银杏黄酮苷和萜类内酯含量的季节变化

以银杏（Ginkgo biloba L.)2年生实生苗和大树为试材,分析根、茎和叶中银杏黄酮苷及萜类内酯含量的季节变化规律。银杏叶中萜类内酯含量从春季起逐渐增加,至夏末秋初达最高值,随后逐渐减少;根和茎中萜类内酯含量的季节变化与叶中相类似,但在冬季休眠期维持较高含量,进入春季伴随叶的萌发生长降低到全年的最低点。银杏茎中萜类内酯含量最低,相当于叶含量的1／3和根含量的1／2。叶中白果内酯含量在总萜类内酯中所占比例较高,而在根和茎中所占比例则较低。随着树龄增加,银杏叶萜类内酯含量下降,这可能与萜类内酯合成能力下降有关。银杏黄酮苷含量在春季幼叶中最高,夏季和秋季相对较低且变化不明显;长枝叶中槲皮素较多,而短枝叶中山柰黄素较多。对不同季节和不同部位的不同成分含量的相关机理进行了讨论。     

银杏愈伤组织的形成及其中黄酮类化合物的产生

单一激素种类对银杏叶片,叶柄和幼茎愈伤组织的诱导中以NAA的效果最佳,2,4－D次之,6－BA最差,除胚乳外,胚,幼苗的胚根,子叶,幼茎,叶片和叶柄,以及成年树的嫩茎,叶片和叶柄各外植体在本试验条件下都能诱发愈伤组织,其中胚,子叶和叶柄的愈伤组织形成频率均可达到100．0％,叶片和幼茎在光照下的愈伤组织诱导频率比黑暗中的略高,而叶柄和胚根则相反,MS和DCR两种培养基都适合银杏幼苗叶片及叶柄愈伤组织的诱导,两者之间不存显著性差异,测得光照培养的3个组织系（ST1,ST2,ST3)中均含银杏黄酮甙元槲皮素,山柰素和异鼠李素,总含量分别为干重的0．35％,0．29％和0．14％,而黑暗中培养的这3个愈伤组织系则没有银杏黄酮的产生。     

杜仲黄酮指纹图谱研究

采用HPLC指纹图谱对杜仲黄酮和银杏黄酮进行了标示,并将杜仲黄酮、银杏黄酮和沙棘黄酮HPLC指纹图谱进行了比较,发现杜仲黄酮和银杏黄酮很相近;杜仲黄酮、银杏黄酮和沙棘黄酮三者均富含槲皮素。研究认为,杜仲黄酮具有广泛的应用前景和较大的开发价值。     

生态条件对银杏叶黄酮积累的影响

以千年银杏（ＧｉｎｋｇｏｂｉｌｏｂａＬ．）古树叶片为材料对不同生境与银杏叶黄酮积累的关系进行了主成分分析和多元回归分析,结果表明影响银杏叶黄酮积累的重要生境因子是纬度、日照百分率、年降雨量和年平均温度。４因子的多元逐步线性回归方程对百年以上银杏叶的黄酮含量预测具良好的拟合效果。曲线回归分析表明,在纬度２８°１９′±２°３４′Ｎ或３８°６′±２°３４′Ｎ,年降水量７６２．３±１１４．５ｍｍ,日照百分率３５．３％±６．３％,年平均温度１５．９５±２．１５℃的条件下,最利于银杏叶黄酮的积累。但有利于叶内黄酮积累的生态条件并非银杏生长发育的最适条件。因此认为,选择有一定逆境胁迫的次适宜环境建立银杏采叶园,有利于提高叶内黄酮含量。     

迎春花中黄酮类化合物含量的研究

以75％乙醇为溶剂,索氏提取器提取,采用比色法和HPLC法研究了迎春花中黄酮类化合物的含量。以芦丁为对照物,比色法测定出其花、叶、茎中总黄酮的含量分别为13．7％、16．2％、6．77％．RSD分别为2．63％、2．57％、2．96％;以槲皮素、山奈素、异鼠李素为对照物,HPLC法测定出其花、叶、茎中的黄酮总量,以甙元计分别为4．205％、3．371％、1．703％,RSD分别为1．76％、1．58％、1．98％。     

