红色系中原牡丹品种花色苷和黄酮的含量分析

采用HPLC-DAD-MS技术分析了红色系中原牡丹(24个品种)花中的主要花色苷类化合物,对各类花色苷的相对含量和分布特点进行了分析,测定了花色苷和黄酮的总含量.共检测到5种花色苷,分别为矢车菊-3,5-O-二葡萄糖苷、天竺葵-3,5-O-二葡萄糖苷、芍药-3,5-O-二葡萄糖苷、矢车菊-3-O-葡萄糖苷、芍药-3-O-葡萄糖苷.依据所含花色苷的类别及相对含量,将红色系中原牡丹分为四大类,分别为Pn、Cy类;Pn、Cy＞Pg类;Pn、Pg类;Pn、Pg＞Cy类;多数品种花色苷含量为(1～10)×10-2%,黄酮含量在0.6%～2.0%;不同品种中,花色苷含量越高,助色系数越低.     

七个江南牡丹品种花瓣中类黄酮物质分析

以“凤尾”、“凤丹白”、“西施”、“粉莲”、“昌红”、“呼红”和“云芳”7个江南牡丹品种为试材,利用UPLC-PDA和UPLC-Q-TOF MS技术对盛花期花瓣中类黄酮组分进行了定性和定量分析.结果表明:江南牡丹花瓣中共检测到15种类黄酮组分,其中花色苷4种:矢车菊素-3,5-二葡萄糖苷、矢车菊素-3-葡萄糖苷、芍药花素-3,5-二葡萄糖苷、芍药花素-3-葡萄糖苷；黄酮7种:木犀草素单糖苷(六碳糖)、木犀草素二糖苷(甲基五碳糖+六碳糖)、芹黄素单糖苷(六碳糖)、芹黄素二糖苷(五碳糖+六碳糖)、芹黄素二糖苷(甲基五碳糖+六碳糖)、金圣草黄素单糖苷(六碳糖)、金圣草黄素二糖苷(六碳糖+甲基五碳糖)；黄酮醇4种:槲皮素单糖苷(六碳糖)、槲皮素二糖苷(六碳糖+六碳糖)、山奈酚二糖苷(六碳糖+六碳糖)、异鼠李素二糖苷(六碳糖+六碳糖)；江南牡丹花瓣中主要的花色苷为芍药花素-3,5-二葡萄糖苷和矢车菊素-3,5-二葡萄糖苷,主要的黄酮为芹黄素糖苷,即芹黄素单糖苷(六碳糖)、芹黄素二糖苷(五碳糖+六碳糖)、芹黄素二糖苷(甲基五碳糖+六碳糖),黄酮醇为山奈酚糖苷,即山奈酚二糖苷(六碳糖+六碳糖).     

观赏向日葵的花色多样性及其与花青苷的关系

 利用英国皇家园艺学会比色卡(RHSCC)和分光色差仪测定法将36份观赏向日葵的花色分为两大类,即黄色系和红色系。红色系向日葵的花色变异较小;黄色系向日葵的花色变异较大。黄色系向日葵又可分为柠檬黄色和橙黄色两个亚类。高效液相色谱法（HPLC）的分析结果表明,试验材料中含有的花青苷共有9种,但这9种花青苷中只有其中的一种在所有样品中均能检测到。对红色向日葵花瓣的花青苷提取液进行多级质谱分析发现,花青苷元类型主要是矢车菊素,其糖苷类型主要是和葡萄糖、鼠李糖和/或阿拉伯糖结合的配糖体;而在纯黄色的向日葵中未检测到这些花青苷,说明矢车菊类花青苷是红色向日葵舌状花显现红色的化学基础。     

