紫锥菊多倍体诱导与鉴定

本实验以紫锥菊愈伤组织上刚分化出的不定芽为材料,用不同浓度秋水仙素溶液对其进行诱导,确定最佳处理浓度和处理时间,并对诱导的多倍体与二倍体进行形态、显微、染色体及过氧化氢酶(CAT)活性的比较鉴定.结果表明:0.025%秋水仙素浓度处理24h的诱导效果最好,诱导率达42.85%;多倍体植株叶片肥厚、根粗壮,气孔面积极显著地大于二倍体植株,染色体数从24～28条不等,CAT平均酶活性是二倍体的2.1倍.     

紫锥菊的组织培养和植株再生

1植物名称紫锥菊(Echinacea purpurea),又名紫松果菊. 2材料类别无菌苗的叶柄切段. 3培养条件(1)芽诱导及增殖培养基:MS 6-BA 1.0mg·L-1(单位下同) NAA 0.5 3%蔗糖 0.7%琼脂,pH 6.0;(2)生根培养基:B5大量元素 MS微量元素、铁盐、有机物 NAA 1.0 2%蔗糖 0.9%琼脂,pH 6.0.培养温度为(25 1)℃,光照12 h·d-1,光照度1500～2000 lx.     

藏红花愈伤组织诱导及其细胞培养的研究

藏红花幼叶愈伤组织的诱导频率在ＭＳ,Ｂ５和Ｗｈｉｔｅ三种培养基上均高达９８％;幼叶和芽的诱导率差异不大,高达９９％;不同时期的叶片差异较大,以幼叶诱导为佳;球茎诱导率为近８０％;激素配比以２．４－Ｄ２．０ｍｇ／Ｌ,ＢＡＰ０．１～０．５ｍｌ／Ｌ为宜。在继代培养阶段,ＭＳ比Ｂ５和Ｗｈｉｔｅ更适合细胞的快速生长繁殖;并且以叶片的愈伤组织生长较快,芽次之,球茎最慢,适合于细胞生长的激素配比为ＮＡＡ２．０～３．０ｍｇ／Ｌ,ＢＡＰ０．５～１．０ｍｇ／Ｌ为宜。     

何首乌愈伤组织的诱导

研究了外植体、光暗条件、植物激素等因子对何首乌愈伤组织诱导的影响 ,以及 6 -BA和何首乌组织提取液对愈伤组织增殖的影响。结果表明 :茎段的愈伤组织诱导优于带侧芽茎段和叶片 ;光照有利于愈伤组织的诱导 ;4因子 4水平的正交实验表明 ,对何首乌茎段愈伤组织诱导的影响 2 ,4 -D >6 -BA >IBA >IAA ,最佳激素搭配是 :MS +2 ,4 -D 2mg L +6 -BA 1mg L +IBA1mg L或MS +2 ,4 -D 2mg L +6 -BA 2mg L +IAA 1mg L ;MS培养基中附加 6 -BA或组织提取液均促进愈伤组织的增殖。     

紫锥菊挥发性成分分析研究

采用气-质(GC-MS)联用技术结合顶空固相微萃取(HS-SPME)与传统共水蒸馏法,对紫锥菊植物干花与干根中挥发性成分进行分析方法研究。GC-MS(配N ist2005标准质谱库)从传统方法提取的紫锥菊干根挥发油中分离分析出188个峰,初步鉴定出68种化合物,从聚二甲基硅氧烷(PDMS 7,100μm)及聚丙烯酸酯(PA 85μm)三种固相微萃取涂层吸附的紫锥菊干花挥发性成分中分别分离分析出27、118、105个峰,各自鉴定出1、22、23种化合物。对不同处理方法及紫锥菊植物不同部位所得数据进行了相识性与差异性的比较,所建分析方法及所得结果为了解紫锥菊植物体挥发性有效成分提供了参考。     

