药用植物巴西吐根的引种繁殖及栽培初探

吐根作为治疗痢疾的特效药而著名,主要具有祛痰、催吐和抗阿米巴痢疾的作用。我国目前对吐根药材的需求全部依赖于进口,国内未有大规模种植,为此,该文对原产巴西的吐根进行了引种繁殖及栽培研究。结果表明:(1)云南省西双版纳傣族自治州景洪市的气候条件能满足吐根的正常生长发育的需求,可作为吐根的引种地。(2)吐根的分根和茎下段可用于扦插繁殖,其发根率和存活率分别100.0%和100.0%、68.0%和75.0%,前者优于后者。(3)用20 mg·L^-1的IAA或IBA浸泡1 h,吐根茎下段的扦插成活率为90.0%或88.0%,均显著高于对照,可用于提高吐根茎下段的扦插成活率。(4)采用分根繁殖的吐根植株在植株生长和药材外观性状较好,单株总根体积较高,吐根一年生植株平均株高为10.66 cm,两年生平均株高为16.54 cm,一年生植株根总体积为2.71 mL,两年生根总体积为3.54 mL。(5)吐根栽培基质可用腐殖土:椰糠体积比为4:1。(6)在吐根的年周期生长中,地径1—3月和9—11月增长明显,株高7—11月增长明显,叶片长和宽3—9月增长明显,根据这些特点,可科学制定相应的水肥管理措施。该研究结果可为吐根的引种繁殖和栽培提供一定的科学参考。     

Anthraquinones in cell suspension cultures of Morinda citrifolia

From cell suspension cultures of Morinda citrifolia five known anthraquinones, rubiadin, lucidin, morindone, lucidin-3-β-primeveroside and morindone-6-β-primeveroside, and seven new anthraquinones were isolated. Six of the seven new quinones were characterized as 2-methyl-3,5,6-trihydroxyanthraquinone, 3-hydroxymorindone, 5,6-dihydroxylucidin, 2-methyl-3,5,6-trihydroxyanthraquinone-6-β-primeveroside, 3-hydroxymorindone-6-β-primeveroside and 5,6-dihydroxylucidin-3-β-primeveroside, respectively.     

Anthocyanins of Cephaelis, Cynomorium, Euterpe, Lavatera and Pinanga

The two malonylated pigments, malonylmalvin and malvidin 3-malonylglucoside, were identified in petals of Lavatera maritima, which belongs to the Malvaceae, a family known to synthesise such pigments. Zwitterionic anthocyanins could not be detected in four other newly examined sources and common unacylated pigments were recorded. Thus, the fruits of the palms Euterpe edulis and Pinanga polymorpha have a mixture of cyanidin 3-glucoside and cyanidin 3-rutinoside, while the fruit of Cephaelis subcoriacea is coloured by cyanidin 3-glucoside. The latter pigment was also obtained from the reddish brown inflorescence of the parasitic plant Cynomorium coccineum.     

Partial purification and characterization of geissoschizine dehydrogenase from suspension cultures of Catharanthus roseus

The characterization and partial purification of geissoschizine dehydrogenase from Catharanthus roseus cell suspension cultures are described. The 35-fold purified enzyme removes the 21α-hydrogen of geissoschizine in a NADP⁺-dependent reaction. NAD⁺, FAD or FMN cannot act as cofactors for the dehydrogenation. Structurally related indole alkaloids are not dehydrogenated. In comparison to enzymes of the ajmalicine pathway, geissoschizine dehydrogenase shows an extremely low specific activity.     

Diethylene glycol disulfide from castor bean cell suspension cultures

Diethylene glycol disulfide was isolated from castor bean cell suspension cultures. Incubation of suspension cultures with Na₂³⁵SO₄ resulted in the incorporation of radioactivity into the isolated diethylene glycol disulfide. Diethylene glycol disulfide was detected in cells (430 nmol/g cells) and in cell-free growth medium (41.5 nmol/ml).     

Biotransformation of thebaine by cell suspension cultures of Papaver somniferum cv. Marianne

Thebaine is biotransformed to neopine by cell suspension cultures of Papaver somniferum cv. Marianne grown in O-B5 medium. Results of precursor stu     

Phenolic synthesis in Perilla cell suspension cultures

Suspension cultures of Perilla ocymoides accumulate caffeic acid, both in free and ester forms, as the only phenylpropanoid end metabolite. Increased levels of growth substances influenced the levels of PAL activity and phenolic accumulation so that cytokinin stimulated, while auxin repressed both parameters. The regulatory role of caffeyl compounds is discussed in relation to their accumulation during the early exponential phase of culture growth.     

