UPLC-TOF-MS快速测定不同核用银杏品种银杏叶中银杏内酯和白果内酯含量

为比较鄂西南不同核用银杏品种叶片中银杏内酯和白果内酯含量,采用超高压液相色谱-飞行时间质谱(UPLC-TOF-MS)联用方法测定微量银杏内酯和白果内酯含量,分析柱为ZORBAX C18(2.1×50 mm,1.8μm)色谱柱,流动相为乙腈-0.1%甲酸水溶液,采用梯度洗脱,飞行时间质谱作为检测器,选择离子采集方式。方法测定银杏内酯和白果内酯的线性范围为0.2~4μg/m L,相关系数为0.9996以上,相对标准偏差小于1.65%。测定结果表明,在四个核用银杏品种中,恩银23号具有较高的银杏内酯和白果内酯含量,同时白果内酯和银杏内酯A、B及C总含量比例较佳,可以作为叶用银杏发展的优选品种。     

银杏萜内酯的分布与矮壮素对其生物合成的调节

银杏萜内酯分为银杏内酯A、B、C、J、M(ginkgolide A、B、C、J、M)和白果内酯(bilobalide),主要存在于银杏叶与根内,近年的研究指出银杏萜内酯分别在银杏叶和根中生物合成[1],Cartayrade等人[2]通过叶片生根实验发现生根叶片的银杏萜内酯含量显著高于未生根叶,因而认为银杏萜内酯是在根部合成,然后运输到叶中积累,目前对此还缺乏进一步的研究报道.     

高效液相色谱分析银杏萜内酯的含量

用高效液相色谱法测定银杏叶及提取物中银杏内酯中Ａ、Ｂ、Ｃ和白果内酯的含量。采用Ｃ１８分离柱,示差检测器,甲醇：水（３３：６７）为流动相,方法回收率平均达９７％以上,变异系数（ＣＶ）为２．１％。     

溶剂萃取银杏萜内酯的因子研究

采用不同溶剂对银杏叶提取物进行萃取,实验表明;乙酸乙酯、丁酮、环已酮、乙醚等对萜内脂的选择性逐次减小.通过调节pH在8～9,增加3%～5%盐离子等方法改善相密度,乙酸乙酯的萃取率在90%以上,萜内酯的含量可富集到35%～40%.     

银杏叶中银杏内酯B及白果内酯的分离鉴定

前文中作者已报道从我国特有植物银杏(Ginkgo biloba L.)的叶子中分到银杏内酯A及C(ginkgolide A,C),本文报道从银杏叶中分到的另外二个结晶,经理化性质、薄层层析、红外光谱及质谱分析,鉴定结晶Ⅲ为银杏内酯B(ginkgolide B),结晶Ⅳ为白果内酯(bilobalide),后者为国内首次分离。     

刺槐叶绿素荧光特性的研究

用L I-6400光合仪对黄土高原常用造林树木刺槐的叶绿素荧光动力学曲线、淬灭分析、荧光光曲线以及荧光AC I曲线等生理特性进行测定和对比分析.结果表明:随着光照时间的加大,刺槐的荧光参数ETR、qP和N PQ逐渐上升并在21 m in左右达到稳定,与光合同步.最大荧光产量F m在叶片转入黑暗后逐渐上升,在44 m in左右趋于稳定;N PQ则开始下降,在35 m in左右达到稳定.当光强度>200μm o l.m-2.s-1时,Ph iPS2和Ph iCO2呈线性正相关,而当光强度<200μm o l.m-2.-s 1时,Ph iPS2和Ph iCO2呈线性负相关.CO2浓度和刺槐的荧光光合速率、Ph iPS2、Ph iCO2值存在着一定的正相关关系.     

高效液相色谱分析银杏萜内酯的含量

用高效液相色谱法测定银杏叶及提取物中银杏内酯Ａ、Ｂ、Ｃ和白果内酯的含量。采用Ｃ１８分离柱,示差检测器,甲醇：水（３３：６７）为流动相,方法回收率平均达９７％以上,变异系数（ＣＶ）为２．１％。     