火龙果愈伤组织诱导与植株再生

为了建立火龙果愈伤组织诱导与植株再生体系,以火龙果茎段、幼苗和子叶为外植体进行离体培养试验。结果表明：茎段诱导愈伤组织的最优培养基为1／2MS＋2,4-D2．0mg·L^-1＋6-BAO．5mg·L^-1,诱导子叶愈伤组织的最适培养基是1／2MS＋2,4-D2．0mg·L^-1＋6-BA1．0mg·L^-1,诱导愈伤组织分化的最优培养基为1／2MS＋6-BA4．0mg·L^-1＋NAA0．5mg·L^-1,最佳生根培养基为1／2MS＋6．BA1mg·L^-1＋NAA0-3mg·L^-1。     

罗布麻愈伤组织诱导及植株再生

以罗布麻(Apocynum venetum L.)当年的成熟种子和5周龄的幼苗叶片为外植体,研究了不同激素组合、暗培养对愈伤组织及植株再生的影响.结果表明,幼苗作外植体诱导愈伤的最佳培养基为添加1.0 mg/L 6-BA 0.2 mg/L IBA的MS培养基;继代培养中1.0 mg/L 6-BA与0.2 mg/L IBA组合愈伤致密而生长迅速,长时间培养硬化的愈伤组织可用添加0.5 mg/L 6-BA和0.1 mg/L IBA培养基和初期暗培养获得大量质地疏松、增殖迅速的愈伤组织;再生苗诱导以0.5 mg/L 6-BA 0.2 mg/L IBA组合为佳;1/2MS附加NAA 0.6 mg/L为适宜的生根培养基,初步建立了罗布麻离体再生体系.     

秦艽体细胞愈伤组织诱导及继代研究

本文报道了秦艽取叶、幼苗为外植体诱导愈伤组织试验。结果表明,取幼苗６７－Ⅴ培养基,２４－Ｄ２５ｍｇ／Ｌ,ＮＡＡ０５ｍｇ／Ｌ,诱导频率最优,可高达１００％。继代培养,以６７％培养基,ＮＡＡ０５ｍｇ／Ｌ,２４－Ｄ（１－１５）ｍｇ／Ｌ最优     

紫锥菊的生药学研究

采用来源鉴定、性状鉴定、显微及理化鉴定的方法研究紫锥菊的生药学内容,为制定药材质量标准、研究、开发和利用紫锥菊的药用资源提供了理论依据。     

紫锥菊单咖啡酰酒石酸和菊苣酸提取工艺优化

采用正交试验设计,选取乙醇体积浓度百分比、提取温度、提取时间、抗氧化剂用量等因素,优化紫锥菊Echinacea purpurea单咖啡酰酒石酸和菊苣酸的加热回流提取工艺,并考察加入抗氧化剂对提取效果的影响。结果表明,优化的提取条件是以25%乙醇,在80 ℃回流提取90 min。抗氧化剂用量对提取效果影响不显著。优化后的加热回流提取条件对紫锥菊单咖啡酰酒石酸和菊苣酸的提取均适用,提取中无需加入抗氧化剂。     

Elicitation of secondary metabolism in Echinacea purpurea L. by gibberellic acid and triazoles

In vitro bioreactor production of Echinacea purpurea L. was used to facilitate the screening of compounds capable of eliciting increased secondary metabolite production. Based on previous experience with various bioreactors, the Southern Sun Liquid Lab Rocker was selected for this study as it produces healthy, vigorous, whole plants. The focus of the present study was to quantify the concentration of the medicinally important secondary metabolites caftaric acid and cichoric acid and to study the effect of growth regulators on the production of these compounds in this system. Both marker compounds were produced at levels that compared favorably to field grown plants. Application of gibberellic acid (GA₃), paclobutrazol, uniconazole and a combined treatment of GA₃ with paclobutrazol to the in vitro plants generally increased the concentration of caftaric acid in the roots with little effect on the concentration in the shoots. The concentration of cichoric acid was higher in the roots of treated plants than in the roots of control plants, but lower in corresponding shoots. The present study highlights the use of in vitro production of whole plants as a model system for studying the regulation of plant secondary metabolism in a controlled environment.     