Quinine and quinidine production by cinchona leaf,root and unorganized cultures

Serially propagated Cinchona ledgeriana and C. succirubra (Rubiaceae) leaf, root and unorganized suspension cultures established from germinated seeds were studied for quinine and quinidine production. Leaf organ cultures were grown and subcultured in Murashige and Skoog's Revised Tobacco Medium supplemented with benzyladenine; root organ cultures were grown on the same medium supplemented with indolebutyric acid; and unorganized suspension cultures were grown on the same medium supplemented with 2,4-dichlorophenoxyacetic acid and benzyladenine. On a dry weight basis, leaf organ cultures of C. ledgeriana contained 0.06 % quinine and 0.05 % quinidine and of C. succirubra contained 0.04 % quinine and 0.04 % quinidine. No quinine and quinidine were detected in either root organ or unorganized suspension cultures.     

Production and composition of extracellular polysaccharide from cell suspension cultures of Mentha

A suspension culture of Mentha was established from callus which formed on the tips of young shoots of a Mentha hybrid (M. arvenis × M. spicata). Changes in growth parameters during a culture cycle were recorded. The general appearance of cells during division and growth, including the changes in cell form, was also represented.Suspension-cultured cells of Mentha hybrid released a large amount of extracellular polysaccharides (ECP) mainly at the logarithmic phase of the growth cycle. The ECP contained galacturonic acid as major components and arabinose, galactose, glucose, xylose, rhamnose and mannose as minor components. The ratio of the uronic acid content to total sugar content in the ECP was below 40% at day 7, but increased up to 90% at day 21. The relative contents of xylose and glucose in the ECP decreased during the culture period, while the arabinose content increased and those of rhamnose, mannose and galactose remained constant.The IR spectrum suggested that the ECP were low-methoxylated pectic polysaccharides. The presence of lignin and related compounds in the ECP was not detected. The protein content of the ECP was about 10% and the main amino acids were alanine, proline, hydroxyproline, valine, asparticacid and serine, in that order.     

岩黄连细胞生长与营养物质消耗的动态学研究

在岩黄连细胞悬浮培养过程中,对培养液pH值,碳源、氮源和磷酸盐含量,以及细胞生物量和生物碱含量进行测定,分析其动态变化过程。结果显示培养液pH值在培养初期降低,后逐渐升高;碳源在培养过程中逐渐被利用,磷酸盐和氮源在培养中期几乎耗尽,其中磷酸盐的消耗速率最快;悬浮细胞的生长周期为20 d左右,第18天细胞鲜重和干重达最大,而第21天脱氢卡维丁和小檗碱的含量最高,分别为8.22mg/L和4.31mg/L。结果表明营养物质(碳、氮和磷)的吸收与细胞生长以及生物碱的合成密切相关,营养元素的相对消耗速率为磷>氮>碳,推测氮和磷是影响岩黄连细胞培养的主要因素。     

Production of aroma compounds from strawberry cell suspension cultures by addition of precursors

The potential of using short-chain fatty acids and -keto-acid as precursors for the production of typical fruit-type aroma compounds by strawberry cell suspension cultures was investigated. Analysis of the headspace by gas chromatography revealed that supplemented strawberry cell suspension cultures were capable of producing low concentrations of ethyl butyrate and butyl butyrate, and converting -ketovalerate to butanal and butanol. No aroma compounds were produced in unsupplemented or heat-treated cell suspension cultures. The results indicated that esterase, decarboxylase, and alcohol dehydrogenase might exist in strawberry cell cultures. Increasing temperature, illumination and addition of mannitol favoured the production of butyl butyrate. No difference was found between one- and two-week-old cultures in the ability to convert precursors to corresponding aroma compounds.     

Biosynthesis of anthraquinones and related compounds in Galium mollugo cell suspension cultures

From Galium mollugo cell suspension cultures, 1,4-dihydroxy-3-prenyl-2-naphtholic acid methyl ester diglucoside was isolated along with anthraquinones and mollugin. Production of the diglucoside was much increased by administering 2-succinylbenzoate to the cultures. The incorporation of 2-succinylbenzoate into lucidin-3-primeveroside, mollugin and the diglucoside in the mode so far proposed for rubiaceous anthraquinones was verified by administration of ¹³C-labelled 2-succinylbenzoate to the cell cultures.     

