Composition and properties of purified phenolics preparations obtained from an extract of industrial blackcurrant (Ribes nigrum L.) pomace

Summary

An extract obtained from industrial blackcurrant pomace was purified and fractionated on an RP C18 column, resulting in three phenolics preparations of different composition. The phenolics in the preparations were quantified by HPLC, their anti-oxidant capacity was determined, and the preparations were subjected to HPLC-MS analysis in order to identify the bioactive compounds present. The purified preparations selected for study were an anthocyanin-rich preparation, a myricetin and quercetin glycosides-containing preparation, and an aglycon-containing preparation. All three preparations were characterised by having a high content of polyphenols. The anthocyanin-rich preparation was the most concentrated in polyphenols [> 50% (w/w)], followed by the aglycon-containing preparation [> 20% (w/w)], and finally the glycosides-containing preparation [> 9% (w/w)]. The preparations were also characterised by their high anti-oxidant capacity, which was > 3,100 µM TEAC g^?1 (TEAC = Trolox Equivalent Anti-oxidant Capacity) for all preparations. The HPLC-MS study confirmed that the anthocyanin-rich preparation was composed of the following anthocyanins: delphinidin-3-glucoside, delphinidin-3-rutinoside, cyanidin-3-glucoside, and cyanidin-3-rutinoside. The aglycon preparation contained four aglycons, of which myricetin and quercetin were predominant, and kaempferol and isorhamnetin were present in lower amounts. The glycosides-containing preparation was found to be the most interesting, since it contained myricetin, quercetin, kaempferol, and isorhamnetin glycosides. Myricetin and quercetin galactosides were also detected in blackcurrant extract for the first time. Moreover, the presence of nitrile-containing compounds, two acylated anthocyanins, and one auron-type compound was detected.     

‘紫红1 号’红肉苹果果肉抗氧化性及花色苷分析

 ‘紫红1号’是新疆红肉野苹果[Malus sieversii f. neidzwetzkyana（Dieck）Langenf.]与‘富士’苹果杂交的F₁代中的一株果肉全红的株系。以其为试材,测定其果肉的花色苷、多酚、类黄酮含量及其抗氧化能力,结果表明其果肉花色苷含量为228.3 mg · kg^-1 FW,总酚含量为2 523 mg · kg^-1 FW,类黄酮含量为2 514 mg · kg^-1 FW,其抗氧化能力（ferric reducing antioxidant power,FRAP）为13.39 μmol · g^-1。利用液质联用（UPLC-PAD-/MS/MS）鉴定了红肉苹果果肉花色苷的主要组分有9种,其中4种分别为矢车菊3–O–葡萄糖苷、矢车菊3–O–半乳糖苷、矢车菊3–O–木糖苷、矢车菊3–O–阿拉伯糖苷,其中矢车菊3–O–半乳糖苷占73.37%,另外5种暂不能确定其结构。本研究表明花色苷的稳定性,发现该色素在pH 1 ~ 3,40 ℃以下比较稳定,高温加速花色苷的降解。花色苷对光照敏感,暴露在室内自然光下15 d其残留率为41.53%。     

Sorption isotherms for osmo-convectively-dried and osmo-freeze-dried apple,sour cherry,and blackcurrant

Summary

The aim of our work was to measure sorption isotherms on freeze-dried and convectively-dried fruits (apple cv. Idared; sour cherry cv. English Morello; blackcurrant cv. Tiben), previously osmotically dehydrated in fructo-oligosaccharide solution, or concentrated apple juice. Isotherms were fitted using the Guggenheim-Anderson-de Boer Model. In none of the cases studied was isotherm non-continuity in the vicinity of the initial value of a_w observed. All isotherms, classified as type III, demonstrated an increase in the equilibrium water content, along with an increase in water activity. A higher water content was observed in lyophilised material compared to material dried by convection. The water content in the monolayer (100 g^?1 dry matter) ranged from 12.0 g for dried apple, to 17.0 g for dried sour cherry. These values can be considered as optimal in order to ensure safe storage conditions. The dried fruits should therefore be kept in a water activity range of 0.45 – 0.54 for lyophilised, and 0.46 – 0.63 for convectively-dried material.     

