菊花不同花色品种中花青素苷代谢分析

应用高效液相色谱和多级质谱联用技术（HPLC-ESI-MSn）,分析菊花（Chrysanthemum×morifolium）白色、粉色、红色、紫色、红紫色和墨色6个色系共计82个品种中花青素苷合成过程的中间产物和最终产物,发现从白色、粉色、红色、紫色、红紫色到墨色花青素苷含量快速增加,分别为4.68、111.60、366.89、543.56、1220.36和2674.95μg·g-1,不同色系间花青素苷的含量差异显著（P〈0.01）,花青素苷含量越高花色越深;墨色菊花品种中总类黄酮含量显著高于其它花色品种（P〈0.01）,其它不同色系间总类黄酮含量差异不显著（P〉0.05）;随着菊花花色变深,从柚皮素分支到圣草酚的代谢流,以及从圣草酚分支到矢车菊素苷的代谢流比例增加。花青素苷成分分析发现：菊花中只含有矢车菊素苷类化合物。根据花青素苷代谢成分分析结果绘制了菊花中花青素苷代谢路径图,即在菊花类黄酮代谢途径中只存在矢车菊素苷代谢分支途径;菊花不同色系在柚皮素和圣草酚2个关键代谢分支点上向不同方向代谢流的分配比例不同,造成花青素苷产物含量不同,导致不同花色。以上研究结果为菊花花色改良的分子育种提供了理论依据。     

基于花青素代谢培育蓝色花卉的研究进展

花色是植物吸引昆虫传播花粉的主要因素,对于植物在自然界的生存必不可少,也是观赏植物最重要的性状之一。在蓬勃发展的花卉产业中,色彩各异花卉的培育,可以弥补自然花色的匮乏,但是令人垂涎的蓝色花比较难培育。花色的多样性主要是由花青素及其衍生物的种类和含量等因素决定的,飞燕草色素的合成是形成蓝色花的关键因素,许多植物体内缺少合成飞燕草色素的结构基因。近年来,利用基因工程技术培育蓝色花的研究也时有报道。文中以常见的观赏植物为例,基于花青素代谢调控,从影响飞燕草色素合成的关键因素和不同分子改良途径培育蓝色花等几个方面对植物花朵呈色的机制进行了综述,并展示不同分子育种策略可能在其他领域的应用,为其他植物或经济作物的色泽改良如彩色棉蓝色纤维的培育等提供参考和技术支持。     

蔷薇属植物花青素苷的测定及其对花瓣呈色的影响

通过目测法及比色卡进行表型观测并选取8种不同花色的蔷薇属植物作为研究对象,利用分光色差仪测定花色表型,采用高效液相色谱质谱联用技术对花青素苷进行定性定量分析,探讨蔷薇属植物花色与花青素苷之间的关系。结果表明,8种蔷薇属植物材料首次检测出8种花青素苷成分,分别为矢车菊素-3-（咖啡酰基）-葡萄糖苷（Cy3CafG）、矢车菊素-3-O-半乳糖苷（Cy3Gal）、芍药素-3-（咖啡酰基）-葡萄糖苷（Pn3CafG）、矢车菊素-3-（顺式-咖啡酰基）-二甲基葡萄糖苷（Cy3(cis Caf)DmG）、矢车菊素-3-（反式-咖啡酰基）-二甲基葡萄糖苷（Cy3(trans Caf)DmG）、矢车菊素-3-二甲基-葡萄糖苷（Cy3DmG）、芍药素-3-（顺式-咖啡酰基）-芸香苷（Pn3(cis Caf)Ru）、芍药素-3-（反式-咖啡酰基）-芸香苷（Pn3(trans Caf)Ru）。8种花青素苷的生物修饰类型包含半乳糖糖基化、甲基化修饰和咖啡酰基化修饰3种,这3种生物修饰类型均在蔷薇属植物中首次报道。花青素苷与CIELab参数相关性结果显示Cy3CafG、Cy3(cis Caf)DmG与花瓣红度（...     

