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Identical amino acid sequence of ferredoxin from Datura metel and D. innoxia.

The complete amino acid sequence of the respective [2Fe-2S] ferredoxins from Datura metel (D. alba) and D. innoxia, have been determined by automated Edman degradation of the entire Cm-protein and of the peptides obtained by tryptic digestion and CNBr treatment. The ferredoxins from the two plants had an identical amino acid sequence, suggesting a very close taxonomic relationship between the two. This is the first report for the identical primary structure of Fds from different species of higher plants.

Amino Acid Sequence↗

Biosynthetic studies on the tropane alkaloid hyoscyamine in Datura stramonium; hyoscyamine is stable to in vivo oxidation and is not derived from littorine via a vicinal interchange process.

The conversion of littorine to hyoscyamine has been investigated by feeding deuterium labelled (RS)-[2-(2)H]-, [3, 3-(2)H(2)]-, [2, 3, 3-(2)H(3)]- phenyllactic acids to transformed root cultures of Datura stramonium. Isolation and GC-MS analyses of the isotope incorporation into the resultant hyoscyamine does not support the involvement of a vicinal interchange process operating during the isomerisation of littorine to hyoscyamine. Additionally a metabolism study with [1'-13C, 3', 3'-(2)H(2)]-hyoscyamine has established that the alkaloid is metabolically stable at C-3' with no evidence for a reversible in vivo oxidation process to the corresponding aldehyde. The data do not support an S-adenosy-L-methionine (SAM 5)/co-enzyme-B(12) mediated process for the isomerisation of littorine to hyoscyamine.

Atropine↗

Specificities of the enzymes of N-alkyltropane biosynthesis in Brugmansia and Datura.

The enzymes N-methylputrescine oxidase (MPO), the tropine-forming tropinone reductase (TRI), the pseudotropine-forming tropinone reductase (TRII), the tropine:acyl-CoA transferase (TAT) and the pseudotropine:acyl-CoA transferase (PAT) extracted from transformed root cultures of Datura stramonium and a Brugmansia candida x aurea hybrid were tested for their ability to accept a range of alternative substrates. MPO activity was tested with N-alkylputrescines and N-alkylcadaverines as substrates. TRI and TRII reduction was tested against a series of N-alkylnortropinones, N-alkylnorpelletierines and structurally related ketones as substrates. TAT and PAT esterification tests used a series of N-substituted tropines, pseudotropines, pelletierinols and pseudopelletierinols as substrates to assess the formation of their respective acetyl and tigloyl esters. The results generally show that these enzymes will accept alien substrates to varying degrees. Such studies may shed some light on the overall topology of the active sites of the enzymes concerned.

Datura stramonium↗

Simultaneous multi-element detection of metal ions bound to a Datura innoxia material.

An on-line detection scheme has been developed for the determination of metal ion affinities for binding to a plant-based substrate. This involves monitoring the effluent of a column packed with cell-wall fragments from the plant Datura innoxia for 27 different elements simultaneously by coupling the column to an ICP emission spectrometer. Previously accepted procedures for removing native metal ions from biological materials by washing the material with a pH 2 solution were found to be insufficient for this material. Measurable amounts of Na, Mg, Al, Ca, Mn, Fe, Ni, Cr, Zn, Cd, Pb, Ba, Sr, and Si were all detected in an effluent from the introduction of 1.0M HCl following washing the material in a pH 2 solution. Metal ion breakthrough curves for Cd(2+), Zn(2+), Ni(2+), Cu(2+), and Pb(2+) were found to exhibit an affinity order of Pb(2+)>Cu(2+)>>Zn(2+) congruent with Cd(2+)>Ni(2+) for an equimolar mixture of these metal ions. This configuration also enabled the displacement of metal ions to be detected as the breakthrough curve for a subsequent metal ion was monitored. Comparison of Ni and Zn binding indicates a simple ion exchange model is insufficient to explain sequential binding of these metal ions.