银杏雄株叶片萜内酯类化合物的HPLC检测分析

目的:银杏叶萜内酯类化合物具重要的药用与保健开发利用价值.方法:在银杏最佳采叶期随机采取银杏叶片,经超声波70%乙醇水溶液提取、石油醚与正己烷萃取,再经ADS-17大孔树脂柱与酸性氧化铝柱纯化,以水∶甲醇∶四氢呋喃=7∶2∶1(V/V)为流动相,采用Waters ODS C18 柱 (250mm ×416mm,5μm)进行HPLC分析:0.6ml/min流速、219nm检测波长、26℃检测温度、15μl进样量.结果: GA、GB、GC和BB分别在23～460μg/ml、21～420μg/ml、22～440μg/ml和27～540μg/ml范围内呈线性相关,最小检出量0.5μg,回收率90.74%～102.38%,RSD 1.63%～4.2%.GBE3得率1%,GA 、GB、 GC 、BB占GBE3的12 55%,超过GBE761的6% 国际标准.结论:该HPLC检测分析技术与方法适用于银杏叶片萜内酯类化合物的提取、纯化和检测.     

NaCl胁迫下交替呼吸途径对叶绿素含量及其荧光特性的影响

以菜豆幼苗作为试验材料,分析了NaCl胁迫下交替呼吸对叶绿素含量以及叶绿素荧光特性变化特征的影响,以探讨交替呼吸途径在逆境下的生理学作用以及植物在盐胁迫下光系统Ⅱ(PSⅡ)的调节作用机制。结果表明:(1)随着NaCl胁迫浓度(0、100、200、300mmol/L)的增高,菜豆幼苗叶片叶绿素含量显著下降,叶片光系统Ⅱ(PSⅡ)潜在最大光化学量子效率(Fv/Fm)、光适应下最大光化学效率(Fv′/Fm′)、PSⅡ光适应下实际光化学效率[Y(Ⅱ)]和光化学荧光猝灭(qP)与对照相比均显著性下降,而非光化学猝灭(NPQ)较对照组显著增加,同时交替呼吸容量在NaCl胁迫下也显著上升。(2)与单独NaCl胁迫相比,在NaCl胁迫下施加交替呼吸的抑制剂水杨基氧肟酸(SHAM)会导致菜豆幼苗叶片叶绿素含量、Fv/Fm、Fv′/Fm′、Y(Ⅱ)和qP进一步显著下降、NPQ进一步显著增加。研究认为,NaCl胁迫导致菜豆叶片光系统Ⅱ光化学效率下降和光能耗散增加,交替呼吸途径可有效缓解NaCl胁迫下菜豆叶绿素含量的减少以及光系统Ⅱ光化学反应效率的下降。     

基于叶绿素荧光参数的小麦叶片叶绿素相对含量估算方法

叶绿素含量是植物学和农业相关研究领域常用的生理指标.叶绿素含量和叶片光合功能密切相关,但是现有的叶绿素含量的测定方法无法实现叶绿素含量和光合功能的同步测定和关联分析.为解决该问题,本研究通过测定35个小麦品种旗叶的SPAD值和叶绿素荧光诱导动力学曲线,分别使用不同时间的快速叶绿素荧光动力学曲线的荧光值,以及33个常用荧...     

银杏愈伤组织诱导的多因子正交试验研究

采用正交试验设计法,研究了NAA、KT、2,4-D、蔗糖浓度和不同外植体类型等因素对银杏愈伤组织诱导的影响。结果表明:不同外植体类型对银杏愈伤组织诱导率影响最大,KT和NAA其次,2,4-D和蔗糖浓度最小。银杏愈伤组织诱导最佳培养基为MS+NAA 0.5 mg·L-1+KT 0.5 mg·L-1+蔗糖40 g·L-1,最佳外植体为茎段,其愈伤组织诱导率可达100%。     

银杏毛状根褐化机制的初步研究

该实验依据现有发根农杆菌侵染法,以银杏无菌苗叶片为外植体,对农杆菌诱导银杏毛状根的实验操作进行优化,并观察记录银杏毛状根继代生长情况,测定继代不同天数的毛状根组织内多酚含量、多酚合成相关酶基因和POD基因相对表达量,以探讨银杏毛状根褐化的生理机制。结果显示:(1)银杏农杆菌侵染最适预培养时间为3d、菌种为R1601、共培养基为White、侵染时间为30min,优化侵染操作后的银杏毛状根诱导率可达72.36%;银杏毛状根继代生长的最适IBA浓度为0.5mg/L。(2)银杏毛状根初始发根时颜色为乳白色,多在伤口处簇生,伸长生长时无向地性,不分叶正背面都能在伤口处诱导出根,且在除菌培养20d时毛状根长可达0.4～0.8cm;继代培养25d时毛状根为白色,继代35d时部分毛状根从根部至中部开始转黄,继代45d时大部分毛状根由白转黄,根部至中部开始有褐化出现。(3)继代35d时毛状根的PAL、C4 H、4CL和POD基因表达量分别为25d的68%、44%、51%、52%;继代45d时PAL、C4 H、4CL、POD基因表达量分别为35d的19%、75%、86%、105%。(4)继代35d、45d的银杏毛状根总酚类含量分别为17.14和13.78mg/g。研究发现,PAL基因表达量变化与其下游的两个关键酶基因(C4 H、4CL)表达量变化不一致,且PAL基因表达量的大幅降低并未对下游C4 H、4CL基因表达有明显影响,说明PAL基因表达量变化对其下游合成速率影响不大,可能不是银杏毛状根苯丙氨酸代谢途径的限速酶;而C4 H与4CL基因表达量变化趋势高度一致,说明C4H应是该途径中的限速酶之一;推测继代前期可能是由PPO作为关键酶参与毛状根褐化反应,导致后期毛状根生长受抑制,POD基因表达水平随之上升,POD参与褐化反应。     