耐寒睡莲花瓣中花青素苷组成及其与花色的关系

睡莲(Nymphaea spp.)为多年生水生观赏花卉。以耐寒睡莲不同花色的119个栽培品种为材料, 利用高效液相色谱(HPLC-DAD)和液质联用技术(HPLC-ESI-MSⁿ)测定了其花瓣中的花青素苷成分。采用英国皇家园艺学会比色卡(RHSCC)和国际照明委员会(CIE)制定的CIEL^*a^*b^*表色系统测量了57个品种的花色, 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: 耐寒睡莲花瓣中含有14种花青素苷, 其中飞燕草素-3-半乳糖-5-乙酰-半乳糖苷(Dp3Ga5acetylGa)、飞燕草素-3-鼠李糖-(1→2)-半乳糖苷(Dp3Rh(1→2)Ga)、矢车菊素-3-半乳糖-(1→2)-半乳糖苷(Cy3Ga(1→2)Ga)、矢车菊素-3-乙酰-半乳糖-(1→2)-半乳糖苷(Cy3acetylGa(1→2)Ga)、矢车菊素-3-没食子酰-半乳糖苷(Cy3galloylGa)、飞燕草素-3-乙酰-葡萄糖苷(Dp3acetylG)、飞燕草素-3-葡萄糖苷(Dp3G)和矢车菊素-3-半乳糖-半乳糖-半乳糖苷(Cy3GaGaGa)8个组分在耐寒睡莲中为首次报道。Dp3Ga、Dp3galloylGa、Cy3Ga(1→2)Ga和Cy3galloylGa是决定耐寒睡莲呈色的关键花青素苷。     

葡萄花青素苷生物合成研究进展

花青素苷是一类重要的天然色素物质,是植物主要呈色物质之一,并在人类健康保护方面发挥越来越重要的作用。同时,它又是一类复杂性状,不仅与遗传基因相关,还与外界温度、湿度、光质及栽培措施等因素息息相关。葡萄花青素苷是近年来花青素苷研究的热点,主要从葡萄花青素苷的结构多样性和合成途径等方面展开简要综述,以期为葡萄花青素苷复杂性状的遗传机理和调控作用机制与分子育种及葡萄优质早熟栽培研究上提供有益的信息。     

环境因子调控植物花青素苷合成及呈色的机理

花青素苷(anthocyanin)是决定被子植物花、果实和种皮等颜色的重要色素之一。花青素苷的合成与积累过程往往与植物发育过程密切相关, 由内外因子共同控制。环境因子通过诱导植物体内花青素苷合成途径相关基因的表达来调控花青素苷的呈色反应。该文追踪了国内外相关研究, 认为光是影响花青素苷呈色的主要环境因子之一, 光质和光强均能在一定程度上影响花青素苷的合成, 其中光质起着更为关键的作用; 低温能诱导花青素苷的积累, 高温则会加速花青素苷的降解;不同的糖类物质均能影响花青素苷的合成, 大部分结构基因和调节基因的表达均受糖调控。关于花发育与花青素苷呈色的关系、观赏植物花色对环境因子的响应以及花青素苷抵御逆境的机理尚待深入研究。因此, 综合考察花发育与植物花青素苷合成及其呈色之间的关系, 特别是光周期对花发育的影响导致花青素苷合成及呈色的机理是花色研究的一个重要课题。利用环境因子调控花色将会极大地提高花卉的观赏价值。     

蓝亚麻花瓣中类黄酮化合物及代谢途径分析

花色是观赏植物重要的观赏性状之一,而类黄酮是其主要的呈色物质。该研究以蓝亚麻花瓣为研究对象,将蓝亚麻开花过程分为5个阶段,并用高效液相色谱—光电二极管阵列检测技术(HPLC-PAD)和高效液相色谱—电喷雾离子化—质谱连用技术(HPLC-ESI-MS)分析不同开花阶段花瓣中类黄酮化合物的成分和含量。结果表明:蓝亚麻花瓣中积累飞燕草素苷、矢车菊素苷和锦葵素苷,未检测到天竺葵素苷,其中以酰基化的飞燕草素苷为主要呈色物质;而总花青素苷含量在第2阶段达到最高。根据花青素苷终产物和类黄酮中间代谢产物推定了蓝亚麻花瓣中类黄酮代谢途径,其中以F3'5'H所引导的分支途径占优势,其主要原因可能是F3'5'H酶活高于F3'H。     

