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Sample records for poppy papaver somniferum

  1. Opium Poppy (Papaver somniferum).

    PubMed

    Chitty, Julie A; Allen, Robert S; Larkin, Philip J

    2006-01-01

    The genetic transformation of opium poppy, Papaver somniferum, offers the opportunity to study the mechanisms involved in the regulation of benzylisoquinoline and morphinan alkaloid biosynthesis. The development of an efficient transformation protocol for opium poppy has allowed us to transform a range of genotypes from all around the world, including previously recalcitrant high-yielding commercial Australian cultivars. The method involves Agrobacterium tumefaciens infection of hypocotyl explants, followed by the production of antibiotic or herbicide resistant embryogenic callus, the subsequent induction of somatic embryos and development into normal plants. The use of different selective agents, binary vectors, and poppy genotypes has demonstrated the robustness and reliability of this protocol in the production of many hundreds of confirmed transgenic poppies.

  2. Chemiluminescence detection of opium poppy (Papaver somniferum) alkaloids.

    PubMed

    Francis, Paul S; Adcock, Jacqui L; Costin, Jason W; Purcell, Stuart D; Pfeffer, Frederick M; Barnett, Neil W

    2008-11-01

    A review with 98 references. The determination of the opium poppy (Papaver somniferum) alkaloids and their semi-synthetic derivatives has important applications in industrial process monitoring, clinical analysis and forensic science. Liquid-phase chemiluminescence reagents such as tris(2,2'-bipyridyl)ruthenium(II) and acidic potassium permanganate exhibit remarkable sensitivity and complementary selectivity for many P. somniferum alkaloids, which has been exploited in the development of a range of analytical procedures using flow analysis, high-performance liquid chromatography, capillary electrophoresis and microfluidic instrumentation.

  3. Opium poppy mosaic virus, a new umbravirus isolated from Papaver somniferum in New Zealand.

    PubMed

    Tang, Joe; Lebas, Bénédicte; Liefting, Lia; Veerakone, Stella; Wei, Ting; Ward, Lisa

    2016-01-01

    A novel virus, tentatively named "opium poppy mosaic virus" (OPMV), was isolated from Papaver somniferum (opium poppy) with leaf mosaic and mottling symptoms in Auckland, New Zealand, in 2006. The virus was mechanically transmitted to herbaceous plants of several species, in which it induced local and/or systemic symptoms. No virus particles were observed by electron microscopy in the diseased P. somniferum or any of the symptomatic herbaceous plants. The complete genomic sequence of 4230 nucleotides contains four open reading frames (ORF) and is most closely related (59.3 %) to tobacco bushy top virus, a member of the genus Umbravirus. These data suggest that OPMV is a new umbravirus.

  4. Opium poppy mosaic virus, a new umbravirus isolated from Papaver somniferum in New Zealand.

    PubMed

    Tang, Joe; Lebas, Bénédicte; Liefting, Lia; Veerakone, Stella; Wei, Ting; Ward, Lisa

    2016-01-01

    A novel virus, tentatively named "opium poppy mosaic virus" (OPMV), was isolated from Papaver somniferum (opium poppy) with leaf mosaic and mottling symptoms in Auckland, New Zealand, in 2006. The virus was mechanically transmitted to herbaceous plants of several species, in which it induced local and/or systemic symptoms. No virus particles were observed by electron microscopy in the diseased P. somniferum or any of the symptomatic herbaceous plants. The complete genomic sequence of 4230 nucleotides contains four open reading frames (ORF) and is most closely related (59.3 %) to tobacco bushy top virus, a member of the genus Umbravirus. These data suggest that OPMV is a new umbravirus. PMID:26514844

  5. Head group specificity of phospholipase D isoenzymes from poppy seedlings (Papaver somniferum L.).

    PubMed

    Oblozinsky, M; Ulbrich-Hofmann, R; Bezakova, L

    2005-02-01

    The biocatalytical potential of two new phospholipase D (PLD) isoenzymes from poppy seedlings (Papaver somniferum L.), PLD-A and PLD-B, was examined by comparing their activities in phospholipid transformation. Both enzymes showed the same ratio in rates of hydrolysis [phosphatidylcholine (PC):phosphatidylglycerol (PG):phosphatidylserine:phosphatidylinositol = 1:0.5:0.3:0.1] and were inactive towards phosphatidylethanolamine (PE). PLD-A did not catalyze head group exchange whereas PLD-B showed a high transphosphatidylation potential in the conversion of PC into PG and PE. This enzyme also catalyzed the transesterification of octadecylphosphocholine into octadecylphosphoglycerol or octadecylphosphoethanolamine.

  6. Identification of conserved micro-RNAs and their target transcripts in opium poppy (Papaver somniferum L.).

    PubMed

    Unver, Turgay; Parmaksiz, Iskender; Dündar, Ekrem

    2010-07-01

    Micro-RNAs (miRNA) are regulatory non-coding class of small RNAs functioning in many organisms. Using computational approaches we have identified 20 conserved opium poppy (Papaver somniferum L.) miRNAs belonging to 16 miRNA families in Expressed Sequence Tags (EST) database. The existence of ESTs suggested that the miRNAs were expressed in P. somniferum. Lengths of mature miRNAs varied from 20 to 23 nucleotides located at the different positions of precursor RNAs. Uracil was found to be a dominant nucleotide in both poppy pre-miRNA sequences (31.28 +/- 7.06% of total nucleotide composition) and in the first position at the 5' end of the mature poppy miRNAs. We have applied quantitative real-time PCR (qRT-PCR) assays to compare and validate expression levels of selected P. somniferum miRNAs and their target transcripts. As a result, some of the predicted miRNAs and their target genes were found to be differentially expressed in P. somniferum leaf and root tissues. A meaningful correlation between three of the four analyzed pairs of miRNAs and their target transcript expression levels was detected. Additionally, using these predicted miRNAs as queries, 41 potential target mRNAs were found in National Center for Biotechnology Information (NCBI) protein-coding nucleotide (mRNA) database of all plant species. Some of the target mRNAs were found to be transcription factors regulating plant development, morphology, and flowering time. Other target mRNAs of identified conserved miRNAs involve in metabolic processes, signal transduction, and stress responses. This study reports the first identification of opium poppy miRNAs.

  7. Crystallization and preliminary X-ray diffraction analysis of salutaridine reductase from the opium poppy Papaver somniferum.

    PubMed

    Higashi, Yasuhiro; Smith, Thomas J; Jez, Joseph M; Kutchan, Toni M

    2010-02-01

    The opium poppy Papaver somniferum is the source of the narcotic analgesics morphine and codeine. Salutaridine reductase (SalR; EC 1.1.1.248) reduces the C-7 keto group of salutaridine to the C-7 (S)-hydroxyl group of salutaridinol in the biosynthetic pathway that leads to morphine in the opium poppy plant. P. somniferum SalR was overproduced in Escherichia coli and purified using cobalt-affinity and size-exclusion chromatography. Hexagonal crystals belonging to space group P6(4)22 or P6(2)22 were obtained using ammonium sulfate as precipitant and diffracted to a resolution of 1.9 A.

  8. Agrobacterium rhizogenes-mediated transformation of opium poppy, Papaver somniferum l., and California poppy, Eschscholzia californica cham., root cultures.

    PubMed

    Park, S U; Facchini, P J

    2000-06-01

    An efficient protocol for the establishment of transgenic opium poppy (Papaver somniferum L.) and California poppy (Eschscholzia californica Cham.) root cultures using A. grobacterium rhizogenes is reported. Five strains of A. rhizogenes were tested for their ability to produce hairy roots on wounded opium poppy seedlings and California poppy embryogenic calli. Three of the strains induced hairy root formation on both species, whereas two others either caused the growth of tumorigenic calli or produced no response. To characterize the putative transgenic roots further, explant tissues were co-cultivated with the most effective A: rhizogenes strain (R1000) carrying the pBI121 binary vector. Except for the co-cultivation medium, all formulations included 50 mg l(-1) paromomycin to select for transformants and 200 mg l(-1) timentin to eliminate the Agrobacterium. Four weeks after infection, paromomycin-resistant roots appeared on 92-98% of explants maintained on hormone-free medium. Isolated hairy roots were propagated in liquid medium containing 1.0 mg l(-1) indole-3-acetic acid to promote rapid growth. Detection of the neomycin phosphotransferase gene, high levels of beta-glucuronidase (GUS) transcripts and enzyme activity, and GUS histochemical localization confirmed the integrative transformation of root cultures. Transgenic roots grew faster than wild-type roots, and California poppy roots grew more rapidly than those of opium poppy. With the exception of a less compact arrangement of epidermal cells and more root hairs, transformed roots of both species displayed anatomical features and benzylisoquinoline alkaloid profiles that were virtually identical to those of wild-type roots. Transgenic root cultures of opium poppy and California poppy are a simple, reliable and well-defined model system to investigate the molecular and metabolic regulation of benzylisoquinoline alkaloid biosynthesis, and to evaluate the genetic engineering potential of these important

  9. Disentangling Peronospora on Papaver: phylogenetics, taxonomy, nomenclature and host range of downy mildew of opium poppy (Papaver somniferum) and related species.

    PubMed

    Voglmayr, Hermann; Montes-Borrego, Miguel; Landa, Blanca B

    2014-01-01

    Based on sequence data from ITS rDNA, cox1 and cox2, six Peronospora species are recognised as phylogenetically distinct on various Papaver species. The host ranges of the four already described species P. arborescens, P. argemones, P. cristata and P. meconopsidis are clarified. Based on sequence data and morphology, two new species, P. apula and P. somniferi, are described from Papaver apulum and P. somniferum, respectively. The second Peronospora species parasitizing Papaver somniferum, that was only recently recorded as Peronospora cristata from Tasmania, is shown to represent a distinct taxon, P. meconopsidis, originally described from Meconopsis cambrica. It is shown that P. meconopsidis on Papaver somniferum is also present and widespread in Europe and Asia, but has been overlooked due to confusion with P. somniferi and due to less prominent, localized disease symptoms. Oospores are reported for the first time for P. meconopsidis from Asian collections on Papaver somniferum. Morphological descriptions, illustrations and a key are provided for all described Peronospora species on Papaver. cox1 and cox2 sequence data are confirmed as equally good barcoding loci for reliable Peronospora species identification, whereas ITS rDNA does sometimes not resolve species boundaries. Molecular phylogenetic data reveal high host specificity of Peronospora on Papaver, which has the important phytopathological implication that wild Papaver spp. cannot play any role as primary inoculum source for downy mildew epidemics in cultivated opium poppy crops.

  10. Disentangling Peronospora on Papaver: Phylogenetics, Taxonomy, Nomenclature and Host Range of Downy Mildew of Opium Poppy (Papaver somniferum) and Related Species

    PubMed Central

    Voglmayr, Hermann; Montes-Borrego, Miguel; Landa, Blanca B.

    2014-01-01

    Based on sequence data from ITS rDNA, cox1 and cox2, six Peronospora species are recognised as phylogenetically distinct on various Papaver species. The host ranges of the four already described species P. arborescens, P. argemones, P. cristata and P. meconopsidis are clarified. Based on sequence data and morphology, two new species, P. apula and P. somniferi, are described from Papaver apulum and P. somniferum, respectively. The second Peronospora species parasitizing Papaver somniferum, that was only recently recorded as Peronospora cristata from Tasmania, is shown to represent a distinct taxon, P. meconopsidis, originally described from Meconopsis cambrica. It is shown that P. meconopsidis on Papaver somniferum is also present and widespread in Europe and Asia, but has been overlooked due to confusion with P. somniferi and due to less prominent, localized disease symptoms. Oospores are reported for the first time for P. meconopsidis from Asian collections on Papaver somniferum. Morphological descriptions, illustrations and a key are provided for all described Peronospora species on Papaver. cox1 and cox2 sequence data are confirmed as equally good barcoding loci for reliable Peronospora species identification, whereas ITS rDNA does sometimes not resolve species boundaries. Molecular phylogenetic data reveal high host specificity of Peronospora on Papaver, which has the important phytopathological implication that wild Papaver spp. cannot play any role as primary inoculum source for downy mildew epidemics in cultivated opium poppy crops. PMID:24806292

  11. Genetic analysis of capsule and its associated economic traits in opium poppy (Papaver somniferum L.).

    PubMed

    Kumar, Birendra; Patra, Nirmal Kumar

    2010-01-01

    Four single crosses (VG20 x SGE48, SGE48 x SG35II, VG26 x SG35II, and SG35II x VG20) in opium poppy (Papaver somniferum L.) were analyzed to study the gene actions involved in the inheritance of quantitative traits, namely plant height, branches/plant, capsules/plant, peduncle length, capsule index, stigmatic rays, straw yield/plant, and morphine content. Simple additive, dominance, and epistatic genetic components were found to be significant for inheritance pattern. Dominance effect (h) was higher than additive effect (d). Digenic interaction indicated the prevalence of dominance x dominance (l) followed by additive x dominance (j) type epistasis. The significance of dominance (h) and dominance x dominance (l) indicated duplicate epistasis for all the traits and crosses except SG35II x VG20 for stigmatic rays. Biparental mating followed by recurrent selection involving desired recombinants may be utilized to improve the component traits.

  12. Expression and activity of cell-wall-degrading enzymes in the latex of opium poppy, Papaver somniferum L.

    PubMed

    Pilatzke-Wunderlich, I; Nessler, C L

    2001-03-01

    The alkaloid-rich latex of the opium poppy, Papaver somniferum L., is valued as a source of pharmaceuticals including thebaine, codeine, and morphine, but is also harvested for heroin production. The poppy laticifer system develops through the gradual disappearance of the common walls between differentiating laticifer elements throughout the plant. Gene homologues for cell-wall-degrading enzymes were found during random sequencing of an opium poppy latex cDNA library. RNA gel blot analysis of cellulase, polygalacturonase beta-subunit, 1,3-beta-glucanase, and xyloglucan endotransglycosylase homologues showed their expression was not limited to laticifers. In contrast, poppy gene homologues to pectin methylesterase (PME), pectin acetylesterase (PAE) and pectate lyase (PL) where all highly expressed and latex-specific. Enzyme assays confirmed the presence of PME, PAE, and PL activities in latex serum. The abundance of transcripts encoding pectin-degrading enzymes in latex suggests that these enzymes may play an important role in laticifer development.

  13. Atomic structure of salutaridine reductase from the opium poppy (Papaver somniferum).

    PubMed

    Higashi, Yasuhiro; Kutchan, Toni M; Smith, Thomas J

    2011-02-25

    The opium poppy (Papaver somniferum L.) is one of the oldest known medicinal plants. In the biosynthetic pathway for morphine and codeine, salutaridine is reduced to salutaridinol by salutaridine reductase (SalR; EC 1.1.1.248) using NADPH as coenzyme. Here, we report the atomic structure of SalR to a resolution of ∼1.9 Å in the presence of NADPH. The core structure is highly homologous to other members of the short chain dehydrogenase/reductase family. The major difference is that the nicotinamide moiety and the substrate-binding pocket are covered by a loop (residues 265-279), on top of which lies a large "flap"-like domain (residues 105-140). This configuration appears to be a combination of the two common structural themes found in other members of the short chain dehydrogenase/reductase family. Previous modeling studies suggested that substrate inhibition is due to mutually exclusive productive and nonproductive modes of substrate binding in the active site. This model was tested via site-directed mutagenesis, and a number of these mutations abrogated substrate inhibition. However, the atomic structure of SalR shows that these mutated residues are instead distributed over a wide area of the enzyme, and many are not in the active site. To explain how residues distal to the active site might affect catalysis, a model is presented whereby SalR may undergo significant conformational changes during catalytic turnover.

  14. Atomic Structure of Salutaridine Reductase from the Opium Poppy (Papaver somniferum)

    SciTech Connect

    Higashi, Yasuhiro; Kutchan, Toni M.; Smith, Thomas J.

    2011-11-18

    The opium poppy (Papaver somniferum L.) is one of the oldest known medicinal plants. In the biosynthetic pathway for morphine and codeine, salutaridine is reduced to salutaridinol by salutaridine reductase (SalR; EC 1.1.1.248) using NADPH as coenzyme. Here, we report the atomic structure of SalR to a resolution of {approx}1.9 {angstrom} in the presence of NADPH. The core structure is highly homologous to other members of the short chain dehydrogenase/reductase family. The major difference is that the nicotinamide moiety and the substrate-binding pocket are covered by a loop (residues 265-279), on top of which lies a large 'flap'-like domain (residues 105-140). This configuration appears to be a combination of the two common structural themes found in other members of the short chain dehydrogenase/reductase family. Previous modeling studies suggested that substrate inhibition is due to mutually exclusive productive and nonproductive modes of substrate binding in the active site. This model was tested via site-directed mutagenesis, and a number of these mutations abrogated substrate inhibition. However, the atomic structure of SalR shows that these mutated residues are instead distributed over a wide area of the enzyme, and many are not in the active site. To explain how residues distal to the active site might affect catalysis, a model is presented whereby SalR may undergo significant conformational changes during catalytic turnover.

  15. Molecular characterization of the salutaridinol 7-O-acetyltransferase involved in morphine biosynthesis in opium poppy Papaver somniferum.

    PubMed

    Grothe, T; Lenz, R; Kutchan, T M

    2001-08-17

    Salutaridinol 7-O-acetyltransferase (EC ) catalyzes the conversion of the phenanthrene alkaloid salutaridinol to salutaridinol-7-O-acetate, the immediate precursor of thebaine along the morphine biosynthetic pathway. We have isolated a cDNA clone that corresponds to the internal amino acid sequences of the native enzyme purified from a cell suspension culture of opium poppy Papaver somniferum. The recombinant enzyme acetylated the 7-hydroxyl moiety of salutaridinol in the presence of acetyl-CoA. The apparent K(m) value for salutaridinol was determined to be 9 microm and 54 microm for acetyl-CoA. The gene transcript was detected in extracts from Papaver orientale and Papaver bracteatum in addition to P. somniferum. Genomic DNA gel blot analysis indicated that there is likely a single copy of this gene in the P. somniferum genome. The amino acid sequence of salutaridinol 7-O-acetyltransferase is most similar (37% identity) to that of deacetylvindoline acetyltransferase of Catharanthus roseus. Salutaridinol 7-O-acetyltransferase is the second enzyme specific to morphine biosynthesis for which we have isolated a cDNA. Taken together with the other cDNAs cloned encoding norcoclaurine 6-O-methyltransferase, (S)-N-methylcoclaurine 3'-hydroxylase, the cytochrome P-450 reductase, and codeinone reductase, significant progress has been made toward accumulating genes of this pathway to enable the end goal of a biotechnological production of morphinan alkaloids.

  16. [Biosynthesis of poppy isoquinoline alkaloids in nature and in vitro culture. 1. Opium poppy (Papaver somniferum L.)].

    PubMed

    Kunakh, V A; Katsan, V A

    2003-01-01

    The analysis of literature data on production of isoquinoline alkaloids by Papaver somniferum L. plants and cell cultures has been made. The relationship of morphinane alkaloids biosynthesis with the processes of tissue and cell differentiation are discussed. The information on enzymes and pathways of regulation of morphine and sanguinarine biosynthesis are presented. The data on sanguinarine and morphine physiological role are analyzed.

  17. Two uncommon phospholipase D isoenzymes from poppy seedlings (Papaver somniferum L.).

    PubMed

    Oblozinsky, Marek; Schoeps, Regina; Ulbrich-Hofmann, Renate; Bezakova, Lydia

    2003-03-17

    Phospholipase D (PLD) has been detected in seedlings of Papaver somniferum L. cv. Lazúr (Papaveraceae). Purification of the enzyme revealed the existence of two forms of PLD (named as PLD-A and PLD-B). The two enzymes strongly differ in their catalytic properties. The pH optima were found at pH 8.0 for PLD-A and at pH 5.5 for PLD-B. While both enzymes show hydrolytic activity toward phosphatidylcholine (PC) and phosphatidyl-p-nitrophenol (PpNP), PLD-B only was able to catalyze the exchange of choline in PC by glycerol. Both enzymes were activated by Ca(2+) ions with an optimum concentration of 10 mM. In contrast to PLDs from other plants, PLD-B was still more activated by Zn(2+) ions with an optimum concentration of 5 mM. The apparent molecular masses of PLD-A and PLD-B, derived from sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), were estimated to be 116.4 and 114.1 kDa. N-terminal protein sequencing indicated N-terminal blockage in both cases. The isoelectric points were found to be 8.7 for PLD-A and 6.7 for PLD-B. Both enzymes were shown to be N-linked glycoproteins. This paper is the first report on PLD in poppy and indicates some important differences of the two enzyme forms to other PLDs known so far.

  18. Investigation of abiogenic stress-induced alterations in the level of secondary metabolites in poppy plants (Papaver somniferum L.).

    PubMed

    Szabó, Beáta; Lakatos, A; Koszegi, T; Botz, L

    2008-12-01

    We aimed to understand the effects of water stress on the alkaloid production in various developmental stages of poppy plants and the effect of stress on the alkaloids content in the capsules. Three stages of the life cycle of Papaver somniferum L. were selected in our studies: Rosette, Flowering and Lancing developmental stages. Four types of water conditions were examined: Control, Withdrawal of Water, 50% Water Supply and Inundation. The morphological monitoring, results of Relative Water Content and proline content were used as indicators of stress. The result of the measurements in poppy leaves show that the secondary metabolites dramatically respond to these stress conditions. The constant water supply was beneficial for the accumulation of alkaloids in the capsules.

  19. The roles of latex and the vascular bundle in morphine biosynthesis in the opium poppy, Papaver somniferum.

    PubMed

    Weid, Marion; Ziegler, Jörg; Kutchan, Toni M

    2004-09-21

    The opium poppy, Papaver somniferum, is one of mankind's oldest medicinal plants. Opium poppy today is the commercial source of the narcotic analgesics morphine and codeine. Along with these two morphinans, opium poppy produces approximately eighty alkaloids belonging to various tetrahydrobenzylisoquinoline-derived classes. It has been known for over a century that morphinan alkaloids accumulate in the latex of opium poppy. With identification of many of the enzymes of alkaloid biosynthesis in this plant, biochemical data suggested involvement of multiple cell types in alkaloid biosynthesis in poppy. Herein the immunolocalization of five enzymes of alkaloid formation in opium poppy is reported: (R,S)-3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase central to the biosynthesis of tetrahydroisoquinoline-derived alkaloids, the berberine bridge enzyme of the sanguinarine pathway, (R,S)-reticuline 7-O-methyltransferase specific to laudanosine formation, and salutaridinol 7-O-acetyltransferase and codeinone reductase, which lead to morphine. In capsule and stem, both O-methyltransferases and the O-acetyltransferase are found predominantly in parenchyma cells within the vascular bundle, and codeinone reductase is localized to laticifers, the site of morphinan alkaloid accumulation. In developing root tip, both O-methyltransferases and the O-acetyltransferase are found in the pericycle of the stele, and the berberine bridge enzyme is localized to parenchyma cells of the root cortex. Laticifers are not found in developing root tip, and, likewise, codeinone reductase was not detected. These results provide cell-specific localization that gives a coherent picture of the spatial distribution of alkaloid biosynthesis in opium poppy.

  20. Analysis of volatile compounds and triglycerides of seed oils extracted from different poppy varieties (Papaver somniferum L.).

    PubMed

    Krist, Sabine; Stuebiger, Gerald; Unterweger, Heidrun; Bandion, Franz; Buchbauer, Gerhard

    2005-10-19

    Poppy seed oil (Oleum Papaveris Seminis) is used for culinary and pharmaceutical purposes, as well as for making soaps, paints, and varnishes. Astonishingly, hardly anything was yet known about the volatile compounds of this promising comestible. Likewise, there are no current published data about the triglyceride (TAG) composition of poppy seed oils available. In this investigation solid-phase microextraction (SPME) with DVB/Carboxen/PDMS Stable-Flex fiber was applied to the study of volatile compounds of several seed oil samples from Papaver somniferum L. (Papaveraceae). 1-Pentanol (3.3-4.9%), 1-hexanal (10.9-30.9%), 1-hexanol (5.3-33.7%), 2-pentylfuran (7.2-10.0%), and caproic acid (2.9-11.5%) could be identified as the main volatile compounds in all examined poppy seed oil samples. Furthermore, the TAG composition of these oils was analyzed by MALDI-ReTOF- and ESI-IT-MS/MS. The predominant TAG components were found to be composed of linoleic, oleic, and palmitic acid, comprising approximately 70% of the oils. TAG patterns of the different poppy varieties were found to be very homogeneous, showing also no significant differences in terms of the applied pressing method of the plant seeds.

  1. Characterization of proteins in latex of the opium poppy (Papaver somniferum) using two-dimensional gel electrophoresis and microsequencing.

    PubMed

    Decker, G; Wanner, G; Zenk, M H; Lottspeich, F

    2000-10-01

    The opium poppy (Papaver somniferum) belongs to the group of latex-containing plants. Latex is the milky-like fluid within laticifer cells. In this study, poppy latex was analyzed with respect to ultrastructure, alkaloid, and protein content. The main goal of this project was the examination of the proteins by two-dimensional gel electrophoresis. In a proteomics approach, we investigated two main fractions of the latex, namely the cytosolic serum and the sedimented fraction containing the alkaloid-accumulating vesicles. Of the serum, representing the protein-rich part of the latex, 75 spots were analyzed by internal peptide microsequencing, followed by a database searching. For 69 proteins a function could be assigned due to homology to known proteins, whereas six spots could not be identified. Furthermore, codeinone reductase, a representative of the specific enzyme system in morphine biosynthesis, could be detected within the cytosolic serum fraction. In the vesicle-containing pellet, 23 protein spots were analyzed. An attempt was also made to separate the vesicle pellet by density centrifugation, followed by investigation of the alkaloid content, ultrastructure, and protein pattern. This study describes the first database of soluble proteins present in the latex of P. somniferum

  2. Molecular cloning and functional expression of codeinone reductase: the penultimate enzyme in morphine biosynthesis in the opium poppy Papaver somniferum.

    PubMed

    Unterlinner, B; Lenz, R; Kutchan, T M

    1999-06-01

    The narcotic analgesic morphine is the major alkaloid of the opium poppy Papaver somniferum. Its biosynthetic precursor codeine is currently the most widely used and effective antitussive agent. Along the morphine biosynthetic pathway in opium poppy, codeinone reductase catalyzes the NADPH-dependent reduction of codeinone to codeine. In this study, we have isolated and characterized four cDNAs encoding codeinone reductase isoforms and have functionally expressed them in Escherichia coli. Heterologously expressed codeinone reductase-calmodulin-binding peptide fusion protein was purified from E. coli using calmodulin affinity column chromatography in a yield of 10 mg enzyme l-1. These four isoforms demonstrated very similar physical properties and substrate specificity. As least six alleles appear to be present in the poppy genome. A comparison of the translations of the nucleotide sequences indicate that the codeinone reductase isoforms are 53% identical to 6'-deoxychalcone synthase from soybean suggesting an evolutionary although not a functional link between enzymes of phenylpropanoid and alkaloid biosynthesis. By sequence comparison, both codeinone reductase and 6'-deoxy- chalcone synthase belong to the aldo/keto reductase family, a group of structurally and functionally related NADPH-dependent oxidoreductases, and thereby possibly arise from primary metabolism.

  3. Exploiting expressed sequence tag databases for the development and characterization of gene-derived simple sequence repeat markers in the opium poppy (Papaver somniferum L.) for forensic applications.

    PubMed

    Lee, Eun Jung; Jin, Gang Nam; Lee, Kyung Lyong; Han, Myun Soo; Lee, Yang Han; Yang, Moon Sik

    2011-09-01

    Simple sequence repeat (SSR) markers in the opium poppy (Papaver somniferum L.) were identified from an expressed sequence tag (EST) database comprised of 20,340 sequences. In total, 2780 SSR-containing sequences were identified. The most frequent microsatellite had an AT/TA motif (37%). Twenty-two opium poppy EST-SSR markers were presently developed and polymorphisms of six markers (psom 2, 4, 12, 13, 17, and 22) were utilized in 135 individuals under narcotic control investigation. An average of three alleles per locus (range: 2-5 alleles) with a mean heterozygosity of 0.167 was detected. Six loci identified 29 unique profiles in 135 individuals. The EST-SSR markers exhibited small degrees of genetic differentiation (fixation index = 0.727, p < 0.001). Other variable markers will be needed to facilitate the forensic identification of the opium poppy for future cases. To determine the potential for cross-species amplification, six markers were tested in five Papaver genera species and two Eschscholzia genera. The psom 4 and psom 17 primer pair was transferable. This is the first study to report SSR markers of the opium poppy.

  4. Genetic studies on collar rot resistance in opium poppy (Papaver somniferum L.).

    PubMed

    Trivedi, Mala; Dhawan, Om Prakash; Tiwari, Rajesh Kumar; Sattar, Abdul

    2005-01-01

    The collar rot disease has been reported recently and occurs at the 10-12-leaf stage of plants of opium poppy. Infected plants topple down and dry prematurely due to fast rotting at the collar region. The inoculum for this study was multiplied on the cornmeal-sand culture. Genetic ratios were calculated by the chi-square test. Inheritance studies on this disease show a monogenic pattern of segregation with the ratio of 3 : 1 at F2, 1 : 2 : 1 at F3 and 1 : 1 at the backcross. Such genetic ratios clearly indicate that a single recessive gene (rs-1) is responsible for disease resistance in opium poppy. The inference drawn on the basis of the present study will be a great help in the future breeding programme of opium poppy for collar rot resistance.

  5. Virus-induced gene silencing is an effective tool for assaying gene function in the basal eudicot species Papaver somniferum (opium poppy).

    PubMed

    Hileman, Lena C; Drea, Sinéad; Martino, Gemma; Litt, Amy; Irish, Vivian F

    2005-10-01

    Virus-induced gene silencing (VIGS) is an attractive method for assaying gene function in species that are resistant to conventional genetic approaches. However, VIGS has been shown to be effective in only a few, closely related plant species. Tobacco rattle virus (TRV), a bipartite RNA virus, has a wide host range and so in principle could serve as an efficient vector for VIGS in a diverse array of plant species. Here we show that a vector based on TRV sequences is effective at silencing the endogenous phytoene desaturase (PapsPDS) gene in Papaver somniferum (opium poppy). We show that this vector does not compromise the growth or reproduction of poppy and the plants did not display viral symptoms. The silencing of PapsPDS resulted in a significant reduction in PapsPDS mRNA and a concomitant photobleached phenotype. The ability to rapidly assay gene function in P. somniferum will be valuable in manipulation of the opiate pathway in this pharmaceutically important species. We suggest that our vacuum infiltration method used to deliver TRV-based vectors into poppy is a promising approach for expanding VIGS to diverse angiosperm species in which traditional delivery methods fail to induce VIGS. Furthermore, these studies demonstrate the utility of TRV-VIGS for probing gene function in a basal eudicot species that is phylogenetically distant from model plant species.

  6. Endophytic colonisation of opium poppy, Papaver somniferum, by an entomopathogenic Beauveria bassiana strain.

    PubMed

    Quesada-Moraga, E; Landa, B B; Muñoz-Ledesma, J; Jiménez-Diáz, R M; Santiago-Alvarez, C

    2006-05-01

    Beauveria bassiana strain EABb 04/01-Tip isolated from stem-borer larvae of Timaspis papaveris (Hymenoptera: Cynipidae), a serious pest of opium poppy in Spain, was shown to be able to become established endophytically in this pharmaceutical crop. Microbiological, molecular and light and electron microscopic methods were used to study fungal colonisation and to describe its mode of penetration. After inoculation with a foliar spray of conidia, microbiological methods showed 100% of plants examined 24, 48, 72 and 144 h after treatment to be colonised endophytically by the fungus, although the percentage of previously surface sterilised leaf pieces showing fungal growth was 100% at 24 and 48 h, and 80 and 75% at 72 and 144 h after treatment, respectively. The fungus was also observed in leaf pieces obtained from newly formed leaves, indicating that it could spread from treated leaves to leaves formed after fungal application. For molecular studies, a polymerase chain reaction (PCR) protocol was used to amplify the ITS1-5.8S-ITS2 regions of the rDNA of the plant and the fungus. This procedure allowed the detection of the fungus on the surface of the leaves and also endophytically, but only at 72 h after treatment. A nucleotide BLAST search revealed that the ITS1-5.8S-ITS2 sequence of strain EABb 04/01-Tip showed 100% homology with a similar sequence from Cordyceps bassiana. SEM images revealed that although numerous conidia were observed on the leaf surface, few germinated and penetrated. Intracellular colonisation by B. bassiana was not observed, but hyphae were detected growing into the xylem vessels. The fungus was found to colonise 40.5 +/- 4.3% of seedlings (with two cotyledons and the two first real leaves) from seeds dressed with a fungal spore suspension. These results may have implications in the biological control of T. papaveris, including the possible systemic protection of the plant against this cynipid.

  7. Isolation and identification of unique marker compounds from the Tasmanian poppy Papaver somniferum N. Implications for the identification of illicit heroin of Tasmanian origin.

    PubMed

    Odell, Luke R; Skopec, Jana; McCluskey, Adam

    2008-03-01

    Tasmanian opium accounts for 25% of the world's legal supply of opium straw, and in 1998-99 sufficient numbers of flower pods (66,013) to manufacture ca 500 kg of heroin were stolen. Whilst the heroin signature program has been developed to determine the origin of heroin from other key producers, no such signature currently exists for Tasmanian derived heroin. Tasmanian poppies contain a unique alkaloid, oripavine, which is the source of 'marker' impurities in illicit heroin produced from Tasmanian poppy straw. Treatment of oripavine (500mg) under Thiboumery and Mohr heroin processing conditions, followed by simple evaporative workup afforded 613 mg of a dark orange residue, which upon extensive chromatographic purification yielded oripavine 3-acetate (2) 22 mg; 3-acetyl-N-acetyldesthebaine (3) 35 mg; 3-acetyl-6-methoxy-4,5-epoxyphenanthrene (4) 5.8 mg; 3,4-diacetyl-6-methoxyphenanthrene (5) 27 mg; and 3,4,6-methoxy-5-[2(N-methylacetamido)]ethylphenanthrene (6) 52 mg. Compounds (2-6) are derived from oripavine and are unique to heroin derived from the Tasmanian poppy Papaver somniferum N. Analysis of illicit heroin samples seized from Turkey, Pakistan, Columbia and Myanmar did not reveal any of the aforementioned marker compounds. We have, however, identified four of these marker compounds (3-6) in seized heroin samples from Australia suggesting that they are of Tasmanian origin. Complete details of the isolation and identification of these compounds are provided.

  8. Invertase in immobilized cells of Papaver somniferum L.

    PubMed

    Stano, J; Nemec, P; Bezáková, L; Kovács, P; Kákoniova, D; Neubert, K; Lisková, D

    1997-03-01

    Papaver somniferum L., (opium poppy) cells were after permeabilization in Tween 80 immobilized by glutaraldehyde without any carrier. Cells immobilized by cross-linking performed the hydrolysis of sucrose. The immobilized cells were characterized by high invertase activity and appropriate physico-mechanical properties.

  9. Gene actions for yield and its attributes and their implications in the inheritance pattern over three generations in opium poppy (Papaver somniferum L.).

    PubMed

    Mishra, Brij K; Mishra, R; Jena, S N; Shukla, Sudhir

    2016-09-01

    The gene actions for yield and its attributes and their inheritance pattern based on five parameter model have been explored in four single crosses (NBIHT-5 × NBIHT-6, NBIHT-5 × NBMHT-1, NBMHT-1 × NBIHT-6 and NBMHT-2 × NBMHT-1) obtained using thebaine rich pure lines of opium poppy (Papaver somniferum L.) for three consecutive generations. All the traits showed nonallelic mode of interaction, however, dominance effect (h) was more pronounced for all the traits except thebaine and papaverine. The dominance × dominance (l) effects were predominant over additive × additive (i) for all traits in all the four crosses except for papaverine. The seed and opium yield, and its contributing traits inherited quantitatively. The fixable gene effects (d) and (i) were lower in magnitude than nonfixable (h) and (l) gene effects. The estimates of heterosis were also higher in comparison to the respective parents which suggested preponderance of dominance gene action for controlling most of the traits. The phenotypic coefficient of variation was marginally higher than those of genotypic coefficient of variation for all the traits. The traits thebaine, narcotine, morphine and opium yield had high heritability coupled with high genetic advance. The leaf number, branches per plant and stem diameter showed positive correlation with opium and seed yields. The selection of plants having large number of leaves, branches and capsules with bigger size would be advantageous to enhance the yield potential. PMID:27659342

  10. Gene actions for yield and its attributes and their implications in the inheritance pattern over three generations in opium poppy (Papaver somniferum L.).

    PubMed

    Mishra, Brij K; Mishra, R; Jena, S N; Shukla, Sudhir

    2016-09-01

    The gene actions for yield and its attributes and their inheritance pattern based on five parameter model have been explored in four single crosses (NBIHT-5 × NBIHT-6, NBIHT-5 × NBMHT-1, NBMHT-1 × NBIHT-6 and NBMHT-2 × NBMHT-1) obtained using thebaine rich pure lines of opium poppy (Papaver somniferum L.) for three consecutive generations. All the traits showed nonallelic mode of interaction, however, dominance effect (h) was more pronounced for all the traits except thebaine and papaverine. The dominance × dominance (l) effects were predominant over additive × additive (i) for all traits in all the four crosses except for papaverine. The seed and opium yield, and its contributing traits inherited quantitatively. The fixable gene effects (d) and (i) were lower in magnitude than nonfixable (h) and (l) gene effects. The estimates of heterosis were also higher in comparison to the respective parents which suggested preponderance of dominance gene action for controlling most of the traits. The phenotypic coefficient of variation was marginally higher than those of genotypic coefficient of variation for all the traits. The traits thebaine, narcotine, morphine and opium yield had high heritability coupled with high genetic advance. The leaf number, branches per plant and stem diameter showed positive correlation with opium and seed yields. The selection of plants having large number of leaves, branches and capsules with bigger size would be advantageous to enhance the yield potential.

  11. Trace and major element levels in rats after oral administration of diesel and biodiesel derived from opium poppy (Papaver somniferum L.) seeds.

    PubMed

    Aksoy, Laçine; Sözbilir, Nalan Bayşu

    2015-10-01

    The study investigated the toxic effects of diesel and biodiesel derived from opium poppy (Papaver somniferum L.) oil seeds on the trace and major elements in kidney, lung, liver, and serum of rats. By the end of 21 days, trace and major element concentrations in kidney, lung, and liver tissues and the serum were measured using inductively coupled plasma-optical emission spectroscopy. We observed that trace and major element levels in kidney, lung, and liver tissues and the serum changed. Especially, important differences were detected in trace and major element concentrations in kidney and lung tissues. In kidney tissue, the concentration differences of calcium, sodium, and zinc (Zn) were found between diesel and biodiesel groups. In lung tissue, the concentration differences of cadmium, lithium, magnesium, manganese, and Zn were found between diesel and biodiesel groups. Among the significant findings, Zn concentration in serum and liver tissue of diesel and biodiesel were different from control (p < 0.05). However, the metal levels of biodiesel group were similar to control group. Due to lesser toxicity of biodiesel, it could be considered as an alternate fuel.

