贯叶金丝桃叶中分泌细胞团的超微结构

随着贯叶金丝桃（Hypericum perforatum L.)叶中分泌细胞团的发育,其细胞中质体的数量和体积逐渐增大,但一些质体局部出现解体,大量的深色管状结构和小泡出现在退化质体的周围,有些小泡与液泡融合,并将其内容物释放至液泡中,导致液泡中出现大量的多泡结构,多膜结构和嗜锇滴。同时,高尔基体分泌小泡进入液泡。然而,当分泌细胞团发育成熟后,分泌细胞被含有灰色均匀的分泌物（金丝桃素）的大液泡所占据,嗜锇滴消失。表明嗜锇滴可能是金丝桃素的前体物,来源于退化的质体。出现于质体和嗜锇滴之间的内质网和高尔基体可能也参与了金丝桃素前体物的合成和细胞内的转运。     

贯叶连翘愈伤组织中分泌细胞群的发生与金丝桃素类物质的积累

贯叶连翘(Hypericum perforatum L．)是一种传统草药,在欧洲被广泛用于治疗抑郁症。其重要的活性成分,金丝桃素类物质储存在茎、叶和花瓣的分泌细胞团中。本文应用组织化学及电子显微镜技术,研究体外培养的贯叶连翘叶肉细胞脱分化产生愈伤组织以及细胞发育过程中金丝桃素类物质的积累、运输的情况,进一步探讨细胞的生长发育与次生代谢产物的关系。发现金丝桃素类物质产生于愈伤组织培养后期,在愈伤组织表面所形成的分泌细胞群中,最初在细胞质中形成,之后运输至液泡中积累,内质网参与了金丝桃素类物质的合成过程。这些结果为利用组织培养技术提高金丝桃素类物质含量提供了理论基础和依据。     

贯叶连翘的分泌结构及其与金丝桃素积累的关系

贯叶连翘（ＨｙｐｅｒｉｃｕｍｐｅｒｆｏｒａｔｕｍＬ．）地上器官分布着分泌细胞球（黑色腺点）、分泌囊（半透明腺点）和分泌道（半透明腺条）３类内部分泌结构。分泌细胞球在茎、叶和花器官中均有分布,由２层鞘细胞包围多个分泌细胞构成实心的分泌细胞团。分泌囊主要分布于叶片中,分泌道则分布于花器官中,它们都是由１～２层切向扁平细胞围绕圆形或长形腔道构成,腔道的贮存物为精油。利用组织化学方法,结合荧光显微镜观察,证实金丝桃素类物质是由分泌细胞球（黑色腺点）所合成和积累的。通过用戊二醛和锇酸固定样品的显微和超微结构观察,发现金丝桃素类物质积累在成熟腺体分泌细胞的中央大液泡中,细胞周围浓厚的细胞质中分布着大量小液泡和高尔基体、内质网等细胞器。在此基础上对金丝桃素类物质的积累过程进行了初步探讨     

贯叶连翘分泌结构的发育及其内含物积累的研究

应用半薄切片和组织化学观察结果表明,贯叶连翘植物内具有分泌囊和分泌细胞团,叶片,萼片,花瓣中都存在2种分泌结构,分泌囊尚存在于果壁内,而分泌细胞团还在花药和茎棱中 ,2种分泌结构均起源于幼叶的基本分生组织内,其原始细胞形态相似,都垂周分裂为2子细胞,接着分别以不同分裂方式形成原始细胞群,并分化出鞘细胞,以后分 囊原始细胞群以裂生方式发育成由1层鞘细胞,1层上皮细胞包围着分泌腔组成的分泌囊,而分泌细胞团原始细胞群则继续增加细胞数目和体积发育成2－4层鞘细胞包围紧密排列的分泌细胞组成的分泌细胞团,其中给终未出现分泌腔,组织[化学试验表明,前者产生和贮存油类物质,而后者产生和贮存金丝桃素类物质。     

贯叶连翘的分泌结构及其与金线桃素积累的关系

贯叶连翘地上器官分布着分泌细胞球（黑色腺点）、分泌囊（半透明腺点）和分泌道（半透明腺条）３类内部分泌细胞,分泌细胞球在茎,叶和花器官中均有分布,由２层鞘细胞包围多个分泌细胞的构成实心的分泌细胞团。     

茉莉酸甲酯(MeJA)对贯叶连翘悬浮细胞生长和贯叶金丝桃素产量的影响

培养基中添加100 μm01·L-1MeJA后,贯叶连翘悬浮细胞生物量增加量和贯叶金丝桃素(HF)的产量分别是未经MeJA处理的1.3倍和1.73倍;培养3 d时,培养基中添加100 μmol·L-1 MeJA能提高HF产量,是未经MeJA处理的2.4倍.     