紫锥菊提取物及酚酸化合物体外抗血小板聚集活性研究

为探究紫锥菊提取物及酚酸化合物体外抗血小板聚集活性,体外抗血小板聚集实验采用Born比浊法,以聚集抑制率和半数抑制浓度为指标评价。同时采用分子对接方法,选择凝血因子V(F5)、凝血因子VIII(F8)及凝血因子XI(F11)与酚酸类化合物进行虚拟对接,研究其抗血小板聚集的分子作用靶点。结果表明化合物S-1～S-10及其提取物对体外ADP诱导的血小板聚集有抑制作用,抑制率呈浓度依赖性,S-6与靶点的结合位点更多,选择性更强。紫锥菊中酚酸类化合物及其提取物在体外均显现出抗ADP诱导的血小板聚集活性,为紫锥菊体内研究提供依据。     

Effects of phenytoin and Echinacea purpurea extract on proliferation and apoptosis of mouse embryonic palatal mesenchymal cells

Cleft palate is one of the most common birth defects. Several environment factors are involved in the disorder, such as smoking, vitamin deficiency and teratogens. We investigated the teratogenic agent phenytoin and extract of the immunostimulant Echinacea purpurea in the etiology of cleft palate associated with the proliferation and apoptosis of mouse embryonic palatal mesenchymal (MEPM) cells. We measured the effects of phenytoin, E. purpurea extract, and the mixture of phenytoin and E. purpurea extract on the cell viability of MEPM cells by CCK‐8 assay and on the proliferation and apoptosis of MEPM cells by BrdU labeling assay, flow cytometry, and TUNEL assay. Exposure to phenytoin for 24 h inhibited cell proliferation and increased cell apoptosis of MEPM cells, and E. purpurea extract had the reverse effect. Importantly, treatment with the mixture of phenytoin and E. purpurea extract increased the proliferation and decreased the apoptosis of MEPM cells as compared with treatment with phenytoin alone. The teratogenic effect of phenytoin on cleft palate is associated with the proliferation and apoptosis of MEPM cells, and E. purpurea extract may have a protective effect. J. Cell. Biochem. 112: 1311–1317, 2011. © 2011 Wiley‐Liss, Inc.     

青叶胆愈伤组织的诱导与液体继代培养

以青叶胆(Swertia mileensis)幼叶为外植体,在添加不同生长调节剂组合的MS培养基上进行愈伤组织诱导。结果表明,最有效的生长调节剂组合为IAA 1.0 mg/L+NAA 1.0 mg/L+KT 1.0 mg/L和NAA 4.0 mg/L+KT 2.0 mg/L+GA₃ 1.0 mg/L,其诱导率分别为96.6%和96.0%。使用以上两种生长调节剂组合进行愈伤组织的液体继代培养,10 d可使愈伤组织增殖率分别达257.3%和310.2%。     

Floral nectar production and nectary anatomy and ultrastructure of Echinacea purpurea (Asteraceae)

BACKGROUND AND AIMS: In spite of the impressive species diversity in the Asteraceae and their widespread appeal to many generalist pollinators, floral-nectary ultrastructure in the family has rarely been investigated. To redress this, a study using Echinacea purpurea, a plant of horticultural and nutraceutical value, was undertaken. Nectar secretion of disc florets was compared with floral nectary ultrastructure taking into account nectar's potential impact upon the reproductive success of this outcrossing species. METHODS: Micropipette collections of nectar in conjunction with refractometry were used to determine the volume and nectar-sugar quantities of disc florets throughout their phenology, from commencement of its production to cessation of secretion. Light, scanning-electron and transmission-electron microscopy were utilized to examine morphology, anatomy and ultrastructure of nectaries of the disc florets. KEY RESULTS: Florets were protandrous with nectar being secreted from anthesis until the third day of the pistillate phase. Nectar production per floret peaked on the first day of stigma receptivity, making the two innermost whorls of open florets most attractive to foraging visitors. Modified stomata were situated along the apical rim of the collar-like nectary, which surrounds the style base and sits on top of the inferior ovary. The floral nectary was supplied by phloem only, and both sieve elements and companion cells were found adjacent to the epidermis; the latter participated in the origin of some of the precursor cells that yielded these specialized cells of phloem. Companion cells possessed wall ingrowths (transfer cells). Lobed nuclei were a key feature of secretory parenchyma cells. CONCLUSIONS: The abundance of mitochondria suggests an eccrine mechanism of secretion, although dictyosomal vesicles may contribute to a granulocrine process. Phloem sap evidently is the main contributor of nectar carbohydrates. From the sieve elements and companion cells, an apoplastic route via intercellular spaces and cell walls, leading to the pores of modified stomata, is available. A symplastic pathway, via plasmodesmata connecting sieve elements to companion, parenchyma and epidermal cells, is also feasible. Uncollected nectar was reabsorbed, and the direct innervation of the nectary by sieve tubes potentially serves a second important route for nectar-sugar reclamation. Microchannels in the outer cuticle may facilitate both secretion and reabsorption.     