花毛茛和银莲花花瓣中花青素苷组成及含量与其花色的关系

以10个不同花色的花毛茛（Ranunculus asiaticus）和4个不同花色的银莲花（Anemone cathayensis）为材料，采用目视测色法，RHSCC 比色卡比色法，色差仪（CR-400）测定花瓣的花色表型，利用双光束紫外—可见光分光光度计（TU-1901）、高效液相色谱—电喷雾离子化—质谱连用技术（HPLC–ESI–MS）对花瓣中花青素苷的成分及结构进行测定，运用多元线性回归方法分析花色与花青素苷组成之间的关系。结果表明：8个积累花青素苷的花毛茛品种花瓣中检测到5种花青素苷元，即天竺葵素、矢车菊素、飞燕草素、芍药素和锦葵素，这些花青素苷元进一步修饰形成了15种花青素苷。红色系和粉色系品种以芍药素和矢车菊素为主要成分，黄色系品种以天竺葵素或类胡萝卜素为主要成分；经分析矢车菊素苷和天竺葵素苷含量均与亮度L*值呈显著正相关，其中天竺葵素苷的积累对花瓣亮度的贡献更大。银莲花4个品种花瓣中共检测出6种花青素苷元，分别为矮牵牛素、天竺葵素、矢车菊素、飞燕草素、芍药素和锦葵素，它们通过不同糖基化和酰基化修饰形成了20种花青素苷，呈现出丰富的花青素苷多样性。     

欧洲报春细胞液pH对花色形成的作用机理初探

为研究欧洲报春花瓣细胞液p H对花色形成的作用机理,以粉、红、白、黄、蓝5个花色欧洲报春的花瓣为材料,采用便携式色差仪、平面型pH计以及UPLC技术对其花色值(CIE L*a*b*)、细胞液p H值及花色素的成分及含量进行测定。结果表明,细胞液pH值以红色花瓣最低(5.07),粉、白、黄色花瓣次之,蓝色花瓣最高(6.33)。红、粉、蓝色品种花瓣中花青苷含量较高,白色和黄色品种花瓣中类黄酮含量最高。同时花瓣细胞液pH值与蓝色品种特有的5种花青苷(飞燕草素3–葡萄糖苷、报春色素苷、矮牵牛–7–甲氧基–3,5–二葡萄糖苷、锦葵素–3–葡糖苷和报春色素–4’–丙二酸酯)的含量呈显著正相关。红、粉和蓝色品种花瓣花色素提取液的颜色随着外界缓冲液pH的增大而逐渐变浅,并最终变为黄色;在可见光区的最大吸收波长发生红移。通过分析可能调控欧洲报春pH的6个基因表达量及其与花瓣细胞液pH值的相关性,发现PvPH4在蓝色花瓣中显著高表达,并与花瓣细胞液p H值呈显著正相关;其编码的蛋白质保守结构域是属于Ca~(2+)/H~+逆向转运超家族(Cha Asuperfamily)的caca2,并且蛋白质三级结构建模显示其与液泡钙离子转运体相似度最高(39%)。因此推测PvPH4通过编码Ca~(2+)/H~+转运跨膜蛋白,影响花瓣细胞液p H值,进而影响花色。     

变异紫斑牡丹红色叶色素与抗氧化活性分析

以紫斑牡丹和变异紫斑牡丹的叶片为试材,采用高效液相色谱、紫外-可见分光光度法、薄层色谱对变异紫斑牡丹红色叶和紫斑牡丹绿色叶的色素进行定性定量分析;采用1,1-二苯基-2-三硝基苯肼自由基和2,2’-联氮-双-(3-乙基苯并噻唑啉-6-磺酸)二铵盐自由基分析2种叶片的体外抗氧化活性。结果表明:变异紫斑牡丹红色叶的颜色发生变化主要是芍药花素-3,5-二葡糖苷和天竺葵素-3,5-二葡糖苷含量上升与叶绿素含量下降的共同作用结果;变异紫斑牡丹红色叶中单体酚含量(绿原酸、芦丁、儿茶素、表二茶素、二氢杨梅酮、对香豆酸和二氢槲皮素)比紫斑牡丹绿色叶中的高;变异紫斑牡丹红色叶较紫斑牡丹的绿色叶相比有较好的抗氧化能力,具有潜在药用价值。     