Azalea indica flower colour as related to the parameters pH,anthocyanins and flavonol co-pigments

The influence of epidermal tissue pH on flower colour was studied in 104 cultivars of Azalea indica (Rhododendron simsii Planch.). Based on H.C.C. visual colour number, cultivars were classified in 4 colour groups: purple; carmine red; red; white.A method for pH determination using microvolumes (30 μl) on a microelectrode has been perfected. Within each colour group, flower colour was determined within a mean pH environment of 3.04 ± 0.06. Epidermal pH was approximately 0.3 units lower than the pH of the expressed sap of entire petals. Purple colours occurred with co-pigmented malvidin or delphinidin glycosides. Carmine red and red colours were produced by the same cyanidin glycosides that in the case of carmine red were co-pigmented with flavonol glycosides. The lower the content of anthocyanins (cyanidin glycosides) and the higher that of flavonols, the bluer the red (higher H.C.C. number; bathochromic shift of about 20 nm). A carmine red flower colour (± 58 B) may also occur when delphinidin glycosides occur in the absence of flavonol glycosides (pH < 4).Prospects for obtaining a blue flower colour are, because of low epidermal pH, very small.     

The Measurement of Sap Tension in the Petioles of Apple,Raspberry and Black Currant Leaves

The pressure chamber technique has been used for rapid measurement of sap tension in the petioles of apple, raspberry and blackcurrant leaves in the field.

When the soil moisture tension was relatively low, the diurnal trend in sap tension in the petioles of apple leaves followed very closely the relative evaporation rate.     

MdMYB10对苹果果皮苯丙氨酸代谢的影响

MdMYB10对苹果果皮花青苷合成起重要作用,但对苯丙氨酸代谢途径中其他酚类物质合成是否起作用尚不清楚。从‘粉红女士’苹果（Malus ? domestica‘Pink Lady’）果皮中克隆了MdMYB10,分别构建其过表达载体pCAMBIA2301-MdMYB10和沉默载体pTRV2-MdMYB10,在套袋苹果果皮下注射农杆菌诱导MdMYB10过表达和沉默,分析MdMYB10对解袋后苹果果皮苯丙氨酸代谢的影响。结果表明,在过表达MdMYB10的果皮组织中,MdMYB10表达量上调了19.62倍,MdPAL、MdCHS、MdF3H、MdDFR、MdANS、MdUFGT、MdFLS、MdANR均有不同程度的上调,只有MdLAR无显著变化;沉默MdMYB10的果皮组织中,MdMYB10表达量降低了87%;MdCHS、MdF3H、MdDFR、MdANS和MdUFGT均有不同程度的下调,其中MdANS和MdUFGT下调最多,分别下调83%和82%。在过表达MdMYB10的果皮中,花青苷含量升高了3.05倍,槲皮素–3–O–半乳糖苷、表儿茶素、原花青素B2、绿原酸、对香豆酸含量均有不同程度升高,根皮苷含量无明显变化;在沉默MdMYB10的果皮中,花青苷含量降低了83%,槲皮素–3–O–半乳糖苷和表儿茶素含量分别降低58%和52%,原花青素B2、根皮苷含量无明显变化,而绿原酸含量升高了24%,对香豆酸含量升高了21%。可见,MdMYB10不仅调控苹果果皮中花青苷合成,同时还调控苯丙氨酸代谢中绿原酸、对香豆酸、槲皮素–3–O–半乳糖苷、表儿茶素和原花青素B2等物质的合成,但对根皮苷的形成作用不明显。     

Fruit quality in ‘Cripp's Pink’ apple,especially colour,as affected by preharvest sprays of aminoethoxyvinylglycine and ethephon

‘Cripp's Pink’ apple grown in Western Australia often develops poor colour at commercial harvest resulting in economic losses. To determine if fruit colour could be improved without advancing ripening, ‘Cripp's Pink’ apple fruit on trees were sprayed with aminoethoxyvinylglycine (AVG) alone, ethephon alone, or AVG followed by ethephon. The experiments were conducted at two different locations in Western Australia in 2002 and 2003. Fruit sprayed with AVG alone had retarded colour development at harvest. However, ethephon applied after AVG enhanced percent red blush, anthocyanin concentration and reduced chlorophyll concentration in the fruit skin in both locations. These fruit had similar colour to those treated with ethephon alone. Internal ethylene concentration and fruit firmness were unaffected by the different treatments in 2002. However, in 2003 AVG with or without ethephon reduced internal ethylene concentration and maintained firmness compared to ethephon alone. In conclusion, AVG treatment alone delayed colour development and ripening of ‘Cripp's Pink’, while AVG application 5 weeks before harvest followed by an ethephon application 2 weeks later enhanced red colour at commercial harvest. This is, therefore, an effective tool for improving colour of ‘Cripp's Pink’ apples at commercial harvest without adversely affecting other fruit quality attributes.     