  12. Genetic parameters and correlations of collar rot resistance with important biochemical and yield traits in opium poppy (Papaver somniferum L.).

    PubMed

    Trivedi, Mala; Tiwari, Rajesh K; Dhawan, Om P

    2006-01-01

    Collar rot, caused by Rhizoctonia solani Kühn, is one of the most severe fungal diseases of opium poppy. In this study, heritability, genetic advance and correlation for 10 agronomic, 1 physiological, 3 biochemical and 1 chemical traits with disease severity index (DSI) for collar rot were assessed in 35 accessions of opium poppy. Most of the economically important characters, like seed and capsule straw yield per plant, oil and protein content of seeds, peroxidase activity in leaves, morphine content of capsule straw and DSI for collar rot showed high heritability as well as genetic advance. Highly significant negative correlation between DSI and seed yield clearly shows that as the disease progresses in plants, seed yield declines, chiefly due to premature death of infected plants as well as low seed and capsule setting in the survived population of susceptible plants. Similarly, a highly significant negative correlation between peroxidase activity and DSI indicated that marker-assisted selection of disease-resistant plants based on high peroxidase activity would be effective and survived susceptible plants could be removed from the population to stop further spread.

  13. Functional characterization of a novel benzylisoquinoline O-methyltransferase suggests its involvement in papaverine biosynthesis in opium poppy (Papaver somniferum L).

    PubMed

    Pienkny, Silke; Brandt, Wolfgang; Schmidt, Jürgen; Kramell, Robert; Ziegler, Jörg

    2009-10-01

    The benzylisoquinoline alkaloids are a highly diverse group of about 2500 compounds which accumulate in a species-specific manner. Despite the numerous compounds which could be identified, the biosynthetic pathways and the participating enzymes or cDNAs could be characterized only for a few selected members, whereas the biosynthesis of the majority of the compounds is still largely unknown. In an attempt to characterize additional biosynthetic steps at the molecular level, integration of alkaloid and transcript profiling across Papaver species was performed. This analysis showed high expression of an expressed sequence tag (EST) of unknown function only in Papaver somniferum varieties. After full-length cloning of the open reading frame and sequence analysis, this EST could be classified as a member of the class II type O-methyltransferase protein family. It was related to O-methyltransferases from benzylisoquinoline biosynthesis, and the amino acid sequence showed 68% identical residues to norcoclaurine 6-O-methyltransferase. However, rather than methylating norcoclaurine, the recombinant protein methylated norreticuline at position seven with a K(m) of 44 mum using S-adenosyl-l-methionine as a cofactor. Of all substrates tested, only norreticuline was converted. Even minor changes in the benzylisoquinoline backbone were not tolerated by the enzyme. Accordingly, the enzyme was named norreticuline 7-O-methyltransferase (N7OMT). This enzyme represents a novel O-methyltransferase in benzylisoquinoline metabolism. Expression analysis showed slightly increased expression of N7OMT in P. somniferum varieties containing papaverine, suggesting its involvement in the partially unknown biosynthesis of this pharmaceutically important compound.

  14. Herbicidal treatments for control of Papaver somniferum L.

    PubMed

    Horowitz, M

    1980-01-01

    Fifty-five commercially available herbicides were evaluated for possible use to destroy illicit opium poppy crops (Papaver somniferum). In the first stage, herbicides were sprayed on poppy plants grown in containers. The following compounds killed poppy plants: (a) herbicides with typical foliar activity--amitrole, bromoxynil, 2,4-D, glyphosate, ioxynil and paraquat; and (b) herbicides with root and foliar activity--the triazines ametryn, atrazine, metribuzin, prometryn, simazine and terbutryn; the substituted ureas benzthiazuron, chloroxuron, diuron, fluometuron, linuron, methabenzthiazuron, neburon and phenobenzuron; and the miscellaneous compounds karbutilate, methazole, oxadiazon and pyrazon. Severe but sublethal injury was caused by cycloate, EPTC, molinate, pobulate, cacodylate + MSMA, ethofumesate, perfluidone and phenmedipham. Abnormal development of vegetative or reproductive parts of the plant was induced by benefin, butralin, dinitramine, pendimethalin, trifluralin, diphenamid, napropamide, dalapon and propham. Efficient herbicides with negligible persistence in soil at the doses applied were evaluated on poppy plants in the field at various stages of growth. Small plants were severely injured by 2,4-D, killed rapidly by bromoxynil, ioxynil, paraquat (in mixture + diquat), and more slowly by glyphosate and metribuzin. The resistance to herbicides increased with the age of the poppy plant. Severe damage with partial kill of developed plants was obtained with bromoxynil, ioxynil, glyphosate, and paraquat + diquat; the last treatment produced the fastest effect.

  15. Herbicidal treatments for control of Papaver somniferum L.

    PubMed

    Horowitz, M

    1980-01-01

    Fifty-five commercially available herbicides were evaluated for possible use to destroy illicit opium poppy crops (Papaver somniferum). In the first stage, herbicides were sprayed on poppy plants grown in containers. The following compounds killed poppy plants: (a) herbicides with typical foliar activity--amitrole, bromoxynil, 2,4-D, glyphosate, ioxynil and paraquat; and (b) herbicides with root and foliar activity--the triazines ametryn, atrazine, metribuzin, prometryn, simazine and terbutryn; the substituted ureas benzthiazuron, chloroxuron, diuron, fluometuron, linuron, methabenzthiazuron, neburon and phenobenzuron; and the miscellaneous compounds karbutilate, methazole, oxadiazon and pyrazon. Severe but sublethal injury was caused by cycloate, EPTC, molinate, pobulate, cacodylate + MSMA, ethofumesate, perfluidone and phenmedipham. Abnormal development of vegetative or reproductive parts of the plant was induced by benefin, butralin, dinitramine, pendimethalin, trifluralin, diphenamid, napropamide, dalapon and propham. Efficient herbicides with negligible persistence in soil at the doses applied were evaluated on poppy plants in the field at various stages of growth. Small plants were severely injured by 2,4-D, killed rapidly by bromoxynil, ioxynil, paraquat (in mixture + diquat), and more slowly by glyphosate and metribuzin. The resistance to herbicides increased with the age of the poppy plant. Severe damage with partial kill of developed plants was obtained with bromoxynil, ioxynil, glyphosate, and paraquat + diquat; the last treatment produced the fastest effect. PMID:6905769

  16. (R,S)-Reticuline 7-O-methyltransferase and (R,S)-norcoclaurine 6-O-methyltransferase of Papaver somniferum - cDNA cloning and characterization of methyl transfer enzymes of alkaloid biosynthesis in opium poppy.

    PubMed

    Ounaroon, Anan; Decker, Gabriele; Schmidt, Jürgen; Lottspeich, Friedrich; Kutchan, Toni M

    2003-12-01

    S-Adenosyl-L-methionine:(R,S)-reticuline 7-O-methyltransferase converts reticuline to laudanine in tetrahydrobenzylisoquinoline biosynthesis in the opium poppy Papaver somniferum. This enzyme activity has not yet been detected in plants. A proteomic analysis of P. somniferum latex identified a gel spot that contained a protein(s) whose partial amino acid sequences were homologous to those of plant O-methyltransferases. cDNA was amplified from P. somniferum RNA by reverse transcription PCR using primers based on these internal amino acid sequences. Recombinant protein was then expressed in Spodoptera frugiperda Sf9 cells in a baculovirus expression vector. Steady-state kinetic measurements with one heterologously expressed enzyme and mass spectrometric analysis of the enzymatic products suggested that this unusual enzyme is capable of carrying through sequential O-methylations on the isoquinoline and on the benzyl moiety of several substrates. The tetrahydrobenzylisoquinolines (R)-reticuline (4.2 sec(-1) mm(-1)), (S)-reticuline (4.5 sec(-1) mm(-1)), (R)-protosinomenine (1.7 sec(-1) mm(-1)), and (R,S)-isoorientaline (1.4 sec(-1) mm(-1)) as well as guaiacol (5.9 sec(-1) mm(-1)) and isovanillic acid (1.2 sec(-1) mm(-1)) are O-methylated by the enzyme with the ratio kcat/K m shown in parentheses. A P. somniferum cDNA encoding (R,S)-norcoclaurine 6-O-methyltransferase was similarly isolated and characterized. This enzyme was less permissive, methylating only (R,S)-norcoclaurine (7.4 sec(-1) mm(-1)), (R)-norprotosinomenine (4.1 sec(-1) mm(-1)), (S)-norprotosinomenine (4.0 sec(-1) mm(-1)) and (R,S)-isoorientaline (1.0 sec(-1) mm(-1)). A phylogenetic comparison of the amino acid sequences of these O-methyltransferases to those from 28 other plant species suggests that these enzymes group more closely to isoquinoline biosynthetic O-methyltransferases from Coptis japonica than to those from Thalictrum tuberosum that can O-methylate both alkaloid and phenylpropanoid substrates.

  17. Transformation of opium poppy (Papaver somniferum L.) with antisense berberine bridge enzyme gene (anti-bbe) via somatic embryogenesis results in an altered ratio of alkaloids in latex but not in roots.

    PubMed

    Frick, Susanne; Chitty, Julie A; Kramell, Robert; Schmidt, Jürgen; Allen, Robert S; Larkin, Philip J; Kutchan, Toni M

    2004-12-01

    The berberine bridge enzyme cDNA bbe from Papaver somniferum L. was transformed in antisense orientation into seedling explants of the industrial elite line C048-6-14-64. In this way, 84 phenotypically normal To plants derived from embryogenic callus cultures were produced. The selfed progeny of these 84 plants yielded several T1 plants with an altered alkaloid profile. One of these plants T1-47, and its siblings T2-1.2 and T2-1.5 are the subject of the present work. The transformation of these plants was evaluated by PCR, and northern and Southern hybridisation. The transgenic plants contained one additional copy of the transgene. The alkaloid content in latex and roots was determined with HPLC and LC-MS. We observed an increased concentration of several pathway intermediates from all biosynthetic branches, e.g., reticuline, laudanine, laudanosine, dehydroreticuline, salutaridine and (S)-scoulerine. The transformation altered the ratio of morphinan and tetrahydrobenzylisoquinoline alkaloids in latex but not the benzophenanthridine alkaloids in roots. The altered alkaloid profile is heritable at least to the T2 generation. These results are the first example of metabolic engineering of the alkaloid pathways in opium poppy and, to our knowledge, the first time that an alkaloid biosynthetic gene has been transformed into the native species, followed by regeneration into a mature plant to enable analyses of the effect of the transgene on metabolism over several generations.

  18. Determination of alkaloids in capsules, milk and ethanolic extracts of poppy (Papaver somniferum L.) by ATR-FT-IR and FT-Raman spectroscopy.

    PubMed

    Schulz, Hartwig; Baranska, Malgorzata; Quilitzsch, Rolf; Schütze, Wolfgang

    2004-10-01

    Fourier transform (FT) infrared spectroscopy using a diamond composite ATR crystal and NIR-FT-Raman spectroscopy techniques were applied for the simultaneous identification and quantification of the most important alkaloids in poppy capsules. Most of the characteristic Raman signals of the alkaloids can be identified in poppy milk isolated from unripe capsules. But also poppy extracts present specific bands relating clearly to the alkaloid fraction. Raman spectra obtained by excitation with a Nd:YAG laser at 1064 nm show no disturbing fluorescence effects; therefore the plant tissue can be recorded without any special preparation. The used diamond ATR technique allows to measure very small sample amounts (5-10 microL or 2-5 mg) without the necessity to perform time-consuming pre-treatments. When applying cluster analysis a reliable discrimination of "low-alkaloid" and "high-alkaloid" poppy single-plants can be easily achieved. The examples presented in this study provide clear evidence of the benefits of Raman and ATR-IR spectroscopy in efficient quality control, forensic analysis and high-throughput evaluation of poppy breeding material.

  19. Detection of a true breeding homeotic gene mutant Pps-1 with partially petaloid sepals in opium poppy (Papaver somniferum L.) and its genetic behavior.

    PubMed

    Dhawan, Om Prakash; Dubey, Mukesh Kumar; Khanuja, Suman Preet Singh

    2007-01-01

    A spontaneous true breeding homeotic gene mutant Pps-1 with distinct partial petaloid sepals was detected in the population of downy mildew (DM)-resistant elite accession I-14 during our studies for the identification of disease resistance sources in opium poppy. The trait was found to be stable and inherited truly in the subsequent generations. Genetic studies were carried out through systematic reciprocal crosses with the parental wild-type genotype I-14, and segregation pattern of phenotypic characteristics in F(1) and F(2) populations clearly indicated single recessive nuclear gene control of the mutant character. The studies have demonstrated that the mutant phenotype is due to mutations at the Pps-1 locus that possibly corresponds to B-class function (according to ABC model) with negative control function. The mutant Pps-1 being single-whorl homeotic mutant might greatly help in providing insight into mechanisms of flower development in opium poppy.

  20. Isoquinoline alkaloid production by transformed cultures of Papaver somniferum.

    PubMed

    Yoshimatsu, K; Shimomura, K

    2001-01-01

    Three clones of transformed cultures of opium poppy (Papaver somniferum L.) were established by infection with Agrobacterium rhizogenes MAFF 03-01724. MAFF clone 1 being capable of forming somatic embryos was selected and its growth and isoquinoline alkaloid production was investigated. The illumination, temperature and nutrient medium composition greatly affected growth, cell morphology and alkaloid accumulation. The MAFF clone 1 cultured in Root Culture medium in the dark at 22 degrees C accumulated a high quantity of sanguinarine (652 micrograms/g dry weight) though the growth was poor (4.4 fold as fresh weight basis after 2 months of culture). The MAFF clone 1 cultured in a quarter macro salt strength Woody Plant medium under 14 h/day light at 22 degrees C developed into plantlets and accumulated significant quantity of codeine (648 micrograms/g dry wt) together with papaverine, noscapine, and sanguinarine. This clone was applied to a rotating drum fermenter (2 L working volume), and ca. 0.3 mg codeine and 0.06 mg sanguinarine were obtained after 4 weeks of culture. One quarter of the codeine produced was found in the culture medium.

  1. [Regulatory role of elements in the formation and accumulation of alkaloids in Papaver somniferum L. seedlings].

    PubMed

    Lovkova, M Ia; Buzuk, G N; Sokolova, S M

    2006-01-01

    The effects of 12 elements on the formation and accumulation of isoquinolines were studied in opium poppy seedlings (Papaver somniferum L.). Three types of concentration dependences of the effects of the elements under study were determined. The elements served as activators (Co, Mo, W, Cr, and Cu) or inhibitors of these processes (B, Fe, V, Mn, and Ca). The molecular mechanisms of action of these regulators are discussed. The optimum concentrations of Co and Mo were tested under combined treatment conditions with these elements.

  2. Comparative macroarray analysis of morphine containing Papaver somniferum and eight morphine free Papaver species identifies an O-methyltransferase involved in benzylisoquinoline biosynthesis.

    PubMed

    Ziegler, Jörg; Diaz-Chávez, María Luisa; Kramell, Robert; Ammer, Christian; Kutchan, Toni M

    2005-10-01

    Benzylisoquinoline alkaloids constitute a group of about 2,500 structures and are mainly produced by plants of the order Ranunculales. But only the opium poppy, Papaver somniferum, and Papaver setigerum are able to produce morphine. In this study, we started to investigate by gene expression analysis the molecular basis for this exceptional biosynthetic ability. A sequencing project from P. somniferum seedlings was initiated using a method based on the amplified fragment length polymorphism technique that resulted in 849 UniGenes. These cDNAs were analysed on macroarrays for differential expression between morphine-containing P. somniferum plants and eight other Papaver species, which accumulate other benzylisoquinolines instead of morphine. Three cDNAs showing increased expression in P. somniferum compared to all the other Papaver species were identified. Whereas two showed no significant homology to any known protein, one putatively encoded an O-methyltransferase. Analysis of substrate specificity of the heterologously expressed protein and mass spectrometric identification of the enzymatic products identified this protein as S-adenosyl-L-methionine:(R,S)-3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase (EC 2.1.1.116). Unlike other O-methyltransferases of different positional specificities implicated in benzylisoquinoline metabolism, the enzyme only accepted tetrahydroxylated tetrahydrobenzylisoquinolines as substrates; methylation was tolerated only at the 6-hydroxy position.

  3. Dendryphion penicillatum and Pleospora papaveracea, Destructive Seedborne Pathogens and Potential Mycoherbicides for Papaver somniferum.

    PubMed

    O'Neill, N R; Jennings, J C; Bailey, B A; Farr, D F

    2000-07-01

    ABSTRACT Dendryphion penicillatum and Pleospora papaveracea were isolated from blighted Papaver somniferum and Papaver bracteatum plants grown in growth chambers and the field in Beltsville, MD. The etiology of the diseases was determined, and the fungi are being investigated as potential mycoherbicides to control the narcotic opium poppy plant. P. papaveracea is known to be a highly destructive seedborne pathogen of Papaver somniferum, causing seedling blight, leaf blight, crown rot, and capsule rot. Single conidia and ascospores were isolated and cultures established from naturally infested seed and diseased foliage and pods of opium poppy from Iran, Colombia, Venezuela, Sweden, India, and the United States (Maryland and Washington). Mycelia and conidia of P. papaveracea and D. penicillatum produced on necrotic leaf tissues appear morphologically similar, and the fungi were previously considered to be anamorph and teleomorph. However, no anamorph/teleomorph connection could be established, and the fungi appear to be distinct taxa. P. papaveracea produced conidia, mature pseudothecia, and chlamydospores in vitro and on infected stems. D. penicillatum produced conidia, microsclerotia, and macronematous conidiophores. Although both fungi were pathogenic to three poppy cultivars, conidial inoculum from P. papaveracea cultures was more virulent than conidial inoculum from D. penicillatum. Eight-week-old plants became necrotic and died 8 days after inoculation with a conidial suspension of P. papaveracea at 2 x 10(5) spores per ml. Disease severity was significantly enhanced by inoculum formulations that contained corn oil, by higher conidial inoculum concentrations, and by increased wetness periods. Symptoms on plants inoculated with either pathogen included leaf and stem necrosis, stem girdling, stunting, necrotic leaf spots, and foliar and pod blight. Inoculated seedlings exhibited wire stem, damping-off, and root rot. Conidia, and less frequently pseudothecia, of P

  4. Involvement of lipoxygenase in elicitor-stimulated sanguinarine accumulation in Papaver somniferum suspension cultures.

    PubMed

    Holková, Ivana; Bezáková, Lýdia; Bilka, František; Balažová, Andrea; Vanko, Marián; Blanáriková, Vítazoslava

    2010-01-01

    The involvement of lipoxygenase (LOX, EC 1.13.11.12) in elicitor-induced opium poppy defense response was investigated. Papaver somniferum L. suspension cultures were treated with abiotic elicitor methyl jasmonate (MJ), fungal elicitor (Botrytis cinerea homogenate) and phenidone (specific inhibitor of LOX) to determine the involvement of this enzyme in production of sanguinarine, the major secondary metabolite of opium poppy cultures. P. somniferum suspension cultures responded to elicitor treatment with strong and transient increase of LOX activity followed by sanguinarine accumulation. LOX activity increased in elicited cultures, reaching 9.8 times of the initial value at 10 h after MJ application and 2.9 times after B. cinerea application. Sanguinarine accumulated to maximal levels of 169.5 ± 12.5 μg g⁻¹ dry cell weight in MJ-elicited cultures and 288.0 ± 10.0 μg g⁻¹ dry cell weight in B. cinerea-elicited cultures. The treatment of cells with phenidone before elicitor addition, significantly reduced sanguinarine production. The relative molecular weight of P. somniferum LOX (83 kDa) was estimated by using immunobloting and its pH optimum was shown to be pH 6.5.

  5. New dimeric morphine from opium poppy (Papaver somuniferum) and its physiological function.

    PubMed

    Morimoto, Satoshi; Suemori, Kazunari; Taura, Futoshi; Shoyama, Yukihiro

    2003-07-01

    Together with bismorphine A (1), a new compound bismorphine B (2) was identified in the wounded capsules of Papaver somniferum. On the basis of analyses of the various spectral data, bismorphine B (2) was determined as a novel morphinan alkaloid, in which two morphine units are coupled through a biphenyl ether bond. When the physiological function of bismorphine B (2) in opium poppy was investigated, it was evident that this alkaloid more effectively cross-links cell wall polysaccharide pectins than bismorphine A (1) and that such cross-linking reaction leads to resistance against hydrolysis by pectinase.

  6. Gas chromatographic/mass spectrometric detection of narcotine, papaverine, and thebaine in seeds of Papaver somniferum.

    PubMed

    Paul, B D; Dreka, C; Knight, E S; Smith, M L

    1996-12-01

    In addition to codeine and morphine, three more compounds: narcotine (noscapine), papaverine, and thebaine were found in Indian and Netherlands poppy seeds (Papaver somniferum L). The compounds were detected by a GC/MS technique and the identities were confirmed by comparing retention times and ion ratios with the known references. The concentrations of codeine, morphine, thebaine, papaverine, and narcotine were 44, 167, 41, 67, and 230 micrograms/g in Indian poppy seeds, and were 1.8, 39, 1.0, 0.17, 0.84 micrograms/g in Netherlands poppy seeds, respectively. Because these compounds may be urinary products after poppy seed consumption, the lowest detectable concentrations of codeine, morphine, thebaine, papaverine, and narcotine in urine are of interest and were found to be 4, 4, 5, 0.4, and 4 ng/ml, respectively. The detection of urinary narcotine, papaverine, or thebaine may be utilized to differentiate poppy seed consumption from illicit codeine, morphine, or heroin use.

  7. Comparative analysis of transcription factor gene families from Papaver somniferum: identification of regulatory factors involved in benzylisoquinoline alkaloid biosynthesis.

    PubMed

    Agarwal, Parul; Pathak, Sumya; Lakhwani, Deepika; Gupta, Parul; Asif, Mehar Hasan; Trivedi, Prabodh Kumar

    2016-05-01

    Opium poppy (Papaver somniferum L.), known for biosynthesis of several therapeutically important benzylisoquinoline alkaloids (BIAs), has emerged as the premier organism to study plant alkaloid metabolism. The most prominent molecules produced in opium poppy include narcotic analgesic morphine, the cough suppressant codeine, the muscle relaxant papaverine and the anti-microbial agent sanguinarine and berberine. Despite several health benefits, biosynthesis of some of these molecules is very low due to tight temporal and spatial regulation of the genes committed to their biosynthesis. Transcription factors, one of the prime regulators of secondary plant product biosynthesis, might be involved in controlled biosynthesis of BIAs in P. somniferum. In this study, identification of members of different transcription factor gene families using transcriptome datasets of 10 cultivars of P. somniferum with distinct chemoprofile has been carried out. Analysis suggests that most represented transcription factor gene family in all the poppy cultivars is WRKY. Comparative transcriptome analysis revealed differential expression pattern of the members of a set of transcription factor gene families among 10 cultivars. Through analysis, two members of WRKY and one member of C3H gene family were identified as potential candidates which might regulate thebaine and papaverine biosynthesis, respectively, in poppy.

  8. Comparative analysis of transcription factor gene families from Papaver somniferum: identification of regulatory factors involved in benzylisoquinoline alkaloid biosynthesis.

    PubMed

    Agarwal, Parul; Pathak, Sumya; Lakhwani, Deepika; Gupta, Parul; Asif, Mehar Hasan; Trivedi, Prabodh Kumar

    2016-05-01

    Opium poppy (Papaver somniferum L.), known for biosynthesis of several therapeutically important benzylisoquinoline alkaloids (BIAs), has emerged as the premier organism to study plant alkaloid metabolism. The most prominent molecules produced in opium poppy include narcotic analgesic morphine, the cough suppressant codeine, the muscle relaxant papaverine and the anti-microbial agent sanguinarine and berberine. Despite several health benefits, biosynthesis of some of these molecules is very low due to tight temporal and spatial regulation of the genes committed to their biosynthesis. Transcription factors, one of the prime regulators of secondary plant product biosynthesis, might be involved in controlled biosynthesis of BIAs in P. somniferum. In this study, identification of members of different transcription factor gene families using transcriptome datasets of 10 cultivars of P. somniferum with distinct chemoprofile has been carried out. Analysis suggests that most represented transcription factor gene family in all the poppy cultivars is WRKY. Comparative transcriptome analysis revealed differential expression pattern of the members of a set of transcription factor gene families among 10 cultivars. Through analysis, two members of WRKY and one member of C3H gene family were identified as potential candidates which might regulate thebaine and papaverine biosynthesis, respectively, in poppy. PMID:26108744

  9. [Effect of a fungal elicitor on levels of sanguinarine and polyphenoloxidase activity in a suspension culture of Papaver somniferum L].

    PubMed

    Balazová, A; Bilka, F; Blanáriková, V; Psenák, M

    2002-07-01

    The opium poppy (Papaver somniferum L.) is still a source for isolation of codeine and morphine. Cell cultures from this plant lose their ability to produce morphinans. Their major alkaloid is sanguinarine. The elicitation of the opium poppy cell cultures by fungal preparation lead to a nine-fold increase in the content of sanguinarine. The specific activity of polyphenoloxidase (PPO) was three-times higher in the elicited compared to the nonelicited cells. Two isoforms of PPO (Mr 63 kDa, 41 kDa) were identified in opium poppy cell cultures by PAGE. The number of PPO isoforms was not affected by elicitation. Phenyl-Sepharose CL-4B was used for affinity purification of PPO. In a single purification step the specific activity of PPO was enriched 14-fold.

  10. Maintenance of High Photosynthetic Rates in Mesophyll Cells Isolated from Papaver somniferum1

    PubMed Central

    Paul, John S.; Bassham, James A.

    1977-01-01

    The establishment and maintenance of high rates of photosynthetic CO2 incorporation in mesophyll cells of Papaver somniferum (opium poppy) depend on a regime of dark and light periods immediately following isolation, as well as carefully adjusted conditions of isolation. Analysis of the incorporation pattern of 14CO2 by the isolated cells indicates an initial “stress-response” period of approximately 20 hours characterized by increased respiratory-type metabolism and diminished photosynthesis. Under the favorable regime, this period is followed by rapid recovery and the reinstatement of a metabolic state strikingly similar to that of intact leaves in which the initial rate of CO2 incorporation is between 110 and 175 μmoles CO2 fixed per mg chlorophyll per hour. The photosynthetic viability of these cells can be maintained for up to 80 hours. Images PMID:16660183

  11. Increasing morphinan alkaloid production by over-expressing codeinone reductase in transgenic Papaver somniferum.

    PubMed

    Larkin, Philip J; Miller, James A C; Allen, Robert S; Chitty, Julie A; Gerlach, Wayne L; Frick, Susanne; Kutchan, Toni M; Fist, Anthony J

    2007-01-01

    Only plants of the Papaver genus (poppies) are able to synthesize morphinan alkaloids, and cultivation of P. somniferum, opium poppy, remains critical for the production and supply of morphine, codeine and various semi-synthetic analgesics. Opium poppy was transformed with constitutively expressed cDNA of codeinone reductase (PsCor1.1), the penultimate step in morphine synthesis. Most transgenic lines showed significant increases in capsule alkaloid content in replicated glasshouse and field trials over 4 years. The morphinan alkaloid contents on a dry weight basis were between 15% and 30% greater than those in control high-yielding genotypes and control non-transgenic segregants. Transgenic leaves had approximately 10-fold greater levels of Cor transcript compared with non-transgenic controls. Two cycles of crossing of the best transgenic line into an elite high-morphine genotype resulted in significant increases in morphine and total alkaloids relative to the elite recurrent parent. No significant changes in alkaloid profiles or quantities were observed in leaf, roots, pollen and seed.

  12. Microbe associated molecular patterns from rhizosphere bacteria trigger germination and Papaver somniferum metabolism under greenhouse conditions.

    PubMed

    Bonilla, A; Sarria, A L F; Algar, E; Muñoz Ledesma, F J; Ramos Solano, B; Fernandes, J B; Gutierrez Mañero, F J

    2014-01-01

    Ten PGPR from different backgrounds were assayed on Papaver somniferum var. Madrigal to evaluate their potential as biotic elicitors to increase alkaloid content under the rationale that some microbe associated molecular patterns (MAMPs) are able to trigger plant metabolism. First, the 10 strains and their culture media at two different concentrations were tested for their ability to trigger seed germination. Then, the best three strains were tested for their ability to increase seedling growth and alkaloid levels under greenhouse conditions. Only three strains and their culture media enhanced germination. Then, germination enhancing capacity of these best three strains, N5.18 Stenotrophomonas maltophilia, Aur9 Chryseobacterium balustinum and N21.4 Pseudomonas fluorescens was evaluated in soil. Finally, the three strains were applied on seedlings at two time points, by soil drench or by foliar spray. Photosynthesis was measured, plant height was recorded, capsules were weighted and alkaloids analyzed by HPLC. Only N5.18 delivered by foliar spray significantly increased plant height coupled to an increase in total alkaloids and a significant increase in opium poppy straw dry weight; these increases were supported by a better photosynthetic efficiency. The relative contents of morphine, thebaine, codeine and oripavine were affected by this treatment causing a significant increase in morphine coupled to a decrease in thebaine, demonstrating the effectivity of MAMPs from N5.18 in this plant species. Considering the increase in capsule biomass and alkaloids together with the acceleration of germination, strain N5.18 appears as a good candidate to elicit plant metabolism and consequently, to increase productivity of Papaver somniferum.

  13. Identification of a secretory phospholipase A2 from Papaver somniferum L. that transforms membrane phospholipids.

    PubMed

    Jablonická, Veronika; Mansfeld, Johanna; Heilmann, Ingo; Obložinský, Marek; Heilmann, Mareike

    2016-09-01

    The full-length sequence of a new secretory phospholipase A2 was identified in opium poppy seedlings (Papaver somniferum L.). The cDNA of poppy phospholipase A2, denoted as pspla2, encodes a protein of 159 amino acids with a 31 amino acid long signal peptide at the N-terminus. PsPLA2 contains a PLA2 signature domain (PA2c), including the Ca(2+)-binding loop (YGKYCGxxxxGC) and the catalytic site motif (DACCxxHDxC) with the conserved catalytic histidine and the calcium-coordinating aspartate residues. The aspartate of the His/Asp dyad playing an important role in animal sPLA2 catalysis is substituted by a serine residue. Furthermore, the PsPLA2 sequence contains 12 conserved cysteine residues to form 6 structural disulfide bonds. The calculated molecular weight of the mature PsPLA2 is 14.0 kDa. Based on the primary structure PsPLA2 belongs to the XIB group of PLA2s. Untagged recombinant PsPLA2 obtained by expression in Escherichia coli, renaturation from inclusion bodies and purification by cation-exchange chromatography was characterized in vitro. The pH optimum for activity of PsPLA2 was found to be pH 7, when using mixed micelles of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and Triton X-100. PsPLA2 specifically cleaves fatty acids from the sn-2 position of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine and shows a pronounced preference for PC over phosphatidyl ethanolamine, -glycerol and -inositol. The active recombinant enzyme was tested in vitro against natural phospholipids isolated from poppy plants and preferably released the unsaturated fatty acids, linoleic acid and linolenic acid, from the naturally occurring mixture of substrate lipids.

  14. Identification of a secretory phospholipase A2 from Papaver somniferum L. that transforms membrane phospholipids.

    PubMed

    Jablonická, Veronika; Mansfeld, Johanna; Heilmann, Ingo; Obložinský, Marek; Heilmann, Mareike

    2016-09-01

    The full-length sequence of a new secretory phospholipase A2 was identified in opium poppy seedlings (Papaver somniferum L.). The cDNA of poppy phospholipase A2, denoted as pspla2, encodes a protein of 159 amino acids with a 31 amino acid long signal peptide at the N-terminus. PsPLA2 contains a PLA2 signature domain (PA2c), including the Ca(2+)-binding loop (YGKYCGxxxxGC) and the catalytic site motif (DACCxxHDxC) with the conserved catalytic histidine and the calcium-coordinating aspartate residues. The aspartate of the His/Asp dyad playing an important role in animal sPLA2 catalysis is substituted by a serine residue. Furthermore, the PsPLA2 sequence contains 12 conserved cysteine residues to form 6 structural disulfide bonds. The calculated molecular weight of the mature PsPLA2 is 14.0 kDa. Based on the primary structure PsPLA2 belongs to the XIB group of PLA2s. Untagged recombinant PsPLA2 obtained by expression in Escherichia coli, renaturation from inclusion bodies and purification by cation-exchange chromatography was characterized in vitro. The pH optimum for activity of PsPLA2 was found to be pH 7, when using mixed micelles of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and Triton X-100. PsPLA2 specifically cleaves fatty acids from the sn-2 position of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine and shows a pronounced preference for PC over phosphatidyl ethanolamine, -glycerol and -inositol. The active recombinant enzyme was tested in vitro against natural phospholipids isolated from poppy plants and preferably released the unsaturated fatty acids, linoleic acid and linolenic acid, from the naturally occurring mixture of substrate lipids. PMID:27473012

  15. [Characteristics of Papaver somniferum L. cv. ikkanshu cultivated in Izu].

    PubMed

    Iida, O; Sekine, T; Inoue, O; Yoshimatsu, K; Shimomura, K

    2000-01-01

    The seeds of Papaver somniferum L. cv. Ikkanshu were sown in November (Autumn sowing: AS) and March (Spring sowing: SS) in a field at Izu Experimental Station for Medicinal Plants of National Institute of Health Sciencs, and both AS and SS plants were cultivated to investigate their growth, opium yield and alkaloid content in the opium. Growing periods from the sowing to the opium harvest were approximately six months for AS plants and three months for SS plants. Sizes of plants and capsules in AS were bigger than those in SS, reflecting their growth period. Opium yields per an are in AS and SS were 212.09 g and 142.03 g, respectively. The opium was able to be collected four times in the AS plants though the SS plants ceased to exude opium after the second incision. Therefore higher yield of opium in AS plants seems to be attributed to an amount of opium in the third and fourth incision. Average morphine content in the total opium was 15.61% in AS plants and 15.04% in SS plants, and the estimated amounts of morphine per an are in AS and SS plants were 33.16 g and 21.38 g, respectively.

  16. Development and validation of a reversed-phase HPLC method for determination of alkaloids from Papaver somniferum L. (Papaveraceae).

    PubMed

    Acevska, Jelena; Dimitrovska, Aneta; Stefkov, Gjoshe; Brezovska, Katerina; Karapandzova, Marija; Kulevanova, Svetlana

    2012-01-01

    An HPLC method for the separation of six target alkaloids from Papaver somniferum L. (morphine, codeine, oripavine, thebaine, papaverine, and noscapine) was developed, optimized, and validated. The chromatographic behavior of these alkaloids was investigated using a reversed-phase chromatography at acidic and alkaline pH. The effects of ion-pairing agents, pH value of the mobile phase, concentration of the buffer components, mobile phase organic modifier, and column temperature were studied. Regardless of the large differences in their pKa values, all alkaloids were separated within a close retention window, and good peak shape was achieved for each of the six alkaloids. The proposed method has adequate selectivity, linearity, accuracy, precision, and reproducibility and is applicable for poppy straw.

  17. Optimization of Ultrasound-Assisted Extraction of Morphine from Capsules of Papaver somniferum by Response Surface Methodology

    PubMed Central

    Bulduk, Ibrahim; Gezer, Bahdışen; Cengiz, Mustafa

    2015-01-01

    In this study, amount of morphine from poppy capsules (Papaver somniferum) was investigated using ultrasonic assisted extraction (UAE). Response surface methodology was used to estimate effective experimental conditions on the content extraction of poppy capsules. For this purpose, solvent/solid ratio (10–20 mL/500 mg sample), pH (1–13), time (30–60 min), and temperature (30–50°C) were chosen as experimental variables. The affected response is extraction recovery values for morphine from poppy straw. For interpreting the relationship between experimental factors and response, a design table was established with combinations of three different concentrations levels of this compound in 29 trials. The second order quadratic model gave a satisfactory description of the experimental data. In our study, R-Squared (0.96), Adj-R-Squared (0.92), and Pred R-Squared (0.78) values for extraction yield display good accuracy of the derived model. The predicted optimal conditions for the highest morphine level (3.38 mg morphine/500 mg-sample) were found at 19.99 mL solvent/500 mg solid ratio, 59.94 min extraction time, 1.10 pH, and 42.36°C temperature. In the optimal extraction conditions, the experimental values are very close to the predicted values. Consequently, the response surface modeling can be achieved sufficiently to predict extraction yield from poppy straw by ultrasound assisted extraction. PMID:25861273

  18. Optimization of Ultrasound-Assisted Extraction of Morphine from Capsules of Papaver somniferum by Response Surface Methodology.

    PubMed

    Bulduk, Ibrahim; Gezer, Bahdışen; Cengiz, Mustafa

    2015-01-01

    In this study, amount of morphine from poppy capsules (Papaver somniferum) was investigated using ultrasonic assisted extraction (UAE). Response surface methodology was used to estimate effective experimental conditions on the content extraction of poppy capsules. For this purpose, solvent/solid ratio (10-20 mL/500 mg sample), pH (1-13), time (30-60 min), and temperature (30-50°C) were chosen as experimental variables. The affected response is extraction recovery values for morphine from poppy straw. For interpreting the relationship between experimental factors and response, a design table was established with combinations of three different concentrations levels of this compound in 29 trials. The second order quadratic model gave a satisfactory description of the experimental data. In our study, R-Squared (0.96), Adj-R-Squared (0.92), and Pred R-Squared (0.78) values for extraction yield display good accuracy of the derived model. The predicted optimal conditions for the highest morphine level (3.38 mg morphine/500 mg-sample) were found at 19.99 mL solvent/500 mg solid ratio, 59.94 min extraction time, 1.10 pH, and 42.36°C temperature. In the optimal extraction conditions, the experimental values are very close to the predicted values. Consequently, the response surface modeling can be achieved sufficiently to predict extraction yield from poppy straw by ultrasound assisted extraction.

  19. Optimization of Ultrasound-Assisted Extraction of Morphine from Capsules of Papaver somniferum by Response Surface Methodology.

