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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↗

Metabolism of the environmental estrogen bisphenol A by plant cell suspension cultures.

The metabolism of the environmental estrogen bisphenol A (BPA) was studied in heterotrophic plant cell suspension cultures of soybean (Glycine max), wheat (Triticum aestivum), foxglove (Digitalis purpurea), and thorn apple (Datura stramonium), which were regarded as metabolic model systems for intact plants. Three main metabolic routes of BPA were observed in the tissues. Most of the radioactivity found in the cell extracts consisted of carbohydrate conjugates of BPA amounting to about 85% (foxglove), 80% (wheat), 7% (soybean) and 15% (thorn apple) of applied 14C. The second main route was formation of non-extractable residues. Portions detected were low in foxglove (3.9% of applied 14C), moderate in wheat (13.5%), high in thorn apple (27.4%) and soybean (49.4%). With thorn apple, BPA derived bound residues were preponderantly resistant towards acid treatment; only traces of BPA were released. The third route was the formation of a highly polar, presumably polymeric material detected in media of soybean and thorn apple (29.3% and 36.0% of applied 14C, respectively). The mechanism of its formation remained unknown. In thorn apple, this highly polar material was formed extremely rapidly, and was considerably stable. Only traces of BPA were liberated by hydrolytic treatment with cellulase or acid. During hydrolysis experiments with glycoside fractions, non-extractable residues and highly polar materials, low amounts of presumably primary metabolites of BPA (up to 6% of applied 14C) were detected besides the parent compound; their chemical structures remained unclear.

Benzhydryl Compounds↗

Toxicity of cypermethrin in Labeo rohita fingerlings: biochemical, enzymatic and haematological consequences.

The effect of exposure to sub-lethal concentrations of cypermethrin, a synthetic pyrethroid pesticide, on biochemical parameters of muscle, blood and enzyme activities in brain, liver and kidney of the Indian major carp, Labeo rohita was studied. The sub-lethal exposure studies were done for up to 45 days at 1/10 and 1/50 of 96 h LC(50) of cypermethrin. The 96 h LC(50) was found to be 0.139 ppm. RNA levels decreased while DNA levels were elevated. Acid phosphatase was unchanged while alkaline phosphatase was depleted. Brain acetylcholinesterase activity was decreased significantly (P<0.05) over a period of 45 days at both cypermethrin concentrations. Lactate dehydrogenase activity in brain and liver was elevated, but inhibited in kidney. Succinate dehydrogenase and ATPase activities were depleted in brain, kidney and liver. There was a decrease in serum protein level over control at both concentrations of the pyrethroid. Blood glucose level and total leucocytes were elevated compared with controls at either concentration from day 15 to day 45. Haemoglobin percentage and total erythrocytes decreased in both sub-lethal concentrations. Extracts of the herb Datura stramonium were effective in countering the toxicity of this pesticide. Our data suggest that sub-lethal exposure of cypermethrin alters the biochemical, haematological parameters and enzymes of organs tissue and exert stress on the fish. Plant extracts may be useful in counteracting some of these effects.

Animals↗

Molecular cloning and functional identification of a plant ornithine decarboxylase cDNA.

A cDNA for a plant ornithine decarboxylase (ODC), a key enzyme in putrescine and polyamine biosynthesis, has been isolated from root cultures of the solanaceous plant Datura stramonium. Reverse transcription-PCR employing degenerate oligonucleotide primers representing conserved motifs from other eukaryotic ODCs was used to isolate the cDNA. The longest open reading frame potentially encodes a peptide of 431 amino acids and exhibits similarity to other eukaryotic ODCs, prokaryotic and eukaryotic arginine decarboxylases (ADCs), prokaryotic meso-diaminopimelate decarboxylases and the product of the tabA gene of Pseudomonas syringae cv. tabaci. Residues involved at the active site of the mouse ODC are conserved in the plant enzyme. The plant ODC does not possess the C-terminal extension found in the mammalian enzyme, implicated in rapid turnover of the protein, suggesting that the plant ODC may have a longer half-life. Expression of the plant ODC in Escherichia coli and demonstration of ODC activity confirmed that the cDNA encodes an active ODC enzyme. This is the first description of the primary structure of a eukaryotic ODC isolated from an organism where the alternative ADC routine to putrescine is present.