Adsorption↗

Distribution of hyoscyamine and scopolamine in Datura stramonium.

The production of hyoscyamine and scopolamine in Datura stramonium has been investigated in the different plant parts, at different stages of their life cycle. Maximum contents were found in the stems and leaves of young plants, hyoscyamine being always the predominant component.

Atropine↗

Steroidal compounds from in vitro regenerated shoots of Datura metel.

Regeneration of adventitious shoots from young leaves of Datura metel is described. Shoot buds developed on MS medium with 2 mg/l and elongated on hormone-free solid basal medium. The microshoots failed to produce alkaloids, but a number of steroidal compounds were detected. The C(28) sterol 3beta,24xi-dihydroxy-ergosta-5,25-dienolide, regarded as the precursor of withanolides and related steroidal lactones, was identified. The withanolide 12-deoxywithastramonolide was also detected.

Benzyl Compounds↗

Evaluation of analgesic effect of Datura fastuosa leaves and seed extracts.

The aqueous extracts of Datura fastuosa leaves and seeds were evaluated for the analgesic effect on acetic acid-induced writhing and hot plate reaction in mice. According to the results, D. fastuosa leaves and seeds extracts at oral doses of 400 and 800 mg/kg are effective as analgesic. The analgesic activity of leaf extract is reduced by naloxone but not that of seed extract.

Acetic Acid↗

Studies on antimycotic properties of Datura metel.

The hexane, chloroform, acetone and methanolic fractions of Datura metel L. were investigated for antifungal properties using pathogenic species of Aspergillus. The chloroform fraction was found to be endowed with antifungal activity. The minimum inhibitory concentration (MIC) of chloroform fraction of D. metel L. was 625.0 microg ml(-1) against all the three species of Aspergillus, i.e. A. fumigatus, A. flavus and A. niger, by microbroth dilution and percent spore germination inhibition assays. The MIC by disc diffusion assay was observed to be 12.5 microg disc(-1). The chloroform fraction of D. metel, when investigated for potency, turned to be 9.2 times less active than amphotericin B. However, it was important to note that the cytotoxicity of chloroform fraction in vitro was 117.8 times less than amphotericin B.

Animals↗

Healing potential of Datura alba on burn wounds in albino rats.

Datura alba Nees (Solanaccae) is popular all over the world for its medicinal uses in asthma, muscle spasm, whooping cough, hemorrhoids, skin ulcers, etc. In India, it is widely used traditionally for the relief of rheumatism and other painful affections. Ayurveda and Siddha practitioners use oil based preparations of this plant from ancient days to till date for all types of wounds. Hence, the present study was chosen to evaluate its scientific validity. The alcohol extract of the D. alba leaves were investigated for the evaluation of its healing efficiency on burn wound models in rats. The crude alcohol extract and one of the fractions exhibited antimicrobial effect against all the pathogens studied. A 10% (w/w) formulation of alcoholic extract was topically applied on thermal wounds. Complete wound closure was observed within 12 days in treated rats. The effect produced by the ointment, in terms of wound contracting ability, wound closure time, tissue regeneration at the wound site and histopathological characteristics were significant in treated rats. Collagen, hexosamine and gelatinase expressions were also well correlative with the healing pattern observed. The present study thus provides a scientific rationale for the traditional use of this plant in the management of wounds.

Animals↗

Investigation of the effect of space environment on the contents of atropine and scopolamine in Datura metel by capillary zone electrophoresis.

The seeds of Datura metel were carried aboard a retrievable satellite and exposed to space environment. The effects of space environment (weightlessness and ionizing radiation) on the contents of atropine and scopolamine in D. metel were investigated by using an effective capillary zone electrophoresis (CZE) method, which employed 50 mmol/l phosphate buffer (pH 8) containing 10% (v/v) tetrahydrofuran as the running buffer. The results showed that the contents of atropine and scopolamine varied to some extent, and the earth-control group has the lowest content of atropine. However, the variation of atropine and scopolamine contents in three groups was not obvious based on t-test. At the same time, the optimization of the separation was discussed in detail and the two compounds were completely separated within 10 min with satisfactory repeatability and calibration linearity.