    PubMed

    Bulduk, Ibrahim; Gezer, Bahdışen; Cengiz, Mustafa

    2015-01-01

    In this study, amount of morphine from poppy capsules (Papaver somniferum) was investigated using ultrasonic assisted extraction (UAE). Response surface methodology was used to estimate effective experimental conditions on the content extraction of poppy capsules. For this purpose, solvent/solid ratio (10-20 mL/500 mg sample), pH (1-13), time (30-60 min), and temperature (30-50°C) were chosen as experimental variables. The affected response is extraction recovery values for morphine from poppy straw. For interpreting the relationship between experimental factors and response, a design table was established with combinations of three different concentrations levels of this compound in 29 trials. The second order quadratic model gave a satisfactory description of the experimental data. In our study, R-Squared (0.96), Adj-R-Squared (0.92), and Pred R-Squared (0.78) values for extraction yield display good accuracy of the derived model. The predicted optimal conditions for the highest morphine level (3.38 mg morphine/500 mg-sample) were found at 19.99 mL solvent/500 mg solid ratio, 59.94 min extraction time, 1.10 pH, and 42.36°C temperature. In the optimal extraction conditions, the experimental values are very close to the predicted values. Consequently, the response surface modeling can be achieved sufficiently to predict extraction yield from poppy straw by ultrasound assisted extraction. PMID:25861273

  20. Genetic and Phenotypic Analyses of a Papaver somniferum T-DNA Insertional Mutant with Altered Alkaloid Composition.

    PubMed

    Kawano, Noriaki; Kiuchi, Fumiyuki; Kawahara, Nobuo; Yoshimatsu, Kayo

    2012-02-02

    The in vitro shoot culture of a T-DNA insertional mutant of Papaver somniferum L. established by the infection of Agrobacterium rhizogenes MAFF03-01724 accumulated thebaine instead of morphine as a major opium alkaloid. To develop a non-narcotic opium poppy and to gain insight into its genetic background, we have transplanted this mutant to soil, and analyzed its alkaloid content along with the manner of inheritance of T-DNA insertion loci among its selfed progenies. In the transplanted T0 primary mutant, the opium (latex) was found to be rich in thebaine (16.3% of dried opium) by HPLC analysis. The analyses on T-DNA insertion loci by inverse PCR, adaptor-ligation PCR, and quantitative real-time PCR revealed that as many as 18 copies of T-DNAs were integrated into a poppy genome in a highly complicated manner. The number of copies of T-DNAs was decreased to seven in the selected T3 progenies, in which the average thebaine content was 2.4-fold that of the wild type plant. This may indicate that the high thebaine phenotype was increasingly stabilized as the number of T-DNA copies was decreased. In addition, by reverse transcription PCR analysis on selected morphine biosynthetic genes, the expression of codeine 6-O-demethylase was clearly shown to be diminished in the T0 in vitro shoot culture, which can be considered as one of the key factors of altered alkaloid composition.

  1. Identification and expression analyses of MYB and WRKY transcription factor genes in Papaver somniferum L.

    PubMed

    Kakeshpour, Tayebeh; Nayebi, Shadi; Rashidi Monfared, Sajad; Moieni, Ahmad; Karimzadeh, Ghasem

    2015-10-01

    Papaver somniferum L. is an herbaceous, annual and diploid plant that is important from pharmacological and strategic point of view. The cDNA clones of two putative MYB and WRKY genes were isolated (GeneBank accession numbers KP411870 and KP203854, respectively) from this plant, via the nested-PCR method, and characterized. The MYB transcription factor (TF) comprises 342 amino acids, and exhibits the structural features of the R2R3MYB protein family. The WRKY TF, a 326 amino acid-long polypeptide, falls structurally into the group II of WRKY protein family. Quantitative real-time PCR (qRT-PCR) analyses indicate the presence of these TFs in all organs of P. somniferum L. and Papaver bracteatum L. Highest expression levels of these two TFs were observed in the leaf tissues of P. somniferum L. while in P. bracteatum L. the espression levels were highest in the root tissues. Promoter analysis of the 10 co-expressed gene clustered involved in noscapine biosynthesis pathway in P. somniferum L. suggested that not only these 10 genes are co-expressed, but also share common regulatory motifs and TFs including MYB and WRKY TFs, and that may explain their common regulation.

  2. Identification and expression analyses of MYB and WRKY transcription factor genes in Papaver somniferum L.

    PubMed

    Kakeshpour, Tayebeh; Nayebi, Shadi; Rashidi Monfared, Sajad; Moieni, Ahmad; Karimzadeh, Ghasem

    2015-10-01

    Papaver somniferum L. is an herbaceous, annual and diploid plant that is important from pharmacological and strategic point of view. The cDNA clones of two putative MYB and WRKY genes were isolated (GeneBank accession numbers KP411870 and KP203854, respectively) from this plant, via the nested-PCR method, and characterized. The MYB transcription factor (TF) comprises 342 amino acids, and exhibits the structural features of the R2R3MYB protein family. The WRKY TF, a 326 amino acid-long polypeptide, falls structurally into the group II of WRKY protein family. Quantitative real-time PCR (qRT-PCR) analyses indicate the presence of these TFs in all organs of P. somniferum L. and Papaver bracteatum L. Highest expression levels of these two TFs were observed in the leaf tissues of P. somniferum L. while in P. bracteatum L. the espression levels were highest in the root tissues. Promoter analysis of the 10 co-expressed gene clustered involved in noscapine biosynthesis pathway in P. somniferum L. suggested that not only these 10 genes are co-expressed, but also share common regulatory motifs and TFs including MYB and WRKY TFs, and that may explain their common regulation. PMID:26600674

  3. Rice salT promoter is activated in Papaver somniferum and Nicotiana tabacum transgenic cells in the absence of exogenous ABA.

    PubMed

    Elleuch; Belbahri; Boetti; David; Thomassetb; David

    2001-01-01

    With the aim of modifying secondary metabolism in Opium poppy (Papaver somniferum) and tobacco (Nicotiana tabacum) cells, gene transfer was performed using the sam1 gene from Arabidopsis thaliana under the control of the salT promoter. This promoter is induced by ABA in rice and in tobacco and we have shown that it is also induced in poppy cells (gus gene). Putatively transformed poppy and tobacco cell lines with the sam1 gene were obtained. In the absence of exogenous inducer we noticed the expression of the transgene resulting in a significant increase of SAM-S activity in all tested transformants of poppy and in half the transgenic tobacco cell lines tested. Addition of ABA to the culture medium failed to enhance the expression of the transgene in both species and resulted in a decrease of the sam1 gene expression in some cell lines. Since the salT promoter is induced by exogenous ABA in both species (gus reporter gene), we suggest a partial sam1 transgene inactivation in certain cell lines. These results show that the efficiency of a regulatory sequence may be different when fused with a reporter gene (gus) compared to fusion with a gene belonging to the housekeeping family (sam1).

  4. Production of Pharmaceuticals from Papaver Cultivars In Vitro

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A methodology to clonally proliferate Iranian poppy (Papaver bracteatum Lindl.) and opium poppy (P. somniferum L.) shoots is presented employing an in vitro hydroponics system (i.e., automated plant culture system (APCS)). Temperature had a profound effect on growth and alkaloid production after 8-...

  5. Comparative transcriptome analysis using high papaverine mutant of Papaver somniferum reveals pathway and uncharacterized steps of papaverine biosynthesis.

    PubMed

    Pathak, Sumya; Lakhwani, Deepika; Gupta, Parul; Mishra, Brij Kishore; Shukla, Sudhir; Asif, Mehar Hasan; Trivedi, Prabodh Kumar

    2013-01-01

    The benzylisoquinoline alkaloid papaverine, synthesized in low amount in most of the opium poppy varieties of Papaver somniferum, is used as a vasodilator muscle relaxant and antispasmodic. Papaverine biosynthesis remains controversial as two different routes utilizing either (S)-coclaurine or (S)-reticuline have been proposed with uncharacterized intermediate steps. In an attempt to elucidate papaverine biosynthesis and identify putative genes involved in uncharacterized steps, we carried out comparative transcriptome analysis of high papaverine mutant (pap1) and normal cultivar (BR086) of P. somniferum. This natural mutant synthesizes more than 12-fold papaverine in comparison to BR086. We established more than 238 Mb transcriptome data separately for pap1 and BR086. Assembly of reads generated 127,342 and 106,128 unigenes in pap1 and BR086, respectively. Digital gene expression analysis of transcriptomes revealed 3,336 differentially expressing unigenes. Enhanced expression of (S)-norcoclaurine-6-O-methyltransferase (6OMT), (S)-3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase (4'OMT), norreticuline 7-O-methyltransferase (N7OMT) and down-regulation of reticuline 7-O-methyltransferase (7OMT) in pap1 in comparison to BR086 suggest (S)-coclaurine as the route for papaverine biosynthesis. We also identified several methyltransferases and dehydrogenases with enhanced expression in pap1 in comparison to BR086. Our analysis using natural mutant, pap1, concludes that (S)-coclaurine is the branch-point intermediate and preferred route for papaverine biosynthesis. Differentially expressing methyltransferases and dehydrogenases identified in this study will help in elucidating complete biosynthetic pathway of papaverine. The information generated will be helpful in developing strategies for enhanced biosynthesis of papaverine through biotechnological approaches.

  6. [Biosynthesis of poppy isoquinoline alkaloids in nature and in vitro culture. 2. Bracteum poppy (Papaver bracteatum Lindl.)].

    PubMed

    Kunakh, V A; Katsan, V A

    2004-01-01

    Literature data and the data of the author's investigations on production of isoquinoline alkaloids by Papaver bracteatum Lindl. have been analyzed. Information on the methods of regulation and cell localization of morphine and sanguinarine biosynthesis is presented. The works studying differentiation processes in tissue cultures of bracteum poppy and relationship thereof with thebaine biosynthesis have been analyzed. Possible mechanism determining the induction of somatic embryos development and thebaine biosynthesis in the culture in vitro are proposed.

  7. Systemic protection of Papaver somniferum L. against Iraella luteipes (Hymenoptera: Cynipidae) by an endophytic strain of Beauveria bassiana (Ascomycota: Hypocreales).

    PubMed

    Quesada-Moraga, E; Muñoz-Ledesma, F J; Santiago-Alvarez, C

    2009-06-01

    The poppy stem gall wasp, Iraella luteipes (Thompson) (Hymenoptera: Cynipidae), is one of the main pests of the opium poppy, Papaver somniferum L., an economically important pharmaceutical crop cultivated worldwide. In a previous study, we obtained from I. luteipes larvae a strain of the entomopathogenic fungus Beauveria bassiana (Ascomycota: Hypocreales) that can become established endophytically in opium poppy plants. A field experiment was conducted to study the ability of this B. bassiana strain to provide systemic protection against damage by I. luteipes in opium poppy in southern Spain for three seasons. Conidial suspensions were applied as seed dressings, leaf sprays, or soil sprays. The effect of the treatment was studied by harvesting fully ripened plants and dissecting I. luteipes larvae from the stem. The effect of treatment on growth and yield was also evaluated. Emergence of I. luteipes adults was not uniform over the 3 yr, with important differences exhibited in the duration of the emergence period, although the flight peaks tended to occur in mid-late April. B. bassiana seed dressings, leaf sprays at the fourth true-leaf stage, and soil sprays were not significantly different in their ability to reduce the number of larvae per plant compared with the controls, with percentage reductions of 36.5-58.5, 64.4-73.4, and 51.9-57.2% in 2005, 2006, and 2007, respectively. Even though the population level of I. luteipes increased over the 3 yr, the efficacy of the fungal inoculation in reducing the larval population was maintained throughout the study period. No significant differences between inoculation methods were detected in the percentage of leaf pieces showing fungal growth when placed on B. bassiana selective medium, with mean values in the range of 10-15% for the three seasons. Leaf pieces from controls did not exhibit any sign of B. bassiana growth when placed on B. bassiana-selective medium. Neither adverse effects on growth and yield nor

  8. [The floral meristem undetermination mutation in Papaver somniferum L.: spontaneous phenotypic variation in ontogeny].

    PubMed

    Beliaeva, R G

    2008-01-01

    A new morphogenetic mutation of the shoot, floral meristem undetermination, was found in Papaver somniferum L. with monocarpic shoot. The expression of the DFM (determination of floral meristem) gene, which limits the proliferative activity of stem cells in the floral meristem, was affected. The mutation displayed spontaneous phenotypic instability in ontogeny, variation in the mutant character expression on different flowers of the same plant in the same genotypic environment. The mutation phenotype varied from no expression or formation of individual phyllomes in the center of the primary ovary to formation of a new flower and a new capsule with viable seeds.

  9. Wound Induced Tanscriptional Regulation of Benzylisoquinoline Pathway and Characterization of Wound Inducible PsWRKY Transcription Factor from Papaver somniferum

    PubMed Central

    Singh, Seema; Phukan, Ujjal J.; Gupta, M. M.; Shanker, Karuna; Shukla, Rakesh Kumar

    2013-01-01

    Wounding is required to be made in the walls of the green seed pod of Opium poppy prior exudation of latex. To withstand this kind of trauma plants regulate expression of some metabolites through an induced transcript level. 167 unique wound-inducible ESTs were identified by a repetitive round of cDNA subtraction after 5 hours of wounding in Papaver somniferum seedlings. Further repetitive reverse northern analysis of these ESTs revealed 80 transcripts showing more than two fold induction, validated through semi-quantitative RT-PCR & real time expression analysis. One of the major classified categories among identified ESTs belonged to benzylisoquinoline transcripts. Tissue specific metabolite analysis of benzylisoquinoline alkaloids (BIAs) in response to wounding revealed increased accumulation of narcotine and papaverine. Promoter analysis of seven transcripts of BIAs pathway showed the presence of W-box cis-element with the consensus sequence of TGAC, which is the proposed binding site for WRKY type transcription factors. One of the Wound inducible ‘WRKY’ EST isolated from our subtracted library was made full-length and named as ‘PsWRKY’. Bacterially expressed PsWRKY interacted with the W-box element having consensus sequence TTGACT/C present in the promoter region of BIAs biosynthetic pathway genes. PsWRKY further activated the TYDC promoter in yeast and transiently in tobacco BY2 cells. Preferential expression of PsWRKY in straw and capsule and its interaction with consensus W-box element present in BIAs pathway gene transcripts suggest its possible involvement in the wound induced regulation of BIAs pathway. PMID:23382823

  10. Wound induced tanscriptional regulation of benzylisoquinoline pathway and characterization of wound inducible PsWRKY transcription factor from Papaver somniferum.

    PubMed

    Mishra, Sonal; Triptahi, Vineeta; Singh, Seema; Phukan, Ujjal J; Gupta, M M; Shanker, Karuna; Shukla, Rakesh Kumar

    2013-01-01

    Wounding is required to be made in the walls of the green seed pod of Opium poppy prior exudation of latex. To withstand this kind of trauma plants regulate expression of some metabolites through an induced transcript level. 167 unique wound-inducible ESTs were identified by a repetitive round of cDNA subtraction after 5 hours of wounding in Papaver somniferum seedlings. Further repetitive reverse northern analysis of these ESTs revealed 80 transcripts showing more than two fold induction, validated through semi-quantitative RT-PCR & real time expression analysis. One of the major classified categories among identified ESTs belonged to benzylisoquinoline transcripts. Tissue specific metabolite analysis of benzylisoquinoline alkaloids (BIAs) in response to wounding revealed increased accumulation of narcotine and papaverine. Promoter analysis of seven transcripts of BIAs pathway showed the presence of W-box cis-element with the consensus sequence of TGAC, which is the proposed binding site for WRKY type transcription factors. One of the Wound inducible 'WRKY' EST isolated from our subtracted library was made full-length and named as 'PsWRKY'. Bacterially expressed PsWRKY interacted with the W-box element having consensus sequence TTGACT/C present in the promoter region of BIAs biosynthetic pathway genes. PsWRKY further activated the TYDC promoter in yeast and transiently in tobacco BY2 cells. Preferential expression of PsWRKY in straw and capsule and its interaction with consensus W-box element present in BIAs pathway gene transcripts suggest its possible involvement in the wound induced regulation of BIAs pathway.

  11. Intoxication due to Papaver rhoeas (Corn Poppy): Five Case Reports.

    PubMed

    Günaydın, Yahya Kemal; Dündar, Zerrin Defne; Çekmen, Bora; Akıllı, Nazire Belgin; Köylü, Ramazan; Cander, Başar

    2015-01-01

    Introduction. In this paper, we aimed to present five Papaver rhoeas intoxication cases, which is very rare in the literature. Case 1. A 35-year-old female patient was admitted to our emergency room with the complaints of nausea, restlessness, and dyspnea developing 3 hours after eating Papaver rhoeas. On physical examination, her general condition was moderate; she was conscious and the vital findings were normal. The pupils were myotic. She was transferred to the toxicology intensive care unit as she experienced a generalized tonic clonic seizure lasting for three minutes. Case 2. A 41-year-old female patient was brought to our emergency room by 112 ambulance as she had contractions in her arms and legs, unconsciousness, and foam coming from her mouth two hours after Papaver rhoeas ingestion. On physical examination, she was confused, the pupils were myotic, and she was tachycardic. Arterial blood gases analysis revealed lactic acidosis. Case 3. A 38-year-old female patient was admitted to our emergency room with complaints of nausea and vomiting two hours after ingestion of Papaver rhoeas. Her physical examination and tests were normal. Case 4. A 34-year-old male patient was admitted to our emergency room with complaints of numbness and loss of power in his arms and legs one hour after Papaver rhoeas ingestion. He was hospitalized at the toxicology intensive care unit for follow-up and treatment. Dyspnea and bradycardia developed on the follow-up. The oxygen saturation without oxygen support was 90%. ECG revealed sinus bradycardia. The cardiac enzymes did not increase. Case 5. A 42-year-old female patient was brought to our emergency room by 112 ambulance with contractions in her arms and legs and unconsciousness two hours after Papaver rhoeas ingestion. On her physical examination, she was confused and the pupils were myotic. Arterial blood gases analysis revealed lactic acidosis. Conclusion. All patients were followed up for a few days and then discharged from

  12. Intoxication due to Papaver rhoeas (Corn Poppy): Five Case Reports.

    PubMed

    Günaydın, Yahya Kemal; Dündar, Zerrin Defne; Çekmen, Bora; Akıllı, Nazire Belgin; Köylü, Ramazan; Cander, Başar

    2015-01-01

    Introduction. In this paper, we aimed to present five Papaver rhoeas intoxication cases, which is very rare in the literature. Case 1. A 35-year-old female patient was admitted to our emergency room with the complaints of nausea, restlessness, and dyspnea developing 3 hours after eating Papaver rhoeas. On physical examination, her general condition was moderate; she was conscious and the vital findings were normal. The pupils were myotic. She was transferred to the toxicology intensive care unit as she experienced a generalized tonic clonic seizure lasting for three minutes. Case 2. A 41-year-old female patient was brought to our emergency room by 112 ambulance as she had contractions in her arms and legs, unconsciousness, and foam coming from her mouth two hours after Papaver rhoeas ingestion. On physical examination, she was confused, the pupils were myotic, and she was tachycardic. Arterial blood gases analysis revealed lactic acidosis. Case 3. A 38-year-old female patient was admitted to our emergency room with complaints of nausea and vomiting two hours after ingestion of Papaver rhoeas. Her physical examination and tests were normal. Case 4. A 34-year-old male patient was admitted to our emergency room with complaints of numbness and loss of power in his arms and legs one hour after Papaver rhoeas ingestion. He was hospitalized at the toxicology intensive care unit for follow-up and treatment. Dyspnea and bradycardia developed on the follow-up. The oxygen saturation without oxygen support was 90%. ECG revealed sinus bradycardia. The cardiac enzymes did not increase. Case 5. A 42-year-old female patient was brought to our emergency room by 112 ambulance with contractions in her arms and legs and unconsciousness two hours after Papaver rhoeas ingestion. On her physical examination, she was confused and the pupils were myotic. Arterial blood gases analysis revealed lactic acidosis. Conclusion. All patients were followed up for a few days and then discharged from

  13. Comparative analysis of Papaver somniferum genotypes having contrasting latex and alkaloid profiles.

    PubMed

    Chaturvedi, Nidarshana; Singh, Mridula; Shukla, Ashutosh K; Shasany, Ajit K; Shanker, Karuna; Lal, Raj K; Khanuja, Suman P S

    2014-07-01

    Papaver somniferum produces therapeutically useful benzylisoquinoline alkaloids (BIAs) like papaverine, thebaine, codeine, and morphine that accumulate in its capsular latex. Morphine is a potent analgesic but is also abused as a narcotic, which has increased the demand for non-narcotic thebaine that can be converted into various analgesics. To curtail the narcotic menace, many distinct genotypes of the plant have been developed that are deficient in morphine and/or latex. Sujata is one such latex-less low alkaloid-producing variety developed from the alkaloid-rich gum harvest variety Sampada. Its utility for gene prospecting and studying differential gene regulation responsible for its low alkaloid, nutritive seed oil, and latex-less phenotype has been exploited in this study. BIA profiling of Sujata and Sampada capsules at the early and late stages indicated that except for thebaine, Sujata had a depressed alkaloid phenotype as compared to Sampada. Comparative transcript-based analysis of the two genotypes was carried out in the early stage capsule (higher thebaine) using subtractive hybridization and microarray. Interrogation of a P. somniferum array yielded many differentially expressing transcripts. Their homology-based annotation classified them into categories--latex related, oil/lipid related, alkaloid related, cell wall related, and others. These leads will be useful to characterize the highly sought after Sujata phenotype.

  14. Comparative qualitative and quantitative determination of alkaloids in narcotic and condiment Papaver somniferum cultivars.

    PubMed

    Frick, Susanne; Kramell, Robert; Schmidt, Jürgen; Fist, Anthony J; Kutchan, Toni M

    2005-05-01

    In the present study morphinan, tetrahydrobenzylisoquinoline, benzo[c]phenanthridine, and phthalideisoquinoline alkaloids were determined qualitatively and quantitatively by HPLC and LC-MS analysis in tissues of the Tasmanian Papaver somniferum L. elite cultivar C048-6-14-64. The data were compared with the results from the low-morphine cultivar "Marianne". In the elite cultivar, 91.2% of the latex alkaloids consist of the three pharmaceutically most valuable alkaloids: morphine, codeine, and thebaine. In the root system, the major alkaloids are sanguinarine/10-hydroxysanguinarine and dihydrosanguinarine/10-hydroxydihydrosanguinarine. In the stems and leaves of C048-6-14-64, the same alkaloids were measured as in the latex. In the stems, a gradient in relative total alkaloid content from the top downward toward the roots was observed. The concentration of morphine was decreasing toward the roots, whereas an increasing gradient from the upper to the lower stem parts was detected for codeine. The relative total alkaloid concentration in leaves remained constant; no gradient was observed. The cultivar "Marianne" displayed a shifted pattern of alkaloid accumulation and reduced levels of total alkaloid. In the condiment cultivar, 80.5% of the alkaloids of the latex consisted of the two phthalideisoquinoline alkaloids narcotoline and noscapine. Only 18.8% of the relative total alkaloid content were morphinan alkaloids. In contrast to the narcotic cultivar, in which the benzo[c]phenanthridines in roots dominated over the morphinan and tetrahydrobenzylisoquinoline alkaloids, the concentration of benzo[c]phenanthridines in "Marianne" was similar to that of morphinan and tetrahydrobenzylisoquinoline alkaloids. These data suggest a differential alkaloid regulation in each cultivar of P. somniferum.

  15. [Variability of flavonol contents during floral morphogenesis in Papaver somniferum L].

    PubMed

    Beliaeva, R G; Evdokimova, L I

    2004-01-01

    We studied the contents of flavonols (kaempferol and quercetin) in the meristem of vegetative and generative apices of the main plant shoot in floral Papaver somniferum mutants, as well as in the normal plants at successive stages of flower development. Five stages of flower development were distinguished. Flavonols (kaempferol and quercetin) were present in all flower organs at all stages of floral morphogenesis we studied. However, their contents and distribution in different organs and at different stages of flower development markedly varied. No significant differences were found in the contents of flavonols in the meristems of vegetative and generative apices of the main shoot in the lines of floral mutants, as well as between the lines with different amounts of vegetative phytomeres. In the plants with normal flower structure, the contents of flavonols (kaempferol + quercetin) sharply increased with the beginning of differentiation of flower organs, i.e. from stage 3, to reach a maximum in the open flower, when gametogenesis is terminated and fertilization takes place. The level of flavonol contents in the petals (upper part) and stamen was at a maximum at all stages of flower development, while that in the gynaecium was at a minimum. The kaempferol: quercetin ratio shifted towards quercetin at successive stages of flower development, most significantly in the stamens. The involvement of flavonols in the regulation of floral morphogenesis at stages of flower organs differentiation and functioning is discussed.

  16. [Construction and tobacco transformation of COR and BBE genes hairpin RNA vector of Papaver somniferum].

    PubMed

    Lu, Yan-Mei; Zhang, Jin-Wen; Wei, Yu-Jie; Wei, Gui-Min; Zhang, Yan-Hong; Gao, Yi-Feng

    2013-07-01

    The gene expressions of codeinone reductase (COR) and berberine bridge enzyme (BBE) in Papaver somniferum were blocked by RNA hairpin of RNA interference (RNAi). The complete sequences of COR and BBE genes were cloned by reverse transcription-polymerase chain reaction (RT-PCR), the results of homology comparison revealed that the cloned COR and BBE genes had high homology with the other gene family members reported in the GenBank. The target sequences of COR and BBE genes were screened in accordance with the design principle of RNAi, a 643 bp fusion gene was obtained by the method of overlapping PCR, then plant expression vector ihpRNA was constructed based on intermediate vector pHANNIBAL and plant expression vector pCEPSPS. With that 78 transgenic plants were obtained through Agrobacterium-mediated and 17 positive plants were screened by PCR, that could initially indicate that the target fragments of COR and BBE gene had been integrated into tobacco genome.

  17. [Influence of fertilizers on yield and yield components of opium from Papaver somniferum Linn].

    PubMed

    Kumagai, Takeo; Hatakeyama, Yoshio; Yoshimatsu, Kayo; Shimomura, Koichiro

    2003-01-01

    The effect of fertilizer on the yield and yield components of opium from Papaver somniferum L.cv.Ikkanshu were investigated from 1993 to 1995. The weight of opium per one capsule was tendency to increase in the order of peat moss > pulverized charcoal > compound fertilizer, but the differences among the results were not significant. Furthermore, no difference in the capsule weight per 100 m2 and the yields of opium and alkaloids per 100 m2 were detected. The significant difference was only observed when the data were compared among the weights and yields of different years. The difference among the yield of opium was mainly due to the weight of opium per one capsule. The weight of opium per one capsule showed a high correlation with capsule husk weight. The alkaloids contents in opium obtained with a different fertilizer application showed no difference. Morphine content at the first lancing was about 11%, and the value decreased with the order of lancing. On the contrary, the codeine and the thebaine content did not change during lancing and the value were 6-7% and 3%, respectively. The papaverine and the noscapine content decreased in the order of lancing.

  18. [Influence of fertilizers on yield and yield components of opium from Papaver somniferum Linn].

    PubMed

    Kumagai, Takeo; Hatakeyama, Yoshio; Yoshimatsu, Kayo; Shimomura, Koichiro

    2003-01-01

    The effect of fertilizer on the yield and yield components of opium from Papaver somniferum L.cv.Ikkanshu were investigated from 1993 to 1995. The weight of opium per one capsule was tendency to increase in the order of peat moss > pulverized charcoal > compound fertilizer, but the differences among the results were not significant. Furthermore, no difference in the capsule weight per 100 m2 and the yields of opium and alkaloids per 100 m2 were detected. The significant difference was only observed when the data were compared among the weights and yields of different years. The difference among the yield of opium was mainly due to the weight of opium per one capsule. The weight of opium per one capsule showed a high correlation with capsule husk weight. The alkaloids contents in opium obtained with a different fertilizer application showed no difference. Morphine content at the first lancing was about 11%, and the value decreased with the order of lancing. On the contrary, the codeine and the thebaine content did not change during lancing and the value were 6-7% and 3%, respectively. The papaverine and the noscapine content decreased in the order of lancing. PMID:14740399

  19. [Identification of Papaver somni ferum L. and Papaver rhoeas using DSWT-FTIR-RBFNN].

    PubMed

    Zhang, Chang-jiang; Cheng, Cun-gui

    2009-05-01

    Infrared spectra of Papaver somniferum L. and Papaver rhoeas were obtained directly, quickly and accurately by Fourier transform infrared spectroscopy (FTIR)with OMNI sampler. Discrete stationary wavelet transform was used to extrude local region of infrared spectra of Papaver somniferum L. and Papaver rhoeas. The difference of infrared spectra between Papaver somniferum L. and Papaver rhoeas was extruded. Accurate identification rate is improved greatly. One dimensional discrete stationary wavelet transform was implemented to the infrared spectra of Papaver somniferum L. and Papaver rhoeas. The difference between Papaver somniferum L. and Papaver rhoeas was observed at all scales in wavelet domain. Two scales, at which the difference between Papaver somniferum L and Papaver rhoeas is the most obvious, were selected to extract the features of Papaver somniferum L. and Papaver rhoeas. A feature vector including eight feature parameters was constructed. The feature vector was input to RBFNN for training in order to accurately identify Papaver somniferum L. and Papaver rhoeas. In experiment, the authors used one hundred and twenty-eight couples of data of Papaver somniferum L. and Papaver rhoeas (including seventy-eight couples of training samples and fifty couples of testing samples). The experimental results show that it is effective to apply discrete stationary wavelet transform on the basis of FTIR to identify the Papaver somniferum L. and Papaver rhoeas. The accurate identification rate of Papaver somniferum L and Papaver rhoeas is 99.8% and 99.9% respectively.

  20. Thin-layer chromatography-densitometry and liquid chromatography analysis of alkaloids in leaves of Papaver somniferum under stress conditions.

    PubMed

    Szabó, Beata; Lakatos, Agnes; Kõszegi, Tamás; Kátay, György; Botz, L

    2005-01-01

    The effect of stress conditions on the concentrations of secondary metabolites were examined during various developmental stages of Papaver somniferum plants. P. somniferum plants were grown in laboratory conditions (Budakalász). The experiment consisted of 22 treatments. Significantly different alkaloid contents can be observed under different stress conditions. In general, the alkaloid contents of plants are very low; therefore, a highly sensitive and reliable method has to be developed for analysis. The amount of alkaloids was measured by 2 separation and detection techniques. Accuracy of the thin-layer chromatography method for quantitative analysis is limited. Without purification of samples the background is too noisy. Column liquid chromatography is a sensitive and relatively inexpensive method that allows precise quantitative determination of the alkaloid content.

  1. One step green synthesis of silver nano/microparticles using extracts of Trachyspermum ammi and Papaver somniferum.

    PubMed

    Vijayaraghavan, K; Nalini, S P Kamala; Prakash, N Udaya; Madhankumar, D

    2012-06-01

    A novel biosynthesis route for silver nanoparticles (Ag-NPs) was attempted in this present investigation using aqueous extracts of Trachyspermum ammi and Papaver somniferum. The main constituents in T. ammi are thymol, p-cymene and γ-terpinene, while P. somniferum consists of morphine and codeine. The essential oil in T. ammi was found to be a good reducing agent than the alkaloids present in P. somniferum for the formation of biocompatible Ag-NPs. The effectiveness of both the extracts was investigated by using same dosage of extract in the synthesis of silver nanoparticle. The results showed that for the same dosage of extracts the T. ammi synthesized various size triangular shaped nanoparticles measuring from 87 nm, to a fewer nanoparticles having a size of 998 nm diagonally. P. somniferum resulted in almost spherical shaped particle ranging in size between 3.2 and 7.6 μm diagonally. Future research based on synthesis of size specific nanoparticle based on the optimization of reaction condition would be an interesting area.

  2. Accumulation dynamics and some cytogenetical tests at Chelidonium majus and Papaver somniferum callus under the magnetic liquid effect

    NASA Astrophysics Data System (ADS)

    Pavel, A.; Trifan, M.; Bara, I. I.; Creanga, D. E.; Cotae, C.

    1999-07-01

    In this paper we studied the capacity of magnetic liquids which are biocompatible (as previous literature reports showed) to influence germination, de-differentiation and re-differentiation processes in vitro, as well as to influence both the mitotic index and the chromosomal aberration frequency. In vitro cultures of two species of pharmaceutical interest, Chelidonium majus L. and Papaver somniferum L., on media supplemented with various concentrations of magnetic liquid, for various explant types, revealed a small but significant effect of the petroleum magnetic liquid on the callus accumulation and induced a considerable rate of chromosomal aberrations.

  3. Investigation of a substrate-specifying residue within Papaver somniferum and Catharanthus roseus aromatic amino acid decarboxylases.

    PubMed

    Torrens-Spence, Michael P; Lazear, Michael; von Guggenberg, Renee; Ding, Haizhen; Li, Jianyong

    2014-10-01

    Plant aromatic amino acid decarboxylases (AAADs) catalyze the decarboxylation of aromatic amino acids with either benzene or indole rings. Because the substrate selectivity of AAADs is intimately related to their physiological functions, primary sequence data and their differentiation could provide significant physiological insights. However, due to general high sequence identity, plant AAAD substrate specificities have been difficult to identify through primary sequence comparison. In this study, bioinformatic approaches were utilized to identify several active site residues within plant AAAD enzymes that may impact substrate specificity. Next a Papaver somniferum tyrosine decarboxylase (TyDC) was selected as a model to verify our putative substrate-dictating residues through mutation. Results indicated that mutagenesis of serine 372 to glycine enables the P. somniferum TyDC to use 5-hydroxytryptophan as a substrate, and reduces the enzyme activity toward 3,4-dihydroxy-L-phenylalanine (dopa). Additionally, the reverse mutation in a Catharanthus roseus tryptophan decarboxylase (TDC) enables the mutant enzyme to utilize tyrosine and dopa as substrates with a reduced affinity toward tryptophan. Molecular modeling and molecular docking of the P. somniferum TyDC and the C. roseus TDC enzymes provided a structural basis to explain alterations in substrate specificity. Identification of an active site residue that impacts substrate selectivity produces a primary sequence identifier that may help differentiate the indolic and phenolic substrate specificities of individual plant AAADs.

  4. Species identification of Papaver by metabolite profiling.

    PubMed

    Choe, Sanggil; Kim, Suncheun; Lee, Chul; Yang, Wonkyung; Park, Yuran; Choi, Hwakyung; Chung, Heesun; Lee, Dongho; Hwang, Bang Yeon

    2011-09-10

    Papaver somniferum L. and Papaver setigerum D.C. are controlled as opium poppy in Korea because they contain narcotic substances such as morphine and codeine. It is one of the critical issues whether the plants similar to opium poppy in shape are controlled plants or not. There are more than 110 species in the genus Papaver worldwide and about 10 species in Korea. As the morphological features of some species are very similar and the alkaloid contents and the ratios among the major alkaloids vary even within the same species, it is often difficult to identify the exact species by the morphological features and/or major alkaloids analysis. To develop a new method that uses metabolite profiling for species discrimination between P. somniferum, Papaver rhoeas and P. setigerum, the gas chromatography/mass spectrometry (GC-MS) data of the alkaline extract were processed with in-house Microsoft Visual Basic(®) modules and the chemical information was analyzed through multivariate statistical analyses such as Hierarchical cluster analysis (HCA), principal component analysis (PCA) and discriminant analysis (DA). The GC-MS results combined with multivariate analysis demonstrated that the metabolite profiling was an efficient technique for the classification and this method will provide a powerful tool for the identification of Korean Papaver species.

  5. Rapid determination of Papaver somniferum alkaloids in process streams using monolithic column high-performance liquid chromatography with chemiluminescence detection.

    PubMed

    Costin, Jason W; Lewis, Simon W; Purcell, Stuart D; Waddell, Lucy R; Francis, Paul S; Barnett, Neil W

    2007-07-30

    We have combined high-performance liquid chromatography (HPLC) separations using a monolithic column with acidic potassium permanganate and tris(2,2'-bipyridyl)ruthenium(II) chemiluminescence detection in a rapid and highly sensitive method to monitor the process of extracting opiate alkaloids from Papaver somniferum. Due to the high flow rates allowed with the monolithic column and the inherent selectivity of the chemiluminescence reactions, the four predominant alkaloids--morphine, codeine, oripavine and thebaine--were determined in less than 2 min. The results obtained with numerous process samples compared favourable with those of the standard HPLC methodology. Limits of detection were 1x10(-10) M, 5x10(-10) M, 5x10(-10) M and 1x10(-9) M, for morphine, codeine, oripavine and thebaine, respectively.

  6. [Morphogenetic effects of the interaction of floral mutations petal-sepal and anther in petal in Papaver somniferum l].

    PubMed

    Beliaeva, P G

    2001-01-01

    The morphogenetic effects of the interaction of the floral mutations petal-sepal and anther in petal in Papaver somniferum L. with a monocarpic shoot were studied. During analysis of the mutations controlled by the genes ptsp and Ant, no plants of the double-mutant class were found in the second generation, in which microsporangia form on the corolla sepal structures. The ratio of phenotypic classes obtained in the experiment corresponds to that inheritance, when the genetic control of mutant characters is realized by nonallele nonlinked genes Ant and ptsp upon epistatic interaction of these genes. These data were confirmed by analysis of the genotypes of F2 plants from the phenotypic class petal-sepal, which include plants that carry both mutant genes Ant and ptsp. The results obtained suggest that the gene Ant, which controls the formation of microsporangia in the corolla metameres, is not expressed in the presence of a mutation of the gene ptsp; i.e., microsporangia are not formed in tissues with photosynthesizing cells. It is evident that the development of microsporangia is determined by the level of a product of the gene Ptsp. The role of flavonols (quercetin), inhibitors of photosynthesis, as a mechanism of regulation of activity of the genes controlling morphogenesis of the corolla elements and differentiation of microsporangia, is discussed.

  7. Hairy root induction of Papaver somniferum var. album, a difficult-to-transform plant, by A rhizogenes LBA 9402.

    PubMed

    Le Flem-Bonhomme, V; Laurain-Mattar, D; Fliniaux, M A

    2004-03-01

    Two strains of Agrobacterium rhizogenes (15834, LBA 9402) and one Agrobacterium tumefaciens strain [GV 3101 (PMP90RK, p35SGUS-2)] and four culture media were tested and compared for their ability to induce hairy root formation on wounded Papaver somniferum L. hypocotyls. Five weeks after the infection with A. rhizogenes LBA 9402, hairy roots appeared on 80% of the hypocotyls maintained in the hormone-free liquid medium. Six hairy-root cultures were established. Transformation was confirmed by polymerase chain reaction analysis. One clone was analysed for its alkaloid production. The total alkaloid content was higher in the transformed roots (0.46+/-0.06% DW) than in the untransformed roots (0.32+/-0.05% DW). The transformed roots accumulated three times more codeine (0.18+/-0.02% DW) than intact roots (0.05+/-0% DW). Moreover, morphine (0.255+/-0.03% DW) and sanguinarine (0.014+/-0% DW) were found in the liquid culture medium.

  8. [Study of degradation of flavonols in the mutants of poppy (Papaver somniferum L.)].