‘金冠’苹果与其无锈芽变的果皮性状比较和防锈技术研究

 为了阐明‘金冠’苹果果锈形成的时间与原因,探索防止果锈形成的方法,以‘金冠’及 其无锈芽变品种‘丰帅’为试材,利用体视显微镜和扫描电镜观察了果面果锈形成过程,测定了果皮色 素和次生代谢产物组分和含量。结果表明,花后4 周‘金冠’苹果果面尚无肉眼可见的果锈时,表皮细 胞蜡质层已经开裂、脱落;花后5 周表皮细胞角质层大量开裂,细胞壁木栓化;花后6 周,大量果锈出 现在果实表面。据此推断,花后4 ~ 6 周是‘金冠’苹果果锈形成的关键时期。另一方面,‘丰帅’果皮 叶绿素和类萝卜素含量显著低于‘金冠’,而总黄酮和绿原酸含量显著高于‘金冠’,特别是在花后4 ~ 6 周。花后4 周利用0.05 ~ 5 mmol · L-1 外源绿原酸处理,可以显著降低‘金冠’果皮果锈指数,且对果实 品质没有不良影响。因而,适当浓度外源绿原酸可望用于苹果防锈实践。     

Effect of sucrose and mannitol on the accumulation of health-promoting compounds and the activity of metabolic enzymes in broccoli sprouts

The contents of ascorbic acid, glucosinolates, sulforaphane, anthocyanins, total phenolics, the activity of myrosinase and phenylalanine ammonialyase (PAL) and the antioxidant activity of broccoli sprouts grown under 88 mM and 176 mM of sucrose and mannitol were investigated. The results showed that the contents of sulforaphane, ascorbic acid and anthocyanins in broccoli sprouts were significantly increased after treatment with 88 mM of sucrose compared with the control. The contents of glucosinolates and total phenolics, the activity of PAL and the antioxidant activity in broccoli sprouts treated with 176 mM sucrose were also significantly increased, whereas the activity of myrosinase was significantly reduced. On the other hand, the contents of glucosinolate, sulforaphane and total phenolics in broccoli sprouts were also significantly increased after treatment with 176 mM of mannitol, although the contents of sulforaphane were markedly reduced compared to those treated with 176 mM of sucrose. Sucrose might induce the production of health-promoting compounds through its role of signaling, generating osmotic pressure or serving as a substrate. These results indicate that sucrose treatment could improve the nutritional value of broccoli, and the sprouts growing under adequate concentration of sucrose could benefit our diet by producing more health-promoting compounds.     

The influence of exposure to light on the phenolic content of ‘Fuji’ apple

In this study, the influence of changes in fruit lighting on individual and total phenols in ‘Fuji’ apple, as well as color development was studied. Content levels of eight quercetin glycosides, five anthocyanins, two catechins and a hydroxycinnamic acid in the skin of apples were analyzed, using high-performance liquid chromatography, tandem mass spectrometry (HPLC-MS). Total phenol, chlorophyll and carotenoid contents were determined by spectrometry. The purpose of this study was to compare content levels of those compounds in apple skin of fruit grown in different parts of the tree canopy, under and outside of the hail net. Lighting of fruit was measured during the last month before harvest. The lowest values were measured in the inner fruit and higher ones in the outer parts of the canopy, while the highest values were measured in fruit growing at the top of the tree. The hail net had no influence on the decrease of lighting in comparison to the control. Light conditions in the tree canopy influenced lower content levels of quercetin glycosides and most anthocyanins in the fruit skin in the inner part of the tree canopy, whereas fruit from the canopy top contained the highest levels of quercetin glycosides and cyanidin glycosides. Catechin, epicatechin and chlorogenic acid content levels in apple skin were independent of fruit position in the canopy or hail net usage, except for chlorogenic acid, where the content level was higher in cases when the orchard was covered with a hail net. Fruit from the top and outer parts of the canopy had a darker and redder coloration than inner fruit, while no influence of canopy position on chlorophyll and carotenoids was detected. Since quercetin glycosides and cyanidin glycosides are influential in red skin color development, better coloration of fruit from the outer and top canopy was observed. More intensive lighting stimulated a higher content level of flavonoids and, consequently, better coloration, which is an important factor in fruit quality.     

Effects of high temperature on flower colour and anthocyanin content in pink flower genotypes of greenhouse chrysanthemum (Chrysanthemum morifolium Ramat.)