Atropine↗

[Production of gastrodin through biotransformation of p-hydroxybenzaldehyde by cell suspension cultures of Datura tatula L].

The conversion of exogenous p-hydroxybenzaldehyde to p-hydroxy-methyl-phenol-beta-D-glucoside (gastrodin) was studied by using cell suspension culture of Datura tatula L. The chemical structure of this synthesized gastrodin was identified based on the spectral analysis and chemical evidence. The conversion procedure of p-hydroxybenzaldehyde into gastrodin by D. tatula L. cell suspension cultures was established. The synthesized gastrodin (II) was isolated from the ferment liquor and identified by spectral analysis. At the same time, the p-hydroxybenzyl alcohol (I) converted through biotransformation of p-hydroxybenzaldehyde by cell suspension cultures of D. tatula L. was also isolated and identified. The efficiency of glucosylation of p-hydroxybenzaldehyde was remarkably enhanced by adding salicylic acid (0.1 mg/L) and keeping the lower pressure (0.001MPa) in 25L airlift loop bioreactor. The biotransformation of exogenous p-hydroxybenzaldehyde to gastrodin by cell suspension culture of D. tatula L. is a promising approach.

Benzaldehydes↗

Kinetic study of littorine rearrangement in Datura innoxia hairy roots by (13)C NMR spectroscopy.

The kinetics of tropane alkaloid biosynthesis, particularly the isomerization of littorine into hyoscyamine, were studied by analyzing the kinetics of carbon-13 ((13)C) in metabolites of Datura innoxia hairy root cultures fed with labeled tropoyl moiety precursors. Both littorine and hyoscyamine were the major alkaloids accumulated, while scopolamine was never detected. Feeding root cultures with (RS)-phenyl[1,3-(13)C(2)]lactic acid led to (13)C spin-spin coupling detected on C-1' and C-2' of the hyoscyamine skeleton, which validated the intramolecular rearrangement of littorine into hyoscyamine. Label from phenyl[1-(13)C]alanine or (RS)-phenyl[1,3-(13)C(2)]lactic acid was incorporated at higher levels in littorine than in hyoscyamine. Initially, the apparent hyoscyamine biosynthesized rate (v(app)()hyo = 0.9 micromol (13)C.flask(-1).d(-1)) was lower than littorine formation (v(app)()litto = 1.8 micromol (13)C.flask(-1).d(-1)), suggesting that the isomerization reaction could be rate limiting. The results obtained for the kinetics of littorine biosynthesis were in agreement with the role of this compound as a direct precursor of hyoscyamine biosynthesis.

Alkaloids↗

The complex-type oligosaccharide binding lectin Datura stramonium agglutinin detects type II A muscle fibres in the branchial biceps from man and cat.

Complex-type oligosaccharides were detected in the sarcoplasm of muscle fibres from cat and human biceps using lectins and anticarbohydrate antibodies. The lectin Datura stramonium agglutinin strongly stained type II A fibres as identified by myosin ATPase activity after alkaline and acid preincubation. In contrast, all muscle fibres showed a moderate coarse granular staining after incubation with Tetracarpidum conophorum agglutinin and Telfairia occidentalis agglutinin which recognize tri-antennary complex glycans poorly bound by D. stramonium agglutinin. Strong sarcoplasmic staining in all muscle fibres was obtained after incubation with an antibody against branched N-acetyllactosamine structure while an antibody against binary 2 --> 3 sialyllactosamine glycans failed to detect the muscle fibres. Treatment of the muscle sections with sialidase prior to incubation with D. stramonium agglutinin did not influence the lectin staining pattern. Staining of blots from electrophoretically separated muscle proteins obtained by homogenization, solubilization and centrifugation of small muscle pieces showed D. stramonium agglutinin binding to a number of bands ranging from 200 kDa to 30 kDa. No D. stramonium agglutinin positive bands were observed in blots from separated mitochondrial proteins while blots from sarcoplasmic reticulum separated by electrophoresis stained many bands in the range from 200 kDa to 30 kDa. It may be concluded that all muscle fibres in human and cat biceps hold intracellular non-sialylated complex-type oligosaccharides and further, that a specific tri-antennary complex-type glycoform is strongly expressed in type II A fibres as recognized by D. stramonium agglutinin. These results indicate a different glycosylation of certain myofibrillar-associated proteins in muscle fibre types.

