木立芦荟组织培养pH分化特性及快速繁殖的研究

以木立芦荟的叶片、叶鞘、带腋芽的茎段为外植体进行试管培养 ,结果叶鞘和茎段可诱导形成愈伤组织 ,腋芽直接萌生。经试验筛选出各培养阶段最适宜的培养基为 :( 1 )愈伤组织诱导 ,MS BA 2 .5mg/L NAA0 .1 5mg/L ;( 2 )腋芽萌生 ,MS BA 2 .0mg/L NAA 0 .1 5mg/L ;( 3 )丛生芽分化及继代 ,MS BA 2 .0mg/L NAA0 .1 0mg/L ;( 4)生根 ,MS BA 0 .3～ 0 .5mg/L IBA 0 .2mg/L 活性碳 0 .5%。研究还发现 ,培养基酸碱度对木立芦荟组织培养分化效果的影响非常显著。     

激素对洋桔梗植株再生的影响及生根培养的研究

以MS为基本培养基 ,附加不同浓度的 6 BA、KT、NAA和IBA诱导洋桔梗叶片外植体的再生植株。结果表明 :MS +6 BA 0 .5～ 1 .0mg/L(单位下同 ) +NAA 0 .2和MS +6 BA 0 .5～ 1 .0 +IBA 0 .2培养基都能诱导外植体产生愈伤组织 ,但 6 BA的浓度必须小于 1 .0mg/L ,否则会导致组织的严重玻璃化 ;MS +KT 1 .0～2 .0 +NAA 0 .2或MS +KT 1 .0～ 2 .0 +IBA 0 .2培养基也能诱导外植体产生愈伤组织 ,愈伤组织出现的时间较早且质地较好 ,适合分化。继代培养时 ,MS培养基中仅加 6 BA 0 .5mg/L或KT 0 .5mg/L ,即能获得较高的分化率。生根培养研究中 ,培养液为 1 /2MS+5 0g/L糖 +IBA 2mg/L的前处理 ,生根效果较好 ,生根率接近基质生根培养的生根率。     

袋鼠花的组织培养和快速繁殖

1　植物名称　袋鼠花 (Anigozanthosflavidus) ,又名袋鼠藤。2　材料类别　茎尖、幼嫩的茎段。3　培养条件　以MS为基本培养基。丛生芽诱导培养基 :( 1 )MS + 6 BA 2mg·L- 1 (单位下同 ) +NAA 0 .2 ;( 2 )MS + 6 BA 1 .5 +NAA 0 .2 ;( 3)MS+ 6 BA 0 .5 +IBA 0 .2。增殖培养基 :( 4 )MS + 6 BA 1 +NAA 0 .2。生根培养基 :( 5 ) 1 / 3MS +IBA0 .1～ 0 .5 + 0 .7%琼脂粉 + 0 .5 %活性碳 + 1 .5 %蔗糖 ;( 6) 1 / 2MS +NAA 0 .5。除培养基 ( 5 )外 ,其它均加 3%的蔗糖。培养基 ( 1 )～ ( 4 )琼脂含量均为0 .7%。pH 5 .8。培养…     

吊钟花的组织培养技术研究

以吊钟花(Enkianthus quinqueflorus Lour.)茎尖及带腋芽的茎段为外植体进行组织培养.结果表明,改良B5培养基(B5大量元素和钙盐+MS有机物、铁盐、微量元素)最有利于吊钟花的培养,外植体在改良B5+2,4D- 1 mg L-1的培养基中,愈伤组织诱导率可达100%.在含BA 1～2 m L-1+NAA0.1～0.5 mg L-1培养基中,可诱导产生不定芽.继代培养以改良B5+BA 1 mg L-1+NAA0.5 mg L-1培养基的增殖系数最高.生根培养基以1/2MS+IBA2 mg L-1为最佳,生根率可达80%以上.试管苗移栽成活率为90%以上.     