    PubMed

    Rat'kin, A V; Evdokimova, L I; Zhanaeva, T A

    2003-01-01

    We studied flavonol-degrading activity of cell-free extracts from petals of the flower color and structure mutants. The relationship between degradation of flavonols (kaempferol, quercetin, and myricetin) and biosynthesis of anthocyanins has been revealed. The highest flavonol-degrading activity has been revealed in white flower mutants towards all substrates, particularly, quercetin. The mutations inhibiting synthesis of an anthocyanin pelargonidin provide for synthesis of various quantities of cyanidin in the petals. The flavonol-degrading activity considerably increases with the content of cyanidin. A similar relationship has been revealed in the mutants synthesizing both cyanidin and pelargonidin. The plants accumulating considerable quantities of pelargonidin in their petals have accordingly higher flavonol-degrading activity and predominantly hydrolyze kaempferol. The plants forming additional pods in their flower (pistillody) have higher flavonol-degrading activity as compared to the anther-in-petal and doubleness mutants.

  9. A Papaver somniferum 10-gene cluster for synthesis of the anticancer alkaloid noscapine.

    PubMed

    Winzer, Thilo; Gazda, Valeria; He, Zhesi; Kaminski, Filip; Kern, Marcelo; Larson, Tony R; Li, Yi; Meade, Fergus; Teodor, Roxana; Vaistij, Fabián E; Walker, Carol; Bowser, Tim A; Graham, Ian A

    2012-06-29

    Noscapine is an antitumor alkaloid from opium poppy that binds tubulin, arrests metaphase, and induces apoptosis in dividing human cells. Elucidation of the biosynthetic pathway will enable improvement in the commercial production of noscapine and related bioactive molecules. Transcriptomic analysis revealed the exclusive expression of 10 genes encoding five distinct enzyme classes in a high noscapine-producing poppy variety, HN1. Analysis of an F(2) mapping population indicated that these genes are tightly linked in HN1, and bacterial artificial chromosome sequencing confirmed that they exist as a complex gene cluster for plant alkaloids. Virus-induced gene silencing resulted in accumulation of pathway intermediates, allowing gene function to be linked to noscapine synthesis and a novel biosynthetic pathway to be proposed.

  10. Metabolic engineering of morphinan alkaloids by over-expression of codeinone reductase in transgenic hairy roots of Papaver bracteatum, the Iranian poppy.

    PubMed

    Sharafi, Ali; Sohi, Haleh Hashemi; Mousavi, Amir; Azadi, Pejman; Khalifani, Bahman Hosseini; Razavi, Khadijeh

    2013-03-01

    Papaver bracteatum has a high content of thebaine. It is used as an alternative to P. somniferum for the production of benzylisoquinoline alkaloid. Papaver bracteatum was genetically engineered to over-express codeinone reductase gene in hairy root cultures. Transcript level of the codeinone reductase gene in transgenic hairy root lines increased up to ten- and 24-fold in comparison with hairy roots without CodR over-expression and wild type roots, respectively. Codeine was produced at (0.04 % dry wt) and morphine was at (0.28 % dry wt) in the transgenic hairy root lines. Papaver bracteatum hairy roots expressing CodR gene thus have a high potential to produce morphinan alkaloids.

  11. [Determination of papaverine in seeds of Papaver somniferum L. and soup of chafing dish by high performance liquid chromatography-fluorescence detection].

    PubMed

    Pi, Li; Hu, Fengzu; Shi, Zhixian

    2005-11-01

    A method of reversed-phase high performance liquid chromatography coupled with fluorescence detection was developed for the separation and determination of papaverine in seeds of Papaver somniferum L. and soup of chafing dish. The liquid chromatographic experiment was carried out under the following conditions: the detector was a fluorescence with excitation wavelength at 285 nm and emission wavelength at 355 nm. An RP-C18 column (250 mm x 4.6 mm i.d., 5 microm) was used. The mobile phase was a mixture of methanol and 0.02 mol/L ammonium acetate (70: 30, v/v). The flow rate was 0.8 mL/min. A linear range was obtained from 0.1 ng to 0.1 microg with a good correlation. The lowest detection limit was 0.02 ng. The average recovery of seeds for Papaver somniferum L. was 99.0% - 100.8%. The method is rapid, accurate, sensitive and suitable for the separation and determination of papaverine in foods.

  12. Two highly homologous phospholipase D isoenzymes from Papaver somniferum L. with different transphosphatidylation potential.

    PubMed

    Lerchner, Alexandra; Mansfeld, Johanna; Schäffner, Ines; Schöps, Regina; Beer, Helge K; Ulbrich-Hofmann, Renate

    2005-12-15

    The genes of two phospholipase D (PLD) isoenzymes, PLD1 and PLD2, from poppy seedlings (2829 and 2828 bp) were completely sequenced. The two genes have 96.9% identity in the encoding region and can be assigned to the alpha-type of plant PLDs. The corresponding amino acid sequences do not contain any signal sequences. One Asn-glycosylation site, six and two phosphorylation sites for protein kinase C and tyrosine kinase, respectively, and two phosphatidylinositol-4,5-bisphosphate binding motifs could be identified. Like in most plant PLDs, two HKD motifs and one C2 domain are present. PLD1 and PLD2 have ten and nine cysteine residues. The two enzymes were expressed in E. coli and purified to homogeneity by Ca2+ ion-mediated hydrophobic interaction chromatography. The Ca2+ ion concentration needed for carrier binding of the two enzymes in chromatography as well as for optimum activity was found to be considerably higher (>100 mM) than with other alpha-type plant PLDs. Although PLD1 and PLD2 differ in eleven amino acids only, they showed remarkable differences in their transphosphatidylation activity. Two amino acid exchanges within and near the first HKD motif contribute to this difference as shown by the A349E/E352Q-variant of PLD2.

  13. Evolution of morphine biosynthesis in opium poppy.

    PubMed

    Ziegler, Jörg; Facchini, Peter J; Geissler, René; Schmidt, Jürgen; Ammer, Christian; Kramell, Robert; Voigtländer, Susan; Gesell, Andreas; Pienkny, Silke; Brandt, Wolfgang

    2009-01-01

    Benzylisoquinoline alkaloids (BIAs) are a group of nitrogen-containing plant secondary metabolites comprised of an estimated 2500 identified structures. In BIA metabolism, (S)-reticuline is a key branch-point intermediate that can be directed into several alkaloid subtypes with different structural skeleton configurations. The morphinan alkaloids are one subclass of BIAs produced in only a few plant species, most notably and abundantly in the opium poppy (Papaver somniferum). Comparative transcriptome analysis of opium poppy and several other Papaver species that do not accumulate morphinan alkaloids showed that known genes encoding BIA biosynthetic enzymes are expressed at higher levels in P. somniferum. Three unknown cDNAs that are co-ordinately expressed with several BIA biosynthetic genes were identified as enzymes in the pathway. One of these enzymes, salutaridine reductase (SalR), which is specific for the production of morphinan alkaloids, was isolated and heterologously overexpressed in its active form not only from P. somniferum, but also from Papaver species that do not produce morphinan alkaloids. SalR is a member of a class of short chain dehydrogenase/reductases (SDRs) that are active as monomers and possess an extended amino acid sequence compared with classical SDRs. Homology modelling and substrate docking revealed the substrate binding site for SalR. The amino acids residues conferring salutaridine binding were compared to several members of the SDR family from different plant species, which non-specifically reduce (-)-menthone to (+)-neomenthol. Previously, it was shown that some of these proteins are involved in plant defence. The recruitment of specific monomeric SDRs from monomeric SDRs involved in plant defence is discussed.

  14. PsAP2 an AP2/ERF family transcription factor from Papaver somniferum enhances abiotic and biotic stress tolerance in transgenic tobacco.

    PubMed

    Mishra, Sonal; Phukan, Ujjal J; Tripathi, Vineeta; Singh, Dhananjay K; Luqman, Suaib; Shukla, Rakesh Kumar

    2015-09-01

    The AP2/ERFs are one of the most important family of transcription factors which regulate multiple responses like stress, metabolism and development in plants. We isolated PsAP2 a novel AP2/ERF from Papaver somniferum which was highly upregulated in response to wounding followed by ethylene, methyl jasmonate and ABA treatment. PsAP2 showed specific binding with both DRE and GCC box elements and it was able to transactivate the reporter genes in yeast. PsAP2 overexpressing transgenic tobacco plants exhibited enhanced tolerance towards both abiotic and biotic stresses . Real time transcript expression analysis showed constitutive upregulation of tobacco Alternative oxidase1a and Myo-inositol-1-phosphate synthase in PsAP2 overexpressing tobacco plants. Further, PsAP2 showed interaction with NtAOX1a promoter in vitro, it also specifically activated the NtAOX1a promoter in yeast and tobacco BY2 cells. The silencing of PsAP2 using VIGS lead to significant reduction in the AOX1 level in P. somniferum. Taken together PsAP2 can directly bind and transcriptionally activate NtAOX1a and its overexpression in tobacco imparted increased tolerance towards both abiotic and biotic stress.

  15. Recessive loci Pps-1 and OM differentially regulate PISTILLATA-1 and APETALA3-1 expression for sepal and petal development in Papaver somniferum.

    PubMed

    Singh, Sharad K; Shukla, Ashutosh K; Dhawan, Om P; Shasany, Ajit K

    2014-01-01

    The involvement of PISTILLATA (PI) and APETALA (AP) transcription factors in the development of floral organs has previously been elucidated but little is known about their upstream regulation. In this investigation, two novel mutants generated in Papaver somniferum were analyzed--one with partially petaloid sepals and another having sepaloid petals. Progeny from reciprocal crosses of respective mutant parent genotypes showed a good fit to the monogenic Mendelian inheritance model, indicating that the mutant traits are likely controlled by the single, recessive nuclear genes named "Pps-1" and "OM" in the partially petaloid sepal and sepaloid petal phenotypes, respectively. Both paralogs of PISTILLATA (PapsPI-1 and PapsPI-3) were obtained from the sepals and petals of P. somniferum. Ectopic expression of PapsPI-1 in tobacco resulted in a partially petaloid sepal phenotype at a low frequency. Upregulation of PapsPI-1 and PapsAP3-1 in the petal and the petal part of partially petaloid sepal mutant and down-regulation of the same in sepaloid petal mutant indicates a differential pattern of regulation for flowering-related genes in various whorls. Similarly, it was found that the recessive mutation OM in sepaloid petal mutant downregulates PapsPI-1 and PapsAP3-1 transcripts. The recessive nature of the mutations was confirmed by the segregation ratios obtained in this analysis.

  16. PsAP2 an AP2/ERF family transcription factor from Papaver somniferum enhances abiotic and biotic stress tolerance in transgenic tobacco.

    PubMed

    Mishra, Sonal; Phukan, Ujjal J; Tripathi, Vineeta; Singh, Dhananjay K; Luqman, Suaib; Shukla, Rakesh Kumar

    2015-09-01

    The AP2/ERFs are one of the most important family of transcription factors which regulate multiple responses like stress, metabolism and development in plants. We isolated PsAP2 a novel AP2/ERF from Papaver somniferum which was highly upregulated in response to wounding followed by ethylene, methyl jasmonate and ABA treatment. PsAP2 showed specific binding with both DRE and GCC box elements and it was able to transactivate the reporter genes in yeast. PsAP2 overexpressing transgenic tobacco plants exhibited enhanced tolerance towards both abiotic and biotic stresses . Real time transcript expression analysis showed constitutive upregulation of tobacco Alternative oxidase1a and Myo-inositol-1-phosphate synthase in PsAP2 overexpressing tobacco plants. Further, PsAP2 showed interaction with NtAOX1a promoter in vitro, it also specifically activated the NtAOX1a promoter in yeast and tobacco BY2 cells. The silencing of PsAP2 using VIGS lead to significant reduction in the AOX1 level in P. somniferum. Taken together PsAP2 can directly bind and transcriptionally activate NtAOX1a and its overexpression in tobacco imparted increased tolerance towards both abiotic and biotic stress. PMID:26319514

  17. Recessive loci Pps-1 and OM differentially regulate PISTILLATA-1 and APETALA3-1 expression for sepal and petal development in Papaver somniferum.

    PubMed

    Singh, Sharad K; Shukla, Ashutosh K; Dhawan, Om P; Shasany, Ajit K

    2014-01-01

    The involvement of PISTILLATA (PI) and APETALA (AP) transcription factors in the development of floral organs has previously been elucidated but little is known about their upstream regulation. In this investigation, two novel mutants generated in Papaver somniferum were analyzed--one with partially petaloid sepals and another having sepaloid petals. Progeny from reciprocal crosses of respective mutant parent genotypes showed a good fit to the monogenic Mendelian inheritance model, indicating that the mutant traits are likely controlled by the single, recessive nuclear genes named "Pps-1" and "OM" in the partially petaloid sepal and sepaloid petal phenotypes, respectively. Both paralogs of PISTILLATA (PapsPI-1 and PapsPI-3) were obtained from the sepals and petals of P. somniferum. Ectopic expression of PapsPI-1 in tobacco resulted in a partially petaloid sepal phenotype at a low frequency. Upregulation of PapsPI-1 and PapsAP3-1 in the petal and the petal part of partially petaloid sepal mutant and down-regulation of the same in sepaloid petal mutant indicates a differential pattern of regulation for flowering-related genes in various whorls. Similarly, it was found that the recessive mutation OM in sepaloid petal mutant downregulates PapsPI-1 and PapsAP3-1 transcripts. The recessive nature of the mutations was confirmed by the segregation ratios obtained in this analysis. PMID:24979593

  18. An assessment of the utility of universal and specific genetic markers for opium poppy identification.

    PubMed

    Lee, Eun J; Hwang, In K; Kim, Nam Y; Lee, Kyung L; Han, Myun S; Lee, Yang H; Kim, Mu Y; Yang, Moon S

    2010-09-01

    The proper identification of illicit plants such as Papaver somniferum L (opium poppy) is important for law enforcement agencies. The identification of opium poppy was presently tested using 10 genetic markers that are universal for all plants or specific to a few poppy plants. The genetic distances of universal markers such as nuclear internal transcribed spacer (ITS), 18S rRNA, plastid rbcL, and trnL-trnF intergenic spacer (IGS) of 14 species included in the Papaveraceae and Fumariaceae family were acquired by sequence comparisons. Both the ITS region and trnL-trnF IGS showed high levels of interspecific divergence. Six Papaver genera-specific markers were developed from coding regions involved in morphine biosynthesis. Three markers (TYDC, NCS, and BBE) produced amplicons only in opium poppy, providing a presence/absence test for opium poppy, while three additional markers (CYP80B1, SAT, and COR) were genus specific. These 10 markers might be useful for the forensic DNA analysis of opium poppy.

  19. Papaverine 7-O-demethylase, a novel 2-oxoglutarate/Fe(2+)-dependent dioxygenase from opium poppy.

    PubMed

    Farrow, Scott C; Facchini, Peter J

    2015-09-14

    Opium poppy (Papaver somniferum) produces several pharmacologically important benzylisoquinoline alkaloids including the vasodilator papaverine. Pacodine and palaudine are tri-O-methylated analogs of papaverine, which contains four O-linked methyl groups. However, the biosynthetic origin of pacodine and palaudine has not been established. Three members of the 2-oxoglutarate/Fe(2+)-dependent dioxygenases (2ODDs) family in opium poppy display widespread O-dealkylation activity on several benzylisoquinoline alkaloids with diverse structural scaffolds, and two are responsible for the antepenultimate and ultimate steps in morphine biosynthesis. We report a novel 2ODD from opium poppy catalyzing the efficient substrate- and regio-specific 7-O-demethylation of papaverine yielding pacodine. The occurrence of papaverine 7-O-demethylase (P7ODM) expands the enzymatic scope of the 2ODD family in opium poppy and suggests an unexpected biosynthetic route to pacodine.

  20. Papaverine 7-O-demethylase, a novel 2-oxoglutarate/Fe(2+)-dependent dioxygenase from opium poppy.

    PubMed

    Farrow, Scott C; Facchini, Peter J

    2015-09-14

    Opium poppy (Papaver somniferum) produces several pharmacologically important benzylisoquinoline alkaloids including the vasodilator papaverine. Pacodine and palaudine are tri-O-methylated analogs of papaverine, which contains four O-linked methyl groups. However, the biosynthetic origin of pacodine and palaudine has not been established. Three members of the 2-oxoglutarate/Fe(2+)-dependent dioxygenases (2ODDs) family in opium poppy display widespread O-dealkylation activity on several benzylisoquinoline alkaloids with diverse structural scaffolds, and two are responsible for the antepenultimate and ultimate steps in morphine biosynthesis. We report a novel 2ODD from opium poppy catalyzing the efficient substrate- and regio-specific 7-O-demethylation of papaverine yielding pacodine. The occurrence of papaverine 7-O-demethylase (P7ODM) expands the enzymatic scope of the 2ODD family in opium poppy and suggests an unexpected biosynthetic route to pacodine. PMID:26264169

  1. Benzylisoquinoline alkaloid biosynthesis in opium poppy.

    PubMed

    Beaudoin, Guillaume A W; Facchini, Peter J

    2014-07-01

    Opium poppy (Papaver somniferum) is one of the world's oldest medicinal plants and remains the only commercial source for the narcotic analgesics morphine, codeine and semi-synthetic derivatives such as oxycodone and naltrexone. The plant also produces several other benzylisoquinoline alkaloids with potent pharmacological properties including the vasodilator papaverine, the cough suppressant and potential anticancer drug noscapine and the antimicrobial agent sanguinarine. Opium poppy has served as a model system to investigate the biosynthesis of benzylisoquinoline alkaloids in plants. The application of biochemical and functional genomics has resulted in a recent surge in the discovery of biosynthetic genes involved in the formation of major benzylisoquinoline alkaloids in opium poppy. The availability of extensive biochemical genetic tools and information pertaining to benzylisoquinoline alkaloid metabolism is facilitating the study of a wide range of phenomena including the structural biology of novel catalysts, the genomic organization of biosynthetic genes, the cellular and sub-cellular localization of biosynthetic enzymes and a variety of biotechnological applications. In this review, we highlight recent developments and summarize the frontiers of knowledge regarding the biochemistry, cellular biology and biotechnology of benzylisoquinoline alkaloid biosynthesis in opium poppy.

  2. Benzylisoquinoline alkaloid biosynthesis in opium poppy.

    PubMed

    Beaudoin, Guillaume A W; Facchini, Peter J

    2014-07-01

    Opium poppy (Papaver somniferum) is one of the world's oldest medicinal plants and remains the only commercial source for the narcotic analgesics morphine, codeine and semi-synthetic derivatives such as oxycodone and naltrexone. The plant also produces several other benzylisoquinoline alkaloids with potent pharmacological properties including the vasodilator papaverine, the cough suppressant and potential anticancer drug noscapine and the antimicrobial agent sanguinarine. Opium poppy has served as a model system to investigate the biosynthesis of benzylisoquinoline alkaloids in plants. The application of biochemical and functional genomics has resulted in a recent surge in the discovery of biosynthetic genes involved in the formation of major benzylisoquinoline alkaloids in opium poppy. The availability of extensive biochemical genetic tools and information pertaining to benzylisoquinoline alkaloid metabolism is facilitating the study of a wide range of phenomena including the structural biology of novel catalysts, the genomic organization of biosynthetic genes, the cellular and sub-cellular localization of biosynthetic enzymes and a variety of biotechnological applications. In this review, we highlight recent developments and summarize the frontiers of knowledge regarding the biochemistry, cellular biology and biotechnology of benzylisoquinoline alkaloid biosynthesis in opium poppy. PMID:24671624

  3. Traditional ecologies of the opium poppy and oral history in rural Turkey.

    PubMed

    Evered, Kyle T

    2011-01-01

    Cultivated in the Eastern Mediterranean region for millennia, the opium poppy (Papaver somniferum) was profoundly significant in the economies, ecologies, cultures, and diets of the peoples of many towns and villages of rural Anatolia. When the United States compelled Turkey to eradicate cultivation of the plant in the early 1970s in order to diminish the flow of heroin into America, farmers were obliged to deal with not only changes in their incomes but also profound changes in their relationships with the land and the state. Although Turkish officials later allowed production to resume in a highly controlled manner for pharmaceutical purposes, significant socioeconomic and ecological dimensions of Turkey's poppy-growing communities were forever changed. Interviewing now-retired poppy farmers, I employ oral history as my primary source of historical evidence to reconstruct these past ecologies and associated social relationships and to give voice to the informants.

  4. Thermooxidative stability of poppy seeds studied by non-isothermal DSC measurements.

    PubMed

    Cibulková, Zuzana; Čertík, Milan; Dubaj, Tibor

    2014-05-01

    Papaver somniferum L. is an important crop cultivated mostly for seed production. Poppy seeds have a high nutritive value and are used as a food and as a source of edible oil. This oil is a rich source of polyunsaturated fatty acids. It is well known that the unsaturated fatty acids easily undergo oxidation reactions, which lead to the reduction of shelf life, nutritional quality, development of unpleasant tastes and odors. The goal of this study was to develop the methodology for testing the stability of poppy seeds using non-isothermal DSC. For the treatment of the experimental data a method based on non-Arrhenian temperature function has been applied and the values the kinetic parameters have been obtained. In order to assess the durability of the commercial poppy seeds, the lengths of induction periods have been calculated.

  5. Traditional ecologies of the opium poppy and oral history in rural Turkey.

    PubMed

    Evered, Kyle T

    2011-01-01

    Cultivated in the Eastern Mediterranean region for millennia, the opium poppy (Papaver somniferum) was profoundly significant in the economies, ecologies, cultures, and diets of the peoples of many towns and villages of rural Anatolia. When the United States compelled Turkey to eradicate cultivation of the plant in the early 1970s in order to diminish the flow of heroin into America, farmers were obliged to deal with not only changes in their incomes but also profound changes in their relationships with the land and the state. Although Turkish officials later allowed production to resume in a highly controlled manner for pharmaceutical purposes, significant socioeconomic and ecological dimensions of Turkey's poppy-growing communities were forever changed. Interviewing now-retired poppy farmers, I employ oral history as my primary source of historical evidence to reconstruct these past ecologies and associated social relationships and to give voice to the informants. PMID:21954490

  6. Genetic and chemical components analysis of Papaver setigerum naturalized in Korea.

    PubMed

    Choe, Sanggil; Lee, Eunjung; Jin, Gang-nam; Lee, Yang Han; Kim, Soo Young; Choi, Hwakyung; Chung, Heesun; Hwang, Bang Yeon; Kim, Suncheun

    2012-10-10

    Of the 110 species of genus Papaver, only Papaver somniferum and P. setigerum are controlled poppies in Korea. All poppy samples share similar morphology therefore it is important to check if they contain controlled substances such as morphine and codeine for forensic purpose. Since the alkaloid content of Papaver plants varies according to their growing stage, chemical components analysis alone is not enough to identify exact species. In 2010, hundreds of poppy plants suspected to be P. somniferum were found in Jeju Island, South Korea. They had a slightly different but overall similar appearance to P. somniferum. Using GC-MS analysis, codeine, rhoeadine, papaverine, protopine, noscapine, setigeridine and trace amounts of morphine were detected in these samples. Although their chemical components were different from what has been described in literatures for P. setigerum, they could be assumed to be P. setigerum based on their morphological features and GC-MS results. Also, chromosome numbers using their seeds showed 2n=44 and the numbers were in accordance with those of P. setigerum. Nucleotide substitution or insertion/deletion of ITS (internal transcribed spacer), 18S rRNA (ribosomal RNA), rbcL (large subunit of ribulose 1,5-bisphosphate carboxylase), trnL-trnF IGS (intergenic spacer), trnL intron and psbA-trnH were assessed as universal genetic markers for P. setigerum. Also, genetic analysis using six target genes involved in the biosynthesis of benzylisoquinoline alkaloids, including TYDC (tyrosine/dopa decarboxylase), SAT (salutaridinol-7-O-acetyltransferase), BBE (berberine bridge enzyme), COR (codeinone reductase), CYP80B1 ((S)-N-methylcoclaurine 3'-hydroxylase) and NCS (norcoclaurine synthase) were tested as Papaver-specific genetic markers by the existence of their PCR products. From the results, the sequences of the 6 universal genetic markers and 6 Papaver-specific genetic markers for P. setigerum were identified and then Genbank accession numbers of

  7. Stereochemical inversion of (S)-reticuline by a cytochrome P450 fusion in opium poppy.

    PubMed

    Farrow, Scott C; Hagel, Jillian M; Beaudoin, Guillaume A W; Burns, Darcy C; Facchini, Peter J

    2015-09-01

    The gateway to morphine biosynthesis in opium poppy (Papaver somniferum) is the stereochemical inversion of (S)-reticuline since the enzyme yielding the first committed intermediate salutaridine is specific for (R)-reticuline. A fusion between a cytochrome P450 (CYP) and an aldo-keto reductase (AKR) catalyzes the S-to-R epimerization of reticuline via 1,2-dehydroreticuline. The reticuline epimerase (REPI) fusion was detected in opium poppy and in Papaver bracteatum, which accumulates thebaine. In contrast, orthologs encoding independent CYP and AKR enzymes catalyzing the respective synthesis and reduction of 1,2-dehydroreticuline were isolated from Papaver rhoeas, which does not accumulate morphinan alkaloids. An ancestral relationship between these enzymes is supported by a conservation of introns in the gene fusions and independent orthologs. Suppression of REPI transcripts using virus-induced gene silencing in opium poppy reduced levels of (R)-reticuline and morphinan alkaloids and increased the overall abundance of (S)-reticuline and its O-methylated derivatives. Discovery of REPI completes the isolation of genes responsible for known steps of morphine biosynthesis.

  8. Stereochemical inversion of (S)-reticuline by a cytochrome P450 fusion in opium poppy.

    PubMed

    Farrow, Scott C; Hagel, Jillian M; Beaudoin, Guillaume A W; Burns, Darcy C; Facchini, Peter J

    2015-09-01

    The gateway to morphine biosynthesis in opium poppy (Papaver somniferum) is the stereochemical inversion of (S)-reticuline since the enzyme yielding the first committed intermediate salutaridine is specific for (R)-reticuline. A fusion between a cytochrome P450 (CYP) and an aldo-keto reductase (AKR) catalyzes the S-to-R epimerization of reticuline via 1,2-dehydroreticuline. The reticuline epimerase (REPI) fusion was detected in opium poppy and in Papaver bracteatum, which accumulates thebaine. In contrast, orthologs encoding independent CYP and AKR enzymes catalyzing the respective synthesis and reduction of 1,2-dehydroreticuline were isolated from Papaver rhoeas, which does not accumulate morphinan alkaloids. An ancestral relationship between these enzymes is supported by a conservation of introns in the gene fusions and independent orthologs. Suppression of REPI transcripts using virus-induced gene silencing in opium poppy reduced levels of (R)-reticuline and morphinan alkaloids and increased the overall abundance of (S)-reticuline and its O-methylated derivatives. Discovery of REPI completes the isolation of genes responsible for known steps of morphine biosynthesis. PMID:26147354

  9. Evolution and diversity of the mechanisms endowing resistance to herbicides inhibiting acetolactate-synthase (ALS) in corn poppy (Papaver rhoeas L.).

    PubMed

    Délye, Christophe; Pernin, Fanny; Scarabel, Laura

    2011-02-01

    We investigated the diversity of mechanisms conferring resistance to herbicides inhibiting acetolactate synthase (ALS) in corn poppy (Papaver rhoeas L.) and the processes underlying the selection for resistance. Six mutant ALS alleles, Arg₁₉₇, His₁₉₇, Leu₁₉₇, Ser₁₉₇, Thr₁₉₇ and Leu₅₇₄ were identified in five Italian populations. Different alleles were found in a same population or a same plant. Comparison of individual plant phenotype (herbicide sensitivity) and genotype (amino-acid substitution(s) at codon 197) showed that all mutant ALS alleles conferred dominant resistance to the field rate of the sulfonylurea tribenuron and moderate or no resistance to the field rate of the triazolopyrimidine florasulam. Depending on the allele, dominant or partially dominant resistance to the field rate of the imidazolinone imazamox was observed. Putative non-target-site resistance mechanisms were also likely present in the populations investigated. The derived Cleaved Amplified Polymorphic Sequence assays targeting ALS codons crucial for herbicide sensitivity developed in this work will facilitate the detection of resistance due to mutant ALS alleles. Nucleotide variation around codon 197 indicated that mutant ALS alleles evolved by multiple, independent appearances. Resistance to ALS inhibitors in P. rhoeas clearly evolved by redundant evolution of a set of mutant ALS alleles and likely of non-target-site mechanisms.

  10. Evolution and diversity of the mechanisms endowing resistance to herbicides inhibiting acetolactate-synthase (ALS) in corn poppy (Papaver rhoeas L.).

    PubMed

    Délye, Christophe; Pernin, Fanny; Scarabel, Laura

    2011-02-01

    We investigated the diversity of mechanisms conferring resistance to herbicides inhibiting acetolactate synthase (ALS) in corn poppy (Papaver rhoeas L.) and the processes underlying the selection for resistance. Six mutant ALS alleles, Arg₁₉₇, His₁₉₇, Leu₁₉₇, Ser₁₉₇, Thr₁₉₇ and Leu₅₇₄ were identified in five Italian populations. Different alleles were found in a same population or a same plant. Comparison of individual plant phenotype (herbicide sensitivity) and genotype (amino-acid substitution(s) at codon 197) showed that all mutant ALS alleles conferred dominant resistance to the field rate of the sulfonylurea tribenuron and moderate or no resistance to the field rate of the triazolopyrimidine florasulam. Depending on the allele, dominant or partially dominant resistance to the field rate of the imidazolinone imazamox was observed. Putative non-target-site resistance mechanisms were also likely present in the populations investigated. The derived Cleaved Amplified Polymorphic Sequence assays targeting ALS codons crucial for herbicide sensitivity developed in this work will facilitate the detection of resistance due to mutant ALS alleles. Nucleotide variation around codon 197 indicated that mutant ALS alleles evolved by multiple, independent appearances. Resistance to ALS inhibitors in P. rhoeas clearly evolved by redundant evolution of a set of mutant ALS alleles and likely of non-target-site mechanisms. PMID:21421378

  11. Enhancement of alkaloid production in opium and California poppy by transactivation using heterologous regulatory factors.

    PubMed

    Apuya, Nestor R; Park, Joon-Hyun; Zhang, Liping; Ahyow, Maurice; Davidow, Patricia; Van Fleet, Jennifer; Rarang, Joel C; Hippley, Matthew; Johnson, Thomas W; Yoo, Hye-Dong; Trieu, Anthony; Krueger, Shannon; Wu, Chuan-yin; Lu, Yu-ping; Flavell, Richard B; Bobzin, Steven C

    2008-02-01

    Genes encoding regulatory factors isolated from Arabidopsis, soybean and corn have been screened to identify those that modulate the expression of genes encoding for enzymes involved in the biosynthesis of morphinan alkaloids in opium poppy (Papaver somniferum) and benzophenanthridine alkaloids in California poppy (Eschscholzia californica). In opium poppy, the over-expression of selected regulatory factors increased the levels of PsCOR (codeinone reductase), Ps4'OMT (S-adenosyl-l-methionine:3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase) and Ps6OMT [(R,S)-norcoclaurine 6-O-methyltransferase] transcripts by 10- to more than 100-fold. These transcriptional activations translated into an enhancement of alkaloid production in opium poppy of up to at least 10-fold. In California poppy, the transactivation effect of regulatory factor WRKY1 resulted in an increase of up to 60-fold in the level of EcCYP80B1 [(S)-N-methylcoclaurine 3'-hydroxylase] and EcBBE (berberine bridge enzyme) transcripts. As a result, the accumulations of selected alkaloid intermediates were enhanced up to 30-fold. The transactivation effects of other regulatory factors led to the accumulation of the same intermediates. These regulatory factors also led to the production of new alkaloids in California poppy callus culture.

  12. Enhancement of alkaloid production in opium and California poppy by transactivation using heterologous regulatory factors.

    PubMed

    Apuya, Nestor R; Park, Joon-Hyun; Zhang, Liping; Ahyow, Maurice; Davidow, Patricia; Van Fleet, Jennifer; Rarang, Joel C; Hippley, Matthew; Johnson, Thomas W; Yoo, Hye-Dong; Trieu, Anthony; Krueger, Shannon; Wu, Chuan-yin; Lu, Yu-ping; Flavell, Richard B; Bobzin, Steven C

    2008-02-01

    Genes encoding regulatory factors isolated from Arabidopsis, soybean and corn have been screened to identify those that modulate the expression of genes encoding for enzymes involved in the biosynthesis of morphinan alkaloids in opium poppy (Papaver somniferum) and benzophenanthridine alkaloids in California poppy (Eschscholzia californica). In opium poppy, the over-expression of selected regulatory factors increased the levels of PsCOR (codeinone reductase), Ps4'OMT (S-adenosyl-l-methionine:3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase) and Ps6OMT [(R,S)-norcoclaurine 6-O-methyltransferase] transcripts by 10- to more than 100-fold. These transcriptional activations translated into an enhancement of alkaloid production in opium poppy of up to at least 10-fold. In California poppy, the transactivation effect of regulatory factor WRKY1 resulted in an increase of up to 60-fold in the level of EcCYP80B1 [(S)-N-methylcoclaurine 3'-hydroxylase] and EcBBE (berberine bridge enzyme) transcripts. As a result, the accumulations of selected alkaloid intermediates were enhanced up to 30-fold. The transactivation effects of other regulatory factors led to the accumulation of the same intermediates. These regulatory factors also led to the production of new alkaloids in California poppy callus culture. PMID:17961129

  13. New Physicochemical Treatment Method of Poppy Seeds for Agriculture and Food Industries

    NASA Astrophysics Data System (ADS)

    Božena, Šerá; Iveta, Gajdová; Michal, Šerý; Petr, Špatenka

    2013-09-01

    Poppy seeds (Papaver somniferum L.) are often attacked by various fungal diseases, and their field germination rate is low. The aim of this experiment was to increase the germination rate and growth of seedlings by modifying the poppy seeds using cold plasma discharge. The seeds were treated in a Plasonic AR-550-M under power of 500 W with argon gas flow of 50 mL/min, oxygen gas flow of 50 mL/min for different time durations in seconds (0, 180, 300, 600, 1800, 3000, 4200, and 5400). The seed germination rate and growth of the seedlings were recorded. Cold plasma positively affected the seed germination rate for time exposure between 180~3000 s. Seedling acceleration on the sixth day of the experiment showed the highest values at exposure 180~600 s. Cold plasma seems to be a good physicochemical way to modify seeds without chemical agronomical application.

  14. A FILAMENTOUS FLOWER orthologue plays a key role in leaf patterning in opium poppy.

    PubMed

    Vosnakis, Nikolaos; Maiden, Afiqah; Kourmpetli, Sofia; Hands, Philip; Sharples, Donna; Drea, Sinéad

    2012-11-01

    The plant-specific YABBY genes were initially defined by their roles in determining abaxial/adaxial cell fate in lateral organs of eudicots, and repressing meristematic genes in differentiating tissues such as leaves. In Arabidopsis thaliana FILAMENTOUS FLOWER (FIL) is also required for inflorescence and floral meristem establishment and flower development in a pathway involving the floral transition and identity genes. Here we describe the characterization of a FIL orthologue from the basal eudicot, Papaver somniferum (the opium poppy), and demonstrate a role for the gene in patterning the highly lobed leaf of the poppy. Silencing of PapsFIL using viral-induced gene silencing resulted in leaves of reduced laminar area, more pronounced margin serration and, in some cases, leaf bifurcation. In contrast, the gene does not appear to affect the development of the flower, and these variations in function are discussed in relation to its taxonomic position as a basal eudicot and its determinate growth habit.

  15. A nested-polymerase chain reaction protocol for detection and population biology studies of Peronospora arborescens, the downy mildew pathogen of opium poppy, using herbarium specimens and asymptomatic, fresh plant tissues.

    PubMed

    Montes-Borrego, Miguel; Muñoz Ledesma, Francisco J; Jiménez-Díaz, Rafael M; Landa, Blanca B

    2009-01-01

    A sensitive nested-polymerase chain reaction (PCR) protocol was developed using either of two primer pairs that improves the in planta detection of Peronospora arborescens DNA. The new protocol represented an increase in sensitivity of 100- to 1,000-fold of detection of the oomycete in opium poppy tissue compared with the detection limit of single PCR using the same primer pairs. The new protocol allowed amplification of 5 to 0.5 fg of Peronospora arborescens DNA mixed with Papaver somniferum DNA. The protocol proved useful for amplifying Peronospora arborescens DNA from 96-year-old herbarium specimens of Papaver spp. and to demonstrate that asymptomatic, systemic infections by Peronospora arborescens can occur in wild Papaver spp. as well as in cultivated opium poppy. Also, the increase in sensitivity of the protocol made possible the detection of seedborne Peronospora arborescens in commercial opium poppy seed stocks in Spain with a high frequency, which poses a threat for pathogen spread. Direct sequencing of purified amplicons allowed alignment of a Peronospora arborescens internal transcribed spacer (ITS) ribosomal DNA (rDNA) sequence up to 730-bp long when combining the sequences obtained with the two primer sets. Maximum parsimony analysis of amplified Peronospora arborescens ITS rDNA sequences from specimens of Papaver dubium, P. hybridum, P. rhoeas, and P. somniferum from different countries indicated for the first time that a degree of host specificity may exist within populations of Peronospora arborescens. The reported protocol will be useful for epidemiological and biogeographical studies of downy mildew diseases as well as to unravel misclassification of Peronospora arborescens and Peronospora cristata, the reported causal agents of the opium poppy downy mildew disease.

  16. Comparative transcript and alkaloid profiling in Papaver species identifies a short chain dehydrogenase/reductase involved in morphine biosynthesis.

    PubMed

    Ziegler, Jörg; Voigtländer, Susan; Schmidt, Jürgen; Kramell, Robert; Miersch, Otto; Ammer, Christian; Gesell, Andreas; Kutchan, Toni M

    2006-10-01

    Plants of the order Ranunculales, especially members of the species Papaver, accumulate a large variety of benzylisoquinoline alkaloids with about 2500 structures, but only the opium poppy (Papaver somniferum) and Papaver setigerum are able to produce the analgesic and narcotic morphine and the antitussive codeine. In this study, we investigated the molecular basis for this exceptional biosynthetic capability by comparison of alkaloid profiles with gene expression profiles between 16 different Papaver species. Out of 2000 expressed sequence tags obtained from P. somniferum, 69 show increased expression in morphinan alkaloid-containing species. One of these cDNAs, exhibiting an expression pattern very similar to previously isolated cDNAs coding for enzymes in benzylisoquinoline biosynthesis, showed the highest amino acid identity to reductases in menthol biosynthesis. After overexpression, the protein encoded by this cDNA reduced the keto group of salutaridine yielding salutaridinol, an intermediate in morphine biosynthesis. The stereoisomer 7-epi-salutaridinol was not formed. Based on its similarities to a previously purified protein from P. somniferum with respect to the high substrate specificity, molecular mass and kinetic data, the recombinant protein was identified as salutaridine reductase (SalR; EC 1.1.1.248). Unlike codeinone reductase, an enzyme acting later in the pathway that catalyses the reduction of a keto group and which belongs to the family of the aldo-keto reductases, the cDNA identified in this study as SalR belongs to the family of short chain dehydrogenases/reductases and is related to reductases in monoterpene metabolism.