钟冠报春苣苔花发育进程中花色素苷含量的变化

采用超高效液相色谱—四级杆飞行时间串联质谱联用技术（UPLC-Q-TOF-MS）测定钟冠报春苣苔（Primulina swinglei）花发育进程中花色素苷在各花器官组织中的分布、含量及比例。共鉴定出17种花色素苷类物质,分别为矢车菊素、芍药花素、飞燕草素、矮牵牛素及锦葵素及其与葡萄糖、芸香糖、乙酰芸香糖等糖苷组合而成的苷元。研究发现,呈紫色的花瓣中蓝紫色的花色素（飞燕草素、矮牵牛素及锦葵素）比例达到64.0%;呈红色的花筒中红粉色的矢车菊素及芍药花素分别占36.7%和20.0%。开花前总花色素苷含量缓慢上升并趋于稳态,该过程中红粉色的花色素占据比例逐渐减小,而蓝紫色花色素所占比例渐趋增加,这可能与花瓣/花筒的比例渐趋增大有关。而花盛开之后,花色素苷含量急剧降低,可能与花色素苷的降解或稀释相关。     

月季种质资源花色多样性及其与花青苷的关系

利用CIE Lab测色体系对227个月季种质资源进行聚类分类,将月季花色均匀地分为7个色系。在a*和b*二维图上,各类花色只有第Ⅲ象限没有分布,即月季中没有蓝色花;根据227个被测品种的彩度值(C*)与亮度值(L*)二维分布图,可将其分为3大类群。大部分种质资源的亮度(L*)与彩度(C*)表现为负相关,分别为以白色和黄色为主的第1类群和以粉色、大部分玫红色、紫红色和橙红色第2类群;但在包括深红色和部分玫红色资源的第3类群中,L*与C*呈正相关关系。HPLC测定结果发现,6种花青苷在101份试验材料中均被检测到,其中含量贡献最大的是矢车菊素–3,5–双葡萄糖苷(Cy3G5G)和天竺葵–3,5–双葡萄糖苷(Pg3G5G);随着花瓣颜色的加深,其花青苷种类和含量也相应地增加,其中第3类群花青苷含量远大于其他两个类群。上述结果表明,月季种质资源存在丰富的花色多样性。进一步的相关性分析发现,6种花青苷总含量(TA)与L*呈负相关关系;与第1、2类群的a*或C*正相关,但在第3类群中呈现负相关。第1类群花青苷与测色参数L*、a*、b*、C*不存在相关性;第2类群中两类Pg花青苷与a*、b*、C*均存在显著正相关,但与L*无显著相关,表明Pg类花青苷只影响花瓣颜色;在第3类群中,只有含量最高的花青苷Cy3G5G与L*、a*、C*存在显著负相关关系,表明Cy3G5G含量的增加既减弱了花瓣亮度也降低了其彩度。     

软枣猕猴桃果肉花色苷关键光响应调节因子AaMYB1的筛选

【目的】从前期‘天源红’不同发育时期果肉转录组测序结果中筛选了与全红型软枣猕猴桃果肉着色相关的14个转录因子基因,鉴定在套袋过程中的表达特征,并筛选关键光响应转录因子。【方法】以全红型软枣猕猴桃品种‘天源红’盛花后8个时期的果肉样品为试材,在盛花后30 d对果实进行套袋处理,以不套袋果实作对照;使用日本柯尼卡美能达可携式色差计CR-400进行色差指标测定,采用超高效液相色谱串联质谱法检测花色苷组分及总含量,利用实时荧光定量PCR技术检测14个转录因子基因的表达量并对其进行相对定量;通过表型、基因表达量与花色苷含量的相关性分析,综合筛选关键光响应转录因子。【结果】随着果实生长发育,果肉颜色均由绿变红,在盛花后120 d红色最深,未套袋果肉红色明显深于套袋果肉,色泽比和色度角的测定结果与表型鉴定结果一致。果肉主要呈色物质是矢车菊素-3-O-半乳糖苷和矢车菊素-3-O-木糖-半乳糖苷,二者与总花色苷含量呈极显著相关;盛花后120 d,矢车菊素-3-O-半乳糖苷和总花色苷含量在套袋与不套袋果肉中差异显著。实时荧光定量PCR结果显示,MYB1在盛花后120 d未套袋果肉中的表达水平显著高于套袋果肉,并且与未套袋果肉矢车菊素-3-O-半乳糖苷和矢车菊素-3-O-木糖-半乳糖苷含量呈显著正相关,与套袋处理果肉不相关。【结论】筛选到软枣猕猴桃花色苷形成响应光照的关键转录因子AaMYB1;套袋处理可能通过抑制AaMYB1的表达从而抑制花色苷(主要是矢车菊素-3-O-半乳糖苷)的合成与积累,从而阻碍果实正常着色。     