三色绿萝的组织培养和植株再生

植物名称:三色绿萝(Epipremnum aureum cv.Tricolor)。材料类别:茎尖、幼嫩的茎段。剪取三色绿萝的顶芽约1cm长,用70％酒精和0.1％升汞表面消毒后,在无菌操作下剥离茎尖约1～2 mm,同时横切茎段1～2mm作为外植体。培养条件:诱导不定芽的培养基:MS+BA 1～2 mg/L(单位下同)+NAA 0.2;诱导愈伤组织培养基:MS+NAA2+BA 0.5;诱导芽分化培养基:MS+BA2+GA_30.1+IAA 0.05;生根培养基:1/2 MS+IBA 1。培养基均为蔗糖3％(生根培养基     

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。     

紫背天葵的组织培养和快速繁殖

1　植物名称　紫背天葵 (Gynurabicolor)。2　材料类别　具芽点的茎段。3　培养条件　基本培养基为MS。芽诱导培养基 :( 1 )MS + 6 BA 1 .5mg·L- 1 (单位下同 ) +NAA0 .1 ;( 2 )MS + 6 BA 2 +NAA 0 .1 +KT 1 ;( 3)MS+ 6 BA 1 .5 +NAA 0 .1 +GA 1。增殖培养基 :( 4 )MS + 6 BA 3+NAA 0 .1 +GA 1。生根培养基 :( 5 )1 /2MS +IBA 0 .5 ;( 6) 1 /2MS +NAA 0 .5 ;( 7)1 /2MS +IBA 0 .5 +NAA 0 .1。上述培养基均含蔗糖 30 g·L- 1 、0 .75 %琼脂 ,pH 5 .8。培养温度为2 3～2 7℃ ,光照度 1 2 0 0～ 1 40 0lx ,光照 1 4h…     

白刺组织培养技术的研究

试验选用唐古特白刺幼嫩茎段和叶片作为材料,研究白刺不同外植体的离体培养技术。结果表明,白刺带芽嫩茎是诱导丛生芽的良好外植体,而叶片是诱导愈伤组织的良好外植体;白刺的最适增殖、壮芽培养基是：MS BA0.5mg／L NAA1．0mg／L GA32．0mg／L;最适生根培养基是：l／2MS IBA0.5mg／L;愈伤组织诱导培养基是：MS 2,4-D0.5～1．0mg／L。     

金鱼草的组织培养

植物名称:金鱼草(Antirrhinum majus)别名龙口花。材料类别:实验材料为橙黄色和红色矮茎、粉红色的中茎。选用杂种F_1后代种苗的第二年萌发的幼芽,用70％的酒精和0.1％的升汞进行表面消毒,切取1cm的小段作为外植体,接种在MS基本培养基上。培养条件:诱导丛生芽的基本培养基为MS培养基。激素处理:(1)BA0.1～2.0mg/L(单位下同)+IBA0.1(BA0.1,0.5,1.0,2.0);(2)BA2.0+NAA0.1。生根培养基:(1)1/2MS+NAA0.02;(2)1/5MS+NAA0.02～0.1。温度为25±2℃,光照度1500lx,每日光照14～16小时。     

趣蝶莲的离体快速繁殖

1　植物名称　趣蝶莲 (Kalanchoesynsepala)。2　材料类别　不定芽、茎尖、带节茎段。3　培养条件　丛生芽诱导培养基 :( 1 )MS + 6 BA2 .0mg·L- 1 (单位下同 ) +NAA 0 .2 ,( 2 )MS +6 BA 1 .0 +NAA 0 .1 ;增殖培养基 :( 3)MS + 6 BA0 .5 +NAA 0 .0 5 ,( 4 )MS + 6 BA 0 .2 +NAA 0 .0 5 ,( 5 )MS + 6 BA 0 .1 +NAA 0 .0 5 ;生根培养基 :( 6)MS ,( 7)MS +IBA 1 .0 +NAA 0 .2 ,( 8)MS +NAA 0 .5。以上培养基均含 30 g·L- 1 蔗糖、0 .7%琼脂 ,pH 5 .5～5 .8。培养温度 ( 2 8± 2 )℃ ,光照度1 5 0 0～ 2 0 0 0lx ,光照…     

The influence of some elicitors on growth and morphogenesis of Hypericum perforatum callus cultures

We studied the effects of elicitors, such as mannan, beta-1,3-glucan, ancymidol, and cork crumbs, on morphogenetic and biosynthetic potencies of shoot cultures of Hypericum perforatum L. In the presence of these elicitors, different morphogenetic structures of H. perforatum callus cultures were formed. A correlation was found between the morphogenetic processes and induction of hypericin and pseudohypericin biosynthesis in the callus cultures.     

贯叶连翘挥发性化学成分研究

采用水蒸气蒸馏法提取中草药贯叶连翘(全草)挥发油。通过气相色谱/质谱联用技术测定了挥发油的化学成分,从中鉴定出75种化合物,其中百里香酚、桉油精、丁香油酚等酚类物质为首次检出。结果表明,贯叶连翘因产地不同、部位不同,其挥发油的成分有较大的差别。     

Identification and quantification of hypericin and pseudohypericin in different Hypericum perforatum L. in vitro cultures.