西昌万寿菊花中化学成分的研究

从万寿菊（Tagetes ereata L．）花中分得8个已知化合物,分别为丁香酸（Syringic acid,1）,3,4二羟基苯甲酸（3,4-dihydroxybenzonic acid,2）,β-香树脂醇（β-amyrin,3）,槲皮万寿菊素（Quercetagetin,4）,Quercetagetin 7-methyl（5）,β-谷甾醇（β-sitosterol,6）,豆甾醇（stigmasterol,7）,正三十四烷（Tetratriacontane,8）。应用光谱法鉴定了所有化合物的结构。     

茄科蔬菜花青素苷分子调控研究进展

花青素苷是一种分布广泛的水溶性色素,不仅赋予了果实多彩的外表,还是天然食用色素的重要来源。近年来有关茄科蔬菜花青素苷的研究逐渐增多,文中从花青素苷结构及其生物合成途径、茄科蔬菜中花青素苷合成代谢的结构基因和调节基因、影响合成的环境因素等方面进行回顾和总结,为进一步阐明茄科蔬菜花青素苷的合成及调控机理、更好利用花青素苷进行果色品质育种的创新提供一些参考。     

丽格海棠花青素苷成分及分布对花色的影响

以红色、红心白边、粉红、玫红、黄色、黄心红边、浅粉和白色8种花色丽的格海棠花瓣为试验材料,采用目视测色法、RHSCC比色法和色差仪测定花瓣表型,通过组织切片法观察花瓣色素细胞的显微结构和分布特点,采用双光束紫外-可见光分光光度计和高效液相色谱-电喷雾离子化-质谱连用技术(HPLC-ESI-MS)测定分析花瓣中花青素苷的成分和含量,为探讨丽格海棠花色的呈色机理和花色育种提供参考。结果显示:(1)丽格海棠的明度L*随花瓣颜色变深而降低,红度a*则表现出相反趋势,红度(a*)和彩度(C*)值与明度(L*)呈显著负相关关系,且a*和C*是影响L*的主要因素。(2)红花品种花瓣色素主要分布于上表皮细胞和海绵组织中;红白花品种花瓣色素主要分布于上下表皮中,且下表皮积累量更多;粉色花和玫红花品种花瓣色素主要分布于上下表皮细胞;黄红花和粉白色花品种花瓣上表皮中含有少量色素,而黄花和白花品种花瓣几乎没有色素积累。各花色丽格海棠花瓣上表皮细胞均为圆锥形,且红花和红白花品种锥形化程度最高,它们花瓣下表皮细胞均呈扁平的长方形。(3)8个丽格海棠品种花瓣中共检测出15种花青素苷,其中10种为芍药素苷,3种为矢车菊素苷,1种为锦葵素苷,1种为飞燕草素苷,酰化花青素苷占多数;红花品种花瓣中总花青素苷含量最高,玫红花品种次之,黄花和白花品种中未检出;除粉红花品种外,其余含花青素苷的品种中芍药素苷含量最高,均占总花青素苷含量的50%以上,是花瓣的主要呈色物质。(4)丽格海棠花瓣中总花青素苷含量与其红度(a*)、彩度(C*)值呈正相关关系、与其L*值呈负相关关系。研究表明,花青素苷的积累有利于丽格海棠花瓣红色化,并影响其花瓣彩度(C*)及明度(L*);色素分布细胞数量和上表皮细胞锥形化明显影响花瓣呈色,且花瓣主要的呈色物质为芍药素苷,酰基化修饰可能影响其明度。     

Accumulation of Anthocyanins and Flavones during Bud and Flower Development in Campanula isophylla Moretti

In the blue flowers of Italian bellflower (Campanula isophyllaMoretti), the formation of anthocyanins progresses from simpleunacylated anthocyanins, delphinidin 3–glucoside and bisdeacylplatyconin,through a series of progressively-acylated and glycosylatedcompounds, including diacylated violdelphin and monodeacylcampanin,to the triacylated campanin. In this study, anthocyanin andflavone contents were very low in buds until a few days beforeanthesis, after which they increased rapidly. Bisdeacylplatyconinand luteolin 7-O -glucoside peaked 2 d before anthesis. Themore complicated luteolin glucosides peaked 2 d after anthesis,slightly preceding monodeacylcampanin and campanin. Total anthocyanincontent peaked approx. 5 d after anthesis followed by a slowdecline. The highest total flavone content was reached at anthesis,after which it remained almost constant, but with some changesin the proportion of individual compounds. In the investigationtwo phenotypes were used, types B and C. Acylation of monodeacylcampaninto campanin is blocked in type B, but not in type C plants.Conversion of bisdeacylplatyconin into acylated anthocyaninswas shown to be slower in type C than in type B plants. Campanula isophylla ; Campanulaceae; Italian bellflower; anthocyanin; flavone; biosynthesis; flower development     

