UV-B radiation affects flavonoids and fungal colonisation in <Emphasis Type="Italic">Fagopyrum esculentum</Emphasis> and <Emphasis Type="Italic">F. tataricum</Emphasis>

In the present study, we have evaluated the effects of increased UV-B radiation that simulates 17% ozone depletion, on fungal colonisation and concentrations of rutin, catechin and quercetin in common buckwheat (Fagopyrum esculentum) and tartary buckwheat (Fagopyrum tataricum). Induced root growth and reduced shoot:root ratios were seen in both of these buckwheat species after enhanced UV-B radiation. There was specific induction of shoot quercetin concentrations in UV-B-treated common buckwheat, whereas there were no specific responses for flavonoid metabolism in tartary buckwheat. Root colonisation with arbuscular mycorrhizal fungi significantly reduced catechin concentrations in common buckwheat roots, and induced rutin concentrations in tartary buckwheat, but did not affect shoot concentrations of the measured phenolics. Specific UV-B-related reductions in the density of microsclerotia were observed in tartary buckwheat, indicating a mechanism that potentially affects fungus-plant interactions. The data support the hypothesis that responses to enhanced UV-B radiation can be influenced by the plant pre-adaptation properties and related changes in flavonoid metabolism.     

荞麦黄酮及其生物合成调控研究进展

荞麦属植物资源丰富,且富含黄酮类成分。通过文献查阅,总结了荞麦黄酮历年研究情况以及热点研究领域。荞麦黄酮研究论文最早发表于1952年,在1952—1999近五十年的时间内,荞麦黄酮的研究论文较少,年发文量少于10篇,荞麦黄酮的研究处于起步阶段。自2000年后,荞麦黄酮逐渐获得更多研究学者的关注,年度发文量逐年上升。近年来,荞麦黄酮研究热点集中在植物学、食品科学技术、农学以及生物化学与分子生物学学科领域中,黄酮抗氧化活性相关的研究论文被引用次数较高,荞麦黄酮的生物活性与营养功能一直备受关注。目前,从荞麦中已经鉴定的黄酮类化合物达80种。槲皮素、山奈酚、木犀草素、鼠李素、异鼠李素、小麦黄素、柚皮素、杨梅素、芹菜素以及橙皮素是荞麦中常见的黄酮苷元结构,芍药色素、花翠素、矢车菊素为荞麦中多见的花青素类型。荞麦黄酮生物合成起源于苯丙烷代谢途径,PAL、CHS、C4H、4CL、CHI、LAR等黄酮合成途径中的多个关键酶基因以及MYB转录因子基因已被克隆鉴定。荞麦MYB转录因子在黄酮生物合成中发挥着重要的诱导调控作用,影响荞麦黄酮合成积累的因素主要有环境因素、植物生长调节剂、生物因素以及品种等,多个因素可相互交叉调控和影响荞麦黄酮的合成。该文通过回顾历年荞麦黄酮研究概况,总结黄酮化合物的种类,归纳黄酮生物合成途径调控机制及主要影响因素,为优质荞麦种植生产和优化提升荞麦产品的营养保健功能奠定理论基础和提供可行方案,为荞麦黄酮的深入研究指明方向。     

苦荞突变体库构建与突变体中芦丁合成相关基因表达分析

苦荞是重要的小杂粮作物之一,营养物质丰富,是天然芦丁的重要来源。突破苦荞育种难题,创制苦荞新种质是目前研究的重要方面。本试验利用甲基磺酸乙酯(EMS)构建了黑丰1号苦荞突变体库,明确了当EMS浓度为1.2%时,诱变效果较好。通过对M1突变株表型观察统计,共获得叶色、叶型、株型、粒型变异单株102株,突变率为3.85%;高效液相色谱技术(HPLC)测定1000株M3材料,获得高芦丁含量突变株系2个和低芦丁突变体株系5个;qRT-PCR对芦丁含量突变体株系中芦丁代谢关键酶基因(CHS、F3H、4CL、FLS、UFGT)进行表达量分析,发现不同株系中上述基因的表达量与芦丁含量相关性不明显,但个别基因如FtFLS基因表达量在高芦丁含量突变体中达到对照的4.55倍。通过突变体的筛选丰富了苦荞基因资源,创新了苦荞新种质,也为苦荞芦丁代谢的分子基础研究提供了材料保证与技术支持。     