紫叶矮樱叶片成色色素分析

以紫叶矮樱叶片为试材,通过薄层层析色谱分析法(TLC)、紫外-可见分光光度法(UV)对其色素种类进行初步推断。通过高效液相色谱法对叶片色素进行进一步验证,并对鉴定出的色素种类进行定量分析。结果表明:紫叶矮樱叶片内含有叶绿素(主要是叶绿素a)、类胡萝卜素,以及矢车菊素(Cyanidin)、天竺葵素、矢车菊素半乳糖苷(Cyanidin-galactoside)以及儿茶素、二氢杨梅酮等单体酚;定量分析发现紫叶矮樱叶片呈色的机制主要是叶绿素与矢车菊素半乳糖苷(Cyanidin-galactoside)和矢车菊素(Cyanidin)2种花青苷共同作用的结果,其相对含量在检测色素总含量中占2.85%,类胡萝卜素、对香豆酸、儿茶素等辅助色素对呈色也起到了一定的辅助作用。     

Analysis of petal anthocyanins to investigate coloration mechanism in herbaceous peony cultivars

Petal coloration and anthocyanin compositions of 41 herbaceous peony cultivars were analyzed. Anthocyanins were identified by high-performance liquid chromatography–electrospray ionization-mass spectrometry (HPLC–ESI-MSⁿ) coupled with photodiode array detection (DAD). Peonidin-3,5-di-O-glucoside (Pn3G5G), pelargonidin-3,5-di-O-glucoside (Pg3G5G), cyanidin-3,5-di-O-glucoside (Cy3G5G), peonidin-3-O-glucoside (Pn3G), and cyanidin-3-O-glucoside (Cy3G) were the five major anthocyanins in herbaceous peony cultivars. Deep purple or reddish purple cultivars contained 4–5 anthocyanins, whereas pink cultivars only contained Cy3G5G and Pn3G5G, and their contents were much lower than those of purple cultivars. According to the chemical structures of three anthocyanidins in association with petal coloration, flowers were classified into three phenotypes: 1. “Pn, Cy, and Pg” (all purple flowers including two pink flowers); 2. “Pn, Cy” (pink flowers); 3 “Pn” (light pink and white flowers). The coloration mechanisms of cultivars with the pink and purple flowers were quite different. Correlations between lightness (L*) and chroma (C*), chromatic component a* and total anthocyanins (TA) value, a* and co-pigmentation index (CI) showed opposite tendencies, whereas L* and TA showed the same tendency in each group. High contents of Pn3G5G and Pg3G5G may responsible for the purple coloration of herbaceous peony cultivars.     

Comparison of anthocyanins in non-blotches and blotches of the petals of Xibei tree peony