Summary

The relationship between the intensity of flower colour and changes in the content of the main anthocyanins under various controlled temperatures was examined in order to clarify the effects of high temperature on flower colouration in six pink flower genotypes of greenhouse chrysanthemum (Chrysanthemum morifolium Ramat.). Poor colouration of flowers was observed at 30°C in all genotypes except ‘Chatoo’. This genotype showed little difference in flower colour between different temperature treatments. The degree of change in flower colour differed depending on the genotype, whereas no clear differences in flower colouring were observed between Summer – Autumn flowering and Autumn-flowering genotypes. All genotypes showed lower contents of the two anthocyanins tested [cyanidin 3-O-(6’’-O-monomalonyl- -glucopyranoside) and cyanidin 3-O-(3’’,6’’-O-dimalonyl- -glucopyranoside)] at higher temperatures. Therefore, flower colour changes were attributable to changes in these two main anthocyanins. Differences in colouration between genotypes and temperature conditions were also detectable in values that were measured using a colorimeter. Changed parameters that were visually verifiable were the a* value, representing the degree of red colour, and the C* value, representing chroma. For ‘Sei-Monako’, which showed visually greater differences between temperature treatments, the a* and C* values were low under high temperature conditions. On the other hand, in ‘Chatoo’, the differences detected by eye and those in a* and C* values between temperature treatments were small. In addition, the present results indicate that mean temperature is more important than either day or night temperature in determining the degree of flower colouration.     

苹果制汁新品种鲁加4号浓缩汁贮藏过程中稳定性的影响因素

以苹果制汁新品种鲁加4号[特拉蒙(Telamon)×新红星(Starkrimson)]的浓缩果汁为试材,研究温度、pH值、糖度和果实成熟度对苹果浓缩汁贮藏过程中稳定性的影响。结果表明,温度对苹果浓缩汁贮藏过程中稳定性影响较大,贮藏温度越高,浓缩汁的稳定性越差;糖度对其影响显著,糖度越高,浓缩汁稳定性越差;pH值和果实成熟度对苹果浓缩汁稳定性也有显著性影响,浓缩汁贮藏1～2周期间,pH值与A420值呈正相关,贮藏3～4周期间,pH值与A420值呈负相关。不同采收期苹果果实出汁率、可溶性固形物含量、多酚含量、浓缩汁酸含量变化不同。果实成熟时采收制得的浓缩汁稳定性比较好。     

草莓和蓝莓果实花青素提取及定量方法的比较

 本研究旨在比较果实花青素提取液的提取效果以及花青素分析方法的准确性。用丙酮-水-甲酸（A）、乙腈-乙酸（B）、乙醇-水-乙酸（C）、甲醇-水-乙酸（D）以及含有盐酸的甲醇（E） 等5种提取液分别提取了草莓和蓝莓果实中的花青素,用比色法和液相色谱法分别测定了花青素的含量。试验结果表明：由于提取液中有机溶剂和酸种类的不同,提取效果差异甚大。提取草莓花青素时,以D的提取效果最好,其次是E、C、A,B的提取量最低,还不到D的1/3。提取蓝莓花青素得到的结果与草莓相似。用比色法分析,D、E、C提取液提取的草莓和蓝莓果实的花青素含量明显低于液相色谱分析的结果, 但B提取液比液相色谱结果高2倍以上,A提取液提取蓝莓时比色法与液相色谱法的结果相近,而提取草莓时比色法比液相色谱结果高出约50%。建议花青素的定量方法应尽量使用液相色谱法,对同一种果实,尤其是以低沸点醇类作提取液时可以用比色法。     

Estimation of the Colour Properties of Apples Varieties Using Neural Network

The consumer acceptance and the quality standard of agricultural products such as apple are determined mostly by their colour. Colour is measured with a colorimeter and quantified using the C.I.E. L^*, a^*, b^* colour space system. It is used commonly by researchers for the classification and identification of apple fruit. To the best of our knowledge, the present study is the first study investigating the prediction of some colour properties of six apple varieties through artificial neural networks (ANN). The apple varieties are ‘Amasya’, ‘Starking’, ‘Granny Smith’, ‘Pink Lady’, ‘Golden Delicious’, ‘Arapk?z?’ and the colour properties are L^* (lightness), a^* (redness), b^* (yellowness), C^* (chroma), h^* (hue angle), CI (chroma index). General Regression Neural Networks (GRNN) and Adaptive Neuro Fuzzy Interface System (ANFIS) structures were employed to predict the colour properties. According to the experimental and simulation results, the proposed ANFIS predictor had a superior performance in prediction of these colour parameters.     

Adventitious shoot regeneration from leaf segments of in vitro cultured shoots of the apple rootstock Jork 9

SUMMARY

Adventitious shoot formation was investigated using leaf segments of in vitro cultured shoots of the apple rootstock Jork 9. Regeneration capacity was influenced by the pretreatment of the mother shoots, macroelements, hormone concentrations, the gelling agent and the carbohydrate source. The highest regeneration rate and most shoots per leaf explant resulted from young leaves on a medium based on MS macroelements supplemented with 22 µM BAP and 0.1 µM_NAA together with sorbitol, at concentrations of 165 mM or 220 mM. Sorbitol was more effective than sucrose, glucose, fructose or a combination of these sugars. A cold and dark pretreatment of the shoots enhanced the formation of adventitious shoots.     