苦荞黄酮转运相关蛋白基因FtAH4L的克隆及表达分析

为研究苦荞黄酮转运相关基因,以苦荞(Fagopyrum tataricum)品种"西荞二号"为材料,克隆到1条质膜H+-ATPase基因(autoinhibited H+-ATPase isoform 4 like,AH4L),将其命名为FtAH4L。通过开放阅读框(ORF)分析,FtAH4L基因cDNA全长3 398bp,开放阅读框2 898bp,编码966个氨基酸残基,理论分子量为109kD,等电点6.48。氨基酸保守基序比对分析表明,AH4L在植物种间较为保守。在茉莉素诱导处理和5种光(白色荧光、LED白光、LED蓝光、LED红光和UV-B)处理芽期苦荞后,采用半定量RT-PCR和AlCl3比色法分析结果表明,茉莉素处理后的苦荞胚轴和子叶中FtAH4L基因表达量与黄酮含量均显著上升,且二者呈正相关关系;5种光对子叶中FtAH4L表达量无显著影响,但均显著增加其黄酮含量;胚轴中,除LED红光外,各种光均显著提高FtAH4L表达量和总黄酮含量,且LED蓝光与UV-B的影响极显著。该研究结果为深入研究FtAH4L基因参与苦荞黄酮转运奠定了基础。     

遮阴对柠檬香茅类黄酮及其合成酶基因差异表达研究

为了解柠檬香茅(Cymbopogon citratu)中类黄酮及其合成酶基因信息,以阳光直射及遮阴环境下生长的柠檬香茅嫩叶为材料,进行代谢组、转录组结合qRT-PCR验证分析。结果表明,柠檬香茅中含有11类共69种黄酮化合物,其中芦丁、去甲基托罗沙黄酮、紫云英苷及葡萄糖醇等类黄酮化合物在遮阴环境下相对含量显著降低;类黄酮生物合成涉及10类酶54个基因,其中类黄酮3''羟化酶(c99177.1)等4个酶基因在遮阴环境下相对表达量显著降低,而异黄酮合成酶(c51975.0)等6个酶基因相对表达量正好相反;其中5个类黄酮合成酶基因在光照及遮阴柠檬香茅中的上下调表达趋势与转录组测序结果中FPKM值变化一致,而二者检测结果中差异表达倍数存在差异。遮阴使柠檬香茅中大多黄酮类化合物相对含量降低,而其合成酶基因上下调表达趋势规律不明显。     

自交可育甜荞、金苦荞、米苦荞不同品系总黄酮、粗蛋白及其蛋白组分含量研究北大核心CSCD

为探究荞麦新品种的营养保健价值,该文对自交可育甜荞、金苦荞、米苦荞共56个不同品系荞麦种子的粗蛋白、总黄酮、蛋白组分含量及其果实性状的变异进行了研究。结果表明:(1)甜荞、金苦荞、米苦荞种子中粗蛋白含量平均值分别为13.19%、15.44%、11.75%,总黄酮含量平均值分别为0.14%、2.50%、2.09%,清蛋白含量的平均值分别为5.22%、6.13%、4.56%,球蛋白含量的平均值分别为1.29%、1.15%、0.91%,醇溶蛋白含量的平均值分别为0.42%、0.58%、0.55%,谷蛋白含量的平均值分别为2.66%、3.36%、2.80%,三种荞麦的蛋白组分均符合清蛋白>谷蛋白>球蛋白>醇溶蛋白。(2)果实性状中,甜荞果实千粒重、果实面积、果实直径的变异系数最大,米苦荞果实周长、果实长宽比、果实长、果实宽和50 mL容重的变异系数最大。(3)相关分析表明甜荞种子粗蛋白含量与果实长宽比、果实长,金苦荞种子粗蛋白含量与果实周长、果实长,米苦荞种子粗蛋白含量与果实宽、总黄酮含量与果实面积、果实宽、果实直径、50 mL容重的相关性均达到了显著或极显著水平。(4)该研究筛选出甜荞(1808-166贵甜2号优系)、金苦荞(多苦74、多苦78)、米苦荞(1906-136黑米荞麦、43-2)等高蛋白含量或高黄酮含量的荞麦品系。该研究结果对荞麦优良品种的选育和荞麦新产品的开发具有一定指导意义。     