鸳鸯茉莉开花过程中花青素组成的变化

为了解鸳鸯茉莉(Brunfelsiaacuminata)花色变化的机理,采用高效液相色谱(HPLC)体系检测其开花过程中花青素组成的变化。结果表明,优化的HPLC体系为:流速为0.8 mL min–1,流动相A为7.5%甲酸乙腈,流动相B为7.5%甲酸水,洗脱程序为0 min,8%A;15 min,18%A;25 min,23%A;45 min,40%A;50 min,8%A。利用优化体系检测到鸳鸯茉莉花瓣中含有锦葵色素-3-O-葡萄糖苷、矮牵牛素葡萄糖苷和飞燕草素葡萄糖苷3种花青苷,其中锦葵色素-3-O-葡萄糖苷的含量最高,飞燕草素葡萄糖苷含量最低,且在花色由深变浅的过程中3种花青苷的含量均降低。因此,鸳鸯茉莉的呈色与这3种花青苷有关,且锦葵色素-3-O-葡萄糖苷起主导作用。     

Molecular Basis for Chemical Evolution of Flavones to Flavonols and Anthocyanins in Land Plants

During the course of evolution of land plants, different classes of flavonoids, including flavonols and anthocyanins, sequentially emerged, facilitating adaptation to the harsh terrestrial environment. Flavanone 3β-hydroxylase (F3H), an enzyme functioning in flavonol and anthocyanin biosynthesis and a member of the 2-oxoglutarate-dependent dioxygenase (2-ODD) family, catalyzes the hydroxylation of (2S)-flavanones to dihydroflavonols, but its origin and evolution remain elusive. Here, we demonstrate that functional flavone synthase Is (FNS Is) are widely distributed in the primitive land plants liverworts and evolutionarily connected to seed plant F3Hs. We identified and characterized a set of 2-ODD enzymes from several liverwort species and plants in various evolutionary clades of the plant kingdom. The bifunctional enzyme FNS I/F2H emerged in liverworts, and FNS I/F3H evolved in Physcomitrium (Physcomitrella) patens and Selaginella moellendorffii, suggesting that they represent the functional transition forms between canonical FNS Is and F3Hs. The functional transition from FNS Is to F3Hs provides a molecular basis for the chemical evolution of flavones to flavonols and anthocyanins, which contributes to the acquisition of a broader spectrum of flavonoids in seed plants and facilitates their adaptation to the terrestrial ecosystem.