Amino Acid Sequence↗

Isolation of the tumor-inducing RNA from oncogenic and nononcogenic Agrobacterium tumefaciens.

Two RNA fractions have been isolated and purified from both oncogenic and nononcogenic strains of Agrobacterium tumefaciens. Both RNAs are capable of inducing the formation of transplantable tumors when introduced at wound sites in stems of Datura stramonium plants. One of these RNA fractions was found to be bound to an RNA-directed DNA polymerase, while the other was associated with the bacterial DNA. Physical evidence suggests that both are single stranded and small in size; linear sucrose gradients show that their size corresponds to a value of 5-6 S. A concentration of 4-5 mug of the RNAs dissolved in 0.01 ml of water is effective in initiating the formation of transplantable tumors in Datura plants.

Centrifugation, Density Gradient↗

Polyamine homeostasis in transgenic plants overexpressing ornithine decarboxylase includes ornithine limitation.

It was reported recently that overexpression of human ornithine decarboxylase (ODC) cDNA in transgenic rice plants resulted in increased steady-state concentration of polyamines, i.e., enough biosynthetic control is invested at this step to enable adjustment of polyamine levels. To investigate critically whether constitutive overexpression of ODC is sufficient to control steady-state polyamine levels, we expressed an ODC cDNA from Datura stramonium in transgenic tobacco plants. Transgenic progeny of self-fertilised primary transformants exhibited increases in ODC activity of 25-fold in leaves and 5-fold in flower buds. However, the increase in putrescine levels was only 1.5- to 2.1-fold in leaves and 1.1- to 1.3-fold in flower buds. Emphatically, no changes to spermidine or spermine steady-state levels or to soluble or insoluble hydroxycinnamic acid-conjugated polyamines were observed. Ornithine feeding to cell suspension cultures derived from the transgenic plants indicated that putrescine accumulation was limited in part by ornithine availability. These results demonstrate that a large increase in the capacity of the tobacco plants to decarboxylate ornithine does not result in a comparable increase in the level of free or conjugated polyamines. Plant polyamine homeostatic mechanisms efficiently accommodate increased ODC activity, suggesting that polyamine biosynthetic control is invested at multiple interdependent steps.

Cells, Cultured↗

Analysis of lectin properties with membrane ultrafiltration and high-pressure liquid chromatography.

A convenient method for the analysis of the binding properties of lectin with fluorogenic sugar chains is described. A lectin (concanavalin A or Datura stramonium agglutinin) was mixed with pyridylaminated sugar chains in buffer and the free chains obtained were isolated by membrane ultrafiltration. The amount of free sugar chains in the filtrate was measured by high-pressure liquid chromatography. The binding constants with the sugar chains, reaction kinetics, and other properties of these lectins were easily investigated. The method is simple and could be used to study the characteristics of any lectin in native form.

Binding Sites↗

The avirulence domain of Cauliflower mosaic virus transactivator/viroplasmin is a determinant of viral virulence in susceptible hosts.

Cauliflower mosaic virus (CaMV) transactivator/viroplasmin (Tav) is a multifunctional protein essential for basic replication of CaMV. It also plays a role in viral pathogenesis in crucifer and solanaceous host plants. Deletion mutagenesis revealed that N- and C-terminal parts of Tav are not essential for CaMV replication in transfected protoplasts. Two deletion mutants having only minimal defects in basic replication were infectious in turnips but only with highly attenuated virulence. This was shown to be due to delayed virus spread within the inoculated leaves and to the upper leaves. Unlike the wild-type virus, the mutant viruses successfully spread locally without inducing a host defense response in inoculated Datura stramonium leaves, but did not spread systemically. These results provide the first evidence that a Tav domain required for avirulence function in solanaceous plants is not essential for CaMV infectivity but has a role in viral virulence in susceptible hosts.

Brassica napus↗

Mutational analysis of bean yellow dwarf virus, a geminivirus of the genus Mastrevirus that is adapted to dicotyledonous plants.