High-quality Fagopyrum esculentum genome provides insights into the flavonoid accumulation among different tissues and self-incompatibility

Common buckwheat (Fagopyrum esculentum) and Tartary buckwheat (Fagopyrum tataricum), the two most widely cultivated buckwheat species, differ greatly in flavonoid content and reproductive mode. Here, we report the first high-quality and chromosome-level genome assembly of common buckwheat with 1.2 Gb. Comparative genomic analysis revealed that common buckwheat underwent a burst of long terminal repeat retrotransposons insertion accompanied by numerous large chromosome rearrangements after divergence from Tartary buckwheat. Moreover, multiple gene families involved in stress tolerance and flavonoid biosynthesis such as multidrug and toxic compound extrusion (MATE) and chalcone synthase (CHS) underwent significant expansion in buckwheat, especially in common buckwheat. Integrated multi-omics analysis identified high expression of catechin biosynthesis-related genes in flower and seed in common buckwheat and high expression of rutin biosynthesis-related genes in seed in Tartary buckwheat as being important for the differences in flavonoid type and content between these buckwheat species. We also identified a candidate key rutin-degrading enzyme gene (Ft8.2377) that was highly expressed in Tartary buckwheat seed. In addition, we identified a haplotype-resolved candidate locus containing many genes reportedly associated with the development of flower and pollen, which was potentially related to self-incompatibility in common buckwheat. Our study provides important resources facilitating future functional genomics-related research of flavonoid biosynthesis and self-incompatibility in buckwheat.     

水母雪莲红色细胞系类黄酮含量和相关基因表达

水母雪莲为中国传统名贵中药,具有散寒除湿、活血通经、抗炎、镇痛等功效,其主要药用成分为类黄酮化合物。以水母雪莲白色系愈伤组织和经低温、高糖、强光诱导得到的红色系为材料,比较不同细胞系中类黄酮活性成分、结构基因和转录因子表达的差异。结果表明,红色系中总黄酮含量为白色系的3.60倍;重要的药用成分为芦丁,其含量达到干重的0.25%,是白色系的2.40倍;红色系中花青苷含量极高,矢车菊素3-O-己糖苷和矢车菊素3-O-琥珀酰己糖苷的含量分别达到干重的0.12%和0.19%;红色系中CHS、F3'H、FNS、FLS、DFR和ANS基因的表达均明显高于白色系;红色系中转录因子MYB、bHLH和WD40的表达也均明显高于白色系,其中MYB的表达量为白色系的19.70倍,说明红色系中转录因子的高水平表达增强了结构基因的表达,进而提高了类黄酮的合成。红色系中bHLH和WD40表达水平相似,而与MYB的表达水平相差很大,推测可能在水母雪莲中bHLH和WD40两种转录因子形成二元复合体后,和MYB共同调控类黄酮合成途径中结构基因的表达。     