The Relationship Between Growth Vigour of Rootstock and Phenolic Contents in Apple (Malus × domestica)

The aim of the study is to determine the effect of different growth vigorous rootstocks on phenolic compounds in leaves of apple. For this purpose it was used the leaves of cultivar ‘Red Chief’ grafted on dwarf (M9), semi-dwarf (M26) and semi-vigorous (MM106) rootstocks. During mid-July, the leaf samples were taken from the middle part of annual shoots. Phenolics of the leaves were determined by HPLC analysis. While significant differences among the rootstocks for p-hydroxybenzoic acid, eriodictyol, ferulic acid and p-coumaric acid were detected, these differences were insignificant for gallic acid and quercetin. It was shown that semi-vigorous rootstocks (MM106) had higher phenolic contents in total than the other two dwarf rootstocks. In addition, apigenin-7-glucoside, chlorogenic acid, caffeic acid, rosmarinic acid, epicatechin, syringic acid, catechin, rutin, resveratrol, hesperidin, naringenin, luteolin, apigenin and acacetin could not to be detected. Data showed that there is the relationship between growth vigour and phenolic contents of apple leaves. Especially, p-hydroxy benzoic acid and p-coumaric acid contents were higher in semi-vigorous rootstock than in dwarf rootstock.     

Molecular Characterization of Some Apple Accessions Using DNA Markers Associated with Fruit Skin and Flesh Colour

Apple (Malus domestica Borkh.) is one of the most widely grown and economically important fruit crops in the world. Skin colour is one of the most important criteria for marketing of apple fruits. Thus, improving fruit colouring is one of the main objectives of apple breeding programs. Even though environmental conditions may affect the colouring of apple, understanding the genetic basis of colouring is important to accelerate the breeding process. In recent years, molecular biology and genetic studies were conducted to explore the molecular basis of colouring in apple. Many genes responsible for the anthocyanin synthesis were identified and their associations with colouring of apple fruit flesh and/or skin were demonstrated. In addition, some DNA markers associated with fruit flesh and skin colour have been developed for screening apple cultivars and hybrids. In this study, 90 apple accessions of international, national, and local decents were selected from the genetic resources collection. Genomic DNA was isolated from leaf tissue of all acession and screened with four different DNA markers associated with fruit flesh and skin colour. Apple accessions in the collection were characterized and their genotypes and genetic potential for fruit colouration were determined by different DNA markers. Additionally, the use of these DNA markers in different apple accessions, well-characterized cultivars and uncharacterized local and national types were investigated.

     

苹果果实中类黄酮化合物的研究进展

 类黄酮是苹果果实中的一类主要多酚类物质, 对人体健康具有多方面的生理功能。已报道的苹果果实类黄酮达34种, 分属黄烷醇、黄酮醇、二氢查耳酮、花青苷和二氢黄酮醇5类。苹果果实中类黄酮的组成和含量因品种和果实部位而异, 通常果肉中的类黄酮种类和含量均明显少于果皮。苹果果实在发育、成熟和贮藏过程中类黄酮含量会发生变化。苹果果实类黄酮的测定普遍采用带二极管阵列检测器或紫外检测器的高效液相色谱仪, 并用质谱仪对类黄酮进行成分确认和结构鉴定。     

苹果树皮中12种酚类化合物的HPLC测定方法

【目的】为了建立高效液相色谱法(HPLC)同时测定苹果树皮中没食子酸、儿茶酚、儿茶素、绿原酸、咖啡酸、表儿茶素、香豆酸、阿魏酸、芦丁、根皮苷、槲皮素、根皮素12种酚类化合物的方法,【方法】选用Eclipse XDB C18色谱柱(150 mm×4.6 mm i.d,5μm),以甲醇和1%乙酸溶液作为流动相进行梯度洗脱,利用VWD检测器,在各种酚类物质的最优吸收波长下进行检测,【结果】得到12种酚类化合物的线性回归方程,相关系数均在0.999以上,检出限(3RSN)分别为0.02、0.02、0.03、0.01、0.02、0.03、0.01、0.01、0.03、0.03、0.05和0.01 mg.L-1。色谱测定方法和包含样品处理方法的加标回收率分别在56.54%～98.83%和90.95%～100.90%。【结论】整个提取和分析测定流程简便、快速、准确,适合苹果树皮中酚类化合物的分析测定。