化,促使花青苷等酚类物质的积累。     

Response of pepper and eggplant to continuous light in relation to leaf chlorosis and activities of antioxidative enzymes

Activities of the oxygen radical-scavenging enzymes were determined in leaves of ‘Kyomidori’ pepper (Capsicum annuum L.) and ‘Senryo’ eggplant (Solanum melongena L.) seedlings under two photoperiod treatments; 12 h light/12 h dark (12/12 h) or with continuous light (24/0 h) for six days. Growing eggplant under continuous light resulted in leaf chlorosis after four days and a sharp decline in the chlorophyll content. In eggplants grown under 24/0 h photoperiod, the activities of antioxidative enzymes; superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) rapidly increased from day two. This was an indication of photooxidative damage. In contrast, leaf chlorosis did not occur in pepper plants grown under continuous light, while the chlorophyll content was even higher in the 24/0 h than in the 12/12 h photoperiod. Continuous light did not enhance the activities of SOD and POD in leaves of pepper. However, the CAT activity increased faster and was significantly higher in pepper than in eggplants grown under continuous illumination. Pepper plants did not appear to suffer from photooxidative damage and thus grew normally under continuous illumination.     

两种叶幕类型‘摩尔多瓦’葡萄套袋果实花色苷代谢的差异

为了研究水平叶幕和直立叶幕所构成的微域环境对套袋葡萄果实花色苷代谢的影响,分别以棚架、篱架代表水平叶幕和直立叶幕,连续两年对棚架和篱架的‘摩尔多瓦’葡萄进行套袋,于果实膨大期开始实时监控果穗袋内微环境的温度和湿度;从转色期至果实成熟,测定不同发育阶段浆果果皮花色苷单体组分与含量,以及果皮花色苷代谢途径相关基因表达量与酶活性,分析果实品质差异。结果表明,2015年、2016年棚架水平叶幕下套袋果实袋内的高温极值及高温比例和湿度波动幅度与篱架直立叶幕套袋相比均有明显降低;连续两年成熟果实还原糖含量分别比篱架套袋的高18.33%和15.41%,且棚架的果实酸度较低;2015年测定棚架套袋果实的单宁及花色苷含量分别比篱架套袋的提高22.32%和35.29%,果实果皮红色度显著提高,而2016年夏季较冷凉,棚架套袋果实的总酚、黄烷醇、类黄酮含量分别比篱架套袋降低10.72%、10.97%和45.04%。2015年花后77 d,棚架套袋果实的花色苷单体含量较高,但到花后91 d时,篱架套袋果实花色苷单体含量补偿性增加。成熟果实果皮中均检测到21种花色苷单体,棚架套袋葡萄花色苷双糖苷化、甲基化及总修饰度均高于篱架套袋果实,但有19种花色苷单体含量低于篱架套袋果实。花后63~105 d,除Vv OMT外,棚架与篱架套袋葡萄花色苷代谢途径中Vv UFGT、Vv LDOX、Vv5GT和Vv PPO基因表达变化趋势一致,但相关酶活性差异较大。由此可见,水平叶幕可降低果实微域环境的温湿度,增加套袋果实品质及花色苷单体修饰程度,但降低花色苷单体含量,并影响花色苷代谢途径中相关酶基因的表达量及活性。