The success of land plants in the colonization of and adaptation to terrestrial ecosystems has been particularly attributed to the emergence and evolution of a unique metabolic capacity that synthesizes diverse specialized metabolites, including flavonoids, a highly polymorphic class of polyphenols (Weng and Chapple, 2010). The flavonoid metabolites have been classified into several subgroups, namely flavanones, dihydroflavonols, flavones, flavonols, flavan-3,4-diols, flavan-3-ols, and anthocyanins, based on their oxidation status and substitution patterns of the core skeleton (Winkel-Shirley, 2001; Martens et al., 2010). Along with the evolution of land plants, different classes of flavonoids emerged (Koes et al., 1994). The basal land plants liverworts produce chalcones, flavanones, and flavones; whereas lycophytes gained the ability to produce proanthocyanidins (Markham, 1984; Koes et al., 1994). Furthermore, both pteridophyta and gymnosperms, while dominated with flavone production, began to produce flavonols (Markham, 1984; Koes et al., 1994). Finally, flavonols and anthocyanins are well represented in angiosperms. Flavonols, which bear a 3-hydroxyl group in the core structure, have been exploited as effective photoprotectants against UV-B radiation (Solovchenko and Schmitz-Eiberger, 2003), as signal providers to symbionts (Hungria et al., 1991), as regulators of the transport of phytohormones (Peer and Murphy, 2007), and as determinants of conditional male fertility (Muhlemann et al., 2018). Anthocyanins, derived from dihydroflavonol, are important for sexual reproduction, acting as attractants for insect pollinators and for animal dispersers of seed (Shimada et al., 2005). It is obvious that a clear chemical evolution trace from chalcones, flavanones, and flavones to flavonols and anthocyanins, occurs across plant phyla. However, the molecular basis for such a chemical evolution remains mysterious.The biosynthesis of flavones and flavonols requires chemical conversion of a common precursor, (2S)-flavanone, and is catalyzed by flavone synthase I (FNS I) and flavanone 3β-hydroxylases (F3Hs), respectively. Both enzymes as well as flavonol synthase (FLS) and anthocyanidin synthase (ANS) belong to a larger enzyme family, the 2-oxoglutarate-dependent dioxygenases (2-ODDs; Farrow and Facchini, 2014). FNS I converts (2S)-flavanone to flavone via desaturation of carbon 2 and 3 of the heterologous ring of flavanone (Gebhardt et al., 2005, 2007), while F3H catalyzes the conversion of (2S)-flavanone to (2R,3R)-dihydroflavonol by hydroxylation of the C-3β position (Supplemental Fig. S1). Subsequently, FLS converts (2R,3R)-dihydroflavonols to their corresponding flavonols, and ANS catalyzes the nonpigmented leucoanthocyanidins (leucopelargonidin, leucocyanidin, and leucodelphinidin) to the pigmented anthocyanidins (pelargonidin, cyanidin, and delphinidin, respectively; Supplemental Fig. S1). These four classes of 2-ODD enzymes phylogenetically form two distinct subgroups, one consisting of F3H and FNS I and the other consisting of FLS and ANS. FNS I and F3H both use flavanone as substrate and exhibit, in general, a relatively narrow substrate specificity (Turnbull et al., 2000; Martens et al., 2003), whereas ANS and FLS display some degree of promiscuity in their substrate preferences and catalytic activities. For example, Arabidopsis (Arabidopsis thaliana) FLS1 is not only capable of converting dihydroflavonols to their corresponding flavonols but also mediates the oxidation of 2S-flavanone (naringenin) to both dihydrokaempferol enantiomers, an activity normally associated with F3H (Prescott et al., 2002). While F3Hs are ubiquitous in vascular plants, FNS Is appear to be confined to the Apiaceae family as well as a few non-Apiaceae species such as rice (Oryza sativa; Lee et al., 2008), maize (Zea mays), and Arabidopsis (Falcone Ferreyra et al., 2015). Prior to the discovery of FNS Is in those non-Apiaceae species, it was assumed that the gene encoding FNS I arose from duplication and mutation of F3H (Martens et al., 2001, 2003; Gebhardt et al., 2005, 2007). However, the FNS Is revealed in both Z. mays and Arabidopsis show very poor sequence similarity with those present in Apiaceae species, which suggests that the evolution of the FNS Is was not as clear-cut as was originally believed. It is likely that the evolution of FNS occurred several times independently. In several cereal crops, such as Z. mays, O. sativa, and wheat (Triticum aestivum), flavones are the major flavonoid substances, which protect the plants during pathogen attack and under biotic or abiotic stress conditions (Righini et al., 2019).Previously, we found that the liverwort Plagiochasma appendiculatum FNS I (which should change to PaFNS I/F2H, according to the function) converted flavanone to 2-hydroxyflavanone and flavone (Han et al., 2014). The dual FNS I and F2H activities of PaFNS I/F2H, together with the fact that its amino acid sequence shares a higher identity with F3Hs than with FNS Is, implicates an evolutionary connection between liverwort FNS Is and seed plant F3Hs. On the other hand, previous in silico analysis failed in identifying any F3H sequences in either the bryophyte Physcomitrium (Physcomitrella) patens or the lycophyte Selaginella moellendorffii, even though both species produce dihydroflavonol-derived metabolites. To identify when and how F3H emerged and evolved to produce a vast variety of flavonoid metabolites, we systematically identified FNS I and F3H homologous sequences from species of different phyla, including liverworts, P. patens, S. moellendorffii, gymnosperms, and angiosperms. Subsequent biochemical characterization revealed that the functionally promiscuous FNS Is widely emerged in the liverworts, which evolved into a dual-function enzyme with both FNS I and F3H activities in both P. patens and S. moellendorffii. Further evolution led to the emergence of F3H with a minor level of FNS I activity in gymnosperm species, while those generated by angiosperm species showed a more specific F3H activity.     

