一种快速有效分析烟草花冠中花青素苷的方法

花青素苷（anthocyanins）成分和含量的分析是花色研究的重要内容之一。该文利用高效液相色谱-电喷雾离子化-质谱技术（HPLC-ESI-MS）, 建立了一种快速有效地分析烟草（Nicotiana tabacum）花冠中花青素苷成分及含量的方法。在保证良好分离效果的基础上, 将分析时间缩短至15分钟, 大大提高了分析速度, 降低了成本, 并成功应用于转基因烟草花冠中花青素苷成分的分析。为不具备高效分析仪器（如超高效液相色谱-串联质谱（UPLC-MS/MS））的一般实验室研究花青素苷合成相关基因在烟草中的异源表达提供了一种有效且实用的分析方法。     

Image analysis and in vivo imaging as tools for investigation of productivity dynamics in anthocyanin-producing cell cultures of Daucus carota

An anthocyanin-producing suspension culture of Daucus carota (L.) cv. Flakkese was used as model system to study secondary metabolite production in cell culture at the individual cell level. An approach was set up in which growth and production of anthocyanins were investigated using a combination of biochemical analysis, image (colour) analysis and in vivo imaging. This novel approach was used to segment the culture in different subpopulations and dissect the productive process in the cell culture grown under two different conditions, known to differ mainly for oxygen supply and mixing intensity (volume of 50 ml or 20 ml in 250 ml flasks). The 20 ml batch cultures gave a higher content and yield of anthocyanins, which depended on a complex balance between events that positively or negatively affected anthocyanin production. A model is proposed in which the different ability of cells to respond to environmental stimuli and stress depends on the different amount of anthocyanins accumulated within cells.     

Anthocyanin Accumulation Mediated by Blue Light and Cytokinin in Arabidopsis Seedlings

It has been reported that pigmentation In plants Is stimulated by light and cytoklnln （CTK）; however, the signaling pathways and the relationship between light and CTK Involved In the regulation of anthocyanln accumulation remain to be elucidated. We Investigated （i） the role of blue light （BL） and CTK In anthocyanln accumulation ; and （ii） the relationship between BL and CTK In wild type （WT） and by4 mutants of Arabidopsis thaiiana. Two-d-old seedlings grown on medium with or without klnetln （KT） or zeatln （ZT） In darkness were Irradiated using BL at different fluence rates for 3 d before the anthocyanln content was determined using a spectrophotometrlc method. Anthocyanln accumulation was strongly Induced by BL In WT seedlings but not In hy4 seedlings, which demonstrated that CRY1 Is the main photoreceptor for BL. Both KT and ZT enhanced the response of the WT seedlings to BL In a dose-dependent manner, whereas they were not sufficient to promote anthocyanln eccumulatlon In darkness. In addition, data from experiments using the hy4 mutant showed that the CTK effect of BL was also CRYl-dependent. The results from experiments with three different treatment programs showed that the relationship between BL and KT In anthocyanln accumulation of Arabidopsis seedlings seems neither muItlpllcatlve nor additive coactlon, but rather Interaction. BL Is necessary for anthocyanln accumulation, and KT might be Involved In the BL signaling pathway.     

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。     

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。     

植物花色素苷生物合成的转录调控

转录调节是植物花色素苷生物合成途径中调节其结构基因表达的重要环节之一。近十多年来,通过调节转录因子的表达激活或抑制有效地调控植物中花色素苷的合成成了众多植物学家研究的重点。现简要介绍了花色素苷的合成运输过程与液泡的沉积扣押、转录因子与调控及转录调节在遗传改良中的应用等方面的研究进展。     

Structure of anthocyanin from the blue petals of Phacelia campanularia and its blue flower color development

The dicaffeoyl anthocyanin, phacelianin, was isolated from blue petals of Phacelia campanularia. Its structure was determined to be 3-O-(6-O-(4'-O-(6-O-(4'-O-beta-d-glucopyranosyl-(E)-caffeoyl)-beta-d-glucopyranosyl)-(E)-caffeoyl)-beta-d-glucopyranosyl)-5-O-(6-O-malonyl-beta-d-glucopyranosyl)delphinidin. The CD of the blue petals of the phacelia showed a strong negative Cotton effect and that of the suspension of the colored protoplasts was the same, indicating that the chromophores of phacelianin may stack intermolecularly in an anti-clockwise stacking manner in the blue-colored vacuoles. In a weakly acidic aqueous solution, phacelianin displayed the same blue color and negative Cotton effect in CD as those of the petals. However, blue-black colored precipitates gradually formed without metal ions. A very small amount of Al(3+) or Fe(3+) may be required to stabilize the blue solution. Phacelianin may take both an inter- and intramolecular stacking form and shows the blue petal color by molecular association and the co-existence of a small amount of metal ions. We also isolated a major anthocyanin from the blue petals of Evolvulus pilosus and revised the structure identical to phacelianin.