银杏叶黄酮提取方法比较

比较不同溶剂提取银杏（ＧｉｎｋｇｏｂｉｌｏｂａＬ．）叶黄酮类化合物的提取效率,从成本效益角度考虑,以７０％乙醇作为提取溶剂更为有利。在分级沉淀中,黄酮含量与蛋白质含量呈正相关,在乙醇提取液中黄酮和蛋白质含量最高;蛋白质的存在有助于提高黄酮的溶解度。乙醇提取液用饱和（ＮＨ４）２ＳＯ４浓缩两次,可使醇相中的黄酮沉淀析出。根据试验结果,提出了银杏叶黄酮的优化提取方法。     

银杏雌株高黄酮种质定向筛选

以46个银杏雌株为试验材料,研究了生育前期和生育后期单株间叶中总黄酮及其组分含量差异及变异,基于聚类分析筛选出高黄酮含量的春茶用、提取黄酮用及两者兼用的优株。主要研究结果如下:(1)生育初期的黄酮及其组分含量显著高于生育后期,两个生育时期的各银杏雌株间的叶中总黄酮及各组分含量均存在极显著性差异。(2)银杏雌株间的叶中总黄酮及各组分含量均存在较大的变异,其中生育前期叶中总黄酮及各组分含量变异系数为2106%～3335%,生育后期为2911%～5013%。(3)生育前期和生育后期的叶中总黄酮含量分别与对应生育期的槲皮素、山奈酚、异鼠李素含量均呈极显著正相关;生育前期和生育后期间叶中总黄酮及其组分含量均呈极显著高度正相关。(4)基于生育前期叶中的总黄酮及3个主要组分含量,筛选出了15、29、30、42、43、44、45、46、47、49、50、48等12个春茶用优株;基于生育后期叶中的总黄酮及3个主要组分含量,筛选出了42、46、43、44、45、47、48、50等8个提取黄酮用优株;基于生育前期和生育后期叶中总黄酮含量,筛选出了29、42、43、44、45、46、47、50、48等9个可作为银杏春茶及黄酮提取的兼用型优株。综上三类分别可作为春茶用、黄酮提取用及兼用型银杏种质,进一步无性系化扩繁、区试或推广。     

银杏叶黄酮甙浸提工艺的研究

对黄酮含量偏低的银杏叶子进行浸提研究,确定浸提剂为50％～60％乙醇水溶液,液固比为8:1～6:1,浸提温度为60～70℃,浸提时间3～1h,两次。黄酮的浸提率在85％以上;选用A-1和A-2型树脂混合吸附,得到银杏叶提取物产品,黄酮含量26％～31％,得率为1.6％～1.9％。该工艺已用于中试生产,成本较低。     

银杏根系甲醇溶提物对离体培养条件下丛枝菌根真菌生长发育的研究

银杏根系甲醇溶提物对离体培养条件下丛枝菌根真菌生长发育的试验结果表明,银杏根系甲醇溶提物对离体条件下丛枝菌根真菌(Glonus mosseae,Gigaspora margarita)生长发育有明显的促进作用,能显著提高孢子萌发率,增加菌丝生长长度。与对照相比,浓度为20％～100％的甲醇溶提物均可显著促进丛枝菌根真菌的生长发育,80％甲醇溶提物效果最明显,银杏根系甲醇溶提物中含丰富的黄酮类化合物,溶提物中黄酮类物质的含量与甲醇洗脱剂的浓度有关,当浓度为80％时,黄酮含量最高,对银杏根系甲醇溶提物中黄酮类化合物含量与丛枝菌根真菌生长发育相关性分析发现,溶提物中黄酮类化合物含量与离体条件下丛枝菌根真菌孢子的生长发育表现为极显著的正相关关系,溶提物中黄酮含量越高,其对丛枝菌根真菌孢子的生长发育促进作用越大。     

细胞破膜对提取植物有效成分得率影响的研究

本文通过真空破膜法对乌头根、银杏叶、红豆杉茎皮及桔子果皮等材料进行细胞破膜处理,然后通过紫外分光光度计、高压液相色谱以及气相色谱等方法分别对其有效成分滇乌碱、银杏黄酮、紫杉醇以及桔皮精油和其精油中柠檬烯等的含量进行了分析。结果表明:滇乌碱得率由0.59％提高到0.81％;银杏叶中黄酮由1.76％提高到1.91％;红豆杉中紫杉醇由0.0139％提高到0.015％,桔皮挥发油由1.310％提高到1.591％,其中柠檬烯的含量由93.26％提高到95.37％。由此可见,对植物样品的细胞膜处理,在提高有效成分提取得率方面具有较显著作用。     