Echinacea purpurea microbiota: bacterial–fungal interactions and the interplay with host and non-host plant species in vitro dual culture

• Important evidence is reported on the antimicrobial and antagonistic properties of bacterial endophytes in Echinacea purpurea and their role in the modulation of plant synthesis of bioactive compounds. Here, endophytic fungi were isolated from E. purpurea, and the dual culture approach was applied to deepen insights into the complex plant–microbiome interaction network.
• In vitro experiments were carried out to evaluate the species specificity of the interaction between host (E. purpurea) and non-host (E. angustifolia and Nicotiana tabacum) plant tissues and bacterial or fungal endophytes isolated from living E. purpurea plants to test interactions between fungal and bacterial endophytes.
• A higher tropism towards plant tissue and growth was observed for both fungal and bacterial isolates compared to controls without plant tissue. The growth of all fungi was significantly inhibited by several bacterial strains that, in turn, were scarcely affected by the presence of fungi. Finally, E. purpurea endophytic bacteria were able to inhibit mycelial growth of the phytopathogen Botrytis cinerea.
• Bacteria and fungi living in symbiosis with wild Echinacea plants interact with each other and could represent a potential source of bioactive compounds and a biocontrol tool.

     

Insulin-like 3 expression and fibrosis induction after intra-testicular injection of magnetic nanoparticles in rat testis and the ameliorative role of Echinacea purpurea extract

The World Health Organization has approved magnetic nanoparticles (MNP) for use as a contrast agent for magnetic resonance imaging or tumor hyperthermia treatment. MNP are toxic over time after intra-testicular injection. A clear strategy to ameliorate the toxic side effects of MNP in normal tissues after medical application has not yet been developed. We used an extract of Echinacea purpurea (EP) as a natural source of antioxidant and free radical scavenging product for detoxification of MNP in testicular tissues. MNP localization in the interstitial area of testicular tissue reduced the expression of insulin-like factor 3 (INSL3) proteins as well as serum testosterone levels. Further, MNP caused accumulation of both collagen and elastin in the interstitial area and increased the thickness of the tunica albuginea. Injection of MNP during administration of EP extract for short periods slightly reduced the toxic side effects of MNP. After extended exposure to EP extract, INSL3 expression and testosterone returned to near control levels. Also, collagen and elastin accumulation caused by MNP was reduced after extended exposure to EP extract. We believe that the ameliorative effect of EP extract is due to its antioxidant properties.     

Regeneration of Echinacea purpurea: Induction of root organogenesis from hypocotyl and cotyledon explants

An in vitro propagation system was developed for Echinacea purpureaL. (purple coneflower), a medicinal plant commonly used in the treatment of colds, flu and related ailments. Root organogenesis from Echinacea purpurea hypocotyl explants was effectively induced by indolebutyric acid. Indoleacetic acid was found to be less effective than indolebutyric acid while treatments with naphthaleneacetic acid were ineffective for induction of root organogenesis. The results of this study have established a micropropagation system for Echinacea purpurea that will provide axenic plant material for further investigations into medicinally active biochemicals and the mass production of high-quality Echinacea purpurea root tissues for the commercial market. This revised version was published online in June 2006 with corrections to the Cover Date.