Gender- and age-related variations in blood viscosity in normal volunteers: A study of the effects of extract of Allium sativum and Ginkgo biloba

This study sought to compare the effects of age and gender on blood viscosity and to appraise the effectiveness of Ginkgo biloba and Allium sativum extracts in reducing blood viscosity. Stage 1: Our sample consisted of 80 male volunteers (40 aged 18-60 and 40 aged 61 and over) and 80 females with the same age profile. Stage 2: We studied 60 male volunteers allocated in groups: placebo, G. biloba, and A. sativum. Stage 3: We studied 25 male volunteers and in the initial, intermediate, and final evaluations, the measures of blood viscosity were repeated. Volunteers were given a clinical evaluation and submitted to laboratory tests. G. biloba led to the highest reduction in blood viscosity compared with placebo and A. sativum. In relation to the use of the two substances, G. biloba and A. sativum, dry extract of G. biloba proved to be more effective in reducing blood viscosity.     

Ginkgo biloba extract protects rat kidney from diabetic and hypoxic damage

Ginkgo biloba extract EGb 761 was studied for its nephroprotective effects in experimentally diabetic and hypoxic rats. Duration of streptozotocin-induced diabetes was 4 months, that of respiratoric hypoxia of the diabetic group 20 min. The daily dose of 100 mg EGb/kg bodyweight started 1 month after induction of the diabetes. EGb reduced diabetes-induced morphological alterations of the kidney such as increase in volume of glomeruli, capillary tufts, urinary space, and thickening of Bowman's capsule basement membrane. Diabetically increased immunostaining of interstitial collagenes of types I, III, and VI was diminished by the EGb extract. EGb reduced the relative total SOD activity from 163% in diabetic kidney to 46%. Additional hypoxia-induced ultrastructural damage was also diminished.     

银杏bHLH91转录因子基因的克隆及表达分析

bHLH转录因子在植物的生长发育、胁迫应答和次生代谢中具有重要的调控作用。该研究通过PCR技术从银杏(Ginkgo biloba)叶中分离得到了一个bHLH基因的cDNA序列,并将其命名为GbbHLH91。序列分析结果显示扩增的GbbHLH91基因cDNA序列长度为1 425 bp,开放阅读框是1 065 bp,编码354个氨基酸,分子量为40.1 kDa,等电点为8.20。系统进化分析结果显示,从用于进化树构建的bHLH蛋白质聚类情况来看,银杏GbbHLH91蛋白与裸子植物油松(Pinus tabuliformis)bHLH蛋白亲缘关系最近,且与被子植物无油樟(Amborella trichopoda)bHLH蛋白相似性达到60%,表明该基因在进化过程中相对比较保守。实时荧光定量PCR分析发现银杏bHLH91基因在银杏的各个组织中均有表达,其中在银杏叶中表达量最高,在根和茎中基因的表达量次之,在银杏雌花和果中表达量较少,在雄花中的表达水平最低;GbbHLH91基因在不同发育时期的银杏叶片中,表达量也存在一定的差异,其中在4月中旬该基因的表达水平达到最高,而后随着叶片的生长发育,该基因的表达水平呈现下降趋势。该研究结果为进一步验证GbbHLH91基因的功能奠定了前期基础。     

Potential of Ginkgo biloba L. leaves in the management of hyperglycemia and hypertension using in vitro models

Leaves from four different Ginkgo biloba L. trees (1 and 2 – females; 3 and 4 – males), grown at the same conditions, were collected during a period of 5 months (from June to October, 2007). Water and 12% ethanol extracts were analyzed for total phenolics content, antioxidant activity, phenolic profile, and the potential in vitro inhibitory effects on α-amylase, α-glucosidase, and Angiotensin I-Converting Enzyme (ACE) enzymes related to the management of diabetes and hypertension. The results indicated a significant difference among the trees in all functional benefits evaluated in the leaf extracts and also found important seasonal variation related to the same functional parameters. In general, the aqueous extracts had higher total phenolic content than the ethanolic extracts. Also, no correlation was found between total phenolics and antioxidant activity. In relation to the ACE inhibition, only ethanolic extracts had inhibitory activity.     