A comparison in non-blotches and blotches of 35 cultivars of Xibei tree peony was investigated to explore flower coloration of Xibei tree peony (the second cultivar group in Chinese tree peony). With high performance liquid chromatography (HPLC) analysis, six anthocyanins including peonidin 3,5-di-O-glucoside (Pn3G5G), peonidin 3-O-glucoside (Pn3G), cyanidin 3,5-di-O-glucoside (Cy3G5G), cyanidin 3-O-glucoside (Cy3G), pelargonidin 3,5-di-O-glucoside (Pg3G5G), and pelargonidin 3-O-glucoside (Pg3G) were detected in non-blotches and blotches of petals. Total anthocyanins (TA), total flavones and flavonols (TF), and the copigmentation index (CI) were also analyzed. Cyanidin-based glycosides, which accumulated abundantly at the basal petal, resulted in blotch formation. Some peculiar cultivars with only one kind of anthocyanins or without anthocyanins in non-blotches were found in this study. For example, ‘Feng Zi Xiu Se’, ‘Ou Duan Si Lian’, and ‘Xi Wang’ contained only Pn3G5G; ‘Bing Shan Cang Yu’ and ‘Jin Bo Dang Yang’ contained only Cy3G; while no anthocyanins were found in ‘Bing Shan Xue Lian’. Several cultivars were rich in Pg-based glycosides, such as ‘Ni Hong Huan Cai’, and ‘Ju Yuan Shao Nv’. These cultivars would be excellent parents for creating new cultivars with novel flower color in the future.     

基于花色表型的牡丹和芍药品种数量分类研究

以38个牡丹和14个芍药为试材,采用色差仪对芍药属品种进行花色表型测定,研究芍药属品种数量分类的情况,以期为芍药属品种鉴定、分类及花色育种提供参考依据。结果表明:聚类分析得到的结果不能科学表征牡丹和芍药花色的分类特点;ISCC-NBS色彩名称表示法对花色的定义更为精确,可将牡丹花色分为黄色、白色、绿色、浅粉色、粉紫色、粉红色、红色、红紫色和红黑色系9类色系;芍药花色分为白色、浅粉色、粉红色、红色和红黑色系5类色系。同时发现芍药属品种花色丰富,且不同色系的芍药属品种花色差异较为显著。     

贴梗海棠花青苷组成及其与花色的关系

 以贴梗海棠[Chaenomeles speciosa（Sweet）Nakai]不同花色的24份种质为材料,调查了其花色在CIE L*a*b*表色系的分布状况,利用高效液相色谱—光电二极管阵列检测（HPLC–DAD）和高效液相色谱—电喷雾离子化—多级质谱联用技术（HPLC–ESI–MSn）定性定量分析其花青苷组分,运用多元线性回归方法分析了花色与花青苷组成之间的关系。结果表明,贴梗海棠花瓣中共含有6种花青苷,分别是矢车菊素–3–O–半乳糖苷（Cy3Ga）、矢车菊素–3–O–葡萄糖苷、天竺葵素–3–O–半乳糖苷、天竺葵素–3–O–（半乳糖葡萄糖苷）[Pg3（Ga-G）]、矢车菊素–3–O–（半乳糖葡萄糖苷）以及矢车菊素–3–O–琥珀酸–阿拉伯糖苷（Cy3SucAra）。Cy3SucAra为首次发现。其中,Cy3Ga、Pg3（Ga-G）和Cy3SucAra是决定贴梗海棠花色的主要色素,这3种色素含量的增加导致花色显著变红。基于花青苷的组成信息,探讨了贴梗海棠的花色改良和蓝色花创制的策略。     

芸薹属5种紫红色蔬菜花青素苷含量及组分分析

采用高效液相色谱-质谱法对芸薹属中红叶芥菜、紫红色大白菜、红菜薹、紫色白菜和紫结球甘蓝新鲜叶片所含花青素苷含量和组成进行鉴定。结果表明,这些蔬菜中花青素苷主要组分为矢车菊素苷,兼有少量的飞燕草素苷和矮牵牛素苷。红叶芥菜、紫红色大白菜、紫色白菜和红菜薹中均含有矢车菊–3–p–香豆酰–丙二酰–葡萄糖苷–5–葡萄糖苷、矢车菊–3–阿魏酰–丙二酰–槐糖苷–5–葡萄糖苷和矢车菊–3–芥子酰–阿魏酰–槐糖苷–5–丙二酰–葡萄糖苷,分别达到总量的46.51%、56.04%、46.38%和68.96%。紫结球甘蓝主要花青素苷成分为矢车菊–3–芥子酰–槐糖苷–5–葡萄糖苷,矢车菊–3–槐糖苷–5–葡萄糖苷和矢车菊–3–p–香豆酰–槐糖苷–5–葡萄糖苷,分别达到总量的41.87%、20.58%和16.02%。花青素苷含量最高的是红叶芥菜,为719.04 μg · g-1 FW;其次是紫红色大白菜,为604.03 μg · g-1 FW;第三是紫结球甘蓝,为264.96 μg · g-1 FW;最低的是红菜薹和紫色白菜,分别为219.07 和130.02 μg · g-1 FW。紫红色大白菜中检测到2 种特有的矢车菊素苷和1 种矮牵牛素苷。     