Animals↗

Gene expression in cadmium-tolerant Datura innoxia: detection and characterization of cDNAs induced in response to Cd2+.

The response of a metal tolerant plant to heavy metal stress involves a number of biochemical pathways. To investigate the overall molecular response of a metal-tolerant plant to heavy-metal exposure, suppressive subtractive hybridization was used to create a library enriched in cadmium-induced cDNAs from cadmium-tolerant Datura innoxia. Two differential screening steps were used to screen the cadmium-induced library resulting in 8 putative cadmium-specific cDNAs out of a pool of 94 clones. Reverse transcriptase polymerase chain reaction was used to confirm that 4 of these 8 clones were cadmium-specific, while the other 4 were induced under heat shock or in the no treatment cells in addition to cadmium exposure. All 8 cDNAs were sequenced and used to search for identification against GenBank. One of the 4 cadmium-specific cDNAs had homology to a sulfur transferase-family protein in Arabidopsis thaliana. The possible link between this result and the heavy-metal response of plants is discussed.

Adaptation, Physiological↗

Accidental mydriasis from exposure to Angel's trumpet (Datura suaveolens).

PURPOSE: To report clinical findings after accidental instillation into the eye of sap from Angel's trumpet (Datura suaveolens). METHODS: We report findings on seven patients who developed sudden onset of unilateral mydriasis. At least three of them also had ipsilateral cycloplegia and one developed transient tachycardia. RESULTS: The symptoms evolved after ocular exposure to sap from Angel's trumpet, a plant containing natural alkaloids with parasympatholytic properties. Six patients were initially unaware of the cause of their symptoms. In these cases, patient history revealed recent contact with Angel's trumpet. CONCLUSION: Accidental ocular instillation of sap from Angel's trumpet should be noted as a cause of sudden onset of mydriasis in otherwise unaffected patients and also of general symptoms like tachycardia.

Adult↗

Purification of the lectin from Datura stramonium.

The lectin from Datura stramonium can be inhibited by oligomers of N-acetylglucosamine. This property was exploited to purify the lectin by affinity chromatography on Sepharosefetuin. The purified lectin is a glycoprotein in having subunits of 40 000 and 45 000 mol.wt.

Acetylglucosamine↗

Evolutionary ecology of Datura stramonium: equal plant fitness benefits of growth and resistance against herbivory.

This study evaluated how natural selection act upon two proposed alternatives of defence (growth and resistance) against natural enemies in a common garden experiment using genetic material (full-sibs) from three populations of the annual plant Datura stramonium. Genetic and phenotypic correlations were used to search for a negative association between both alternatives of defence. Finally, the presence/absence of natural enemies was manipulated to evaluate the selective value of growth as a response against herbivory. Results indicated the presence of genetic variation for growth and resistance (1--relative damage), whereas only population differentiation for resistance was detected. No correlation between growth and resistance was detected either at the phenotypic or the genetic level. Selection analysis revealed the presence of equal fitness benefits of growth and resistance among populations. The presence/absence of natural herbivores revealed that herbivory did not alter the pattern of selection on growth. The results indicate that both strategies of defence can evolve simultaneously within populations of D. stramonium.

Biological Evolution↗