  17. Analysis of fluid extracts obtained from Papaver rhoeas petals contaminated with Papaver bracteatum petals.

    PubMed

    Gambaro, Veniero; Minghetti, Paola; Arnoldi, Sebastiano; Colombo, Maria Laura; Dellʼacqua, Lucia; Casiraghi, Antonella; Guerrini, Katia; Farè, Fiorenza; Roda, Gabriella

    2012-08-01

    In this paper, we report a case of misidentification of medicinal plants involving dried petals of Papaver rhoeas (red poppy) contaminated with Papaver bracteatum (scarlet poppy) petals. Preliminary TLC analysis indicated the presence of thebaine either in the fluid extracts or in the petals. It was therefore necessary to carry out an accurate botanic examination of the plant material, which revealed contamination of the red poppy petals with scarlet poppy petals. Moreover, to confirm the adulteration, we developed and validated an efficient, reversed-phase ion pair HPLC method for determination of the alkaloids specific for the Papaver species. Six petal batches and five commercial fluid extracts were analyzed. Only one petal batch from Iran contained thebaine and its analogue oripavine while the alkaloids typical for the Papaver bracteatum species were identified in all fluid extracts, meaning that they were all prepared with contaminated petals.

  18. Sequence analysis of two new members of the major latex protein gene family supports the triploid-hybrid origin of the opium poppy.

    PubMed

    Nessler, C L

    1994-02-25

    Latex from the opium poppy (Papaver somniferum) contains an abundant group of laticifer-specific, low-molecular-weight polypeptides called the major latex proteins (MLPs). MLPs are encoded by a family of nine genes which can be divided into two distinct subfamilies based on DNA gel blot analysis. This report describes two new members of the MLP gene family (MLP146 and MLP149) that are physically linked and separated by approx. 5.5 kb. Comparison of their nucleotide and deduced amino-acid sequences with those of other MLP indicates that both new MLP belong to the MLP15 subfamily. The organization of the MLP family is consistent with the triploid-hybrid origin of the opium poppy.

  19. Tyrosine aminotransferase contributes to benzylisoquinoline alkaloid biosynthesis in opium poppy.

    PubMed

    Lee, Eun-Jeong; Facchini, Peter J

    2011-11-01

    Tyrosine aminotransferase (TyrAT) catalyzes the transamination of L-Tyr and α-ketoglutarate, yielding 4-hydroxyphenylpyruvic acid and L-glutamate. The decarboxylation product of 4-hydroxyphenylpyruvic acid, 4-hydroxyphenylacetaldehyde, is a precursor to a large and diverse group of natural products known collectively as benzylisoquinoline alkaloids (BIAs). We have isolated and characterized a TyrAT cDNA from opium poppy (Papaver somniferum), which remains the only commercial source for several pharmaceutical BIAs, including codeine, morphine, and noscapine. TyrAT belongs to group I pyridoxal 5'-phosphate (PLP)-dependent enzymes wherein Schiff base formation occurs between PLP and a specific Lys residue. The amino acid sequence of TyrAT showed considerable homology to other putative plant TyrATs, although few of these have been functionally characterized. Purified, recombinant TyrAT displayed a molecular mass of approximately 46 kD and a substrate preference for L-Tyr and α-ketoglutarate, with apparent K(m) values of 1.82 and 0.35 mm, respectively. No specific requirement for PLP was detected in vitro. Liquid chromatography-tandem mass spectrometry confirmed the conversion of L-Tyr to 4-hydroxyphenylpyruvate. TyrAT gene transcripts were most abundant in roots and stems of mature opium poppy plants. Virus-induced gene silencing was used to evaluate the contribution of TyrAT to BIA metabolism in opium poppy. TyrAT transcript levels were reduced by at least 80% in silenced plants compared with controls and showed a moderate reduction in total alkaloid content. The modest correlation between transcript levels and BIA accumulation in opium poppy supports a role for TyrAT in the generation of alkaloid precursors, but it also suggests the occurrence of other sources for 4-hydroxyphenylacetaldehyde.

  20. Recent and projected increases in atmospheric carbon dioxide and the potential impacts on growth and alkaloid production in wild poppy (Papaver setigerum DC.)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In the current study, we quantified changes in the growth and alkaloid content of wild poppy, (P. setigerum) as a function of recent and projected changes in global atmospheric carbon dioxide concentration, [CO2]. The experimental [CO2] values (300, 400, 500 and 600 µmol mol-1) correspond roughly t...

  1. AFLP studies on downy-mildew-resistant and downy-mildew-susceptible genotypes of opium poppy.

    PubMed

    Dubey, Mukesh K; Shasany, Ajit K; Dhawan, Om P; Shukla, Ashutosh K; Khanuja, Suman P S

    2010-04-01

    Downy mildew (DM) caused by Peronospora arborescens, is a serious disease in opium poppy (Papaver somniferum), which has a world-wide spread. The establishment of DM-resistant cultivars appears to be a sustainable way to control the In this paper, we present the results of a study aimed at the identification of amplified fragment length polymorphism (AFLP) markers for DM-resistance in opium poppy. Three opium poppy genotypes (inbred over about 10 years): Pps-1 (DM-resistant), Jawahar-16 (DM-susceptible) and H-9 (DM-susceptible) were crossed in a diallel manner and the F(1) progeny along with the parents were subjected to AFLP analysis of chloroplast (cp) and nuclear DNA with seven and nine EcoRI / MseI primer combinations, respectively. cpDNA AFLP analysis identified 24 Pps-1 (DM-resistant)-specific unique fragments that were found to be maternally inherited in both the crosses, Pps-1 x Jawahar-16 and Pps-1 x H-9. In the case of nuclear DNA AFLP analysis, it was found that 17 fragments inherited from Pps-1 were common to the reciprocal crosses of both (i) Pps-1 and Jawahar-16 as well as (ii) Pps-1 and H-9. This is the first molecular investigation on the identification of polymorphism between DM-resistant and DM-susceptible opium poppy genotypes and development of DM-resistant opium poppy genotypespecific AFLP markers. These AFLP markers could be used in future genetic studies for analysis of linkage to the downy mildew resistance trait.

  2. Characterization of three O-methyltransferases involved in noscapine biosynthesis in opium poppy.

    PubMed

    Dang, Thu-Thuy T; Facchini, Peter J

    2012-06-01

    Noscapine is a benzylisoquinoline alkaloid produced in opium poppy (Papaver somniferum) and other members of the Papaveraceae. It has been used as a cough suppressant and more recently was shown to possess anticancer activity. However, the biosynthesis of noscapine in opium poppy has not been established. A proposed pathway leading from (S)-reticuline to noscapine includes (S)-scoulerine, (S)-canadine, and (S)-N-methylcanadine as intermediates. Stem cDNA libraries and latex extracts of eight opium poppy cultivars displaying different alkaloid profiles were subjected to massively parallel pyrosequencing and liquid chromatography-tandem mass spectrometry, respectively. Comparative transcript and metabolite profiling revealed the occurrence of three cDNAs encoding O-methyltransferases designated as SOMT1, SOMT2, and SOMT3 that correlated with the accumulation of noscapine in the eight cultivars. SOMT transcripts were detected in all opium poppy organs but were most abundant in aerial organs, where noscapine primarily accumulates. SOMT2 and SOMT3 showed strict substrate specificity and regiospecificity as 9-O-methyltransferases targeting (S)-scoulerine. In contrast, SOMT1 was able to sequentially 9- and 2-O-methylate (S)-scoulerine, yielding (S)-tetrahydropalmatine. SOMT1 also sequentially 3'- and 7-O-methylated both (S)-norreticuline and (S)-reticuline with relatively high substrate affinity, yielding (S)-tetrahydropapaverine and (S)-laudanosine, respectively. The metabolic functions of SOMT1, SOMT2, and SOMT3 were investigated in planta using virus-induced gene silencing. Reduction of SOMT1 or SOMT2 transcript levels resulted in a significant decrease in noscapine accumulation. Reduced SOMT1 transcript levels also caused a decrease in papaverine accumulation, confirming the selective roles for these enzymes in the biosynthesis of both alkaloids in opium poppy. PMID:22535422

  3. Characterization of three O-methyltransferases involved in noscapine biosynthesis in opium poppy.

    PubMed

    Dang, Thu-Thuy T; Facchini, Peter J

    2012-06-01

    Noscapine is a benzylisoquinoline alkaloid produced in opium poppy (Papaver somniferum) and other members of the Papaveraceae. It has been used as a cough suppressant and more recently was shown to possess anticancer activity. However, the biosynthesis of noscapine in opium poppy has not been established. A proposed pathway leading from (S)-reticuline to noscapine includes (S)-scoulerine, (S)-canadine, and (S)-N-methylcanadine as intermediates. Stem cDNA libraries and latex extracts of eight opium poppy cultivars displaying different alkaloid profiles were subjected to massively parallel pyrosequencing and liquid chromatography-tandem mass spectrometry, respectively. Comparative transcript and metabolite profiling revealed the occurrence of three cDNAs encoding O-methyltransferases designated as SOMT1, SOMT2, and SOMT3 that correlated with the accumulation of noscapine in the eight cultivars. SOMT transcripts were detected in all opium poppy organs but were most abundant in aerial organs, where noscapine primarily accumulates. SOMT2 and SOMT3 showed strict substrate specificity and regiospecificity as 9-O-methyltransferases targeting (S)-scoulerine. In contrast, SOMT1 was able to sequentially 9- and 2-O-methylate (S)-scoulerine, yielding (S)-tetrahydropalmatine. SOMT1 also sequentially 3'- and 7-O-methylated both (S)-norreticuline and (S)-reticuline with relatively high substrate affinity, yielding (S)-tetrahydropapaverine and (S)-laudanosine, respectively. The metabolic functions of SOMT1, SOMT2, and SOMT3 were investigated in planta using virus-induced gene silencing. Reduction of SOMT1 or SOMT2 transcript levels resulted in a significant decrease in noscapine accumulation. Reduced SOMT1 transcript levels also caused a decrease in papaverine accumulation, confirming the selective roles for these enzymes in the biosynthesis of both alkaloids in opium poppy.

  4. Improved sanguinarine production via biotic and abiotic elicitations and precursor feeding in cell suspensions of latex-less variety of Papaver somniferum with their gene expression studies and upscaling in bioreactor.

    PubMed

    Verma, Priyanka; Khan, Shamshad Ahmad; Mathur, Ajay K; Ghosh, Sumit; Shanker, Karuna; Kalra, Alok

    2014-11-01

    Elicitors play an important role in challenging the plant defense system through plant-environment interaction and thus altering the secondary metabolite production. Culture filtrates of four endophytic fungi, namely, Chaetomium globosum, Aspergillus niveoglaucus, Paecilomyces lilacinus, and Trichoderma harzianum were tested on embryogenic cell suspensions of latex-less Papaver somniferum in dose-dependent kinetics. Besides this, abiotic elicitors salicylic acid, hydrogen peroxide, and carbon dioxide were also applied for improved sanguinarine production. Maximum biomass accumulation (growth index (GI) = 293.50 ± 14.82) and sanguinarine production (0.090 ± 0.008 % dry wt.) were registered by addition of 3.3 % v/v T. harzanium culture filtrate. Interestingly, it was further enhanced (GI = 323.40 ± 25.30; 0.105 ± 0.008 % dry wt.) when T. harzanium culture filtrate was employed along with 50 μM shikimate. This was also supported by real-time (RT) (qPCR), where 8-9-fold increase in cheilanthifoline synthase (CFS), stylopine synthase (STS), tetrahydroprotoberberine cis-N-methyltransferase (TNMT), and protopine 6-hydroxylase (P6H) transcripts was observed. Among abiotic elicitors, while hydrogen peroxide and carbon dioxide registered low level of sanguinarine accumulation, maximum sanguinarine content was detected by 250 μM salicylic acid (0.058 ± 0.003 % dry wt.; GI = 172.75 ± 13.40). RT (qPCR) also confirms the downregulation of sanguinarine pathway on CO2 supplementation. Various parameters ranging from agitation speed (70 rpm), impeller type (marine), media volume (2 l), inoculum weight (100 g), and culture duration (9 days) were optimized during upscaling in 5-l stirred tank bioreactor to obtain maximum sanguinarine production (GI = 434.00; 0.119 ± 0.070 % dry wt.). Addition of 3.3 % v/v T. harzanium culture filtrate and 50-μM shikimate was done on the 6th day of bioreactor run.

  5. Characterization of a flavoprotein oxidase from opium poppy catalyzing the final steps in sanguinarine and papaverine biosynthesis.

    PubMed

    Hagel, Jillian M; Beaudoin, Guillaume A W; Fossati, Elena; Ekins, Andrew; Martin, Vincent J J; Facchini, Peter J

    2012-12-14

    Benzylisoquinoline alkaloids are a diverse class of plant specialized metabolites that includes the analgesic morphine, the antimicrobials sanguinarine and berberine, and the vasodilator papaverine. The two-electron oxidation of dihydrosanguinarine catalyzed by dihydrobenzophenanthridine oxidase (DBOX) is the final step in sanguinarine biosynthesis. The formation of the fully conjugated ring system in sanguinarine is similar to the four-electron oxidations of (S)-canadine to berberine and (S)-tetrahydropapaverine to papaverine. We report the isolation and functional characterization of an opium poppy (Papaver somniferum) cDNA encoding DBOX, a flavoprotein oxidase with homology to (S)-tetrahydroprotoberberine oxidase and the berberine bridge enzyme. A query of translated opium poppy stem transcriptome databases using berberine bridge enzyme yielded several candidate genes, including an (S)-tetrahydroprotoberberine oxidase-like sequence selected for heterologous expression in Pichia pastoris. The recombinant enzyme preferentially catalyzed the oxidation of dihydrosanguinarine to sanguinarine but also converted (RS)-tetrahydropapaverine to papaverine and several protoberberine alkaloids to oxidized forms, including (RS)-canadine to berberine. The K(m) values of 201 and 146 μm for dihydrosanguinarine and the protoberberine alkaloid (S)-scoulerine, respectively, suggested high concentrations of these substrates in the plant. Virus-induced gene silencing to reduce DBOX transcript levels resulted in a corresponding reduction in sanguinarine, dihydrosanguinarine, and papaverine accumulation in opium poppy roots in support of DBOX as a multifunctional oxidative enzyme in BIA metabolism.

  6. Characterization of a Flavoprotein Oxidase from Opium Poppy Catalyzing the Final Steps in Sanguinarine and Papaverine Biosynthesis*

    PubMed Central

    Hagel, Jillian M.; Beaudoin, Guillaume A. W.; Fossati, Elena; Ekins, Andrew; Martin, Vincent J. J.; Facchini, Peter J.

    2012-01-01

    Benzylisoquinoline alkaloids are a diverse class of plant specialized metabolites that includes the analgesic morphine, the antimicrobials sanguinarine and berberine, and the vasodilator papaverine. The two-electron oxidation of dihydrosanguinarine catalyzed by dihydrobenzophenanthridine oxidase (DBOX) is the final step in sanguinarine biosynthesis. The formation of the fully conjugated ring system in sanguinarine is similar to the four-electron oxidations of (S)-canadine to berberine and (S)-tetrahydropapaverine to papaverine. We report the isolation and functional characterization of an opium poppy (Papaver somniferum) cDNA encoding DBOX, a flavoprotein oxidase with homology to (S)-tetrahydroprotoberberine oxidase and the berberine bridge enzyme. A query of translated opium poppy stem transcriptome databases using berberine bridge enzyme yielded several candidate genes, including an (S)-tetrahydroprotoberberine oxidase-like sequence selected for heterologous expression in Pichia pastoris. The recombinant enzyme preferentially catalyzed the oxidation of dihydrosanguinarine to sanguinarine but also converted (RS)-tetrahydropapaverine to papaverine and several protoberberine alkaloids to oxidized forms, including (RS)-canadine to berberine. The Km values of 201 and 146 μm for dihydrosanguinarine and the protoberberine alkaloid (S)-scoulerine, respectively, suggested high concentrations of these substrates in the plant. Virus-induced gene silencing to reduce DBOX transcript levels resulted in a corresponding reduction in sanguinarine, dihydrosanguinarine, and papaverine accumulation in opium poppy roots in support of DBOX as a multifunctional oxidative enzyme in BIA metabolism. PMID:23118227

  7. Systematic silencing of benzylisoquinoline alkaloid biosynthetic genes reveals the major route to papaverine in opium poppy.

    PubMed

    Desgagné-Penix, Isabel; Facchini, Peter J

    2012-10-01

    Papaverine, a major benzylisoquinoline alkaloid in opium poppy (Papaver somniferum), is used as a vasodilator and antispasmodic. Conversion of the initial intermediate (S)-norcoclaurine to papaverine involves 3'-hydroxylation, four O-methylations and dehydrogenation. However, our understanding of papaverine biosynthesis remains controversial more than a century after an initial scheme was proposed. In vitro assays and in vivo labeling studies have been insufficient to establish the sequence of conversions, the potential role of the intermediate (S)-reticuline, and the enzymes involved. We used virus-induced gene silencing in opium poppy to individually suppress the expression of six genes with putative roles in papaverine biosynthesis. Suppression of the gene encoding coclaurine N-methyltransferase dramatically increased papaverine levels at the expense of N-methylated alkaloids, indicating that the main biosynthetic route to papaverine proceeds via N-desmethylated compounds rather than through (S)-reticuline. Suppression of genes encoding (S)-3'-hydroxy-N-methylcoclaurine 4-O-methyltransferase and norreticuline 7-O-methyltransferase, which accept certain N-desmethylated alkaloids, reduced papaverine content. In contrast, suppression of genes encoding N-methylcoclaurine 3'-hydroxylase or reticuline 7-O-methyltransferase, which are specific for N-methylated alkaloids, did not affect papaverine levels. Suppression of norcoclaurine 6-O-methyltransferase transcript levels significantly suppressed total alkaloid accumulation, implicating (S)-coclaurine as a key branch-point intermediate. The differential detection of N-desmethylated compounds in response to suppression of specific genes highlights the primary route to papaverine.

  8. Regulation of the alkaloid biosynthesis by miRNA in opium poppy.

    PubMed

    Boke, Hatice; Ozhuner, Esma; Turktas, Mine; Parmaksiz, Iskender; Ozcan, Sebahattin; Unver, Turgay

    2015-04-01

    Opium poppy (Papaver somniferum) is an important medicinal plant producing benzylisoquinoline alkaloids (BIA). MicroRNAs (miRNAs) are endogenous small RNAs (sRNAs) of approximately 21 nucleotides. They are noncoding, but regulate gene expression in eukaryotes. Although many studies have been conducted on the identification and functions of plant miRNA, scarce researches on miRNA regulation of alkaloid biosynthesis have been reported. In this study, a total of 316 conserved and 11 novel miRNAs were identified in opium poppy using second-generation sequencing and direct cloning. Tissue-specific regulation of miRNA expression was comparatively analysed by miRNA microarray assays. A total of 232 miRNAs were found to be differentially expressed among four tissues. Likewise, 1469 target transcripts were detected using in silico and experimental approaches. The Kyoto Encyclopedia of Genes and Genomes pathway analyses indicated that miRNA putatively regulates carbohydrate metabolism and genetic-information processing. Additionally, miRNA target transcripts were mostly involved in response to stress against various factors and secondary-metabolite biosynthesis processes. Target transcript identification analyses revealed that some of the miRNAs might be involved in BIA biosynthesis, such as pso-miR13, pso-miR2161 and pso-miR408. Additionally, three putatively mature miRNA sequences were predicted to be targeting BIA-biosynthesis genes.

  9. Morphine biosynthesis in opium poppy involves two cell types: sieve elements and laticifers.

    PubMed

    Onoyovwe, Akpevwe; Hagel, Jillian M; Chen, Xue; Khan, Morgan F; Schriemer, David C; Facchini, Peter J

    2013-10-01

    Immunofluorescence labeling and shotgun proteomics were used to establish the cell type-specific localization of morphine biosynthesis in opium poppy (Papaver somniferum). Polyclonal antibodies for each of six enzymes involved in converting (R)-reticuline to morphine detected corresponding antigens in sieve elements of the phloem, as described previously for all upstream enzymes transforming (S)-norcoclaurine to (S)-reticuline. Validated shotgun proteomics performed on whole-stem and latex total protein extracts generated 2031 and 830 distinct protein families, respectively. Proteins corresponding to nine morphine biosynthetic enzymes were represented in the whole stem, whereas only four of the final five pathway enzymes were detected in the latex. Salutaridine synthase was detected in the whole stem, but not in the latex subproteome. The final three enzymes converting thebaine to morphine were among the most abundant active latex proteins despite a limited occurrence in laticifers suggested by immunofluorescence labeling. Multiple charge isoforms of two key O-demethylases in the latex were revealed by two-dimensional immunoblot analysis. Salutaridine biosynthesis appears to occur only in sieve elements, whereas conversion of thebaine to morphine is predominant in adjacent laticifers, which contain morphine-rich latex. Complementary use of immunofluorescence labeling and shotgun proteomics has substantially resolved the cellular localization of morphine biosynthesis in opium poppy.

  10. Regulation of the alkaloid biosynthesis by miRNA in opium poppy.

    PubMed

    Boke, Hatice; Ozhuner, Esma; Turktas, Mine; Parmaksiz, Iskender; Ozcan, Sebahattin; Unver, Turgay

    2015-04-01

    Opium poppy (Papaver somniferum) is an important medicinal plant producing benzylisoquinoline alkaloids (BIA). MicroRNAs (miRNAs) are endogenous small RNAs (sRNAs) of approximately 21 nucleotides. They are noncoding, but regulate gene expression in eukaryotes. Although many studies have been conducted on the identification and functions of plant miRNA, scarce researches on miRNA regulation of alkaloid biosynthesis have been reported. In this study, a total of 316 conserved and 11 novel miRNAs were identified in opium poppy using second-generation sequencing and direct cloning. Tissue-specific regulation of miRNA expression was comparatively analysed by miRNA microarray assays. A total of 232 miRNAs were found to be differentially expressed among four tissues. Likewise, 1469 target transcripts were detected using in silico and experimental approaches. The Kyoto Encyclopedia of Genes and Genomes pathway analyses indicated that miRNA putatively regulates carbohydrate metabolism and genetic-information processing. Additionally, miRNA target transcripts were mostly involved in response to stress against various factors and secondary-metabolite biosynthesis processes. Target transcript identification analyses revealed that some of the miRNAs might be involved in BIA biosynthesis, such as pso-miR13, pso-miR2161 and pso-miR408. Additionally, three putatively mature miRNA sequences were predicted to be targeting BIA-biosynthesis genes. PMID:25735537

  11. Isolation and Characterisation of 11 Polymorphic Microsatellite Markers in Papaver rhoeas L. (Corn Poppy), a Major Annual Plant Species from Cultivated Areas

    PubMed Central

    Kati, Vaya; Le Corre, Valérie; Michel, Séverine; Jaffrelo, Lydia; Poncet, Charles; Délye, Christophe

    2013-01-01

    Papaver rhoeas, an annual plant species in the Papaveraceae family, is part of the biodiversity of agricultural ecosystems and also a noxious agronomic weed. We developed microsatellite markers to study the genetic diversity of P. rhoeas, using an enriched microsatellite library coupled with 454 next-generation sequencing. A total of 13,825 sequences were obtained that yielded 1795 microsatellite loci. After discarding loci with less than six repeats of the microsatellite motif, automated primer design was successful for 598 loci. We tested 74 of these loci for amplification with a total of 97 primer pairs. Thirty loci passed our tests and were subsequently tested for polymorphism using 384 P. rhoeas plants originating from 12 populations from France. Of the 30 loci, 11 showed reliable polymorphism not affected by the presence of null alleles. The number of alleles and the expected heterozygosity ranged from 3 to 7.4 and from 0.27 to 0.73, respectively. A low but significant genetic differentiation among populations was observed (FST = 0.04; p < 0.001). The 11 validated polymorphic microsatellite markers developed in this work will be useful in studies of genetic diversity and population structure of P. rhoeas, assisting in designing management strategies for the control or the conservation of this species. PMID:23263674

  12. Isolation and characterisation of 11 polymorphic microsatellite markers in Papaver rhoeas L. (Corn Poppy), a major annual plant species from cultivated areas.

    PubMed

    Kati, Vaya; Corre, Valérie Le; Michel, Séverine; Jaffrelo, Lydia; Poncet, Charles; Délye, Christophe

    2012-12-24

    Papaver rhoeas, an annual plant species in the Papaveraceae family, is part of the biodiversity of agricultural ecosystems and also a noxious agronomic weed. We developed microsatellite markers to study the genetic diversity of P. rhoeas, using an enriched microsatellite library coupled with 454 next-generation sequencing. A total of 13,825 sequences were obtained that yielded 1795 microsatellite loci. After discarding loci with less than six repeats of the microsatellite motif, automated primer design was successful for 598 loci. We tested 74 of these loci for amplification with a total of 97 primer pairs. Thirty loci passed our tests and were subsequently tested for polymorphism using 384 P. rhoeas plants originating from 12 populations from France. Of the 30 loci, 11 showed reliable polymorphism not affected by the presence of null alleles. The number of alleles and the expected heterozygosity ranged from 3 to 7.4 and from 0.27 to 0.73, respectively. A low but significant genetic differentiation among populations was observed (F(ST) = 0.04; p < 0.001). The 11 validated polymorphic microsatellite markers developed in this work will be useful in studies of genetic diversity and population structure of P. rhoeas, assisting in designing management strategies for the control or the conservation of this species.

  13. Orally active opioid compounds from a non-poppy source.

    PubMed

    Raffa, Robert B; Beckett, Jaclyn R; Brahmbhatt, Vivek N; Ebinger, Theresa M; Fabian, Chrisjon A; Nixon, Justin R; Orlando, Steven T; Rana, Chintan A; Tejani, Ali H; Tomazic, Robert J

    2013-06-27

    The basic science and clinical use of morphine and other "opioid" drugs are based almost exclusively on the extracts or analogues of compounds isolated from a single source, the opium poppy (Papaver somniferum). However, it now appears that biological diversity has evolved an alternative source. Specifically, at least two alkaloids isolated from the plant Mitragyna speciosa, mitragynine ((E)-2-[(2S,3S)-3-ethyl-8-methoxy-1,2,3,4,6,7,12,12b-octahydroindolo[3,2-h]quinolizin-2-yl]-3-methoxyprop-2-enoic acid methyl ester; 9-methoxy coryantheidine; MG) and 7-hydroxymitragynine (7-OH-MG), and several synthetic analogues of these natural products display centrally mediated (supraspinal and spinal) antinociceptive (analgesic) activity in various pain models. Several characteristics of these compounds suggest a classic "opioid" mechanism of action: nanomolar affinity for opioid receptors, competitive interaction with the opioid receptor antagonist naloxone, and two-way analgesic cross-tolerance with morphine. However, other characteristics of the compounds suggest novelty, particularly chemical structure and possible greater separation from side effects. We review the chemical and pharmacological properties of these compounds. PMID:23517479

  14. CYP719B1 is salutaridine synthase, the C-C phenol-coupling enzyme of morphine biosynthesis in opium poppy.

    PubMed

    Gesell, Andreas; Rolf, Megan; Ziegler, Jörg; Díaz Chávez, María Luisa; Huang, Fong-Chin; Kutchan, Toni M

    2009-09-01

    Morphine is a powerful analgesic natural product produced by the opium poppy Papaver somniferum. Although formal syntheses of this alkaloid have been reported, the morphine molecule contains five stereocenters and a C-C phenol linkage that to date render a total synthesis of morphine commercially unfeasible. The C-C phenol-coupling reaction along the biosynthetic pathway to morphine in opium poppy is catalyzed by the cytochrome P450-dependent oxygenase salutaridine synthase. We report herein on the identification of salutaridine synthase as a member of the CYP719 family of cytochromes P450 during a screen of recombinant cytochromes P450 of opium poppy functionally expressed in Spodoptera frugiperda Sf9 cells. Recombinant CYP719B1 is a highly stereo- and regioselective enzyme; of forty-one compounds tested as potential substrates, only (R)-reticuline and (R)-norreticuline resulted in formation of a product (salutaridine and norsalutaridine, respectively). To date, CYP719s have been characterized catalyzing only the formation of a methylenedioxy bridge in berberine biosynthesis (canadine synthase, CYP719A1) and in benzo[c]phenanthridine biosynthesis (stylopine synthase, CYP719A14). Previously identified phenol-coupling enzymes of plant alkaloid biosynthesis belong only to the CYP80 family of cytochromes. CYP719B1 therefore is the prototype for a new family of plant cytochromes P450 that catalyze formation of a phenol-couple.

  15. Death in a legal poppy field in Spain.

    PubMed

    Martínez, María Antonia; Ballesteros, Salomé; Almarza, Elena; Garijo, Joaquín

    2016-08-01

    Opium is a substance extracted from Papaver somniferum L. Opium latex contains morphine, codeine, and thebaine and non-analgesic alkaloids such as papaverine and noscapine. In Spain opium growing is allowed only for scientific or pharmaceutical purposes and harvest is supervised by the Spanish Health Ministry. This work describes a sudden fatality involving opium consumption in a legal poppy field. The toxicological and autopsy findings, previous disease, paraphernalia, and scenario are discussed in order to clarify cause and manner of death. A 32-year-old white caucasian male was found unresponsive in a legal poppy field in the South of Spain. The emergency medical services responded to the scene where he was pronounced dead. The friends explained that the deceased had presented with about 30min of convulsions; in spite of trying to keep his airway tract open they noted that "he stayed airless". According to them the victim suffered from epilepsy. Tools found beside his body consisted of plain wood sticks with a blade razor, a fabric handle, and paper. A comprehensive toxicological screening for abuse and psychoactive drugs was performed in the deceased samples. This included ethanol and volatile analysis by HS-GC-FID in peripheral blood and urine, enzyme immunoassay in urine by CEDIA, and a basic drug screening in all samples (including paraphernalia) by GC-MS using modes full scan for screening/confirmation and selected ion monitoring for quantitation. The peripheral blood, urine, vitreous, and gastric content contained the following concentrations of opiates expressed in mg/L (gastric content additionally also expressed in mg total): 0.10, 7.12, 0.23, and 14.80 (2.81mg total) of thebaine, 0.13, 4.50, 0.13, and 6.60 (1.25mg total) of morphine (free), 0.48, 0.88, 0.17, and 1.50 (0.28mg total) of codeine. These tree opiates were also detected in the tools (paraphernalia) used by the deceased for opium consumption. Other toxicological findings were metabolites of

  16. Death in a legal poppy field in Spain.

    PubMed

    Martínez, María Antonia; Ballesteros, Salomé; Almarza, Elena; Garijo, Joaquín

    2016-08-01

    Opium is a substance extracted from Papaver somniferum L. Opium latex contains morphine, codeine, and thebaine and non-analgesic alkaloids such as papaverine and noscapine. In Spain opium growing is allowed only for scientific or pharmaceutical purposes and harvest is supervised by the Spanish Health Ministry. This work describes a sudden fatality involving opium consumption in a legal poppy field. The toxicological and autopsy findings, previous disease, paraphernalia, and scenario are discussed in order to clarify cause and manner of death. A 32-year-old white caucasian male was found unresponsive in a legal poppy field in the South of Spain. The emergency medical services responded to the scene where he was pronounced dead. The friends explained that the deceased had presented with about 30min of convulsions; in spite of trying to keep his airway tract open they noted that "he stayed airless". According to them the victim suffered from epilepsy. Tools found beside his body consisted of plain wood sticks with a blade razor, a fabric handle, and paper. A comprehensive toxicological screening for abuse and psychoactive drugs was performed in the deceased samples. This included ethanol and volatile analysis by HS-GC-FID in peripheral blood and urine, enzyme immunoassay in urine by CEDIA, and a basic drug screening in all samples (including paraphernalia) by GC-MS using modes full scan for screening/confirmation and selected ion monitoring for quantitation. The peripheral blood, urine, vitreous, and gastric content contained the following concentrations of opiates expressed in mg/L (gastric content additionally also expressed in mg total): 0.10, 7.12, 0.23, and 14.80 (2.81mg total) of thebaine, 0.13, 4.50, 0.13, and 6.60 (1.25mg total) of morphine (free), 0.48, 0.88, 0.17, and 1.50 (0.28mg total) of codeine. These tree opiates were also detected in the tools (paraphernalia) used by the deceased for opium consumption. Other toxicological findings were metabolites of

  17. Quantitative 1H nuclear magnetic resonance metabolite profiling as a functional genomics platform to investigate alkaloid biosynthesis in opium poppy.

    PubMed

    Hagel, Jillian M; Weljie, Aalim M; Vogel, Hans J; Facchini, Peter J

    2008-08-01

    Opium poppy (Papaver somniferum) produces a diverse array of bioactive benzylisoquinoline alkaloids and has emerged as a versatile model system to study plant alkaloid metabolism. The plant is widely cultivated as the only commercial source of the narcotic analgesics morphine and codeine. Variations in plant secondary metabolism as a result of genetic diversity are often associated with perturbations in other metabolic pathways. As part of a functional genomics platform, we used (1)H nuclear magnetic resonance (NMR) metabolite profiling for the analysis of primary and secondary metabolism in opium poppy. Aqueous and chloroform extracts of six different opium poppy cultivars were subjected to chemometric analysis. Principle component analysis of the (1)H NMR spectra for latex extracts clearly distinguished two varieties, including a low-alkaloid variety and a high-thebaine, low-morphine cultivar. Distinction was also made between pharmaceutical-grade opium poppy cultivars and a condiment variety. Such phenotypic differences were not observed in root extracts. Loading plots confirmed that morphinan alkaloids contributed predominantly to the variance in latex extracts. Quantification of 34 root and 21 latex metabolites, performed using Chenomx NMR Suite version 4.6, showed major differences in the accumulation of specific alkaloids in the latex of the low-alkaloid and high-thebaine, low-morphine varieties. Relatively few differences were found in the levels of other metabolites, indicating that the variation was specific for alkaloid metabolism. Exceptions in the low-alkaloid cultivar included an increased accumulation of the alkaloid precursor tyramine and reduced levels of sucrose, some amino acids, and malate. Real-time polymerase chain reaction analysis of 42 genes involved in primary and secondary metabolism showed differential gene expression mainly associated with alkaloid biosynthesis. Reduced alkaloid levels in the condiment variety were associated with the

  18. Plant defense responses in opium poppy cell cultures revealed by liquid chromatography-tandem mass spectrometry proteomics.

    PubMed

    Zulak, Katherine G; Khan, Morgan F; Alcantara, Joenel; Schriemer, David C; Facchini, Peter J

    2009-01-01

    Opium poppy (Papaver somniferum) produces a diverse array of bioactive benzylisoquinoline alkaloids, including the narcotic analgesic morphine and the antimicrobial agent sanguinarine. In contrast to the plant, cell cultures of opium poppy do not accumulate alkaloids constitutively but produce sanguinarine in response to treatment with certain fungal-derived elicitors. The induction of sanguinarine biosynthesis provides a model platform to characterize the regulation of benzylisoquinoline alkaloid pathways and other defense responses. Proteome analysis of elicitor-treated opium poppy cell cultures by two-dimensional denaturing-polyacrylamide gel electrophoresis coupled with liquid chromatography-tandem mass spectrometry facilitated the identification of 219 of 340 protein spots based on peptide fragment fingerprint searches of a combination of databases. Of the 219 hits, 129 were identified through pre-existing plant proteome databases, 63 were identified by matching predicted translation products in opium poppy-expressed sequence tag databases, and the remainder shared evidence from both databases. Metabolic enzymes represented the largest category of proteins and included S-adenosylmethionine synthetase, several glycolytic, and a nearly complete set of tricarboxylic acid cycle enzymes, one alkaloid, and several other secondary metabolic enzymes. The abundance of chaperones, heat shock proteins, protein degradation factors, and pathogenesis-related proteins provided a comprehensive proteomics view on the coordination of plant defense responses. Qualitative comparison of protein abundance in control and elicitor-treated cell cultures allowed the separation of induced and constitutive or suppressed proteins. DNA microarrays were used to corroborate increases in protein abundance with a corresponding induction in cognate transcript levels.

  19. Poppy APETALA1/FRUITFULL orthologs control flowering time, branching, perianth identity, and fruit development.

    PubMed

    Pabón-Mora, Natalia; Ambrose, Barbara A; Litt, Amy

    2012-04-01

    Several MADS box gene lineages involved in flower development have undergone duplications that correlate with the diversification of large groups of flowering plants. In the APETALA1 gene lineage, a major duplication coincides with the origin of the core eudicots, resulting in the euFUL and the euAP1 clades. Arabidopsis FRUITFULL (FUL) and APETALA1 (AP1) function redundantly in specifying floral meristem identity but function independently in sepal and petal identity (AP1) and in proper fruit development and determinacy (FUL). Many of these functions are largely conserved in other core eudicot euAP1 and euFUL genes, but notably, the role of APETALA1 as an "A-function" (sepal and petal identity) gene is thought to be Brassicaceae specific. Understanding how functional divergence of the core eudicot duplicates occurred requires a careful examination of the function of preduplication (FUL-like) genes. Using virus-induced gene silencing, we show that FUL-like genes in opium poppy (Papaver somniferum) and California poppy (Eschscholzia californica) function in axillary meristem growth and in floral meristem and sepal identity and that they also play a key role in fruit development. Interestingly, in opium poppy, these genes also control flowering time and petal identity, suggesting that AP1/FUL homologs might have been independently recruited in petal identity. Because the FUL-like gene functional repertoire encompasses all roles previously described for the core eudicot euAP1 and euFUL genes, we postulate subfunctionalization as the functional outcome after the major AP1/FUL gene lineage duplication event.

  20. Poppy APETALA1/FRUITFULL Orthologs Control Flowering Time, Branching, Perianth Identity, and Fruit Development1[W][OA

    PubMed Central

    Pabón-Mora, Natalia; Ambrose, Barbara A.; Litt, Amy

    2012-01-01

    Several MADS box gene lineages involved in flower development have undergone duplications that correlate with the diversification of large groups of flowering plants. In the APETALA1 gene lineage, a major duplication coincides with the origin of the core eudicots, resulting in the euFUL and the euAP1 clades. Arabidopsis FRUITFULL (FUL) and APETALA1 (AP1) function redundantly in specifying floral meristem identity but function independently in sepal and petal identity (AP1) and in proper fruit development and determinacy (FUL). Many of these functions are largely conserved in other core eudicot euAP1 and euFUL genes, but notably, the role of APETALA1 as an “A-function” (sepal and petal identity) gene is thought to be Brassicaceae specific. Understanding how functional divergence of the core eudicot duplicates occurred requires a careful examination of the function of preduplication (FUL-like) genes. Using virus-induced gene silencing, we show that FUL-like genes in opium poppy (Papaver somniferum) and California poppy (Eschscholzia californica) function in axillary meristem growth and in floral meristem and sepal identity and that they also play a key role in fruit development. Interestingly, in opium poppy, these genes also control flowering time and petal identity, suggesting that AP1/FUL homologs might have been independently recruited in petal identity. Because the FUL-like gene functional repertoire encompasses all roles previously described for the core eudicot euAP1 and euFUL genes, we postulate subfunctionalization as the functional outcome after the major AP1/FUL gene lineage duplication event. PMID:22286183

  1. Isolation and characterization of a cDNA encoding (S)-cis-N-methylstylopine 14-hydroxylase from opium poppy, a key enzyme in sanguinarine biosynthesis.