Molecular phylogeography of Carapichea ipecacuanha,an amphitropical shrub that occurs in the understory of both semideciduous and evergreen forests

The medicinal shrub Carapichea ipecacuanha (ipecac) is an amphitropic species with three disjunct areas of distribution. In the Brazilian Atlantic and Amazonian ranges, the species was associated mostly with the understory of seasonal semideciduous forests, whereas in the Central American–Colombian range, the species occurred in the understory of moist evergreen forests. We examined the phylogeographic structure of ipecac using chloroplast trnT‐trnL and nuclear internal transcribed spacer (ITS) sequences from 120 and 46 specimens, respectively. To complement existing data on root alkaloid profiles, we used high‐performance liquid chromatography to assess the levels of emetine and cephaeline in 33 specimens from the two Brazilian ranges. The three ranges shared neither nuclear nor chloroplast haplotypes. The phylogeographic structures showed an uneven distribution of genetic diversity, sharp breaks and high levels of genetic differentiation among ranges. Our results suggest that the extant populations are descendents of at least four distinct ancestral lineages. The Atlantic ipecacs showed higher levels of genetic diversity than ipecacs from the other two ranges; it is likely that they derive from two ancestral lineages, with long‐term persistence in that region. The Amazonian ipecacs were monomorphic with respect to the ITS and cpDNA sequences, which supports the view that there was a recent expansion from a single parental source after a strong genetic bottleneck. The existence of a fourth distinct lineage is apparent from the high levels of genetic and chemical differentiation that we identified in the Central American–Columbian ipecacs.     

Volatile metabolites from suspension cultures of Taraxacum officinale

A suspension culture of T. officinale (dandelion) was developed and maintained on modified Gamborg's B5 and Murashige and Skoog's media. Volatile metabolites, which collectively had an ‘apple-like’ odour, were released into the head-space air above the cultures. Analysis by GC-MS indicated the presence of acetic acid butyl ester, 2-methyl-1-propanol, n-butyl alcohol, 4-phenyl-1-butanol, 4-hydroxy-4-methyl-2-pentanone, acetic acid, 4-terpineol, β-terpineol and -terpineol.     

Formation of edulinine and furoquinoline alkaloids from quinoline derivatives by cell suspension cultures of Ruta graveolens

The formation of furoquinoline alkaloids and of edulinine, elaborated by cell suspension cultures of Ruta graveolens, was found to occur by way of 4-hydroxy-2-quinolone. Other substrates transformed to furoquinolines included 4-hydroxy- and 4-methoxy-3-(3-methyl-2-butenyl)-2-quinolone, known earlier as natural precursors in studies with whole plants. Involvement of dictamnine as a natural precursor of 8-methoxydictamnine (γ-fagarine) and skimmianine was proved using ¹⁴C-labelled compounds. Edulinine in the cell suspensions was formed from such substrates as 4-hydroxy-N-methyl-2-quinolone, 4-hydroxy-3-(3-methyl-2-butenyl)-N-methyl-2-quinolone and its 4- methyl ether; this is probably the natural biosynthetic sequence. Changes in alkaloid yields were noted upon prolonged subculturing.     

Callus formation from protoplasys of Sorghum cell suspension cultures

Cell suspension cultures have been obtained from three cultivars of Sorghum bicolor L. Moench. Protoplasts readily obtained from these cultures underwent sustained cell division and callus formation.     

Plant regeneration of protoplasts isolated from suspension cultures of Solanum lycopersicoides

A protocol was developed for the isolation, culture and plant regeneration of protoplasts isolated from suspension cultures of Solanum lycopersicoides Dun. (LA 1990). Protoplasts were isolated by an overnight enzyme digestion, further purified by washing in W5 salts solution, and plated in two modified MS protoplast culture media with and without type VII agarose. The addition of agarose to the two culture media did not enhance plating efficiencies and shoot regeneration percentages and in some cases was even inhibitory. Unlike the experience with some other solanaceous species, the deletion of ammonium from the protoplast culture medium was not found to be beneficial. Protoplasts sustained continuous division in the modified MS media and up to 70% of the protoplast-derived calli readily regenerated shoots on MS salts and vitamins medium containing zeatin and GA.     

Production of ferruginol by cell suspension cultures of Salvia miltiorrhiza

The time-course production of ferruginol, a diterpene, was examined during growth of Salvia miltiorrhiza in cell culture. Ferruginol was produced in the lag and stationary phases of growth and so production of this secondary metabolite was inversely related to active cell division. The effects of auxins and light on ferruginol production were also examined.     

Biosynthesis of morindone and alizarin in intact plants and cell suspension cultures of Morinda citrifolia

Chrysophanol (1,8-dihydroxy-3-methylanthraquinone) is an acetate-derived anthraquinone which is substituted in both benzenoid rings. Morindone (1,5,6-trihydroxy-2-methylanthraquinone) is also substituted in both rings but is shown to be derived from shikimic acid via o-succinoylbenzoic acid.