Investigations have been made to develop an efficient protocol for micropropagation allowing to improve hypericin and pseudohypericin productions in Hypericum perforatum L. in vitro cultures. The role of growth regulator treatments has been particularly studied. Three in vitro culture lines with different morphological characteristics were obtained during H. perforatum micropropagation and referred to shoots, calli and plantlets according to their appearance. Multiplication and callogenesis from apical segments from sterile germinated seedlings were obtained on solid MS/B5 culture medium in the presence of N6-benzyladenine (BA) (0.1-5.0 mg/l BA). Regenerative potential of shoots was assessed on medium supplemented with auxins (0.05-1.0 mg/l), indole-3-acetic acid (IAA) or indole-3-butyric acid (IBA). The main goal of the research was to summarize the influence of plant growth regulators on hypericin and pseudohypericin productions in in vitro cultures of Hypericum. A rapid method for naphtodianthrone quantification was developed. The use of a reversed-phase high performance liquid chromatography (HPLC) method with fluorescence detection was used. Identification of the compounds was confirmed by electrospray ionization-mass spectrometry (ESI-MS) with electrospray in negative ion mode [M-H] . Calli, shoots and plantlets of H. perforatum produced hypericin and pseudohypericin. The concentration range of BA from 0.1 to 2.0 mg/l improved the production of hypericin (25-50 microg/g dry mass (DM)) and pseudohypericin (170-350 microg/g DM) in shoots. In callus cultures, BA (4.0-5.0 mg/l) did not changed hypericin contents (15-20 microg/g DM) but influenced pseudohypericin productions (120-180 microg/g DM). In the presence of auxins (IAA and IBA), Hypericum plantlets produced hypericin (30-100 microg/g DM) and pseudohypericin (120-400 microg/g DM). The presence of IAA did not influence naphtodianthrone productions in plantlets, but IBA decreased hypericin and pseudohypericin amounts in plantlets. The specific accumulation of the naphtodianthrones in in vitro cultures was influenced by phytohormonal supplementation of the medium. Results indicated that the production of hypericin and pseudohypericin could be increased by carefully adapted in vitro cultures. Hypericum in vitro cultures represent promising systems for hypericin and pseudohypericin productions.     

NO对金丝桃悬浮细胞生长及金丝桃素生物合成的促进作用研究

一氧化氮 (NO)是近年来发现的一种新型植物信号分子。以硝普钠 (Sodiumnitroprusside ,SNP)为一氧化氮 (NO)的供体 ,研究外源NO对金丝桃悬浮细胞生长及金丝桃素生物合成的影响。试验结果表明 ,金丝桃悬浮细胞在含 0 5和 15 0mmol LSNP的培养基中培养 2 0d后 ,细胞的干重分别为对照组的 140%和50% ;细胞中金丝桃素的含量分别为对照组的 98%和210%。试验结果表明 ,低浓度SNP处理有利于金丝桃悬浮细胞生长 ,而高浓度SNP可以促进金丝桃素的合成。在细胞培养初期 (0d)加入 0.5mmol LSNP并在指数生长后期 (14d)加入15.0mmol LSNP的金丝桃悬浮细胞在培养 2.5d后 ,细胞的干重和金丝桃素的含量分别为对照组的1.4和1.8倍 ,金丝桃素的产量达15.2mg/L ,比对照高3.2倍。SNP对金丝桃悬浮细胞生长及金丝桃素含量的影响可以被NO专一性淬灭剂CPITO(2-4-carboxyphenyl-4 ,4 ,5 ,5-tetramethylimidazoline-1-oxyl-3-oxide)所抑制,说明SNP是通过其分解产物NO影响细胞生长和金丝桃素的合成。试验结果同时表明,在15.0mmol/L的SNP处理下,金丝桃悬浮细胞中的苯丙氨酸解氨酶(PAL)的活性显著升高,推测NO可能通过触发金丝桃悬浮细胞的防卫反应,激活了细胞中金丝桃素的生物合成途径。     

Antioxidant properties of some hydroalcoholic plant extracts with antiinflammatory activity