    PubMed

    Beaudoin, Guillaume A W; Facchini, Peter J

    2013-02-15

    Sanguinarine is a benzo[c]phenenthridine alkaloid with potent antimicrobial properties found commonly in plants of the Papaveraceae, including the roots of opium poppy (Papaver somniferum). Sanguinarine is formed from the central 1-benzylisoquinoline intermediate (S)-reticuline via the protoberberine alkaloid (S)-scoulerine, which undergoes five enzymatic oxidations and an N-methylation. The first four oxidations from (S)-scoulerine are catalyzed by cytochromes P450, whereas the final conversion involves a flavoprotein oxidase. All but one gene in the biosynthetic pathway from (S)-reticuline to sanguinarine has been identified. In this communication, we report the isolation and characterization of (S)-cis-N-methylstylopine 14-hydroxylase (MSH) from opium poppy based on the transcriptional induction in elicitor-treated cell suspension cultures and root-specific expression of the corresponding gene. Along with protopine 6-hydroxylase, which catalyzes the subsequent and penultimate step in sanguinarine biosynthesis, MSH is a member of the CYP82N subfamily of cytochromes P450. The full-length MSH cDNA was expressed in Saccharomyces cerevisiae and the recombinant microsomal protein was tested for enzymatic activity using 25 benzylisoquinoline alkaloids representing a wide range of structural subgroups. The only enzymatic substrates were the N-methylated protoberberine alkaloids N-methylstylopine and N-methylcanadine, which were converted to protopine and allocryptopine, respectively.

  2. Differences in the effect of phosphatidylinositol 4,5-bisphosphate on the hydrolytic and transphosphatidylation activities of membrane-bound phospholipase D from poppy seedlings.

    PubMed

    Oblozinsky, Marek; Bezakova, Lydia; Mansfeld, Johanna; Heilmann, Ingo; Ulbrich-Hofmann, Renate

    2013-08-01

    The hydrolytic activity of phospholipase D (PLD) yielding phosphatidic acid from phosphatidylcholine and other glycerophospholipids is known to be involved in many cellular processes. In contrast, it is not clear whether the competitive transphosphatidylation activity of PLD catalyzing the head group exchange of phospholipids has a natural function. In poppy seedlings (Papaver somniferum L.) where lipid metabolism and alkaloid synthesis are closely linked, five isoenzymes with different substrate and hydrolysis/transphosphatidylation selectivities have been detected hitherto. A membrane-bound PLD, found in microsomal fractions of poppy seedlings, is active at micromolar concentrations of Ca(2+) ions and needs phosphatidylinositol 4,5-bisphosphate (PIP2) as effector in the hydrolysis of phosphatidylcholine (PC). The optimum PIP2 concentration at 1.2 mol% of the concentration of the substrate PC indicates a specific activation effect. Transphosphatidylation with glycerol, ethanolamine, l-serine, or myo-inositol as acceptor alcohols is also activated by PIP2, however, with an optimum concentration at 0.6-0.9 mol%. In contrast to hydrolysis, a basic transphosphatidylation activity occurs even in the absence of PIP2, suggesting a different fine-tuning of the two competing reactions.

  3. Systematic silencing of benzylisoquinoline alkaloid biosynthetic genes reveals the major route to papaverine in opium poppy.

    PubMed

    Desgagné-Penix, Isabel; Facchini, Peter J

    2012-10-01

    Papaverine, a major benzylisoquinoline alkaloid in opium poppy (Papaver somniferum), is used as a vasodilator and antispasmodic. Conversion of the initial intermediate (S)-norcoclaurine to papaverine involves 3'-hydroxylation, four O-methylations and dehydrogenation. However, our understanding of papaverine biosynthesis remains controversial more than a century after an initial scheme was proposed. In vitro assays and in vivo labeling studies have been insufficient to establish the sequence of conversions, the potential role of the intermediate (S)-reticuline, and the enzymes involved. We used virus-induced gene silencing in opium poppy to individually suppress the expression of six genes with putative roles in papaverine biosynthesis. Suppression of the gene encoding coclaurine N-methyltransferase dramatically increased papaverine levels at the expense of N-methylated alkaloids, indicating that the main biosynthetic route to papaverine proceeds via N-desmethylated compounds rather than through (S)-reticuline. Suppression of genes encoding (S)-3'-hydroxy-N-methylcoclaurine 4-O-methyltransferase and norreticuline 7-O-methyltransferase, which accept certain N-desmethylated alkaloids, reduced papaverine content. In contrast, suppression of genes encoding N-methylcoclaurine 3'-hydroxylase or reticuline 7-O-methyltransferase, which are specific for N-methylated alkaloids, did not affect papaverine levels. Suppression of norcoclaurine 6-O-methyltransferase transcript levels significantly suppressed total alkaloid accumulation, implicating (S)-coclaurine as a key branch-point intermediate. The differential detection of N-desmethylated compounds in response to suppression of specific genes highlights the primary route to papaverine. PMID:22725256

  4. Systematic knockdown of morphine pathway enzymes in opium poppy using virus-induced gene silencing.

    PubMed

    Wijekoon, Champa P; Facchini, Peter J

    2012-03-01

    Opium poppy (Papaver somniferum) remains the sole commercial source for several pharmaceutical alkaloids including the narcotic analgesics codeine and morphine, and the semi-synthetic drugs oxycodone, buprenorphine and naltrexone. Although most of the biosynthetic genes have been identified, the post-transcriptional regulation of the morphinan alkaloid pathway has not been determined. We have used virus-induced gene silencing (VIGS) as a functional genomics tool to investigate the regulation of morphine biosynthesis via a systematic reduction in enzyme levels responsible for the final six steps in the pathway. Specific gene silencing was confirmed at the transcript level by real-time quantitative PCR (polymerase chain reaction), and at the protein level by immunoblot analysis using antibodies raised against salutaridine synthase (SalSyn), salutaridine reductase (SalR), salutaridine 7-O-acetyltransferase (SalAT), thebaine 6-O-demethylase (T6ODM), codeinone reductase (COR), and codeine O-demethylase (CODM). In some cases, silencing a specific biosynthetic gene resulted in a predictable accumulation of the substrate for the corresponding enzyme. Reduced SalSyn, SalR, T6ODM and CODM protein levels correlated with lower morphine levels and a substantial increase in the accumulation of reticuline, salutaridine, thebaine and codeine, respectively. In contrast, the silencing of genes encoding SalAT and COR resulted in the accumulation of salutaridine and reticuline, respectively, which are not the corresponding enzymatic substrates. The silencing of alkaloid biosynthetic genes using VIGS confirms the physiological function of enzymes previously characterized in vitro, provides insight into the biochemical regulation of morphine biosynthesis, and demonstrates the immense potential for metabolic engineering in opium poppy.

  5. Manipulating the Biosynthesis of Bioactive Compound Alkaloids for Next-Generation Metabolic Engineering in Opium Poppy Using CRISPR-Cas 9 Genome Editing Technology.

    PubMed

    Alagoz, Yagiz; Gurkok, Tugba; Zhang, Baohong; Unver, Turgay

    2016-01-01

    Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated9 (Cas9) endonuclease system is a powerful RNA-guided genome editing tool. CRISPR/Cas9 has been well studied in model plant species for targeted genome editing. However, few studies have been reported on plant species without whole genome sequence information. Currently, no study has been performed to manipulate metabolic pathways using CRISPR/Cas9. In this study, the type II CRISPR/SpCas9 system was used to knock out, via nonhomologous end-joining genome repair, the 4'OMT2 in opium poppy (Papaver somniferum L.), a gene which regulates the biosythesis of benzylisoquinoline alkaloids (BIAs). For sgRNA transcription, viral-based TRV and synthetic binary plasmids were designed and delivered into plant cells with a Cas9 encoding-synthetic vector by Agrobacterium-mediated transformation. InDels formed by CRISPR/Cas9 were detected by sequence analysis. Our results showed that the biosynthesis of BIAs (e.g. morphine, thebaine) was significantly reduced in the transgenic plants suggesting that 4'OMT2 was efficiently knocked-out by our CRISPR-Cas9 genome editing approach. In addition, a novel uncharacterized alkaloid was observed only in CRISPR/Cas9 edited plants. Thus, the applicabilitiy of the CRISPR/Cas9 system was demonstrated for the first time for medicinal aromatic plants by sgRNAs transcribed from both synthetic and viral vectors to regulate BIA metabolism and biosynthesis. PMID:27483984

  6. Manipulating the Biosynthesis of Bioactive Compound Alkaloids for Next-Generation Metabolic Engineering in Opium Poppy Using CRISPR-Cas 9 Genome Editing Technology.

    PubMed

    Alagoz, Yagiz; Gurkok, Tugba; Zhang, Baohong; Unver, Turgay

    2016-08-03

    Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated9 (Cas9) endonuclease system is a powerful RNA-guided genome editing tool. CRISPR/Cas9 has been well studied in model plant species for targeted genome editing. However, few studies have been reported on plant species without whole genome sequence information. Currently, no study has been performed to manipulate metabolic pathways using CRISPR/Cas9. In this study, the type II CRISPR/SpCas9 system was used to knock out, via nonhomologous end-joining genome repair, the 4'OMT2 in opium poppy (Papaver somniferum L.), a gene which regulates the biosythesis of benzylisoquinoline alkaloids (BIAs). For sgRNA transcription, viral-based TRV and synthetic binary plasmids were designed and delivered into plant cells with a Cas9 encoding-synthetic vector by Agrobacterium-mediated transformation. InDels formed by CRISPR/Cas9 were detected by sequence analysis. Our results showed that the biosynthesis of BIAs (e.g. morphine, thebaine) was significantly reduced in the transgenic plants suggesting that 4'OMT2 was efficiently knocked-out by our CRISPR-Cas9 genome editing approach. In addition, a novel uncharacterized alkaloid was observed only in CRISPR/Cas9 edited plants. Thus, the applicabilitiy of the CRISPR/Cas9 system was demonstrated for the first time for medicinal aromatic plants by sgRNAs transcribed from both synthetic and viral vectors to regulate BIA metabolism and biosynthesis.

  7. Use of Climatic Data to Identify Potential Sites in the United States for Growing Papaver bracteatum as a Pharmaceutical Crop.

    NASA Astrophysics Data System (ADS)

    Neild, R. E.

    1987-09-01

    A procedure utilizing maps in the National Climatic Atlas was used to survey the United States to find areas for domesticating Papaver bracteatum, a wild poppy from the Trans-Caucasus. This poppy has unique properties which make it a possible heroinless source of codeine. Climatic conditions similar to the native habitat and requirements of Papaver bracteatum can be found in areas of western Utah, northern Nevada, southern Idaho, eastern Washington and Oregon, northeastern California and southwestern Wyoming. A detailed climatic analysis was made to evaluate more specific locations within this large area. A site near Medford, Oregon was selected as best suited for developing modern cultural procedures and improving strains to successfully grow crops of Papaver bracteatum.

  8. Hepatotoxicity associated with the ingestion of Papaver Rhoease.

    PubMed

    Gonullu, Hayriye; Karadas, Sevdegul; Dulger, Ahmet Cumhur; Ebinc, Senar

    2014-10-01

    Liver injury appears to be associated with the use of herbal products and wild plants.This paper presents what is to our knowledge the first case report in the world of acute liver toxicity caused by Papaver Rhoease. A 17-year- old woman from eastern Turkey ingested Papaver Rhoease which is known as "poppy flower" among the local inhabitants. Her liver function tests were initially 10- 15 fold higher than normal. She was hospitalised in intensive care unit and on day 2 of hospitalisation, the results of her liver function tests showed about a hundred-fold increase compared to the normal level. Her general status deteriorated and she was transferred to another medical centre for liver transplantation.

  9. 7 CFR 400.677 - Definitions.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... charges (7 CFR 400.116). A delinquent debt does not include debts discharged in bankruptcy and other debts... (cannabis sativa), opium poppies (papaver somniferum), and other drug-producing plants, the planting...

  10. Manipulating the Biosynthesis of Bioactive Compound Alkaloids for Next-Generation Metabolic Engineering in Opium Poppy Using CRISPR-Cas 9 Genome Editing Technology

    PubMed Central

    Alagoz, Yagiz; Gurkok, Tugba; Zhang, Baohong; Unver, Turgay

    2016-01-01

    Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated9 (Cas9) endonuclease system is a powerful RNA-guided genome editing tool. CRISPR/Cas9 has been well studied in model plant species for targeted genome editing. However, few studies have been reported on plant species without whole genome sequence information. Currently, no study has been performed to manipulate metabolic pathways using CRISPR/Cas9. In this study, the type II CRISPR/SpCas9 system was used to knock out, via nonhomologous end-joining genome repair, the 4′OMT2 in opium poppy (Papaver somniferum L.), a gene which regulates the biosythesis of benzylisoquinoline alkaloids (BIAs). For sgRNA transcription, viral-based TRV and synthetic binary plasmids were designed and delivered into plant cells with a Cas9 encoding-synthetic vector by Agrobacterium-mediated transformation. InDels formed by CRISPR/Cas9 were detected by sequence analysis. Our results showed that the biosynthesis of BIAs (e.g. morphine, thebaine) was significantly reduced in the transgenic plants suggesting that 4′OMT2 was efficiently knocked-out by our CRISPR-Cas9 genome editing approach. In addition, a novel uncharacterized alkaloid was observed only in CRISPR/Cas9 edited plants. Thus, the applicabilitiy of the CRISPR/Cas9 system was demonstrated for the first time for medicinal aromatic plants by sgRNAs transcribed from both synthetic and viral vectors to regulate BIA metabolism and biosynthesis. PMID:27483984

  11. Marc Chagall: "Wild Poppies."

    ERIC Educational Resources Information Center

    White, Carolyn

    1987-01-01

    Based on a full-color reproduction of Marc Chagall's painting, "Wild Poppies," the goals of this lesson plan are to introduce students to artist's use of dreams and memories in making art, to communicate the idea that artists include their visual memories of people and things they love in their artwork, and to introduce the concepts of line and…

  12. Self-incompatibility in Papaver: identification of the pollen S-determinant PrpS.

    PubMed

    Poulter, Natalie S; Wheeler, Michael J; Bosch, Maurice; Franklin-Tong, Vernonica E

    2010-04-01

    Many flowering plants are hermaphrodite, posing the problem of self-fertilization and the subsequent loss of the genetic fitness of the offspring. To prevent this, many plants have developed a genetically controlled mechanism called self-incompatibility (SI). When the male and female S-determinants match, self (incompatible) pollen is recognized and rejected before fertilization can occur. In poppy (Papaver rhoeas), the pistil S-determinant (PrsS) is a small secreted protein that interacts with incompatible pollen, initiating a Ca(2+)-dependent signalling network. SI triggers several downstream events, including depolymerization of the cytoskeleton, phosphorylation of two soluble inorganic pyrophosphatases and an MAPK (mitogen-activated protein kinase). This culminates in PCD (programmed cell death) involving several caspase-like activities. The recent discovery of the Papaver pollen S-determinant PrpS marks a significant step forward in the understanding of the Papaver SI system. PrpS encodes a ~20 kDa predicted transmembrane protein which has no homology with known proteins. It is specifically expressed in pollen, linked to the pistil S-determinant, and displays the high polymorphism expected of an S-locus determinant. The present review focuses on the discovery and characterization of PrpS which strongly support the hypothesis that Papaver SI is triggered by the interaction of PrsS and PrpS.

  13. Discrimination among species of Papaver based on the plastid rpl16 gene and the rpl16-rpl14 spacer sequence.

    PubMed

    Hosokawa, Keizo; Shibata, Toshiro; Nakamura, Ikuo; Hishida, Atsuyuki

    2004-01-28

    An attempt was made to discriminate among six species of Papaver (P. bracteatum, P. orientale, P. pseudo-orientale, P. rhoeas, P. setigerum and P. somniferum) by comparing the nucleotide sequences of the plastid rpl16 gene and the rpl16-rpl14 spacer region. Comparison of sequences allowed us to distinguish five species, namely P. bracteatum, P. orientale, P. pseudo-orientale, P. rhoeas and P. setigerum plus P. somniferum from one another, but sequences from P. setigerum and P. somniferum were identical. It is difficult to distinguish between P. bracteatum, P. orientale and P. pseudo-orientale at the vegetative stage of growth. However, our method allowed us to distinguish between these three species and the others using nucleotide sequences and should allow identification of P. bracteatum that has been cultivated illegally in the garden in Japan. Furthermore, P. rhoeas was clearly discriminated from P. setigerum and P. somniferum by reference to the sequence of the rpl16 exon using young seedlings.

  14. Alkaloid biosynthesis in Papaver sp. cells in culture and during organogenesis.

    PubMed

    Alkhimova, O G; Kyrylenko, T K; Vagyn, Y V; Heslop-Harrison, J S

    2001-01-01

    In vitro cell cultures of two Papaver species, P. somniferum and P. bracteatum initiated from mature seeds were screened for their ability to produce alkaloids. Protocols for callus induction, somatic embryogenesis and organogenesis were established. The alkaloid contents were analysed by high-performance-liquid chromatography, thin-layer chromatography and spectrophotometric assays. Undifferentiated callus produced small amounts of sanguinarine, which increased with the degree of tissue differentiation. Embryogenic calli were maintained in culture for more than 2 years, retaining a high regeneration capability. Thin-layer chromatography analysis revealed variations in alkaloid spectrum between parallel cell lines. The morphinan alkaloid, thebaine, was found to be accumulated exclusively in morphogenous strains of P. bracteatum, and morphine was the major alkaloid in the spectrum of P. somniferum dedifferentiated callus. Regenerant plants synthesized thebaine and sanguinarine at the same level as juvenile plants grown from P. bracteatum seeds. We revealed differences in the ability to produce different types of alkaloids: seed-derived plants were able to accumulate thebaine while undifferentiated primary cell cultures produced only sanguinarine. The production of either sanguinarine and morphinan alkaloids are found in regenerants showing that both metabolic pathways were active in young plantlets.

  15. Latex-less opium poppy: cause for less latex and reduced peduncle strength.

    PubMed

    Chaturvedi, Nidarshana; Singh, Sharad K; Shukla, Ashutosh K; Lal, Raj K; Gupta, Madan M; Dwivedi, Upendra N; Shasany, Ajit K

    2014-03-01

    A genotype 'Sujata' developed earlier at CSIR-CIMAP from its parent 'Sampada' is considered to be the latex-less variety of Papaver somniferum. These two genotypes are contrasting in terms of latex and stem strength. Earlier we have carried out microarray analysis to identify differentially expressing genes from the capsules of the two genotypes. In this study, the peduncles of the two genotypes were compared for the anatomy revealing less number of laticifers in the cortex and vascular bundles. One of the important cell wall-related genes (for laccase) from the microarray analysis showing significantly higher expression in 'Sampada' capsule was taken up for further characterization in the peduncle here. It was functionally characterized through transient overexpression and RNAi suppression in 'Sujata' and 'Sampada'. The increase in acid insoluble lignin and total lignin in overexpressed tissue of 'Sujata', and comparable decrease in suppressed tissue of 'Sampada', along with corresponding increase and decrease in the transcript abundance of laccase confirm the involvement of laccase in lignin biosynthesis. Negligible transcript in phloem compared to the xylem tissue localized its expression in xylem tissue. This demonstrates the involvement of P. somniferum laccase in lignin biosynthesis of xylem, providing strength to the peduncle/stem and preventing lodging.

  16. Measurement of some Benzylisoquinoline Alkaloids in Different Organs of Persian Poppy during Ontogenetical Stages.

    PubMed

    Rezaei, Mahdi; Naghavi, Mohammad Reza; Hosseinzadeh, Abdol Hadi; Abbasi, Alireza

    2016-05-01

    Papaver bracteatum, a perennial species, has been known as a rich source of thebaine and a potential alternative to Papaver somniferum for the production of codeine and some semisynthetic antagonist drugs. In this study, ion mobility spectrum (IMS) of the root, leaf, bottom part of stem, upper part of stem, capsule wall, petal, and capsule content during developmental stages of P. bracteatum including annual rosette, perennial rosette, bud initiation, pendulous bud, preflowering, and lancing were investigated. The IMS revealed thebaine, papaverine, and noscapine as the major components of the extracted alkaloids. Based on the results of the study it appears that, at least in part, there is a competition among the biosynthesis pathways of papaverine, noscapine, and morphinan alkaloids from a common source. Root and capsule wall were the most potent organs for extraction of thebaine, while lancing stage was the best developmental stage for thebaine exploitation. However, it seems that total biomass of root and capsule wall plays a key role in the final selection of favorite organ. Although papaverine and noscapine in the stem at preflowering stage had the most quantity, significant amounts were found in the capsule wall. In general, total alkaloid content of leaf was lower than the other plant parts. PMID:27061130

  17. Rapid identification of a narcotic plant Papaver bracteatum using flow cytometry.

    PubMed

    Aragane, Masako; Watanabe, Daisuke; Nakajima, Jun'ichi; Yoshida, Masao; Yoshizawa, Masao; Abe, Tomohiro; Nishiyama, Rei; Suzuki, Jin; Moriyasu, Takako; Nakae, Dai; Sudo, Hiroshi; Sato, Hiroyuki; Hishida, Atuyuki; Kawahara, Nobuo; Makabe, So; Nakamura, Ikuo; Mii, Masahiro

    2014-10-01

    In May 2011, numerous poppy plants closely resembling Papaver bracteatum Lindl., a type of narcotic plant that is illegal in Japan, were distributed directly from several large flower shops or through online shopping throughout Japan, including the Tokyo Metropolitan area. In order to better identify the narcotic plants, the relative nuclear DNA content at the vegetative stage was measured by flow cytometric (FCM) analysis in 3 closely-related species of the genus Papaver section Oxytona, namely P. orientale, P. pseudo-orientale, and P. bracteatum, based on the difference between the chromosome numbers of these species. The results showed that the nuclear DNA content differed between these 3 species, and that most of the commercially distributed plants examined in this study could be identified as P. bracteatum. The remaining plants were P. pseudo-orientale, a non-narcotic plant. In addition, the FCM results for the identification of P. bracteatum completely agreed with the results obtained by the morphological analysis, the inter-genic spacer sequence of rpl16-rpl14 (PS-ID sequence) of chloroplast DNA, and the presence of thebaine. These results clearly indicate the usefulness of FCM analysis for the identification of P. bracteatum plants, including when they are in their vegetative stage. PMID:24952707

  18. Rapid identification of a narcotic plant Papaver bracteatum using flow cytometry.

    PubMed

    Aragane, Masako; Watanabe, Daisuke; Nakajima, Jun'ichi; Yoshida, Masao; Yoshizawa, Masao; Abe, Tomohiro; Nishiyama, Rei; Suzuki, Jin; Moriyasu, Takako; Nakae, Dai; Sudo, Hiroshi; Sato, Hiroyuki; Hishida, Atuyuki; Kawahara, Nobuo; Makabe, So; Nakamura, Ikuo; Mii, Masahiro

    2014-10-01

    In May 2011, numerous poppy plants closely resembling Papaver bracteatum Lindl., a type of narcotic plant that is illegal in Japan, were distributed directly from several large flower shops or through online shopping throughout Japan, including the Tokyo Metropolitan area. In order to better identify the narcotic plants, the relative nuclear DNA content at the vegetative stage was measured by flow cytometric (FCM) analysis in 3 closely-related species of the genus Papaver section Oxytona, namely P. orientale, P. pseudo-orientale, and P. bracteatum, based on the difference between the chromosome numbers of these species. The results showed that the nuclear DNA content differed between these 3 species, and that most of the commercially distributed plants examined in this study could be identified as P. bracteatum. The remaining plants were P. pseudo-orientale, a non-narcotic plant. In addition, the FCM results for the identification of P. bracteatum completely agreed with the results obtained by the morphological analysis, the inter-genic spacer sequence of rpl16-rpl14 (PS-ID sequence) of chloroplast DNA, and the presence of thebaine. These results clearly indicate the usefulness of FCM analysis for the identification of P. bracteatum plants, including when they are in their vegetative stage.

  19. POPPY: Physical Optics Propagation in PYthon

    NASA Astrophysics Data System (ADS)

    Perrin, Marshall; Long, Joseph; Douglas, Ewan; Sivaramakrishnan, Anand; Slocum, Christine

    2016-02-01

    POPPY (Physical Optics Propagation in PYthon) simulates physical optical propagation including diffraction. It implements a flexible framework for modeling Fraunhofer and Fresnel diffraction and point spread function formation, particularly in the context of astronomical telescopes. POPPY provides the optical modeling framework for WebbPSF (ascl:1504.007) and was developed as part of a simulation package for JWST, but is available separately and is broadly applicable to many kinds of imaging simulations.

  20. The rules of drug taking: wine and poppy derivatives in the ancient world. VII. A ritual use of poppy derivatives?

    PubMed

    Nencini, P

    1997-08-01

    Besides fertility, poppies have been used to symbolize sleep, night, and death. Consistent with the agrarian origin of their ritual use, poppies also became a symbol of reincarnation. Several literary and iconographic sources, in particular of the early Roman imperial age, are here interpreted as evidence that poppy derivatives were ingested during mystery rites. The reversible narcotic effects of poppy derivatives should have allowed a "realistic" representation of death and reincarnation, as intended by the Orphic belief of the transmigration of souls.

  1. Self-incompatibility in Papaver: advances in integrating the signalling network.

    PubMed

    Eaves, Deborah J; Flores-Ortiz, Carlos; Haque, Tamanna; Lin, Zongcheng; Teng, Nianjun; Franklin-Tong, Vernonica E

    2014-04-01

    Self-fertilization, which results in reduced fitness of offspring, is a common problem in hermaphrodite angiosperms. To prevent this, many plants utilize SI (self-incompatibility), which is determined by the multi-allelic S-locus, that allows discrimination between self (incompatible) and non-self (compatible) pollen by the pistil. In poppy (Papaver rhoeas), the pistil S-determinant (PrsS) is a small secreted protein which interacts with the pollen S-determinant PrpS, a ~20 kDa novel transmembrane protein. Interaction of matching pollen and pistil S-determinants results in self-recognition, initiating a Ca²⁺-dependent signalling network in incompatible pollen. This triggers several downstream events, including alterations to the cytoskeleton, phosphorylation of sPPases (soluble inorganic pyrophosphatases) and an MAPK (mitogen-activated protein kinase), increases in ROS (reactive oxygen species) and nitric oxide (NO), and activation of several caspase-like activities. This results in the inhibition of pollen tube growth, prevention of self-fertilization and ultimately PCD (programmed cell death) in incompatible pollen. The present review focuses on our current understanding of the integration of these signals with their targets in the SI/PCD network. We also discuss our recent functional expression of PrpS in Arabidopsis thaliana pollen.

  2. Extraction and Functional Properties of Proteins from Pre-roasted and Enzyme Treated Poppyseed (Papaver somniferum L.) Press Cakes.

    PubMed

    Yilmaz, Emin; Emir, Dilek Dündar

    2016-01-01

    In this study, proteins of the defatted meals obtained from cold-pressed poppyseed previously treated (pre-roasting and enzyme against control) were extracted and their compositional and functional properties were determined. Saline-alkaline extraction (pH 11-12, and 0.2-0.6 M NaCI) and isoelectric point (pH 4.0-5.5) precipitation technique showed that seed pre-roasting enhances protein yield while enzyme treatment reduces it. There were 7 bands on SDS-PAGE, and enzyme treated samples were weaker than control. While enzyme treatment decreased denaturation temperatures (T(d)), roasting enhanced the enthalpy change (ΔH) values. Pre-treatments caused a decrease in protein least gelling concentration (LGC) values. Water and oil holding capacities (WHC and OHC) were found lower in enzyme treated and higher in preroasted samples. Similar effects were also determined for emulsifying activity (EA) and emulsion stability (ES) values. While foaming capacity (FC) in treated samples decreased, foam stability (FS) increased oppositely. In conclusion, poppyseed meals can be nutritionally good source for diet protein, and a limited pre-roasting can be very beneficial for enhanced protein extraction yield and desirable functional properties. PMID:26972462

  3. Extraction and Functional Properties of Proteins from Pre-roasted and Enzyme Treated Poppyseed (Papaver somniferum L.) Press Cakes.

    PubMed

    Yilmaz, Emin; Emir, Dilek Dündar

    2016-01-01

    In this study, proteins of the defatted meals obtained from cold-pressed poppyseed previously treated (pre-roasting and enzyme against control) were extracted and their compositional and functional properties were determined. Saline-alkaline extraction (pH 11-12, and 0.2-0.6 M NaCI) and isoelectric point (pH 4.0-5.5) precipitation technique showed that seed pre-roasting enhances protein yield while enzyme treatment reduces it. There were 7 bands on SDS-PAGE, and enzyme treated samples were weaker than control. While enzyme treatment decreased denaturation temperatures (T(d)), roasting enhanced the enthalpy change (ΔH) values. Pre-treatments caused a decrease in protein least gelling concentration (LGC) values. Water and oil holding capacities (WHC and OHC) were found lower in enzyme treated and higher in preroasted samples. Similar effects were also determined for emulsifying activity (EA) and emulsion stability (ES) values. While foaming capacity (FC) in treated samples decreased, foam stability (FS) increased oppositely. In conclusion, poppyseed meals can be nutritionally good source for diet protein, and a limited pre-roasting can be very beneficial for enhanced protein extraction yield and desirable functional properties.

  4. Afghanistan, poppies, and the global pain crisis.

    PubMed

    Clark, Peter A; Sillup, George P; Capo, Joseph A

    2010-03-01

    The World Health Organization has reported that somewhere between 30-86 million people suffer from moderate to severe pain due to cancer, HIV/AIDS, burns, wounds and other illnesses annually and do not have access to proper opiate anesthetics to control the pain [1]. The vast majority of these people live in poor nations where medicinal opiates are either too expensive or not readily available. In this paper, it is argued that access to adequate healthcare is a human right and that adequate healthcare includes management of pain. The solution to this problem may be in Afghanistan, a country now overwhelmed with poverty and war. Afghanistan is the world's leading producer of heroin. The increase in heroin production in Afghanistan has caused the United States and the international community to begin to eradicate Afghanistan's poppy fields leading to increased poverty among poppy farmers. This paper proposed a paradigm that can be implemented in Afghanistan which would allow for Afghan farmers to continue growing their poppy crop for medicinal opiates like morphine for poor nations. The paradigm covers all parameters of medicinal opiates production including licensing, security, cultivation, harvest, and factory production of medicinal opiates. The paradigm proposed is less expensive than eradication, brings honest income to Afghan farmers and the new Afghan nation, and can eventually lead to Afghanistan acquiring a respectable role in the world community. In closing, a full ethical analysis of the paradigm is included to justify the arguments made in the paper. PMID:20190697

  5. Afghanistan, poppies, and the global pain crisis.

    PubMed

    Clark, Peter A; Sillup, George P; Capo, Joseph A

    2010-03-01

    The World Health Organization has reported that somewhere between 30-86 million people suffer from moderate to severe pain due to cancer, HIV/AIDS, burns, wounds and other illnesses annually and do not have access to proper opiate anesthetics to control the pain [1]. The vast majority of these people live in poor nations where medicinal opiates are either too expensive or not readily available. In this paper, it is argued that access to adequate healthcare is a human right and that adequate healthcare includes management of pain. The solution to this problem may be in Afghanistan, a country now overwhelmed with poverty and war. Afghanistan is the world's leading producer of heroin. The increase in heroin production in Afghanistan has caused the United States and the international community to begin to eradicate Afghanistan's poppy fields leading to increased poverty among poppy farmers. This paper proposed a paradigm that can be implemented in Afghanistan which would allow for Afghan farmers to continue growing their poppy crop for medicinal opiates like morphine for poor nations. The paradigm covers all parameters of medicinal opiates production including licensing, security, cultivation, harvest, and factory production of medicinal opiates. The paradigm proposed is less expensive than eradication, brings honest income to Afghan farmers and the new Afghan nation, and can eventually lead to Afghanistan acquiring a respectable role in the world community. In closing, a full ethical analysis of the paradigm is included to justify the arguments made in the paper.

  6. Organic parasite control for poultry and rabbits in British Columbia, Canada.

    PubMed

    Lans, Cheryl; Turner, Nancy

    2011-01-01

    Plants used for treating endo- and ectoparasites of rabbits and poultry in British Columbia included Arctium lappa (burdock), Artemisia sp. (wormwood), Chenopodium album (lambsquarters) and C. ambrosioides (epazote), Cirsium arvense (Canada thistle), Juniperus spp. (juniper), Mentha piperita (peppermint), Nicotiana sp. (tobacco), Papaver somniferum (opium poppy), Rubus spp. (blackberry and raspberry relatives), Symphytum officinale (comfrey), Taraxacum officinale (common dandelion), Thuja plicata (western redcedar) and Urtica dioica (stinging nettle). PMID:21756341

  7. Organic parasite control for poultry and rabbits in British Columbia, Canada

    PubMed Central

    2011-01-01

    Plants used for treating endo- and ectoparasites of rabbits and poultry in British Columbia included Arctium lappa (burdock), Artemisia sp. (wormwood), Chenopodium album (lambsquarters) and C. ambrosioides (epazote), Cirsium arvense (Canada thistle), Juniperus spp. (juniper), Mentha piperita (peppermint), Nicotiana sp. (tobacco), Papaver somniferum (opium poppy), Rubus spp. (blackberry and raspberry relatives), Symphytum officinale (comfrey), Taraxacum officinale (common dandelion), Thuja plicata (western redcedar) and Urtica dioica (stinging nettle). PMID:21756341

  8. Organic parasite control for poultry and rabbits in British Columbia, Canada.

    PubMed

    Lans, Cheryl; Turner, Nancy

    2011-07-14

    Plants used for treating endo- and ectoparasites of rabbits and poultry in British Columbia included Arctium lappa (burdock), Artemisia sp. (wormwood), Chenopodium album (lambsquarters) and C. ambrosioides (epazote), Cirsium arvense (Canada thistle), Juniperus spp. (juniper), Mentha piperita (peppermint), Nicotiana sp. (tobacco), Papaver somniferum (opium poppy), Rubus spp. (blackberry and raspberry relatives), Symphytum officinale (comfrey), Taraxacum officinale (common dandelion), Thuja plicata (western redcedar) and Urtica dioica (stinging nettle).

  9. The poppy seed defense: a novel solution.

    PubMed

    Chen, P; Braithwaite, R A; George, C; Hylands, P J; Parkin, M C; Smith, N W; Kicman, A T

    2014-03-01

    A major toxicological challenge is distinguishing whether morphine in urine, in the absence of 6-monoacetylmorphine (6-MAM), originates from 'street' heroin use or poppy seed ingestion. Manufacturing byproducts from the synthesis of illicit heroin include those that originate from the reaction of acetic anhydride with the alkaloid impurity, thebaine, which undergoes skeletal rearrangement, resulting in compounds with a 2-(N-methylacetamido)ethyl side-chain. The hypothesis that the tertiary amide in this side-chain is resistant to endogenous hydrolysis was supported from in-vitro experiments; a glucuronide metabolite (designated 'ATM4G') was identified that may be used as a marker of 'street' heroin administration. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis for this metabolite was then performed on selected urine specimens from 22 known heroin users, these being negative on routine testing for 6-MAM by gas chromatography-mass spectrometry (GC-MS), using the generally applied reporting threshold of 10 ng/mL, but positive for the presence of morphine. Peaks corresponding to the retention time for the metabolite marker were clearly observed for 16 of the 22 samples, with variations of the ratios of its three dependent ions being within ± 30% of that produced in vitro. Conversely, 6-MAM was detected in only 3 samples, but at concentrations <1 ng/mL. Such a high frequency for the presence of the metabolite marker in urine, in the absence of 6-MAM, is noteworthy and suggests that detection of this metabolite may offer an important advance in forensic toxicology, allowing the development of a new and more definitive test for heroin abuse and thus a potential solution to the so-called 'poppy seed defense'.

  10. Enhanced morphinan alkaloid production in hairy root cultures of Papaver bracteatum by over-expression of salutaridinol 7-o-acetyltransferase gene via Agrobacterium rhizogenes mediated transformation.

    PubMed

    Sharafi, Ali; Hashemi Sohi, Haleh; Mousavi, Amir; Azadi, Pejman; Dehsara, Bahareh; Hosseini Khalifani, Bahman

    2013-11-01

    Papaver bracteatum is an important medicinal plant valued for its high content of thebaine and an alternative to P. somniferum for benzylisoquinoline alkaloid production. Salutaridinol 7-o-acetyltransferase (SalAT) is a key gene in morphinan alkaloids biosynthesis pathway. Over expression of SalAT gene was used for metabolic engineering in P. bracteatum hairy root cultures. Transcript level of the salutaridinol 7-o-acetyltransferase gene in transgenic hairy root lines increased up to 154 and 128 % in comparison with hairy roots without SalAT over expression and wild type roots, respectively. High performance liquid chromatography analysis showed that the transgenic hairy roots relatively improved levels of thebaine (1.28 % dry weight), codeine (0.02 % dry weight) and morphine (0.03 % dry weight) compared to those hairy roots without SalAT over expression. This suggests that P. bracteatum hairy roots expressing the SalAT gene could be potentially used for the production of valuable morphinan alkaloids.

  11. [Guillain-Barre syndrome as a result of poisoning with a mixture of "kompot"(Polish heroin) and drugs].

    PubMed

    Gawlikowski, T; Winnik, L

    2001-01-01

    Guillain-Barré Syndrome is the most common form of polineuropathy. It is suggested that some infectious, immune and allergic factors are involved in developing the syndrome. "Kompot" or "Polish heroine", a domestic product, is produced from poppy straw or juice of poppy head (Papaver somniferum) and it is administered intravenously. "Kompot" shows variable contents of heroine, 6-MAM, 3-MAM, morphine, acetylo-codeine and codeine as well as papaverine, thebaine and narcotine. The case history of a young man dependent on "Polish heroine" who developed the Guillain-Barré Syndrome due to severe intoxication induced by home made heroine, barbiturates and benzodiazepines is described.

  12. [Screening potential DNA barcode regions of genus Papaver].