红掌佛焰苞中花色素与颜色形成的关系

利用切片观察法、显色法和比色法对佛焰苞中花色素的分布情况、组分及含量进行了初步分析。结果表明:红掌佛焰苞中主要含有类黄酮(包括花青素苷、黄酮和黄酮醇)、叶绿素和类胡萝卜素3类色素。大部分品种中类黄酮主要分布于上下表皮细胞中,‘萨维尔’等少数品种的栅栏组织中也有类黄酮分布;而叶绿素和类胡萝卜素则分布在栅栏组织中。各色系均含有类黄酮,红色系比其它色系含有更多的花青素苷,绿色系则含有叶绿素;参试品种中只有棕红色品种‘热带之夜’含有少量类胡萝卜素。花青素苷和叶绿素是佛焰苞显色的主要因素,总体上总花青素苷和叶绿素含量与颜色明度L*呈负相关,而与彩度C*呈正相关,即随着总花青素苷含量的增加,红色和粉红色佛焰苞的红色色调加强,叶绿素含量增加可使绿色和白色佛焰苞的绿色和黄色色调加强。     

Comparison of flower color with anthocyanin composition patterns in evergreen azalea

In evergreen azaleas, major anthocyanins were detected from petals of wild species and cultivars by HPLC analysis. Depending on flower color, all samples were divided into three groups: red, purple or white, using the Japan color standard for horticultural plants. The chromatic components a* and b* values of red group samples showed a convergent distribution, whereas those of purple group samples showed a wider distribution. According to the HPLC analysis, red group samples had two to four major anthocyanins, and those of the purple group had two to six major ones. In contrast, no anthocyanins were detected in the white group petals, although anthocyanidins were detected. These results suggest that the anthocyanin constitution of the purple group flowers is more varied than that of the red group flowers, and this wider variety among purple flowers contributes to extending the diversity of flower color in evergreen azalea.     

Identification and quantification of anthocyanins in different coloured cultivars of ornamental kale (Brassica oleracea L. var. acephala DC)

Anthocyanins are responsible for the colour of many fruits, vegetables, flowers, and coloured-leaved trees. Ornamental kale (Brassica oleracea L. var. acephala DC) is widely cultivated for its colourful inner leaves. To investigate the relationship between the degree of colouration and anthocyanin distribution, content, and composition in ornamental kale, the authors studied the pigment characteristics of five cultivars with different coloured leaves (white, pink, red, purple, and purple-black). Microscopy observation, spectrophotometer, and high-performance liquid chromatography-mass spectrometry (HPLC-MS) analysis of fresh inner leaves revealed that pink, red, and purple colourations were associated with high levels of anthocyanin, while purple-black was the result of the combination of anthocyanins and chlorophyll. In the coloured cultivars, anthocyanins were abundant mainly in the first and second cell layers below the epidermis in both the hypocotyls and inner leaves. No anthocyanin was found in the white-leaved phenotype cultivar. Anthocyanin content increased as leaf colour deepened from pink, red, to purple cultivars, which had little chlorophyll and carotenoid. The authors identified eight anthocyanins in the four coloured cultivars, including one non-acylated, four monoacylated, and three diacylated cyanidin glycosides. Cyanidin-3-(sinapoyl)(feruloyl)-diglucoside-5-glucoside was the most abundant anthocyanin in the four coloured cultivars followed by cyanidin-3-(sinapoyl)-diglucoside-5-glucoside. The analysis of anthocyanin accumulation characterisation provides important information on evaluating colouration patterns in coloured plants, and will be helpful for breeding desired leaf colours in ornamental kale.