Studies on flower color in Chrysanthemum morifolium RAMAT. I. Anthocyanins

Isolation and characterization of anthocyanins from the flowersof two cultivars of Chrysanthemum morifolium RAMAT. are reported.The main pigment is a new glucoside of cyanidin. ¹Cultivated at the Kyoto University Agricultural ExperimentalFarm (Received October 25, 1969; )     

芦蒿花总黄酮提取工艺的研究

采用正交试验,以芦蒿总黄酮得率为考察指标,对影响芦蒿总黄酮提取工艺的因素进行了探讨和研究,得出了芦蒿花总黄酮提取过程的优化条件。     

黄山贡菊花、叶中总黄酮及绿原酸动态积累研究

本文分别采用紫外分光光度法和HPLC法对贡菊花及叶中总黄酮及绿原酸的含量进行分析,研究不同生长时期黄山贡菊花和叶中总黄酮与绿原酸动态积累规律。结果表明:黄山贡菊花中总黄酮和绿原酸含量在中花期均达到最高值,而在采摘期内变化较大;黄山贡菊叶中总黄酮和绿原酸含量随着生长期的延长呈现逐渐上升趋势,并在生长后期总黄酮和绿原酸含量分别出现109.30和7.87 mg/g的峰值。     

紫背天葵中营养成分及总黄酮分析

对食、药两用植物紫背天葵营养成分和总黄酮进行了研究 ,结果显示紫背天葵各种营养成分齐全且丰富 ,其中总氨基酸质量分数达 1 3.0 3% ,必需氨基酸占总氨基酸的 4 4 .1 3%。含人体必需的无机元素 1 2种 ,人体必需的微量元素 7种。植物中正丁醇提取部位的总黄酮含量为 0 .4 1 %。这些研究为人们认识紫背天葵的营养和药用价值 ,充分开发利用祖国的植物资源提供参考     

Changes in the Levels of Endogenous Growth Regulators during Development of the Flowers of Chrysanthemum morifolium

Gibberellin-like substances and an auxin similar to IAA weredetected by bioassays in extracts of flowers of Chrysanthemummorifolium. The activity of these substances was shown to reacha maximum early in the development of the flower when its relativegrowth-rate was at a maximum, and then to decline with the relativegrowth-rate. The leaves of lateral flowering shoots were found to containgibberellins similar to those detected in the flowers whilea different gibberellin, which appeared to decrease in activitywith the age of the shoot, was detected in the stem. An auxinsimilar to indol-3yl-acetic acid (IAA) was also detected inthese stems. Growth-promoting substances were not detected inthe old stems and leaves from the main shoot. Gas-liquid chromatographyrevealed the presence of a number of additional gibberellinsin the flowers. The chemical nature of the growth substances is discussed inrelation to their biological and chromatographic behaviour.     

Isocucurbic Acid Derivatives and Soluble Epoxide Hydroxylase Inhibitors from the Flowers of Chrysanthemum indicum L.

Soluble epoxide hydrolase (sEH) inhibitory activity guided fractionation and isolation of two new isocucurbic acid derivatives ( 1 and 2 ) and nine known compounds ( 3 – 11 ) from the flowers of Chrysanthemum indicum L. Their structures were elucidated on the basis of spectroscopic data interpretation and comparison with those reported in previous studies. Luteolin ( 3 ), acacetin-7-O-β-D-glucopyranoside ( 6 ), and methyl 3,4-di-O-caffeoylquinate ( 10 ) displayed sEH inhibitory activities with IC₅₀ values ranging from 13.7±3.6 to 20.8±0.4 μM. Enzyme kinetic analysis revealed that 3 , 6 , and 10 were non-competitive inhibitors with K_i values of 14.8±0.5, 31.2±0.8, and 3.9±0.2 μM, respectively. Additionally, molecular docking studies indicated compound 10 had the ability to form six hydrogen bonds at sEH active site, resulting binding energy as low as −9.58 Kcal/mol.