低海拔地区黑苦荞酚类物质含量、组成及抗氧化活性研究

为探究黑苦荞的市场利用价值,该研究选择种植于湖北江汉平原低海拔地区的川荞1号和九江苦荞作为材料,分析苦荞籽粒中游离酚、结合酚、总酚、游离黄酮、结合黄酮和总黄酮的含量,利用DPPH自由基法、ABTS自由基法和铁离子还原抗氧化法(FRAP)三种抗氧化测试模型综合评价其体外抗氧化活性,并运用高效液相色谱(HPLC)技术对其酚类物质的组成进行鉴定。结果表明:(1)川荞1号籽粒的总酚和总黄酮含量显著高于九江苦荞,分别为27.38 mg GAE·g~(-1)DW、31.46 mg RE·g~(-1)DW和12.71 mg GAE·g~(-1)DW、14.68 mg RE·g~(-1)DW;其中游离酚与游离黄酮含量显著高于结合酚与结合黄酮含量,均占总酚和总黄酮含量的79%以上,且九江苦荞中结合酚和结合黄酮的含量高于川荞1号。(2)苦荞籽粒中酚类物质主要由芦丁、槲皮素、表儿茶素、山奈酚、山奈酚-3-芸香糖苷和槲皮素-3-O-芸香糖苷-3'-O-吡喃葡萄糖苷等黄酮类化合物组成,其中游离酚以芦丁和槲皮素为主,结合酚以表儿茶素和芦丁为主。(3)苦荞籽粒提取物均具有一定的抗氧化活性,黑苦荞川荞1号游离态DPPH、ABTS和FRAP抗氧化能力值分别为30.14、11.03、18.84 mg TE·g~(-1)DW,高于九江苦荞,而结合态三种抗氧化能力值低于九江苦荞,但黑苦荞川荞1号总抗氧化能力显著高于九江苦荞。在低海拔地区江汉平原,种植的黑苦荞川荞1号籽粒具有较高含量的酚类物质,符合后续的食品加工的生产要求,市场开发前景广阔。     

Flower colour and cytochromes P450

Flavonoids are major constituents of flower colour. Plants accumulate specific flavonoids and thus every species often exhibits a limited flower colour range. Three cytochromes P450 play critical roles in the flavonoid biosynthetic pathway. Flavonoid 3′-hydroxylase (F3′H, CYP75B) and flavonoid 3′,5′-hydroxylase (F3′5′H, CYP75A) catalyze the hydroxylation of the B-ring of flavonoids and are necessary to biosynthesize cyanidin-(red to magenta) and delphinidin-(violet to blue) based anthocyanins, respectively. Pelargonidin-based anthocyanins (orange to red) are synthesized in their absence. Some species such as roses, carnations and chrysanthemums do not have violet/blue flower colour due to deficiency of F3′5′H. Successful expression of heterologous F3′5′H genes in roses and carnations results in delphinidin production, causing a novel blue/violet flower colour. Down-regulation of F3′H and F3′5′H genes has yielded orange petunia and pink torenia colour that accumulate pelargonidin-based anthocyanins. Flavone synthase II (CYP93B) catalyzes the synthesis of flavones that contribute to the bluing of flower colour, and modulation of FNSII gene expression in petunia and tobacco changes their flower colour. Extensive engineering of the anthocyanin pathway is therefore now possible, and can be expected to enhance the range of flower colours.     

Isolation and functional characterization of a novel gene coding for flavonoid 3′-hydroxylase from globe artichoke

Globe artichoke (Cynara cardunculus L. var. scolymus) is rich in flavonoids which contribute to its health-promoting properties. With the aim of understanding the genetic control of flavonoid accumulation in artichoke, we isolated an artichoke full-length cDNA sequence encoding flavonoid 3′-hydroxylase (F3′H), a major enzyme of the flavonoid hydroxylation pattern. In silico studies confirmed that the deduced amino acid sequence of CcF3′H is highly similar to F3′Hs isolated from other Asteraceae. The Northern blot analysis demonstrated that CcF3′H was highly expressed in leaves and in specific parts of the heads. Its expression differed slightly among artichoke cultivars. The overexpression of CcF3′H in tobacco plants led to the accumulation of flavonoids and to an increase of flower colour intensity, thus identifying CcF3′H as promising candidate for genetic engineering. CcF3′H represents the first structural gene of the flavonoid biosynthesis isolated from C. cardunculus, and its characterization sheds light on the accumulation of flavonoids.