    PubMed

    Zhang, Shuang; Liu, Yu-jing; Wu, Yan-sheng; Cao, Ying; Yuan, Yuan

    2015-08-01

    DNA barcoding is an effective technique in species identification. To determine the candidate sequences which can be used as DNA barcode to identify in Papaver genus, five potential sequences (ITS, matK, psbA-trnH, rbcL, trnL-trnF) were screened. 69 sequences were downloaded from Genbank, including 21 ITS sequences, 10 matK sequences, 8 psbA-trnH sequences, 14 rbcL sequences and 16 trnL-trnF sequences. Mega 6.0 was used to analysis the comparison of sequences. By the methods of calculating the distances in intraspecific and interspecific divergences, evaluating DNA barcoding gap and constructing NJ and UPMGA phylogenetic trees. The sequence trnL-trnF performed best. In conclusion, trnL-trnF can be considered as a novel DNA barcode in Papaver genus, other four sequences can be as combination barcode for identification.

  13. [Screening potential DNA barcode regions of genus Papaver].

    PubMed

    Zhang, Shuang; Liu, Yu-jing; Wu, Yan-sheng; Cao, Ying; Yuan, Yuan

    2015-08-01

    DNA barcoding is an effective technique in species identification. To determine the candidate sequences which can be used as DNA barcode to identify in Papaver genus, five potential sequences (ITS, matK, psbA-trnH, rbcL, trnL-trnF) were screened. 69 sequences were downloaded from Genbank, including 21 ITS sequences, 10 matK sequences, 8 psbA-trnH sequences, 14 rbcL sequences and 16 trnL-trnF sequences. Mega 6.0 was used to analysis the comparison of sequences. By the methods of calculating the distances in intraspecific and interspecific divergences, evaluating DNA barcoding gap and constructing NJ and UPMGA phylogenetic trees. The sequence trnL-trnF performed best. In conclusion, trnL-trnF can be considered as a novel DNA barcode in Papaver genus, other four sequences can be as combination barcode for identification. PMID:26677693

  14. Exposure of consumers to morphine from poppy seeds in Hungary.

    PubMed

    Zentai, A; Sali, J; Szeitzné-Szabó, M; Szabó, I J; Ambrus, Á

    2012-01-01

    Poppy seed-containing foods are popular dishes in Hungary and some other Central European countries. The alkaloids of poppy are used in the production of medicines. Poppy seeds used as food may also contain considerable amounts of alkaloids, which raises the question of food safety. Morphine, codeine, thebaine and noscapine concentrations of poppy seed samples from the period 2001-2010 and consumption data from two Hungarian surveys, carried out in 2003 and 2009, were evaluated. Exposure calculations were made for morphine intake by both point estimate and probabilistic methods, and the uncertainty of the calculated values was estimated. The point estimate for the acute consumer exposure, calculated using the 97.5th percentiles of morphine concentration and of poppy seed consumption and taking into account the reduction of morphine content by processing, was 78.64 µg (kg bw)⁻¹ day⁻¹ for adults, and 116.90 µg (kg bw)⁻¹ day⁻¹ for children. Based on probabilistic estimations, the 97.5th and 99th percentile exposures ranged between 18.3-25.4 and 25.6-47.4 µg (kg bw)⁻¹ day⁻¹ for adults, and between 32.9 and 66.4 µg (kg bw)⁻¹ day⁻¹ for children, respectively. As a no observed effect level (NOEL) had not been established, the significance of exposure could not be assessed.

  15. Fatal intoxications in Denmark following intake of morphine from opium poppies.

    PubMed

    Steentoft, A; Kaa, E; Worm, K

    1988-01-01

    In Denmark it is legal to grow opium poppies for the production of poppy seeds and until 1986 for decoration purposes, too. Danish poppy capsules contain 0.3-5 mg morphine per capsule and the content of morphine in opium exuded from the capsules may amount to 24%. This has resulted in misuse as both fresh and dried poppy capsules have been used for the production of "opium tea". During the period 1982-1985, seven casualties occurred among drug addicts in Denmark which were solely or partly caused by these opium poppies. PMID:3227727

  16. Occurrence, genetic control and evolution of non-target-site based resistance to herbicides inhibiting acetolactate synthase (ALS) in the dicot weed Papaver rhoeas.

    PubMed

    Scarabel, Laura; Pernin, Fanny; Délye, Christophe

    2015-09-01

    Non-target-site resistance (NTSR) to herbicides is a major issue for the chemical control of weeds. Whilst predominant in grass weeds, NTSR remains largely uninvestigated in dicot weeds. We investigated the occurrence, inheritance and genetic control of NTSR to acetolactate synthase (ALS) inhibitors in Papaver rhoeas (corn poppy) using progenies from plants with potential NTSR to the imidazolinone herbicide imazamox. NTSR to imazamox was inherited from parents over two successive generations. NTSR to tritosulfuron (a sulfonylurea) was observed in F1 generations and inherited in F2 generations. NTSR to florasulam (a triazolopyrimidine) emerged in F2 generations. Our findings suggest NTSR was polygenic and gradually built-up by accumulation over generations of loci with moderate individual effects in single plants. We also demonstrated that ALS alleles conferring herbicide resistance can co-exist with NTSR loci in P. rhoeas plants. Previous research focussed on TSR in P. rhoeas, which most likely caused underestimation of NTSR significance in this species. This may also apply to other dicot species. From our data, resistance to ALS inhibitors in P. rhoeas appears complex, and involves well-known mutant ALS alleles and a set of unknown NTSR loci that confer resistance to ALS inhibitors from different chemical families. PMID:26259184

  17. Occurrence, genetic control and evolution of non-target-site based resistance to herbicides inhibiting acetolactate synthase (ALS) in the dicot weed Papaver rhoeas.

    PubMed

    Scarabel, Laura; Pernin, Fanny; Délye, Christophe

    2015-09-01

    Non-target-site resistance (NTSR) to herbicides is a major issue for the chemical control of weeds. Whilst predominant in grass weeds, NTSR remains largely uninvestigated in dicot weeds. We investigated the occurrence, inheritance and genetic control of NTSR to acetolactate synthase (ALS) inhibitors in Papaver rhoeas (corn poppy) using progenies from plants with potential NTSR to the imidazolinone herbicide imazamox. NTSR to imazamox was inherited from parents over two successive generations. NTSR to tritosulfuron (a sulfonylurea) was observed in F1 generations and inherited in F2 generations. NTSR to florasulam (a triazolopyrimidine) emerged in F2 generations. Our findings suggest NTSR was polygenic and gradually built-up by accumulation over generations of loci with moderate individual effects in single plants. We also demonstrated that ALS alleles conferring herbicide resistance can co-exist with NTSR loci in P. rhoeas plants. Previous research focussed on TSR in P. rhoeas, which most likely caused underestimation of NTSR significance in this species. This may also apply to other dicot species. From our data, resistance to ALS inhibitors in P. rhoeas appears complex, and involves well-known mutant ALS alleles and a set of unknown NTSR loci that confer resistance to ALS inhibitors from different chemical families.

  18. Self-incompatibility-induced programmed cell death in field poppy pollen involves dramatic acidification of the incompatible pollen tube cytosol.

    PubMed

    Wilkins, Katie A; Bosch, Maurice; Haque, Tamanna; Teng, Nianjun; Poulter, Natalie S; Franklin-Tong, Vernonica E

    2015-03-01

    Self-incompatibility (SI) is an important genetically controlled mechanism to prevent inbreeding in higher plants. SI involves highly specific interactions during pollination, resulting in the rejection of incompatible (self) pollen. Programmed cell death (PCD) is an important mechanism for destroying cells in a precisely regulated manner. SI in field poppy (Papaver rhoeas) triggers PCD in incompatible pollen. During SI-induced PCD, we previously observed a major acidification of the pollen cytosol. Here, we present measurements of temporal alterations in cytosolic pH ([pH]cyt); they were surprisingly rapid, reaching pH 6.4 within 10 min of SI induction and stabilizing by 60 min at pH 5.5. By manipulating the [pH]cyt of the pollen tubes in vivo, we show that [pH]cyt acidification is an integral and essential event for SI-induced PCD. Here, we provide evidence showing the physiological relevance of the cytosolic acidification and identify key targets of this major physiological alteration. A small drop in [pH]cyt inhibits the activity of a soluble inorganic pyrophosphatase required for pollen tube growth. We also show that [pH]cyt acidification is necessary and sufficient for triggering several key hallmark features of the SI PCD signaling pathway, notably activation of a DEVDase/caspase-3-like activity and formation of SI-induced punctate actin foci. Importantly, the actin binding proteins Cyclase-Associated Protein and Actin-Depolymerizing Factor are identified as key downstream targets. Thus, we have shown the biological relevance of an extreme but physiologically relevant alteration in [pH]cyt and its effect on several components in the context of SI-induced events and PCD.

  19. Effect of ground poppy seed as a fat replacer on meat burgers.

    PubMed

    Gök, Veli; Akkaya, Levent; Obuz, Ersel; Bulut, Sait

    2011-12-01

    Poppy seed paste was used in koefte production as an animal fat replacer and efficacy of using poppy seed was investigated. The use of ground poppy seed in formulation of meat burgers had no effect on the moisture content of uncooked meat burgers; however, it did have a significant effect (p<0.05) on the fat content of meat burgers. Ground poppy seed addition significantly affected (p<0.05) cooking yield, moisture retention, and fat retention of meat burgers. Ground poppy seed addition significantly decreased (p<0.05) saturated fatty acid contents as the amount of ground poppy seed increased in meat burgers. A strong significant decrease (p<0.05) on the cholesterol content of meat burgers with ground poppy seed addition was observed. Samples having 20% ground poppy seed had significantly better texture and juiciness score (p<0.05) than any other sample which could be explained by increased moisture retention. Replacing animal fat with ground poppy seed paste is effective and may pave the way for an innovative meat product.

  20. Tall Poppies: Bullying Behaviors Faced by Australian High-Performance School-Age Athletes

    ERIC Educational Resources Information Center

    O'Neill, Maureen; Calder, Angela; Allen, Bill

    2014-01-01

    Little is known about Australian high-performance school-age athletes' experiences as victims of the tall poppy syndrome. Tall poppies are successful individuals bullied by those who are less successful in order to "normalize them." Nineteen current or previous national or international high-performance school-age athletes were…

  1. Socio-climatic Exposure of an Afghan Poppy Farmer

    NASA Astrophysics Data System (ADS)

    Mankin, J. S.; Diffenbaugh, N. S.

    2011-12-01

    Many posit that climate impacts from anthropogenic greenhouse gas emissions will have consequences for the natural and agricultural systems on which humans rely for food, energy, and livelihoods, and therefore, on stability and human security. However, many of the potential mechanisms of action in climate impacts and human systems response, as well as the differential vulnerabilities of such systems, remain underexplored and unquantified. Here I present two initial steps necessary to characterize and quantify the consequences of climate change for farmer livelihood in Afghanistan, given both climate impacts and farmer vulnerabilities. The first is a conceptual model mapping the potential relationships between Afghanistan's climate, the winter agricultural season, and the country's political economy of violence and instability. The second is a utility-based decision model for assessing farmer response sensitivity to various climate impacts based on crop sensitivities. A farmer's winter planting decision can be modeled roughly as a tradeoff between cultivating the two crops that dominate the winter growing season-opium poppy (a climate tolerant cash crop) and wheat (a climatically vulnerable crop grown for household consumption). Early sensitivity analysis results suggest that wheat yield dominates farmer decision making variability; however, such initial results may dependent on the relative parameter ranges of wheat and poppy yields. Importantly though, the variance in Afghanistan's winter harvest yields of poppy and wheat is tightly linked to household livelihood and thus, is indirectly connected to the wider instability and insecurity within the country. This initial analysis motivates my focused research on the sensitivity of these crops to climate variability in order to project farmer well-being and decision sensitivity in a warmer world.

  2. Buprenorphine for cancer pain: is it ready for prime time?

    PubMed

    Prommer, Eric

    2015-12-01

    Buprenorphine (BUP) is a semisynthetic derivative of the opium alkaloid thebaine found in the poppy Papaver somniferum. Its chemical structure contains the morphine structure but differs by having a cyclopropylmethyl group. Buprenorphine is a potent µ opioid agonist. Buprenorphine undergoes extensive first-pass metabolism in the liver and gut. The development of a transdermal BUP formulation in 2001 led to its evaluation in cancer pain. This article provides the practitioner with an update on the current role of BUP in cancer care. It highlights data suggesting effectiveness in various types of cancer pain. The article reviews pharmacology, routes of administration, adverse effects, drug interactions, and cost considerations. PMID:25163678

  3. Effect of poppy seed consummation on the positive results of opiates screening in biological samples.

    PubMed

    Jankovicová, Katarína; Ulbrich, Pavol; Fuknová, Mária

    2009-04-01

    Poppy seed is a popular substance of many traditional Slovak cakes. We can eat quite great amount of it, sometimes more than 50 g. Existing problem in interpreting the results of opiate urine analysis in case of drug abuse arises from the natural occurrence of opiate alkaloids in poppy seed. Interpretation of morphine presence in urine sample is in some cases a problem because morphine present in the urine sample may come from different "sources". The presence of additional, respectively, other opiate in urine sample is significant help when interpreting the presence of morphine. We used poppy seed bought in supermarket for our experiment. Presence of morphine and codeine was determined in poppy seed extracts, whereas the concentration of majority opiate-morphine was 0.9 mg/100 g (9 ppm). This poppy seed was used for two series of experiment-poppy seed consummation, where four persons consumed 100g of poppy seed in the first series and 50 g in the second series. Urine samples were taken in regular 1h intervals where first urine sample was given for testing 3 h after consummation. Concentrations of total opiates were determined in each urine sample by screening examination. Morphine concentrations were determined in selected urine samples using GC/MS with internal standard.

  4. Degradation of morphine in opium poppy processing waste composting.

    PubMed

    Wang, Yin Quan; Zhang, Jin Lin; Schuchardt, Frank; Wang, Yan

    2014-09-01

    To investigate morphine degradation and optimize turning frequency in opium poppy processing waste composting, a pilot scale windrow composting trial was run for 55 days. Four treatments were designed as without turning (A1), every 5 days turning (A2), every 10 days turning (A3) and every 15 days turning (A4). During composting, a range of physicochemical parameters including the residual morphine degradation, temperature, pH, and the contents of total C, total N, total P and total K were investigated. For all treatments, the residual morphine content decreased below the detection limit and reached the safety standards after day 30 of composting, the longest duration of high temperature (⩾50 °C) was observed in A3, pH increased 16.9-17.54%, total carbon content decreased 15.5-22.5%, C/N ratio reduced from 46 to 26, and the content of total phosphorus and total potassium increased slightly. The final compost obtained by a mixture of all four piles was up to 55.3% of organic matter, 3.3% of total nutrient (N, P2O5 and K2O) and 7.6 of pH. A turning frequency of every ten days for a windrow composting of opium poppy processing waste is recommended to produce homogenous compost.

  5. Concentrations of Morphine and Codeine in Paired Oral Fluid and Urine Specimens Following Ingestion of a Poppy Seed Roll and Raw Poppy Seeds

    PubMed Central

    Samano, Kimberly L.; Clouette, Randal E.; Rowland, Barbara J.; Sample, R.H. Barry

    2015-01-01

    Interpretation of opiate drug test results can be challenging due to casual dietary consumption of poppy seeds, which may contain variable opiate content. Opiate concentrations in paired oral fluid (OF), collected with the Oral-Eze® Oral Fluid Collection System, and urine were analyzed after ingestion of poppy seeds from the same source, consumed raw or contained in a roll. In Part 1, 12 individuals consumed equal portions of a poppy seed roll. For Part 2, the same individuals consumed an equivalent quantity of raw poppy seeds, containing ∼3.2 mg of morphine and 0.6 mg of codeine. Specimens were analyzed both by enzyme immunoassay (opiates) and by GC–MS (morphine/codeine). Urinary morphine was between 155–1,408 (roll) and 294–4,213 ng/mL (raw), measured at 2, 4, 6 and 20 h post-ingestion. Urinary codeine concentrations between 140–194 (roll) and 121–664 ng/mL (raw) were observed up to 6 h post-ingestion. Following consumption of raw poppy seeds, OF specimens were positive, above LOQ, from 0.25 to 3.0 h with morphine ranging from 7 to 600 ng/mL and codeine from 8 to 112 ng/mL. After poppy seed roll consumption, morphine concentrations of 7–143 ng/mL were observed up to 1.5 h with codeine detected in only 5.5% of OF specimens and ranging from 8 to 28 ng/mL. Combined with the existing poppy seed literature, these results support previous findings and provide guidance for interpretation of OF opiate testing. PMID:26378141

  6. Concentrations of Morphine and Codeine in Paired Oral Fluid and Urine Specimens Following Ingestion of a Poppy Seed Roll and Raw Poppy Seeds.

    PubMed

    Samano, Kimberly L; Clouette, Randal E; Rowland, Barbara J; Sample, R H Barry

    2015-10-01

    Interpretation of opiate drug test results can be challenging due to casual dietary consumption of poppy seeds, which may contain variable opiate content. Opiate concentrations in paired oral fluid (OF), collected with the Oral-Eze(®) Oral Fluid Collection System, and urine were analyzed after ingestion of poppy seeds from the same source, consumed raw or contained in a roll. In Part 1, 12 individuals consumed equal portions of a poppy seed roll. For Part 2, the same individuals consumed an equivalent quantity of raw poppy seeds, containing ∼3.2 mg of morphine and 0.6 mg of codeine. Specimens were analyzed both by enzyme immunoassay (opiates) and by GC-MS (morphine/codeine). Urinary morphine was between 155-1,408 (roll) and 294-4,213 ng/mL (raw), measured at 2, 4, 6 and 20 h post-ingestion. Urinary codeine concentrations between 140-194 (roll) and 121-664 ng/mL (raw) were observed up to 6 h post-ingestion. Following consumption of raw poppy seeds, OF specimens were positive, above LOQ, from 0.25 to 3.0 h with morphine ranging from 7 to 600 ng/mL and codeine from 8 to 112 ng/mL. After poppy seed roll consumption, morphine concentrations of 7-143 ng/mL were observed up to 1.5 h with codeine detected in only 5.5% of OF specimens and ranging from 8 to 28 ng/mL. Combined with the existing poppy seed literature, these results support previous findings and provide guidance for interpretation of OF opiate testing.

  7. Concentrations of Morphine and Codeine in Paired Oral Fluid and Urine Specimens Following Ingestion of a Poppy Seed Roll and Raw Poppy Seeds.

    PubMed

    Samano, Kimberly L; Clouette, Randal E; Rowland, Barbara J; Sample, R H Barry

    2015-10-01

    Interpretation of opiate drug test results can be challenging due to casual dietary consumption of poppy seeds, which may contain variable opiate content. Opiate concentrations in paired oral fluid (OF), collected with the Oral-Eze(®) Oral Fluid Collection System, and urine were analyzed after ingestion of poppy seeds from the same source, consumed raw or contained in a roll. In Part 1, 12 individuals consumed equal portions of a poppy seed roll. For Part 2, the same individuals consumed an equivalent quantity of raw poppy seeds, containing ∼3.2 mg of morphine and 0.6 mg of codeine. Specimens were analyzed both by enzyme immunoassay (opiates) and by GC-MS (morphine/codeine). Urinary morphine was between 155-1,408 (roll) and 294-4,213 ng/mL (raw), measured at 2, 4, 6 and 20 h post-ingestion. Urinary codeine concentrations between 140-194 (roll) and 121-664 ng/mL (raw) were observed up to 6 h post-ingestion. Following consumption of raw poppy seeds, OF specimens were positive, above LOQ, from 0.25 to 3.0 h with morphine ranging from 7 to 600 ng/mL and codeine from 8 to 112 ng/mL. After poppy seed roll consumption, morphine concentrations of 7-143 ng/mL were observed up to 1.5 h with codeine detected in only 5.5% of OF specimens and ranging from 8 to 28 ng/mL. Combined with the existing poppy seed literature, these results support previous findings and provide guidance for interpretation of OF opiate testing. PMID:26378141

  8. Self-Incompatibility-Induced Programmed Cell Death in Field Poppy Pollen Involves Dramatic Acidification of the Incompatible Pollen Tube Cytosol1[OPEN

    PubMed Central

    Wilkins, Katie A.; Bosch, Maurice; Haque, Tamanna; Teng, Nianjun; Poulter, Natalie S.; Franklin-Tong, Vernonica E.

    2015-01-01

    Self-incompatibility (SI) is an important genetically controlled mechanism to prevent inbreeding in higher plants. SI involves highly specific interactions during pollination, resulting in the rejection of incompatible (self) pollen. Programmed cell death (PCD) is an important mechanism for destroying cells in a precisely regulated manner. SI in field poppy (Papaver rhoeas) triggers PCD in incompatible pollen. During SI-induced PCD, we previously observed a major acidification of the pollen cytosol. Here, we present measurements of temporal alterations in cytosolic pH ([pH]cyt); they were surprisingly rapid, reaching pH 6.4 within 10 min of SI induction and stabilizing by 60 min at pH 5.5. By manipulating the [pH]cyt of the pollen tubes in vivo, we show that [pH]cyt acidification is an integral and essential event for SI-induced PCD. Here, we provide evidence showing the physiological relevance of the cytosolic acidification and identify key targets of this major physiological alteration. A small drop in [pH]cyt inhibits the activity of a soluble inorganic pyrophosphatase required for pollen tube growth. We also show that [pH]cyt acidification is necessary and sufficient for triggering several key hallmark features of the SI PCD signaling pathway, notably activation of a DEVDase/caspase-3-like activity and formation of SI-induced punctate actin foci. Importantly, the actin binding proteins Cyclase-Associated Protein and Actin-Depolymerizing Factor are identified as key downstream targets. Thus, we have shown the biological relevance of an extreme but physiologically relevant alteration in [pH]cyt and its effect on several components in the context of SI-induced events and PCD. PMID:25630437

  9. [Alimentary intake of opioid alkaloids by horses. Hazards due to poppy-containing feeds].

    PubMed

    Hertzsch, R; Emmerich, I U; Lachenmeier, D W; Sproll, C; Monakhova, Y B; Aboling, S; Bachmann, U; Vervuert, I

    2015-01-01

    Opioid alkaloids were identified in the urine of horses during an anti-doping control and in a case of intoxication. In both cases, it was suspected that the horses had ingested poppy-contaminated feed. To verify this suspicion, possible opioid alkaloid sources in Germany were identified through a literature research. Additionally, the contaminated feed was botanically and chemically analysed. The results indicated that both cases were most probably caused by the poppy in the feed. This highlights the previously underestimated risk of an intake of poppy-contaminated feed in horses. Recommendations are formulated for the prevention of positive doping-test results and intoxications by poppy-contaminated feeds in horses. Furthermore, a threshold for morphine in urine samples in competing horses is proposed.

  10. [Alimentary intake of opioid alkaloids by horses. Hazards due to poppy-containing feeds].

    PubMed

    Hertzsch, R; Emmerich, I U; Lachenmeier, D W; Sproll, C; Monakhova, Y B; Aboling, S; Bachmann, U; Vervuert, I

    2015-01-01

    Opioid alkaloids were identified in the urine of horses during an anti-doping control and in a case of intoxication. In both cases, it was suspected that the horses had ingested poppy-contaminated feed. To verify this suspicion, possible opioid alkaloid sources in Germany were identified through a literature research. Additionally, the contaminated feed was botanically and chemically analysed. The results indicated that both cases were most probably caused by the poppy in the feed. This highlights the previously underestimated risk of an intake of poppy-contaminated feed in horses. Recommendations are formulated for the prevention of positive doping-test results and intoxications by poppy-contaminated feeds in horses. Furthermore, a threshold for morphine in urine samples in competing horses is proposed. PMID:25624174

  11. Poppy seed foods and opiate drug testing--where are we today?

    PubMed

    Lachenmeier, Dirk W; Sproll, Constanze; Musshoff, Frank

    2010-02-01

    Seeds of the opium poppy plant are legally sold and widely consumed as food. Due to contamination during harvesting, the seeds can contain morphine and other opiate alkaloids. The objective of this study is to review the toxicology of poppy seed foods regarding influence on opiate drug tests. Computer-assisted literature review resulted in 95 identified references. Normal poppy seed consumption is generally regarded as safe. During food processing, the morphine content is considerably reduced (up to 90%). The possibility of false-positive opiate drug tests after poppy food ingestion exists. There are no unambiguous markers available to differentiate poppy food ingestion from heroin or pharmaceutical morphine use. This is also a problem in heroin-assisted maintenance programs. A basic requirement in such substitution programs is the patients' abstinence from any other drugs, including additional illicit heroin. Also a lack of forensic ingestion trials was detected that consider all factors influencing the morphine content in biologic matrices after consumption. Most studies did not control for the losses during food processing, so that the initial morphine dosage was overestimated. The large reduction of the morphine content during past years raises questions about the validity of the "poppy seed defence." However, a threshold of food use that would not lead to positive drug tests with certainty is currently unavailable. Research is needed to prove if the morphine contents in today's foods still pose the possibility of influencing drug tests. Future trials should consider processing-related morphine losses. PMID:19901868

  12. Effect of Replacing Beef Fat with Poppy Seed Oil on Quality of Turkish Sucuk.

    PubMed

    Gök, Vel

    2015-01-01

    Sucuk is the most popular dry-fermented meat product. Sucuk has a relatively high fat. Poppy seed oil as animal fat replacer was used in Turkish sucuk and effects of its use on sucuk quality were investigated. There was a significant (p<0.5) treatment × ripening time interaction for moisture, pH (p<0.05) and 2-thiobarbituric acid reactive substances (TBARS) values (p<0.01). Increasing poppy seed oil level decreased (p<0.05) TBARS values. Addition of poppy seed oil to the sucuks had a significant effect (p<0.01) on hardness, cohesiveness, gumminess, chewiness and springiness values. Cholesterol content of sucuks decreased (p<0.05) with poppy seed oil addition. Using pre-emulsified poppy seed oil as partial fat replacer in Turkish sucuk decreased cholesterol and saturated fatty acid content, but increased polyunsaturated fatty acids. Poppy seed oil as partial animal fat replacer in Turkish sucuk may have significant health benefits.

  13. Effect of Replacing Beef Fat with Poppy Seed Oil on Quality of Turkish Sucuk

    PubMed Central

    2015-01-01

    Sucuk is the most popular dry-fermented meat product. Sucuk has a relatively high fat. Poppy seed oil as animal fat replacer was used in Turkish sucuk and effects of its use on sucuk quality were investigated. There was a significant (p<0.5) treatment × ripening time interaction for moisture, pH (p<0.05) and 2-thiobarbituric acid reactive substances (TBARS) values (p<0.01). Increasing poppy seed oil level decreased (p<0.05) TBARS values. Addition of poppy seed oil to the sucuks had a significant effect (p<0.01) on hardness, cohesiveness, gumminess, chewiness and springiness values. Cholesterol content of sucuks decreased (p<0.05) with poppy seed oil addition. Using pre-emulsified poppy seed oil as partial fat replacer in Turkish sucuk decreased cholesterol and saturated fatty acid content, but increased polyunsaturated fatty acids. Poppy seed oil as partial animal fat replacer in Turkish sucuk may have significant health benefits. PMID:26761834

  14. Poppy seed foods and opiate drug testing--where are we today?

    PubMed

    Lachenmeier, Dirk W; Sproll, Constanze; Musshoff, Frank

    2010-02-01

    Seeds of the opium poppy plant are legally sold and widely consumed as food. Due to contamination during harvesting, the seeds can contain morphine and other opiate alkaloids. The objective of this study is to review the toxicology of poppy seed foods regarding influence on opiate drug tests. Computer-assisted literature review resulted in 95 identified references. Normal poppy seed consumption is generally regarded as safe. During food processing, the morphine content is considerably reduced (up to 90%). The possibility of false-positive opiate drug tests after poppy food ingestion exists. There are no unambiguous markers available to differentiate poppy food ingestion from heroin or pharmaceutical morphine use. This is also a problem in heroin-assisted maintenance programs. A basic requirement in such substitution programs is the patients' abstinence from any other drugs, including additional illicit heroin. Also a lack of forensic ingestion trials was detected that consider all factors influencing the morphine content in biologic matrices after consumption. Most studies did not control for the losses during food processing, so that the initial morphine dosage was overestimated. The large reduction of the morphine content during past years raises questions about the validity of the "poppy seed defence." However, a threshold of food use that would not lead to positive drug tests with certainty is currently unavailable. Research is needed to prove if the morphine contents in today's foods still pose the possibility of influencing drug tests. Future trials should consider processing-related morphine losses.

  15. Effect of Replacing Beef Fat with Poppy Seed Oil on Quality of Turkish Sucuk.

    PubMed

    Gök, Vel

    2015-01-01

    Sucuk is the most popular dry-fermented meat product. Sucuk has a relatively high fat. Poppy seed oil as animal fat replacer was used in Turkish sucuk and effects of its use on sucuk quality were investigated. There was a significant (p<0.5) treatment × ripening time interaction for moisture, pH (p<0.05) and 2-thiobarbituric acid reactive substances (TBARS) values (p<0.01). Increasing poppy seed oil level decreased (p<0.05) TBARS values. Addition of poppy seed oil to the sucuks had a significant effect (p<0.01) on hardness, cohesiveness, gumminess, chewiness and springiness values. Cholesterol content of sucuks decreased (p<0.05) with poppy seed oil addition. Using pre-emulsified poppy seed oil as partial fat replacer in Turkish sucuk decreased cholesterol and saturated fatty acid content, but increased polyunsaturated fatty acids. Poppy seed oil as partial animal fat replacer in Turkish sucuk may have significant health benefits. PMID:26761834

  16. The hidden habit of the entomopathogenic fungus Beauveria bassiana: first demonstration of vertical plant transmission.

    PubMed

    Quesada-Moraga, Enrique; López-Díaz, Cristina; Landa, Blanca Beatriz

    2014-01-01

    Beauveria bassiana strain 04/01-Tip, obtained from a larva of the opium poppy stem gall wasp Iraella luteipes (Hymenoptera; Cynipidae), endophytically colonizes opium poppy (Papaver somniferum L.) plants and protects them against this pest. The goal of this study was to monitor the dynamics of endophytic colonization of opium poppy by B. bassiana after the fungus was applied to the seed and to ascertain whether the fungus is transmitted vertically via seeds. Using a species-specific nested PCR protocol and DNA extracted from surface-sterilised leaf pieces or seeds of B. bassiana-inoculated opium poppy plants, the fungus was detected within the plant beginning at the growth stage of rosette building and them throughout the entire plant growth cycle (about 120-140 days after sowing). The fungus was also detected in seeds from 50% of the capsules sampled. Seeds that showed positive amplification for B. bassiana were planted in sterile soil and the endophyte was again detected in more than 42% of the plants sampled during all plant growth stages. Beauveria bassiana was transmitted to seeds in 25% of the plants from the second generation that formed a mature capsule. These results demonstrate for the first time the vertical transmission of an entomopathogenic fungus from endophytically colonised maternal plants. This information is crucial to better understand the ecological role of entomopathogenic fungi as plant endophytes and may allow development of a sustainable and cost effective strategy for I. luteipes management in P. somniferum.

  17. The Hidden Habit of the Entomopathogenic Fungus Beauveria bassiana: First Demonstration of Vertical Plant Transmission

    PubMed Central

    Quesada-Moraga, Enrique; López-Díaz, Cristina; Landa, Blanca Beatriz

    2014-01-01

    Beauveria bassiana strain 04/01-Tip, obtained from a larva of the opium poppy stem gall wasp Iraella luteipes (Hymenoptera; Cynipidae), endophytically colonizes opium poppy (Papaver somniferum L.) plants and protects them against this pest. The goal of this study was to monitor the dynamics of endophytic colonization of opium poppy by B. bassiana after the fungus was applied to the seed and to ascertain whether the fungus is transmitted vertically via seeds. Using a species-specific nested PCR protocol and DNA extracted from surface-sterilised leaf pieces or seeds of B. bassiana-inoculated opium poppy plants, the fungus was detected within the plant beginning at the growth stage of rosette building and them throughout the entire plant growth cycle (about 120–140 days after sowing). The fungus was also detected in seeds from 50% of the capsules sampled. Seeds that showed positive amplification for B. bassiana were planted in sterile soil and the endophyte was again detected in more than 42% of the plants sampled during all plant growth stages. Beauveria bassiana was transmitted to seeds in 25% of the plants from the second generation that formed a mature capsule. These results demonstrate for the first time the vertical transmission of an entomopathogenic fungus from endophytically colonised maternal plants. This information is crucial to better understand the ecological role of entomopathogenic fungi as plant endophytes and may allow development of a sustainable and cost effective strategy for I. luteipes management in P. somniferum. PMID:24551242

  18. A microbial biomanufacturing platform for natural and semisynthetic opioids.

    PubMed

    Thodey, Kate; Galanie, Stephanie; Smolke, Christina D

    2014-10-01

    Opiates and related molecules are medically essential, but their production via field cultivation of opium poppy Papaver somniferum leads to supply inefficiencies and insecurity. As an alternative production strategy, we developed baker's yeast Saccharomyces cerevisiae as a microbial host for the transformation of opiates. Yeast strains engineered to express heterologous genes from P. somniferum and bacterium Pseudomonas putida M10 convert thebaine to codeine, morphine, hydromorphone, hydrocodone and oxycodone. We discovered a new biosynthetic branch to neopine and neomorphine, which diverted pathway flux from morphine and other target products. We optimized strain titer and specificity by titrating gene copy number, enhancing cosubstrate supply, applying a spatial engineering strategy and performing high-density fermentation, which resulted in total opioid titers up to 131 mg/l. This work is an important step toward total biosynthesis of valuable benzylisoquinoline alkaloid drug molecules and demonstrates the potential for developing a sustainable and secure yeast biomanufacturing platform for opioids. PMID:25151135

  19. Engineering biosynthesis of the anticancer alkaloid noscapine in yeast

    PubMed Central

    Li, Yanran; Smolke, Christina D.

    2016-01-01

    Noscapine is a potential anticancer drug isolated from the opium poppy Papaver somniferum, and genes encoding enzymes responsible for the synthesis of noscapine have been recently discovered to be clustered on the genome of P. somniferum. Here, we reconstitute the noscapine gene cluster in Saccharomyces cerevisiae to achieve the microbial production of noscapine and related pathway intermediates, complementing and extending previous in planta and in vitro investigations. Our work provides structural validation of the secoberberine intermediates and the description of the narcotoline-4′-O-methyltransferase, suggesting this activity is catalysed by a unique heterodimer. We also reconstitute a 14-step biosynthetic pathway of noscapine from the simple alkaloid norlaudanosoline by engineering a yeast strain expressing 16 heterologous plant enzymes, achieving reconstitution of a complex plant pathway in a microbial host. Other engineered yeasts produce previously inaccessible pathway intermediates and a novel derivative, thereby advancing protoberberine and noscapine related drug discovery. PMID:27378283

  20. A microbial biomanufacturing platform for natural and semisynthetic opioids.

    PubMed

    Thodey, Kate; Galanie, Stephanie; Smolke, Christina D

    2014-10-01

    Opiates and related molecules are medically essential, but their production via field cultivation of opium poppy Papaver somniferum leads to supply inefficiencies and insecurity. As an alternative production strategy, we developed baker's yeast Saccharomyces cerevisiae as a microbial host for the transformation of opiates. Yeast strains engineered to express heterologous genes from P. somniferum and bacterium Pseudomonas putida M10 convert thebaine to codeine, morphine, hydromorphone, hydrocodone and oxycodone. We discovered a new biosynthetic branch to neopine and neomorphine, which diverted pathway flux from morphine and other target products. We optimized strain titer and specificity by titrating gene copy number, enhancing cosubstrate supply, applying a spatial engineering strategy and performing high-density fermentation, which resulted in total opioid titers up to 131 mg/l. This work is an important step toward total biosynthesis of valuable benzylisoquinoline alkaloid drug molecules and demonstrates the potential for developing a sustainable and secure yeast biomanufacturing platform for opioids.

  1. Engineering biosynthesis of the anticancer alkaloid noscapine in yeast.

    PubMed

    Li, Yanran; Smolke, Christina D

    2016-01-01

    Noscapine is a potential anticancer drug isolated from the opium poppy Papaver somniferum, and genes encoding enzymes responsible for the synthesis of noscapine have been recently discovered to be clustered on the genome of P. somniferum. Here, we reconstitute the noscapine gene cluster in Saccharomyces cerevisiae to achieve the microbial production of noscapine and related pathway intermediates, complementing and extending previous in planta and in vitro investigations. Our work provides structural validation of the secoberberine intermediates and the description of the narcotoline-4'-O-methyltransferase, suggesting this activity is catalysed by a unique heterodimer. We also reconstitute a 14-step biosynthetic pathway of noscapine from the simple alkaloid norlaudanosoline by engineering a yeast strain expressing 16 heterologous plant enzymes, achieving reconstitution of a complex plant pathway in a microbial host. Other engineered yeasts produce previously inaccessible pathway intermediates and a novel derivative, thereby advancing protoberberine and noscapine related drug discovery. PMID:27378283

  2. Thebaine in hair as a marker for chronic use of illegal opium poppy substances.

    PubMed

    Lee, Sooyeun; Park, Yonghoon; Han, Eunyoung; Choi, Hwakyung; Chung, Heesun; Oh, Seung Min; Chung, Kyu Hyuck

    2011-01-30

    Opium poppy products are often illegally used for both recreational and medicinal purposes. In order to demonstrate the ingestion of opium poppy substances, morphine, codeine and their metabolites have been identified. However, morphine and codeine also originate from the ingestion of therapeutic drugs. Therefore, thebaine, one of the main opium alkaloids, in hair was suggested as a marker for chronic use of illegal opium poppy substances in the present study. First, thebaine was included in the analyte list of our routine analytical method for the simultaneous quantification of codeine, morphine, norcodeine, normorphine and 6-acetylmorphine (6-AM) in hair, which was fully validated previously. Then, the incorporation of thebaine and other opiates into hair and the effect of hair pigmentation were examined using lean Zucker rats with both dark grey and white hair on the same body. Thebaine was also measured in hair samples from actual cases of opium poppy substance use. Consequently, thebaine in hair was demonstrated as a marker of chronic use of illegal opium poppy substances using an animal study and actual cases. Thebaine and other opiates were successfully measured in pigmented hair from rats that ingested raw opium suspension. Moreover, thebaine identified in hair excluded possibility of ingestion of pharmaceutical opiates in actual cases.

  3. Cloning and characterization of canadine synthase involved in noscapine biosynthesis in opium poppy.

    PubMed

    Dang, Thu-Thuy T; Facchini, Peter J

    2014-01-01

    Noscapine biosynthesis in opium poppy is thought to occur via N-methylcanadine, which would be produced through 9-O-methylation of (S)-scoulerine, methylenedioxy bridge formation on (S)-tetrahydrocolumbamine, and N-methylation of (S)-canadine. Only scoulerine 9-O-methyltransferase has been functionally characterized. We report the isolation and characterization of a cytochrome P450 (CYP719A21) from opium poppy that converts (S)-tetrahydrocolumbamine to (S)-canadine. Recombinant CYP719A21 displayed strict substrate specificity and high affinity (Km=4.63±0.71 μM) for (S)-tetrahydrocolumbamine. Virus-induced gene silencing of CYP719A21 caused a significant increase in (S)-tetrahydrocolumbamine accumulation and a corresponding decrease in the levels of putative downstream intermediates and noscapine in opium poppy plants.