Bean yellow dwarf virus (BeYDV) is an atypical member of the geminivirus genus Mastrevirus that infects dicotyledonous plants. BeYDV DNA contains six open reading frames (ORFs) with the capacity to encode proteins in excess of 10 kDa. Two virion-sense ORFs (V1 and V2) and two complementary-sense ORFs (C1 and C2) have homologues in all mastreviruses, while ORFs C3 and C4 are not conserved. To investigate their functions, each of the ORFs has been truncated by either frame-shifting or the introduction of a stop codon. We demonstrate that an ORF V1 mutant replicated efficiently in Nicotiana tabacum protoplasts but was unable to systemically infect Phaseolus vulgaris and Datura stramonium, consistent with a role for V1 protein in virus movement. However, the mutant was able to systemically infect Nicotiana benthamiana although the onset of symptoms was appreciably delayed in comparison with wild-type virus. Disruption of ORF V2, encoding the coat protein, prevented systemic infection of all three hosts but the mutant replicated in protoplasts. Both ORF C1 and ORF C2 were essential for replication in protoplasts. Modification of the complementary-sense splice donor and acceptor sequences also prevented replication. Removal of the intron prevented systemic infection, although the intronless mutant was able to produce functional replication-associated protein (Rep) and replicated efficiently in protoplasts. ORFs C3 and C4 were not required for systemic infection. Our results indicate that four ORFs are spatially and functionally conserved in mastreviruses that infect both monocotyledonous and dicotyledonous plants.

Base Sequence↗

The distinct disease phenotypes of the common and yellow vein strains of Tomato golden mosaic virus are determined by nucleotide differences in the 3'-terminal region of the gene encoding the movement protein.

In Nicotiana benthamiana, the common strain of the bipartite geminivirus Tomato golden mosaic virus (csTGMV) induces extensive chlorosis whereas the yellow vein strain (yvTGMV) produces veinal chlorosis on systemically infected leaves. In Datura stramonium, csTGMV produces leaf distortion and a severe chlorotic mosaic whereas yvTGMV produces only small chlorotic lesions on systemically infected leaves. Genetic recombination and site-directed mutagenesis studies using infectious clones of csTGMV and yvTGMV have identified a role in symptom production for the gene encoding the movement protein (MP). The MP amino acid at position 272, either valine (csTGMV) or isoleucine (yvTGMV), influenced symptoms in both hosts by inducing an intermediate phenotype when exchanged between the two strains. Exchange of an additional strain-specific MP amino acid at position 288, either glutamine (csTGMV) or lysine (yvTGMV), resulted in the change of symptom phenotype to that of the other strain. In situ hybridization analysis in N. benthamiana demonstrated that there was no qualitative difference in the tissue distribution of the two strains although csTGMV accumulated in higher amounts, suggesting that the efficiency of virus movement rather than distinct differences in tissue specificity of the strains is responsible for the symptom phenotypes.

3' Untranslated Regions↗

Datura lectin is both an anti-mitogen and a co-mitogen acting synergistically with phorbol ester.

The lectins from Datura stramonium, Lycopersicon esculentum, and Solanum tuberosum are structurally related and possess a similar carbohydrate specificity, yet the Datura lectin is mitogenic for human lymphocytes while the other two are not. However, the Datura lectin was found to antagonize blast transformation induced by purified protein derivative (PPD), even within the concentration range at which it was optimally mitogenic on its own. The presence of a submitogenic concentration of the phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA) enormously enhanced the mitogenic activity of Datura lectin. No such synergy was observed, however, between tomato lectin or S. tuberosum agglutinin (STA) and TPA, nor between Datura lectin and the calcium ionophore A 23187.

Calcimycin↗

Protein chemotaxonomy of the solanaceae. VI. Amino acid sequence of ferredoxin from Nicotian tabacum.

The complete amino acid sequence of [2Fe-2S] ferredoxin from Nicotiana tabacum has been determined by automated Edman degradation of the entire Cm-protein and of the peptides obtained by trypsin and Asp-N endoproteinase digestions. This ferredoxin exhibited 9, 10, 8, and 10 differences respectively in its amino acid sequence, when compared with the ferredoxins of Datura stramonium, D. metel, D. arborea, and Physalis alkekengi var.francheti but 17-28 differences for other angiosperms, and 34-37 differences for fern and horsetails. These results are in agreement with the taxonomic position for these plants.