The hydroalcoholic extracts of Calendula officinalis, Hypericum perforatum, Plantago lanceolata and Glycyrrhiza glabra which exhibited different anti-inflammatory activities were evaluated for the possible mode of action by studying their antioxidant potential. In the present study we investigated if standardized hydroalcoholic extracts of plants such as Calendula officinalis, Hypericum perforatum, Plantago lanceolata and Glycyrrhiza glabra produced by Hofigal Stock Company could modulate the respiratory burst of human activated neutrophils, as a consequence of their antioxidant capacity. Their antioxidant properties were measured using a colorimetric assay (Total Antioxidant Status kit). We demonstrated that Hypericum perforatum and Calendula officinalis hydroalcoholic extracts possessed a significant antioxidant activity while Plantago lanceolata and Glycyrrhiza glabra hydroalcoholic extracts had a minor antioxidant status. Using reactive oxygen species-generating systems (OZ-activated human PMN neutrophils), Calendula officinalis and Hypericum perforatum extracts showed strong reactive oxygen species scavenging property, Hypericum perforatum extract exhibing the highest scavenging activity. These results confirm the potential of Calendula officinalis and Hypericum perforatum investigated hydroalcoholic extracts as medicinal remedies to be used in different inflammatory/allergic diseases. These extracts could be a useful tool for obtaining new antioxidant/anti-inflammatory agents.     

Role of flavonoids in controlling the phototoxicity of Hypericum perforatum extracts.

Hypericum perforatum extracts are used mainly as oral antidepressants. Depending on source the extracts contain various amounts of phenylpropanes, flavonol derivates, biflavones, proathocyanidines, xanthones, phloroglucinoles, some amino acids, naphtodianthrones (hypericines) and essential oil constituents. The therapeutic use of Hypericum perforatum extracts however is limited by their phototoxic potential. It was the aim of the present study to investigate the phototoxic potential of 3 Hypericum perforatum extracts from different sources as well as some of its main constituents. In order to systematically study the phototoxic potential we established a modified neutral red assay utilizing an immortalized human keratinocyte cell line (HaCaT cells) as substrate and UVA irradiation. This modified neutral red assay was found to be a simple and reliable method for detecting phototoxic effects of reference agents and plant extracts. The validity of this method was demonstrated with known phototoxic compounds like chloropromazine and psoralenes like 5-MOP. Hypericum perforatum extracts demonstrated cytotoxicity and photocytotoxicity in a dose and UVA-dose dependent manner. Hypericine itself also evoked severe phototoxic effects and was thus identified as the main phototoxic constituent. Among the tested flavonoids quercitrin was found to be cytotoxic, while rutin unexpectedly demonstrated phototoxicity whereas quercitrin was effective to control the phototoxic activity of Hypericum perforatum extracts.     

用贯叶金丝桃残渣栽培香菇的研究

用提取金丝桃素后的贯叶金丝桃残渣和锯木屑作培养基分别栽培香菇 ,比较了由两种培养基栽培香菇的生物学效率、香菇的可溶性蛋白含量和超氧化物歧化酶 (SOD)活力。结果表明 :用贯叶金丝桃残渣栽培的香菇有较高的生物学效率、其香菇可溶性蛋白含量和SOD活力亦高 ,说明贯叶金丝桃残渣适合栽培香菇 ,为栽培香菇开发了一种新的生物资源。     

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。     

Stimulation of the production of hypericins by mannan in Hypericum perforatum shoot cultures

Shoot organ cultures were established from callus derived from anthers of Hypericum perforatum flowers and the effect of elicitors on hypericin and pseudohypericin production in shoot organ cultures was investigated. Mannan stimulated pseudohypericin production up to four fold (0.82 mg/g dry wt) and hypericin production up to two fold (0.04 mg/g dry wt.) beta-1,3-glucan and pectin slightly stimulated pseudohypericin production (ca. two fold), but had no effect on hypericin production. On the other hand, yeast extract showed no stimulatory effect, on either hypericin or pseudohypericin production.     

Phytochemical profiling of New and Old World Hypericum (St. John's Wort) species

Botanical extracts of Hypericum perforatum L. (common St. John's Wort) are used in the USA and in Europe as a treatment for mild to moderate depression, although controversy surrounds the identity of the active constituent(s). RP-HPLC with photodiode array detection was used to separate and quantify nine compounds of pharmacological interest in extracts from 74 taxa of Hypericum native to the Old and New World. Chemical profiles of these constituents may be used to distinguish extracts of H. perforatum from those of other species of Hypericum, and to indicate species that may be of interest for further phytochemical investigation.