不同光照强度对假紫万年青生长和叶绿素荧光参数的影响

为了解光照对假紫万年青(Belosynapsis ciliata)生长的影响,研究了不同光照强度下假紫万年青形态特征和叶绿素荧光参数的变化。结果表明,随着遮阴强度的增加,假紫万年青的自然株高有所增加,节数、茎粗、分枝数和茎长度均减少;而不同光照强度对假紫万年青节间长的影响不明显;其叶绿素a、叶绿素b、叶绿素总含量和叶绿素a/b先增加后又减少,以15.2%NS时达到最大。叶绿素b含量与叶绿素荧光的Fv/Fm呈负相关,Fv/Fm为0.75~0.8。因此,在高光照条件下,假紫万年青匍匐近地生长,随着光照条件的逐渐减弱,趋向直立生长。在屋顶绿化种植的初期采用适度遮阴(35.2%NS)可使其更好的生长,加速成坪,降低管理成本。     

Gas exchange of Ginkgo biloba leaves at different CO2 concentration levels

One and a half year-old Ginkgo saplings were grown for 2 years in 7 litre pots with medium fertile soil at ambient air CO₂ concentration and at 700 μmol mol⁻¹ CO₂ in temperature and humidity-controlled cabinets standing in the field. In the middle of the 2nd season of CO₂ enrichment, CO₂ exchange and transpiration in response to CO₂ concentration was measured with a mini-cuvette system. In addition, the same measurements were conducted in the crown of one 60-year-old tree in the field. Number of leaves/tree was enhanced by elevated CO₂ and specific leaf area decreased significantly.CO₂ compensation points were reached at 75–84 μmol mol⁻¹ CO₂. Gas exchange of Ginkgo saplings reacted more intensively upon CO₂ than those of the adult Ginkgo. On an average, stomatal conductance decreased by 30% as CO₂ concentration increased from 30 to 1000 μmol mol⁻¹ CO₂. Water use efficiency of net photosynthesis was positively correlated with CO₂ concentration levels. Saturation of net photosynthesis and lowest level of stomatal conductance was reached by the leaves of Ginkgo saplings at >1000 μmol mol⁻¹ CO₂. Acclimation of leaf net CO₂ assimilation to the elevated CO₂ concentration at growth occurred after 2 years of exposure. Maximum of net CO₂ assimilation was 56% higher at ambient air CO₂ concentration than at 700 μmol mol⁻¹ CO₂.     

文心兰浅绿条纹突变体的生理生化及叶绿素荧光特性研究

以文心兰浅绿条纹突变体为材料,分析叶片光合色素含量和组成、叶绿素合成前体物质含量以及叶绿素荧光参数的变化,观察突变体叶绿体超微结构的改变,以探寻其叶色变异的生理基础。结果表明:(1)突变体叶绿素a(Chl a)、叶绿素b(Chl b)、类胡萝卜素(Car)和总叶绿素(Chl)含量分别比叶色正常植株显著降低了37.1%、34.0%、30.8%和36.3%。(2)突变体叶绿素生物合成受阻于胆色素原(PBG)到尿卟啉原Ⅲ(UrogenⅢ)的反应步骤。(3)突变体叶绿体发育存在明显的缺陷,基粒数目及基粒片层的垛叠层数明显减少,嗜锇颗粒及囊泡较多。(4)突变体初始荧光(Fo)比正常植株高39%,最大荧光(Fm)、最大光化学效率(Fv/Fm)、PSⅡ有效光化学效率(Fv′/Fm′)和PSⅡ实际光化学效率(ΦPSⅡ)均显著低于正常植株,但光化学淬灭系数(qP)和非光化学淬灭系数(NPQ)与正常植株无显著差异。研究结果说明,文心兰叶绿素生物合成受阻和叶绿体结构发育不良,导致叶绿素的含量下降,致使突变体叶片呈现浅绿条纹,光能利用率降低。