不同整地施肥措施对银杏构件生长及药用成分的影响

研究了田间条件下不同整地施肥措施对银杏构件生长及药用成分的影响．结果表明,对银杏株高生长促进作用最大的措施依次为：施农家肥+间作>爆破整地+间作>施农家肥>爆破整地>间作,提高生长量分别为14．5％、8．6％、5．7％、3．2％和0;对银杏当年新梢生长促进作用最大的措施依次为：施农家肥+间作>爆破整地+间作>间作>施农家肥>爆破整地,提高生长量分别为58．1％、36．6％、33．1％、30．2％和14．0％;不同的整地、施肥和间作措施样地长枝数无显著差异,而短枝数量与总叶数有显著差异;施农家肥加间作措施对提高银杏药用成分含量作用最大,叶片中懈皮素和芦丁含量分别为对照的4．2倍和2．2倍．     

无土栽培下酸铝耦合对银杏生长的胁迫效应

本试验采用U₂₀₇均匀设计方案,通过无土栽培方法研究酸铝耦合处理对银杏幼株生长的效应。结果表明,在pH 4.5～5.5范围内,能耐铝（AlCl₃）浓度可达0.4 mmol/L,而高浓度（0.8～1.2 mmol/L）的铝对银杏有毒害作用;pH高于5.0时,提高培养液中铝浓度对银杏生长无明显影响。总之,酸铝耦合加剧对银杏植株的毒害作用,尤其对根系的伤害更为明显,甚至导致烂根死根,从而使地上部停止生长。     

Oxyanthraquinones and flavonoids from garland chrysanthenum

The stems of the vegetable plant garland chrysanthemum (Chrysanthemum coronarium L.) were shown to contain emodin (in its aglycon and glycoside forms) and chrysophanol. Chrysophanol and chrysazin were isolated from the roots of the plant. Because the pigments identified are derivatives of 1,8-dihydroxyanthroquinone, garland chrysanthemum may be a medicinal plant and have utility as a component of laxative species. The leaves of C. coronarium were shown to be rich in quercetin and its glycosides, rutin and isoquercetin. Taken together, this observation and the known high content of ascorbic acid and carotenoids in the plant suggest that C. coronarium may be useful in preventing cardiac and vascular diseases.     

Identification of Terpene Lactones and Flavonol Glycosides in Preparations Based on Ginkgo Biloba Extract and a New Way of Semi-Quantitative Determination of Flavonol Glycosides by <Superscript>1</Superscript>H NMR Spectroscopy

The composition of terpen lactones and flavonol glycosides of commercial preparation series based on Ginkgo biloba extracts was investigated by ¹H NMR spectroscopy. The content of individual terpen lactones was determined using DMSO-d₆ and acetone-d₆ solvents. The effect of the structure of flavonol glycosides on the signal of the hydroxyl proton at a position 5 of the ring A was examined. A new approach was proposed for semiquantitative determination of the total amount of flavonol glycosides by the integral intensity of this signal, which is a superposition of the singlets in the region of 12.5–12.65 ppm of individual flavonoids in DMSO-d₆. Since the corresponding signals of aglycones (quercetin, kaempferol, isorhamnetin), which are minor components of the Ginkgo biloba extracts, appear separately in a slightly different region (12.45–12.48 ppm), the proposed method can also be used for detecting adulteration of Ginkgo biloba extracts by means of the addition into them of relatively cheap aglycones or rutin as well as for assessment of the content of flavonoids of similar structure in some types of plant raw materials.     