Amino Acid Sequence↗

Poisoning by the use of Datura leaves in a homemade toothpaste.

Datura stramonium and related species are relatively common causes of atropine-like poisoning by ingestion or inhalation. Toxic absorption after mucosal application is evident in 24 h of atropinism sustained by a woman who used a toothpaste mixed with the leaves and flowers of Datura sp., table salt, vinegar and an alcoholic beverage.

Absorption↗

Acute anticholinergic syndrome due to Jimson seed ingestion. Clinical and laboratory observation in six cases.

Ten patients presented with acute anticholinergic syndrome secondary to Jimson seed (Datura stramonium) ingestion. Six of the 10 patients required hospitalization because of hyperpyrexia and severe neurologic derangement. Electroencephalograms recorded immediately after admission showed [1] slow wave activity, and [2] bizarre rhythmical bursts of high-voltage sharp wave activity; both of which rapidly resolved during the next 24 hours, as did the associated clinical findings of hyperreflexia, bilateral dorsiflexor Babinski responses, and decerebrate posturing. Previously unreported elevation of serum glutamic-oxalacetic transaminase and lactic dehydrogenase and prothrombin time prolongation are documented. The pathogenic mechanism accounting for abberation of these laboratory values remains undefined. All patients showed rapid clinical improvement; follow-up neurologic evaluation and electroencephalograms have been within normal limits. Because of the widespread availability and potential abuse of the Jimson seed, the clinical syndrome associated with its toxicity merits recognition.

Acute Disease↗

[Acute intoxication with "sobi-lobi" (Datura). Four cases in Niger].

The datura (Datura stramonium, Datura metel) belongs to a group of plants from the solanacies variety and can be the cause of voluntary intoxication. In Nigher, its flowers and seeds are used for their hallucinogenic properties. Its alcaloïds have an atropine-like effect. We are reporting here the first four cases of voluntary intoxication among teenagers in Niamey (Republic of Niger). The symptoms, which happened to be identical for the four patients, lead us quickly towards the possibility of a collective intoxication. During the questioning, the patients admitted they had smoked the plant's dried flowers, but also they drunk an herbal tea mixed with datura seeds. Symptoms are comparable to that of intoxications caused by atropine. Neurological signs are the main symptoms (agitation, delirium, disorientation, hallucination, mydriasis, retention of urine...). The others noticeable signs are: fever, dry mouth, tachycardia, thirstiness. The evolution was favourable as the patients left the hospital between D2 and D4. Those clinical features are compared to the literature items, which are inexistent in western Africa. The prognosis may be fatal, especially for the child and during massive intoxications meant to be autolystic or toxicomaniac. The recovery when possible leads usually to no sequels however sever the clinical picture is. The consultation of the population enabled us to comprehend that datura is known and consumed for its hallucinogenic proprieties. The therapeutic guidance is described and prevention suggested.

Acute Disease↗

[Jimson weed poisoning].

Datura stramonium abuse causes a potentially lethal anticholinergic intoxication. Today, with the internet widely available, our youth are potentially exposed to partial and quite often dangerous information that systematically disregards the danger of Datura use. The authors suspect that without educational efforts regarding the dark side of Datura use, we shall see a rise in poisoning by this dangerous substance. This review outlines the general management of the intoxication.

Datura stramonium↗

[Pathogenesis and induced virus resistance in tobacco plants affected by tomato spotted wilt virus].

Pathogenecity and virulence of tomato spotted wilt virus isolated from tobacco were studied under the field conditions of the Crimea in the tobacco plants of Immunny 580 variety and Datura stramonium. As to their virulence the obtained 26 isolates were divided into three groups: strongly virulent (severe), middle-virulent and weakly virulent ("mosaic"). It has been shown that strongly virulent isolates interfere with weakly virulent ones and induce the development of nonspecific resistance in tobacco plants, this resistance is also efficient to the tobacco mosaic virus.

Datura stramonium↗