从贯叶连翘叶提取金丝桃素的工艺探讨

对从贯叶连翘中提取金丝桃素的工艺条件进行了优化研究,结果表明,金丝桃素提取较佳工艺为：45℃的水浸泡2h后,加入0.5%NaOH,用75％甲醇在0.05Mpa下78℃提取5次,每次1.5h,溶媒量为5倍,浓缩温度与干燥温度为78℃。     

金丝桃属植物叶中分泌结构的比较解剖学研究

利用整体透明法、石蜡制片法和半薄切片法,对金丝桃属9组43种l亚种1变种的植物叶分泌 结构的类型、形态、结构和分布进行了比较研究。结果表明,分泌结构是金丝桃属植物叶片普遍的结构特征,根据其分泌结构的特征,可划分为分泌细胞团、分泌囊(道)和韧皮部中分泌小管道等3种分泌结构。其中,分泌细胞团按其在叶片的分布可分为叶缘型和散生型;分泌囊按其在叶横切面中的位置可分为栅栏组织型、海绵组织型、居中型(位于栅栏组织型与海绵组织型之间)和横跨叶肉组织型。根据分泌囊和分泌细胞团在该属植物叶中的分布可划分为3种类型：(1)只有分泌囊的类型;(2)只有分泌细胞团的类型;(3)具分泌囊和分泌细胞团的类型。两种主要分泌结构的类型、分布密度、分布位置及其形态等方面在组间和种间均存在—定的差异,对金丝桃属属以下等级的区分具有一定意义。在此基础上,对该属分泌结构的形态演化以及金丝桃属各组间的亲缘关系进行了探讨。     

从贯叶连翘中提取金丝桃素的工艺探讨

对从贯叶连翘中提取金丝桃素的工艺条件进行了优化研究 ,结果表明 ,金丝桃素提取较佳工艺为 :45℃的水浸泡 2 h后 ,加入 0 .5 % Na OH,用 75 %甲醇在 0 .0 5 Mpa下 78℃提取5次 ,每次 1 .5 h,溶媒量为 5倍 ,浓缩温度与干燥温度为 78℃     

贯叶金丝桃组织培养的研究

分别以甘肃天水贯叶金丝桃的幼根、幼茎、幼叶为外植体．在1／2MS培养基上附加各类激素,进行贯叶金丝桃的组培实验。研究发现各外植体的增殖速率由高到低分别为幼茎、幼根、幼叶,且得到贯叶金丝桃组培各阶段的最佳培养基成分。诱导愈伤组织的培养基为1／2MS 1．3～1．6mg／L BA 0．2mg／L NAA;培养基1／2MS 1．3～1．6mg／L BA 0．15mg／L NAA有利于不定芽的形成;诱导不定根的培养基为l／2MS IBA0．5～O．8mg／L 蔗糖2．0％。向1／2MS培养基中添加不同的生长素(IAA,IBA,NAA,2．4-D)．在不同浓度梯度的培养基上进行诱导贯叶金丝桃的愈伤组织及不定根的试验,结果表明：生长素IAA,IBA既可诱导愈伤组织,又可以诱导不定根的产生。生长素NAA,2,4-D可诱导产生愈伤组织,但对不定根的诱导作用较差。     

Toxic Effects of Pb on Anatomy and Hypericin Content in Hypericum perforatum L.

Lead (Pb) is one of the most common heavy metal contaminants in the environment. The present study was therefore undertaken to determine the effects of Pb on structural characteristics and hypericin production in Hypericum perforatum. Mature plants were treated with contaminated soil in seven treatments (75, 150, 300, 600, 800, 1000, and 1500 mg/kg Pb in soil) with three repeats per treatment every 14 days. Maximum observed Pb content in shoot parts was observed in the treatments with 600 and 1500 mg/kg Pb. The Pb concentration in roots was higher than in shoot parts, enhanced with increasing Pb concentration in the soil. In this study, Pb treatment significantly influenced the morphology, anatomy, and hypericin content in the plant. Anatomical characteristics of leaf, stem, and root affected by Pb contamination, as well as scanning electron microscopy (SEM) studies, revealed structural changes in stomata and epicuticular waxes. Under Pb toxicity, anatomical symptoms occurred in leaves, including increase in sizes of epidermal cells, mesophyll tissue, and diameter of stems and roots, as well as amplified vascular bundles and pith area. This, therefore, indicated that metal contamination can change the chemical composition of this plant. Maximum hypericin content was observed in the treatment containing 600 mg/kg Pb in soil, which then decreased.     

Enhancing hypericin production of Hypericum perforatum cell suspension culture by ozone exposure

Accumulation of secondary metabolites is one of the common reactions of plants to ozone exposure in nature. To investigate the effect of ozone on the production of desired compounds of plant cell cultures, we assayed hypericin production of Hypericum perforatum suspension cell cultures treated with different doses of ozone at different culture phases. The results show that hypericin contents of the cells treated with 60 to 180 nL L^?1 ozone are significantly higher than those of the control, showing that ozone exposure may stimulate hypericin synthesis. Hypericin production of the cells treated with ozone at exponential phase is higher than that of lag and stationary phase, which suggests that exponential phase cell cultures are more responsive to ozone exposure than lag and stationary phase cells. The highest hypericin production is obtained by the cells exposed to 90 nL L^?1 ozone at late exponential phase for 3 h, being about fourfold of the control. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011     