温度和土壤水分对银杏叶黄酮类化合物积累的影响

以2年生银杏实生苗为试材,在人工气候室内采用土培盆栽试验方法,研究了温度和土壤水分对银杏叶黄酮类化合物积累的影响.试验设置土壤含水量(W)和温度(T)各3个梯度,W₁、W₂、W₃分别为田间持水量的55%～60%、40%～45%、30%～35%;T₁、T₂、T₃白天和夜间的温度分别为15/5 ℃、25/15 ℃、35/25 ℃.结果表明: T₁温度条件下,各土壤水分处理的银杏叶中的槲皮素、山奈酚、异鼠李素和总黄酮含量普遍高于T₂和T₃,而土壤水分对银杏叶中各种黄酮类化合物积累的影响不显著;银杏叶中黄酮类化合物以山奈酚含量最高,其次为槲皮素和异鼠李素;T₃温度下银杏单株总黄酮产量普遍高于T₂和T₁.在收获前适当采取土壤覆盖和灌水等措施降低种植园的温度,有利于提高银杏叶中黄酮的含量,增加单位面积黄酮的产量.     

Hydroxyanthraquinones and flavonoids of garland chrysanthemum

The stems of the vegetable plant garland chrysanthemum (Chrysanthemum coronarium L.) were shown to contain emodin (in its aglycone and glycoside forms) and chrysophanol. Chrysophanol and chrysazin were isolated from the roots of the plant. Because the pigments identified are derivatives of 1,8-dihydroxyangarland chrysanthemum may be a medicinal plant and have utility as a component of laxative spethroquinone, cies. The leaves ofC. coronarium were shown to be rich in quercetin and its glycosides, rutin and isoquercitrin. Taken together, this observation and the known high content of ascorbic acid and carotenoids in the plant suggest that C.coronarium may be useful in preventing cardiac and vascular diseases.     

Quercetinase pirin makes poliovirus replication resistant to flavonoid quercetin

Flavonoid quercetin and its derivative, methylquercetin, inhibit the replication of poliovirus in several cell lines. Here, we show that replication of poliovirus is inhibited by quercetin and that the extent of this inhibition depends on the intracellular content of pirin, a quercetinase. HeLa cells contain higher content of pirin protein than normal kidney human epithelial (NKE) or 293 cells do. Poliovirus replication in HeLa cells is significantly more resistant to quercetin than its replication in NKE and 293 cells. Overexpression of pirin reduced antiviral inhibitory effect of quercetin, while siRNA-induced suppression of pirin level made poliovirus replication more sensitive to the flavonoid. The results suggest that quercetinase activity of pirin determines the resistance of poliovirus infection to quercetin.     

Potential effets of ginkgo (Ginkgo biloba,L.) on female reproduction

This minireview will briefly outline the basic knowledge concerning the provenance, biological active constituents of ginkgo (Ginkgo biloba, L.) and its general health effects. Ginkgo has been shown to affect female reproductive functions: it can affect ovarian folliculo- and oogenesis, embryogenesis, promote ovarian granulosa cell apoptosis, reduce their proliferation and the release of ovarian hormones. Usually, ginkgo extract mainly suppresses, but its constituents like amifostine, leuprorelin, quercetin and kaempherol can promote ovarian functions. This may indicate the existence of anti-reproductive ginkgo constituent(s), such as ginkgolide B and allopregnenolone which, like ginkgo extract, can promote ovarian cell apoptosis and suppress ovarian follicullogenesis and oogenesis. Ginkgo effects could be mediated by an action on brain functions, ovarian steroidogenesis, oxidative processes, intracellular regulators of ovarian cell proliferation and apoptosis and GABA receptors. Ginkgo and its molecules, ginkgolide B and allopregnenolone can be useful for prevention and treatment of reproduction-related disorders like ovarian cancer, ovarian ischemia and menopausal syndrome. On the other hand, its constituents amifostine, leuprorelin, quercetin and kaempherol could be potentially applicable as biostimulators of female reproductive processes in human and veterinary medicine and animal production. Nevertheless, application of ginkgo is still limited by insufficient or contradictory knowledge concerning its active constituents, characters, targets and mediators of its action and their functional interrelationships. Impact of ginkgo action on reproductive organs other than ovaries remains largely unknown. Addressing these issues with proper animal and clinical studies could help to understand the distinct efficacy and consequences of medical application of ginkgo.     

银杏再生根和不定根的形成及其中银杏内酯的产生

附加０．５ｍｇ／Ｌ２,４－Ｄ的ＤＣＲ２基较适合愈伤组织再生根的形成,子叶上能直接诱发不定根,根培养物的生长速度远比愈伤组织慢,但共合成银杏内酯的能力却较愈伤组织强。