Variation of Bioactive Compounds in Hypericum perforatum Growing in Turkey During Its Phenological Cycle

The present study was conducted to determine phenologic and morphogeneUc variation of hyperlcln, chlorogenlc acid and flavonoids, as rutin, hyperoside, apigenin-7-O-glucoside, quercitrin, quercetin content of Hypericum perforatum L. growing in Turkey. Wild growing plants were harvested at vegetative, floral budding, full flowering, fresh frulUng and mature fruiting stages and dissected into stem, leaf and reproductive tissues and assayed for bioacUve compounds by the High performance liquid chromatography （HPLC） method. Hypericin concentration ranged between 0 and 2.73 mg/g DW, chlorogenic acid 0.00-3.64 mg/g DW, rutin 0.00-3.36 mg/g DW, hyperoside 0.04- 22.42 mg/g DW, quercitrin 0.03-3.46 mg/g DW and quercetin 0.04-1.02 mg/g DW depending on ontogenetic and morphogenetic sampling. Leaves were found to be superior to stems and reproductive parts with regard to phenolic accumulation for all compounds tested while flowers accumulated the highest levels of hypericln. Quercltrln, quercetln and hypericin content in all tissues increased with advancing of developmental stages and reached their highest level during flower ontogenesis. Similarly, chlorogenic acid, hyperoside and apigenin-7-O-glucoside content in different plant parts increased during plant development, however, the highest level was observed at different stages of plant phenology for each tissue. Chlorogenic acid was not detected in stems, leaves and reproductive parts in several stages of plant phenology and its variation during plant growth showed inconsistent manner. In contrast to the other compounds examined, rutin content of stems and leaves decreased with advanc- ing of plant development and the highest level for both tissues was observed at the vegetative stage. However, content of the same compound in reproductive parts was the highest at mature fruiting. The present findings might be useful to obtain increased concentration of these natural compounds.     

贯叶连翘中的新抗真菌黄酮苷

从贯叶连翘( Hypericum perforatum L.)地上部分的80%乙醇提取物中分离得到1个新的黄酮苷和6个已知黄酮类化合物.采用红外光谱、紫外光谱、质谱和核磁共振光谱等波谱技术测得新黄酮苷的结构为6″-O-乙酰基槲皮素-3-O-β- D -阿洛糖苷(1,6″-O-acetyl quercetin 3-O-β- D -alloside),6个已知化合物分别被鉴定为槲皮苷(quercitrin)、金丝桃苷(hyperoside)、蓄苷(avicularin)、芦丁(rutin)、槲皮素(quercetin)和山萘酚(kaemferol).体外抗真菌活性实验表明,新黄酮苷(1)、槲皮苷和槲皮素对植物病原菌 Helminthosporium sativum Pamel King et Bakke具有一定的抑制作用,三者的最小抑制浓度(MIC)分别为:25、50和50 μg/mL.另外,1和槲皮苷对禾赤色镰孢霉( Fusarium graminearum Schw.)的生长也有抑制作用(MIC值均为100 μg/mL).     

Hyperforin accumulates in the translucent glands of Hypericum perforatum

BACKGROUND AND AIMS: Hypericum perforatum contains the therapeutically important compounds hypericin and hyperforin. Hypericin is known to accumulate in the dark glands. This investigation aimed to determine the accumulation site of hyperforin. METHODS: Dark and translucent glands as well as non-secretory tissue in leaves were manually isolated under the microscope. Hyperforin content was quantified by UV HPLC. Secretory structures were surveyed anatomically. KEY RESULTS: The hyperforin content of intact leaves was found to be about 3 mg g(-1) fresh tissue, whereas a content of about 7 mg g(-1) fresh material was found in isolated translucent glands. Hyperforin was found only to occur in minute amounts in dark glands (approx. 0.4 mg g(-1) fresh tissue). In non-secretory tissue no hyperforin was detected. CONCLUSIONS: The accumulation of hyperforin detected in the translucent glands supports the proposed hypothesis that hyperforin is synthesized by the same biosynthetic machinery as monoterpenes in the chloroplasts of cells delimiting the gland.     

贯叶连翘的试管快繁研究

以贯叶连翘的茎节等为外植体 ,进行植株再生和快速繁殖研究。结果表明在BA、NAA、IBA和 2 ,4 -D不同组合的MS或 1/ 2MS培养基上 ,以茎段为外植体的愈伤组织和试管苗易于诱导 ;在 1/ 2MS BA 1～ 1.5 (mg/l) NAA 0 .1～ 0 .5 (mg/l)的培养基中增殖速度快 ,繁殖系数高 ;在 1/ 2MS NAA(IBA) 0 .5～ 1(mg/l)培养基中均能诱导生根。     

贯叶连翘的水培及其代谢产物检测

水培可诱导贯叶连翘组培苗生根能力强,根活力也增加;生根苗在1/6MS培养液中培养6周后的金丝桃素(HP)、假金丝桃素(PHP)和贯叶金丝桃素(HF)含量分别比基质[腐质土 蛭石(1:1)]中培养的提高10.13%、16.00%和61.36%。