  4. The opium poppy as a symbol of sleep in Bertel Thorvaldsen's relief of 1815.

    PubMed

    Tekiner, Halil; Kosar, Muberra

    2016-03-01

    Danish sculptor Bertel Thorvaldsen (1770-1844) is one of the most remarkable representatives of Neoclassicist sculptural art in Europe, which was largely inspired by the classical art and culture of Greek and Roman antiquity. A pair of marble reliefs, Night and Day, exhibited in the Thorvaldsen Museum (Copenhagen), marks the culmination of Thorvaldsen's relief art and is of particular interest to the history of sleep medicine. In the first relief, Night, an angel with her neck bent and eyes closed has two babies in her embrace and seems to be floating down in grief, with an owl hovering behind her. Her hair is also twined with opium poppies, the symbol of sleep and death in antiquity. Our findings suggest that this relief not only indicates a mythological association between the opium poppy and sleep but also has a strong connotation with the poppy's medicinal use for inducing sleep throughout the centuries.

  5. The opium poppy as a symbol of sleep in Bertel Thorvaldsen's relief of 1815.

    PubMed

    Tekiner, Halil; Kosar, Muberra

    2016-03-01

    Danish sculptor Bertel Thorvaldsen (1770-1844) is one of the most remarkable representatives of Neoclassicist sculptural art in Europe, which was largely inspired by the classical art and culture of Greek and Roman antiquity. A pair of marble reliefs, Night and Day, exhibited in the Thorvaldsen Museum (Copenhagen), marks the culmination of Thorvaldsen's relief art and is of particular interest to the history of sleep medicine. In the first relief, Night, an angel with her neck bent and eyes closed has two babies in her embrace and seems to be floating down in grief, with an owl hovering behind her. Her hair is also twined with opium poppies, the symbol of sleep and death in antiquity. Our findings suggest that this relief not only indicates a mythological association between the opium poppy and sleep but also has a strong connotation with the poppy's medicinal use for inducing sleep throughout the centuries. PMID:26210393

  6. Differentiation of opium and poppy straw using capillary electrophoresis and pattern recognition techniques.

    PubMed

    Reid, Raymond G; Durham, David G; Boyle, Susanne P; Low, Ann S; Wangboonskul, Jinda

    2007-12-12

    Opium samples from four different locations and poppy straw from different plant varieties have been assayed using micellar capillary electrophoresis incorporating a sweeping technique. Individual alkaloids (morphine, codeine, papaverine, noscapine, thebaine, oripavine, reticuline and narceine) were quantitatively determined in the different samples by a validated capillary electrophoresis method. Unsupervised pattern recognition of the opium samples and the poppy straw samples using hierarchical cluster analysis (HCA) and principal component analysis (PCA), showed distinct clusters. Supervised pattern recognition using soft independent modelling of class analogy (SIMCA) was performed to show individual groupings and allow unknown samples to be classified according to the models built using the CZE assay results.

  7. Differentiation of opium and poppy straw using capillary electrophoresis and pattern recognition techniques.

    PubMed

    Reid, Raymond G; Durham, David G; Boyle, Susanne P; Low, Ann S; Wangboonskul, Jinda

    2007-12-12

    Opium samples from four different locations and poppy straw from different plant varieties have been assayed using micellar capillary electrophoresis incorporating a sweeping technique. Individual alkaloids (morphine, codeine, papaverine, noscapine, thebaine, oripavine, reticuline and narceine) were quantitatively determined in the different samples by a validated capillary electrophoresis method. Unsupervised pattern recognition of the opium samples and the poppy straw samples using hierarchical cluster analysis (HCA) and principal component analysis (PCA), showed distinct clusters. Supervised pattern recognition using soft independent modelling of class analogy (SIMCA) was performed to show individual groupings and allow unknown samples to be classified according to the models built using the CZE assay results. PMID:18022406

  8. The california poppy (eschscholtzia mexicana) as a copper indicator plant - a new example

    USGS Publications Warehouse

    Chaffee, M.A.; Gale, C.W.

    1976-01-01

    The abundance and distribution of the California poppy (Eschscholtzia mexicana) correlates closely with the copper-rich outcrop of a small porphyry-type deposit in Arizona. Chemical factors are probably more important than physical factors in determining why this species is sometimes found as a copper indicator plant. ?? 1976.

  9. Papaver Rhoeas L. Hydroalcoholic Extract Exacerbates Forced Swimming Test-Induced Depression in Mice

    PubMed Central

    Osanloo, Naser; Najafi-Abedi, Akram; Jafari, Fatemeh; Javid, Farshid; Pirpiran, Mohsen; Memar Jafari, Mohammad-Reza; Mousavi Khosravi, Seyed Ali; Rahimzadeh Behzadi, Mohammad; Ranjbaran, Mina; Sahraei, Hedayat

    2016-01-01

    Introduction: Depression is one of the most frequent psychiatric disorders in the world with occurs with higher incidence in women. In the present study, the effect of water-alcoholic extract of Papaver rhoeas L. on forced swimming test (FST) in Swiss-Webster mice were examined. Methods: We used Swiss-Webster mice (20–25 g) to execute FST on them. The plant extract (1, 10, 30, and 100 mg/kg) was injected to the animals 30 minutes before each session. Fluoxetine (20 mg/kg) was used as standard antidepressant drug. In another group of animals, 30 minutes after extract administration, blood samples were taken from retro-orbital sinus for corticosterone assay. Yet in third group, the drugs were injected to the animals and 30 minutes later, their activities were tested in an open field apparatus. Results: Our experiments showed that the extract efficiently reduced FST time both in male and female mice dose-dependently. This effect was comparable with fluoxetine. In addition, corticosterone assay indicated that plasma corticosterone in animals which received extract was higher than those amounts in fluoxetine and saline controls. Moreover, the animals did not show any motor activity deficit in all doses of the extract and fluoxetine compared to saline control. Conclusion: The extract of Papaver rhoeas can reduce immobility time which is comparable to the effect of fluoxetine. Also the effect of the extract is contrary to its effects on plasma corticosterone level and or animals’ activity. PMID:27563412

  10. Visitor center at the Antelope Valley California Poppy Reserve, Lancaster, California

    SciTech Connect

    Colyer, R.D.; Freeman, S.P.

    1981-01-01

    The Antelope Valley California Poppy Reserve contains the largest remaining stand of the California Poppy (Eschschozia Californica), the state flower of California. To welcome the thousands of people viewing the desert wildflowers each spring, the State of California decided to build a visitor/interpretive center. This building deals primarily with the question of fit; a building's fit aesthetically with its site and the fit of a building's design response to the climate of the site. In this case, both aspects of this question led the client and architects to seek an earth sheltered solution using materials at least metaphorically indigenous to the region. On both a technical and formal level, this building seeks to fit the unique climate and historical heritage of its site.

  11. Plant science. Morphinan biosynthesis in opium poppy requires a P450-oxidoreductase fusion protein.

    PubMed

    Winzer, Thilo; Kern, Marcelo; King, Andrew J; Larson, Tony R; Teodor, Roxana I; Donninger, Samantha L; Li, Yi; Dowle, Adam A; Cartwright, Jared; Bates, Rachel; Ashford, David; Thomas, Jerry; Walker, Carol; Bowser, Tim A; Graham, Ian A

    2015-07-17

    Morphinan alkaloids from the opium poppy are used for pain relief. The direction of metabolites to morphinan biosynthesis requires isomerization of (S)- to (R)-reticuline. Characterization of high-reticuline poppy mutants revealed a genetic locus, designated STORR [(S)- to (R)-reticuline] that encodes both cytochrome P450 and oxidoreductase modules, the latter belonging to the aldo-keto reductase family. Metabolite analysis of mutant alleles and heterologous expression demonstrate that the P450 module is responsible for the conversion of (S)-reticuline to 1,2-dehydroreticuline, whereas the oxidoreductase module converts 1,2-dehydroreticuline to (R)-reticuline rather than functioning as a P450 redox partner. Proteomic analysis confirmed that these two modules are contained on a single polypeptide in vivo. This modular assembly implies a selection pressure favoring substrate channeling. The fusion protein STORR may enable microbial-based morphinan production.

  12. Plant science. Morphinan biosynthesis in opium poppy requires a P450-oxidoreductase fusion protein.

    PubMed

    Winzer, Thilo; Kern, Marcelo; King, Andrew J; Larson, Tony R; Teodor, Roxana I; Donninger, Samantha L; Li, Yi; Dowle, Adam A; Cartwright, Jared; Bates, Rachel; Ashford, David; Thomas, Jerry; Walker, Carol; Bowser, Tim A; Graham, Ian A

    2015-07-17

    Morphinan alkaloids from the opium poppy are used for pain relief. The direction of metabolites to morphinan biosynthesis requires isomerization of (S)- to (R)-reticuline. Characterization of high-reticuline poppy mutants revealed a genetic locus, designated STORR [(S)- to (R)-reticuline] that encodes both cytochrome P450 and oxidoreductase modules, the latter belonging to the aldo-keto reductase family. Metabolite analysis of mutant alleles and heterologous expression demonstrate that the P450 module is responsible for the conversion of (S)-reticuline to 1,2-dehydroreticuline, whereas the oxidoreductase module converts 1,2-dehydroreticuline to (R)-reticuline rather than functioning as a P450 redox partner. Proteomic analysis confirmed that these two modules are contained on a single polypeptide in vivo. This modular assembly implies a selection pressure favoring substrate channeling. The fusion protein STORR may enable microbial-based morphinan production. PMID:26113639

  13. 7 CFR 400.46 - Definitions.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 CFR 12.2, the following definitions are applicable: (a) Controlled substance means any prohibited... (erythroxylum coca), marijuana (cannabis satiua), opium poppies (papauer somniferum), and other...

  14. 7 CFR 400.46 - Definitions.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 CFR 12.2, the following definitions are applicable: (a) Controlled substance means any prohibited... (erythroxylum coca), marijuana (cannabis satiua), opium poppies (papauer somniferum), and other...

  15. 7 CFR 400.46 - Definitions.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 CFR 12.2, the following definitions are applicable: (a) Controlled substance means any prohibited... (erythroxylum coca), marijuana (cannabis satiua), opium poppies (papauer somniferum), and other...

  16. 7 CFR 400.46 - Definitions.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 CFR 12.2, the following definitions are applicable: (a) Controlled substance means any prohibited... (erythroxylum coca), marijuana (cannabis satiua), opium poppies (papauer somniferum), and other...

  17. 7 CFR 400.46 - Definitions.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 CFR 12.2, the following definitions are applicable: (a) Controlled substance means any prohibited... (erythroxylum coca), marijuana (cannabis satiua), opium poppies (papauer somniferum), and other...

  18. Morphine and Codeine in Oral Fluid after Controlled Poppy Seed Administration

    PubMed Central

    Concheiro, Marta; Newmeyer, Matthew N.; da Costa, Jose Luiz; Flegel, Ron; Gorelick, David A.; Huestis, Marilyn A.

    2014-01-01

    Opiates are an important drug class in drug testing programs. Ingestion of poppy seeds containing morphine and codeine can yield positive opiate tests and mislead result interpretation in forensic and clinical settings. Multiple publications evaluated urine opiate concentrations following poppy seed ingestion, but only 2 addressed oral fluid (OF) results; neither provided the ingested morphine and codeine dosage. We administered two 45g raw poppy seed doses, each containing 15.7mg morphine and 3.1mg codeine, 8h apart to 17 healthy adults. All OF specimens were screened by on-site OF immunoassay Draeger DrugTest 5000, and confirmed with OF collected with Oral-Eze® device and quantified by liquid chromatography tandem mass spectrometry (1μg/L morphine and codeine limits of quantification). Specimens (n=459) were collected before and up to 32h after the first dose. All specimens screened positive 0.5h after dosing and remained positive for 0.5-13h at Draeger 20μg/L morphine cutoff. Maximum OF morphine and codeine concentrations (Cmax) were 177 and 32.6μg/L, with times to Cmax (Tmax) of 0.5-1h and 0.5-2.5h post-dose, respectively. Windows of detection after the second dose extended at least 24h for morphine and to 18h for codeine. After both doses, the last morphine positive OF result was 1h with 40μg/L 2004 proposed US Substance Abuse and Mental Health Services Administration cutoff, and 0.5h with 95μg/L cutoff, recently recommended by the Driving Under the Influence of Drugs and Medicines project. Positive OF morphine results are possible 0.5-1h after ingestion of 15.7mg of morphine in raw poppy seeds, depending upon the cutoff employed. PMID:25345619

  19. Poppy seed oil protection of the hippocampus after cerebral ischemia and re-perfusion in rats.

    PubMed

    Cevik-Demirkan, A; Oztaşan, N; Oguzhan, E O; Cil, N; Coskun, S

    2012-11-01

    The brain is highly sensitive to hypoxia; this is true particularly of parts that are crucial for cognitive function. The effects of hypoxia are especially dramatic in the hippocampus. We evaluated the potential protective effects of poppy seed oil on the number of hippocampus cells and the serum antioxidant/oxidant status after cerebral ischemia and re-perfusion (CIR). Eighteen rats were divided into three equal groups. Group 1 served as the control group without CIR. Group 2 received poppy seed oil daily by oral gavage at a dose of 0.4 ml/kg, while group 3 was given 0.4 ml/kg saline solution by oral gavage per day; these treatments were continued for one month. Groups 2 and 3 were subjected to CIR induced by clamps on two points of both of the carotid arteries for 45 min followed by 45 min re-perfusion. There were significant decreases in the number of hippocampus cells between groups 1 and 2, and between groups 1 and 3. The mean cell number in group 2 was not significantly different from that of group 3. The serum nitric oxide levels in CIR groups were elevated significantly compared to controls, and were significantly higher in group 2 than in group 3. The glutathione levels were increased significantly in the poppy seed oil treated group compared to the saline CIR groups. The malondialdehyde levels were markedly increased in group 3 compared to both groups 1 and 2. Our study suggests that poppy seed oil can improve antioxidant defense capacity after CIR, although this treatment did not alter significantly the frequency of cell death.

  20. Acetylation serves as a protective group in noscapine biosynthesis in opium poppy.

    PubMed

    Dang, Thu-Thuy T; Chen, Xue; Facchini, Peter J

    2015-02-01

    We have characterized four sequential enzymes that transform 1-hydroxy-N-methylcanadine to narcotoline hemiacetal, completing our elucidation of noscapine biosynthesis in opium poppy. Two cytochromes P450 catalyze hydroxylations at C13 and C8 on the protoberberine scaffold, the latter step inducing ring opening and the formation of an aldehyde moiety. Acetylation at C13 before C8 hydroxylation introduces a protective group subsequently hydrolyzed by a carboxylesterase, which triggers rearrangement to a cyclic hemiacetal.

  1. Acetylation serves as a protective group in noscapine biosynthesis in opium poppy.

    PubMed

    Dang, Thu-Thuy T; Chen, Xue; Facchini, Peter J

    2015-02-01

    We have characterized four sequential enzymes that transform 1-hydroxy-N-methylcanadine to narcotoline hemiacetal, completing our elucidation of noscapine biosynthesis in opium poppy. Two cytochromes P450 catalyze hydroxylations at C13 and C8 on the protoberberine scaffold, the latter step inducing ring opening and the formation of an aldehyde moiety. Acetylation at C13 before C8 hydroxylation introduces a protective group subsequently hydrolyzed by a carboxylesterase, which triggers rearrangement to a cyclic hemiacetal. PMID:25485687

  2. Integration of deep transcript and targeted metabolite profiles for eight cultivars of opium poppy.

    PubMed

    Desgagné-Penix, Isabel; Farrow, Scott C; Cram, Dustin; Nowak, Jacek; Facchini, Peter J

    2012-06-01

    Recent advances in DNA sequencing technology and analytical mass spectrometry are providing unprecedented opportunities to develop the functional genomics resources required to investigate complex biological processes in non-model plants. Opium poppy produces a wide variety of benzylisoquinoline alkaloids (BIAs), including the pharmaceutical compounds codeine, morphine, noscapine and papaverine. A functional genomics platform to identify novel BIA biosynthetic and regulatory genes in opium poppy has been established based on the differential metabolite profile of eight selected cultivars. Stem cDNA libraries from each of the eight opium poppy cultivars were subjected to 454 pyrosequencing and searchable expressed sequence tag databases were created from the assembled reads. These deep and integrated metabolite and transcript databases provide a nearly complete representation of the genetic and metabolic variances responsible for the differential occurrence of specific BIAs in each cultivar as demonstrated using the biochemically well characterized pathway from tyrosine to morphine. Similar correlations between the occurrence of specific transcripts and alkaloids effectively reveals candidate genes encoding uncharacterized biosynthetic enzymes as shown using cytochromes P450 potentially involved in the formation of papaverine and noscapine.

  3. 1H NMR-based metabolic profiling for evaluating poppy seed rancidity and brewing.

    PubMed

    Jawień, Ewa; Ząbek, Adam; Deja, Stanisław; Łukaszewicz, Marcin; Młynarz, Piotr

    2015-12-01

    Poppy seeds are widely used in household and commercial confectionery. The aim of this study was to demonstrate the application of metabolic profiling for industrial monitoring of the molecular changes which occur during minced poppy seed rancidity and brewing processes performed on raw seeds. Both forms of poppy seeds were obtained from a confectionery company. Proton nuclear magnetic resonance (1H NMR) was applied as the analytical method of choice together with multivariate statistical data analysis. Metabolic fingerprinting was applied as a bioprocess control tool to monitor rancidity with the trajectory of change and brewing progressions. Low molecular weight compounds were found to be statistically significant biomarkers of these bioprocesses. Changes in concentrations of chemical compounds were explained relative to the biochemical processes and external conditions. The obtained results provide valuable and comprehensive information to gain a better understanding of the biology of rancidity and brewing processes, while demonstrating the potential for applying NMR spectroscopy combined with multivariate data analysis tools for quality control in food industries involved in the processing of oilseeds. This precious and versatile information gives a better understanding of the biology of these processes. PMID:26540222

  4. Biosynthesis of Nudicaulins: A (13) CO2 -pulse/chase labeling study with Papaver nudicaule.

    PubMed

    Tatsis, Evangelos C; Eylert, Eva; Maddula, Ravi Kumar; Ostrozhenkova, Elena; Svatoš, Aleš; Eisenreich, Wolfgang; Schneider, Bernd

    2014-07-21

    Nudicaulins are unique alkaloids responsible for the yellow color of the petals of some papaveraceaous plants. To elucidate the unknown biosynthetic origin of the skeleton, a (13) CO2 -pulse/chase experiment was performed with growing Papaver nudicaule plants. (13) C NMR analysis revealed more than 20 multiple (13) C-enriched isotopologues in nudicaulins from the petals of (13) CO2 -labeled plants. The complex labeling pattern was compared with the isotopologue composition of a kaempferol derivative that was isolated from petals of the same (13) CO2 -labeled plants. The deconvolution of the labeling profiles indicated that the nudicaulin scaffold is assembled from products or intermediates of indole metabolism, the phenylpropanoid pathway, and the polyketide biosynthesis. Naringenin-type compounds and tryptophan/tryptamine are potential substrates for the condensation reaction finally generating the aglycone skeleton of nudicaulins.

  5. Morphine and codeine concentrations in human urine following controlled poppy seeds administration of known opiate content.

    PubMed

    Smith, Michael L; Nichols, Daniel C; Underwood, Paula; Fuller, Zachary; Moser, Matthew A; LoDico, Charles; Gorelick, David A; Newmeyer, Matthew N; Concheiro, Marta; Huestis, Marilyn A

    2014-08-01

    Opiates are an important component for drug testing due to their high abuse potential. Proper urine opiate interpretation includes ruling out poppy seed ingestion; however, detailed elimination studies after controlled poppy seed administration with known morphine and codeine doses are not available. Therefore, we investigated urine opiate pharmacokinetics after controlled oral administration of uncooked poppy seeds with known morphine and codeine content. Participants were administered two 45 g oral poppy seed doses 8 h apart, each containing 15.7 mg morphine and 3mg codeine. Urine was collected ad libitum up to 32 h after the first dose. Specimens were analyzed with the Roche Opiates II immunoassay at 2000 and 300 μg/L cutoffs, and the ThermoFisher CEDIA(®) heroin metabolite (6-acetylmorphine, 6-AM) and Lin-Zhi 6-AM immunoassays with 10 μg/L cutoffs to determine if poppy seed ingestion could produce positive results in these heroin marker assays. In addition, all specimens were quantified for morphine and codeine by GC/MS. Participants (N=22) provided 391 urine specimens over 32 h following dosing; 26.6% and 83.4% were positive for morphine at 2000 and 300 μg/L GC/MS cutoffs, respectively. For the 19 subjects who completed the study, morphine concentrations ranged from <300 to 7522 μg/L with a median peak concentration of 5239 μg/L. The median first morphine-positive urine sample at 2000 μg/L cutoff concentration occurred at 6.6 h (1.2-12.1), with the last positive from 2.6 to 18 h after the second dose. No specimens were positive for codeine at a cutoff concentration of 2000 μg/L, but 20.2% exceeded 300 μg/L, with peak concentrations of 658 μg/L (284-1540). The Roche Opiates II immunoassay had efficiencies greater than 96% for the 2000 and 300 μg/L cutoffs. The CEDIA 6-AM immunoassay had a specificity of 91%, while the Lin-Zhi assay had no false positive results. These data provide valuable information for interpreting urine opiate results.

  6. Morphine and Codeine Concentrations in Human Urine following Controlled Poppy Seeds Administration of Known Opiate Content

    PubMed Central

    Smith, Michael L.; Nichols, Daniel C.; Underwood, Paula; Fuller, Zachary; Moser, Matthew A.; LoDico, Charles; Gorelick, David A.; Newmeyer, Matthew N.; Concheiro, Marta; Huestis, Marilyn A.

    2014-01-01

    Opiates are an important component for drug testing due to their high abuse potential. Proper urine opiate interpretation includes ruling out poppy seed ingestion; however, detailed elimination studies after controlled poppy seed administration with known morphine and codeine doses are not available. Therefore, we investigated urine opiate pharmacokinetics after controlled oral administration of uncooked poppy seeds with known morphine and codeine content. Participants were administered two 45g oral poppy seed doses 8h apart, each containing 15.7mg morphine and 3mg codeine. Urine was collected ad libitum up to 32h after the first dose. Specimens were analyzed with the Roche Opiates II immunoassay at 2,000 and 300μg/L cutoffs, and the ThermoFisher CEDIA® Heroin Metabolite (6-acetylmorphine, 6AM) and Lin-Zhi 6AM immunoassays with 10μg/L cutoffs to determine if poppy seed ingestion could produce positive results in these heroin marker assays. In addition, all specimens were quantified for morphine and codeine by GC/MS. Participants (N=22) provided 391 urine specimens over 32h following dosing; 26.6% and 83.4% were positive for morphine at 2,000 and 300μg/L GC/MS cutoffs, respectively. For the 19 subjects who completed the study, morphine concentrations ranged from <300 to 7,522μg/L with a median peak concentration of 5,239μg/L. The median first morphine-positive urine sample at 2,000μg/L cutoff concentration occurred at 6.6h (1.2-12.1), with the last positive from 2.6 to 18h after the second dose. No specimens were positive for codeine at a cutoff concentration of 2,000μg/L, but 20.2% exceeded 300μg/L, with peak concentrations of 658 μg/L (284-1540). The Roche Opiates II immunoassay had efficiencies greater than 96% for the 2000 and 300μg/L cutoffs. The CEDIA 6AM immunoassay had a specificity of 91%, while the Lin-Zhi assay had no false positive results. These data provide valuable information for interpreting urine opiate results. PMID:24887324

  7. Himalayan-Tibetan plateau uplift drives divergence of polyploid poppies: Meconopsis Viguier (Papaveraceae).

    PubMed

    Xie, Hongyan; Ash, Julian E; Linde, Celeste C; Cunningham, Saul; Nicotra, Adrienne

    2014-01-01

    Meconopsis Viguier (Papaveraceae) is an iconic genus of alpine forbs that includes medicinal and ornamental species. This study extends previous phylogenetic analyses of Meconopsis, using ITS sequences representing all the major Meconopsis clades. Phenotypic traits are also analysed for all described species. Our results show that Meconopsis evolved as a ≥ octaploid clade, with considerable interior structure reflecting further changes in ploidy levels as well as phenotypic differentiation. We support the exclusion of a few species as Cathcartia or Papaver, making Meconopsis a Tibetan region clade. Based on average rates of nucleotide substitution in angiosperm herbs we estimate that the Meconopsis clade diverged from the Meconella clade of Papaver approximately 16.6 Ma. This is soon after the 'hard' collision of the Indian continent with Asia caused uplift of the Himalaya and Hengduan ranges, greatly extended the Tibetan plateau, and initiated monsoonal climates. Eight major clades within Meconopsis are well supported and these correspond closely to previously recognised subgenus groups. However, the relationship among the clades is poorly resolved, probably because they diverged rapidly ∼15-11 Ma. Two of these clades are ∼dodecaploid but appear to have originated independently. The eight clades have distinct distributions, variously associated with the Himalaya, the eastern Plateau and Hengduan ranges. Some Meconopsis species were not monophyletic, suggesting that a combination of multilocus molecular and phenotypic traits is required when defining and revising species. PMID:24933008

  8. Himalayan-Tibetan Plateau Uplift Drives Divergence of Polyploid Poppies: Meconopsis Viguier (Papaveraceae)

    PubMed Central

    Xie, Hongyan; Ash, Julian E.; Linde, Celeste C.; Cunningham, Saul; Nicotra, Adrienne

    2014-01-01

    Meconopsis Viguier (Papaveraceae) is an iconic genus of alpine forbs that includes medicinal and ornamental species. This study extends previous phylogenetic analyses of Meconopsis, using ITS sequences representing all the major Meconopsis clades. Phenotypic traits are also analysed for all described species. Our results show that Meconopsis evolved as a ≥ octaploid clade, with considerable interior structure reflecting further changes in ploidy levels as well as phenotypic differentiation. We support the exclusion of a few species as Cathcartia or Papaver, making Meconopsis a Tibetan region clade. Based on average rates of nucleotide substitution in angiosperm herbs we estimate that the Meconopsis clade diverged from the Meconella clade of Papaver approximately 16.6 Ma. This is soon after the ‘hard’ collision of the Indian continent with Asia caused uplift of the Himalaya and Hengduan ranges, greatly extended the Tibetan plateau, and initiated monsoonal climates. Eight major clades within Meconopsis are well supported and these correspond closely to previously recognised subgenus groups. However, the relationship among the clades is poorly resolved, probably because they diverged rapidly ∼15-11 Ma. Two of these clades are ∼dodecaploid but appear to have originated independently. The eight clades have distinct distributions, variously associated with the Himalaya, the eastern Plateau and Hengduan ranges. Some Meconopsis species were not monophyletic, suggesting that a combination of multilocus molecular and phenotypic traits is required when defining and revising species. PMID:24933008

  9. Genetic transformation via somatic embryogenesis to establish herbicide-resistant opium poppy.

    PubMed

    Facchini, Peter J; Loukanina, Natalia; Blanche, Vincent

    2008-04-01

    A reliable genetic transformation protocol via somatic embryogenesis has been developed for the production of fertile, herbicide-resistant opium poppy plants. Transformation was mediated by Agrobacterium tumefaciens using the pCAMBIA3301 vector, which harbors the phosphinothricin acetyltransferase (pat) gene driven by a tandem repeat of the cauliflower mosaic virus (CaMV) 35S promoter and the beta-glucuronidase (gus) structural gene driven by a single copy of the CaMV 35S promoter between left- and right-border sequences. Co-cultivation of explants and A. tumefaciens was performed in the presence of 50 microM ATP and 50 microM MgCl(2). Root explants pre-cultured on callus induction medium were used for transformation. Herbicide-resistant, proliferating callus was obtained from explants on a medium containing both 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BA). Globular embryogenic callus, induced by removal of the BA from the medium, was placed on a hormone-free medium to form somatic embryos, which were converted to plantlets under specific culture conditions. Plantlets with roots were transferred to soil, allowed to mature and set seed. Both pat and gus gene transcripts, and PAT and GUS enzyme activities were detected in the transgenic lines tested. Histochemical localization of GUS activity in T(1) opium poppy plants revealed transgene expression in most tissues of all plant organs. The protocol required 8-12 months to establish transgenic T(1) seed stocks and was developed using a commercial opium poppy cultivar that produces high levels of pharmaceutical alkaloids.

  10. Prickly poppies can get pricklier: ontogenetic patterns in the induction of physical defense traits.

    PubMed

    Hoan, Ryan P; Ormond, Rhys A; Barton, Kasey E

    2014-01-01

    Plant ontogeny is a common source of variation in defense and herbivory. Yet, few studies have investigated how the induction of physical defense traits changes across plant ontogeny. Physical defense traits are costly to produce, and thus, it was predicted that induction as a cost-saving strategy would be particularly favorable for seedlings, leading to ontogenetic declines in the inducibility of these traits. We tested for induction of three different physical defense traits (prickles, latex and leaf toughness) in response to mechanical defoliation and jasmonic acid application using prickly poppies (Argemone glauca and A. mexicana, Papaveraceae) as a model system. Genetic variation in the induction of physical defenses was tested using maternal sib-ships sampled from multiple populations. Both species induced higher densities of laminar prickles, although the magnitude of induction was much higher in the endemic Hawaiian prickly poppy, A. glauca, than in the cosmopolitan A. mexicana. The magnitude of prickle induction was also higher in young compared to older juvenile plant stages in A. glauca, demonstrating a strong role of ontogeny. Neither latex exudation nor leaf toughness was induced in either species. Although significant genetic variation was detected within and among populations for constitutive expression of physical defense traits in Argemone, there was no evidence for genetic variation in the induction of these traits. This study provides the first evidence for the induction of physical defenses in prickly poppies, emphasizing how an ontogenetically explicit framework can reveal new insights into plant defense. Moreover, this study illustrates how sister species comparisons between island vs. continental plants can provide new insights into plant functional and evolutionary ecology, highlighting a fruitful area for future research on more species pairs. PMID:24802133

  11. Prickly Poppies Can Get Pricklier: Ontogenetic Patterns in the Induction of Physical Defense Traits

    PubMed Central

    Hoan, Ryan P.; Ormond, Rhys A.; Barton, Kasey E.

    2014-01-01

    Plant ontogeny is a common source of variation in defense and herbivory. Yet, few studies have investigated how the induction of physical defense traits changes across plant ontogeny. Physical defense traits are costly to produce, and thus, it was predicted that induction as a cost-saving strategy would be particularly favorable for seedlings, leading to ontogenetic declines in the inducibility of these traits. We tested for induction of three different physical defense traits (prickles, latex and leaf toughness) in response to mechanical defoliation and jasmonic acid application using prickly poppies (Argemone glauca and A. mexicana, Papaveraceae) as a model system. Genetic variation in the induction of physical defenses was tested using maternal sib-ships sampled from multiple populations. Both species induced higher densities of laminar prickles, although the magnitude of induction was much higher in the endemic Hawaiian prickly poppy, A. glauca, than in the cosmopolitan A. mexicana. The magnitude of prickle induction was also higher in young compared to older juvenile plant stages in A. glauca, demonstrating a strong role of ontogeny. Neither latex exudation nor leaf toughness was induced in either species. Although significant genetic variation was detected within and among populations for constitutive expression of physical defense traits in Argemone, there was no evidence for genetic variation in the induction of these traits. This study provides the first evidence for the induction of physical defenses in prickly poppies, emphasizing how an ontogenetically explicit framework can reveal new insights into plant defense. Moreover, this study illustrates how sister species comparisons between island vs. continental plants can provide new insights into plant functional and evolutionary ecology, highlighting a fruitful area for future research on more species pairs. PMID:24802133

  12. Morphine and codeine in oral fluid after controlled poppy seed administration.

    PubMed

    Concheiro, Marta; Newmeyer, Matthew N; da Costa, Jose Luiz; Flegel, Ron; Gorelick, David A; Huestis, Marilyn A

    2015-07-01

    Opiates are an important drug class in drug testing programmes. Ingestion of poppy seeds containing morphine and codeine can yield positive opiate tests and mislead result interpretation in forensic and clinical settings. Multiple publications evaluated urine opiate concentrations following poppy seed ingestion, but only two addressed oral fluid (OF) results; neither provided the ingested morphine and codeine dosage. We administered two 45 g raw poppy seed doses, each containing 15.7 mg morphine and 3.1 mg codeine, 8 h apart to 17 healthy adults. All OF specimens were screened by on-site OF immunoassay Draeger DrugTest 5000, and confirmed with OF collected with Oral-Eze® device and quantified by liquid chromatography-tandem mass spectrometry (1 µg/L morphine and codeine limits of quantification). Specimens (n = 459) were collected before and up to 32 h after the first dose. All specimens screened positive 0.5 h after dosing and remained positive for 0.5-13 h at Draeger 20 µg/L morphine cut-off. Maximum OF morphine and codeine concentrations (Cmax ) were 177 and 32.6 µg/L, with times to Cmax (Tmax ) of 0.5-1 h and 0.5-2.5 h post-dose, respectively. Windows of detection after the second dose extended at least 24 h for morphine and to 18 h for codeine. After both doses, the last morphine positive OF result was 1 h with 40 µg/L 2004 proposed US Substance Abuse and Mental Health Services Administration cut-off, and 0.5 h with 95 µg/L cut-off, recently recommended by the Driving under the Influence of Drugs and Medicines project. Positive OF morphine results are possible 0.5-1 h after ingestion of 15.7 mg of morphine in raw poppy seeds, depending on the cut-off employed. PMID:25345619

  13. Morphine and codeine in oral fluid after controlled poppy seed administration.

    PubMed

    Concheiro, Marta; Newmeyer, Matthew N; da Costa, Jose Luiz; Flegel, Ron; Gorelick, David A; Huestis, Marilyn A

    2015-07-01

    Opiates are an important drug class in drug testing programmes. Ingestion of poppy seeds containing morphine and codeine can yield positive opiate tests and mislead result interpretation in forensic and clinical settings. Multiple publications evaluated urine opiate concentrations following poppy seed ingestion, but only two addressed oral fluid (OF) results; neither provided the ingested morphine and codeine dosage. We administered two 45 g raw poppy seed doses, each containing 15.7 mg morphine and 3.1 mg codeine, 8 h apart to 17 healthy adults. All OF specimens were screened by on-site OF immunoassay Draeger DrugTest 5000, and confirmed with OF collected with Oral-Eze® device and quantified by liquid chromatography-tandem mass spectrometry (1 µg/L morphine and codeine limits of quantification). Specimens (n = 459) were collected before and up to 32 h after the first dose. All specimens screened positive 0.5 h after dosing and remained positive for 0.5-13 h at Draeger 20 µg/L morphine cut-off. Maximum OF morphine and codeine concentrations (Cmax ) were 177 and 32.6 µg/L, with times to Cmax (Tmax ) of 0.5-1 h and 0.5-2.5 h post-dose, respectively. Windows of detection after the second dose extended at least 24 h for morphine and to 18 h for codeine. After both doses, the last morphine positive OF result was 1 h with 40 µg/L 2004 proposed US Substance Abuse and Mental Health Services Administration cut-off, and 0.5 h with 95 µg/L cut-off, recently recommended by the Driving under the Influence of Drugs and Medicines project. Positive OF morphine results are possible 0.5-1 h after ingestion of 15.7 mg of morphine in raw poppy seeds, depending on the cut-off employed.

  14. The rules of drug taking: wine and poppy derivatives in the ancient world. I. General introduction.

    PubMed

    Nencini, P

    1997-01-01

    Many believe that the public attitude toward psychotropic drugs is easy to change because it depends upon transient socioeconomic and cultural contingencies. This belief is here tested by analyzing literary sources on the use of wine and poppy derivatives in the Greek and Roman civilizations. As expected, the existence of elaborated myths about the discover of wine supports the view that wine drinking played an important social role in the ancient world. Contingencies in which drinking was considered illicit were carefully selected with the ultimate aim of maintaining sobriety. Rules of drinking were often enforced by law. This first part of the study lists drunkenness outcomes considered particularly harmful by the Ancients.

  15. Chemometric approach for development, optimization, and validation of different chromatographic methods for separation of opium alkaloids.

    PubMed

    Acevska, J; Stefkov, G; Petkovska, R; Kulevanova, S; Dimitrovska, A

    2012-05-01

    The excessive and continuously growing interest in the simultaneous determination of poppy alkaloids imposes the development and optimization of convenient high-throughput methods for the assessment of the qualitative and quantitative profile of alkaloids in poppy straw. Systematic optimization of two chromatographic methods (gas chromatography (GC)/flame ionization detector (FID)/mass spectrometry (MS) and reversed-phase (RP)-high-performance liquid chromatography (HPLC)/diode array detector (DAD)) for the separation of alkaloids from Papaver somniferum L. (Papaveraceae) was carried out. The effects of various conditions on the predefined chromatographic descriptors were investigated using chemometrics. A full factorial linear design of experiments for determining the relationship between chromatographic conditions and the retention behavior of the analytes was used. Central composite circumscribed design was utilized for the final method optimization. By conducting the optimization of the methods in very rational manner, a great deal of excessive and unproductive laboratory research work was avoided. The developed chromatographic methods were validated and compared in line with the resolving power, sensitivity, accuracy, speed, cost, ecological aspects, and compatibility with the poppy straw extraction procedure. The separation of the opium alkaloids using the GC/FID/MS method was achieved within 10 min, avoiding any derivatization step. This method has a stronger resolving power, shorter analysis time, better cost/effectiveness factor than the RP-HPLC/DAD method and is in line with the "green trend" of the analysis. The RP-HPLC/DAD method on the other hand displayed better sensitivity for all tested alkaloids. The proposed methods provide both fast screening and an accurate content assessment of the six alkaloids in the poppy samples obtained from the selection program of Papaver strains.

  16. "Poppies are democracy!" A critical geopolitics of opium eradication and reintroduction in Turkey.

    PubMed

    Evered, Kyle T

    2011-01-01

    Historical scholarship in traditional geopolitics often relied on documents authored by states and by other influential actors. Although much work in the subfield of critical geopolitics thus far has addressed imbalances constructed in official, academic, and popular media due to a privileging of such narratives, priority might also be given to unearthing and bringing to light alternative geopolitical perspectives from otherwise marginalized populations. Utilizing the early-1970s case of the United States' first “war on drugs,” this article examines the geopolitics of opium-poppy eradication and its consequences within Turkey. Employing not only archival and secondary sources but also oral histories from now-retired poppy farmers, this study examines the diffusion of U.S. antinarcotics policies into the Anatolian countryside and the enduring impressions that the United States and Turkish government created. In doing so, this research gives voice to those farmers targeted by eradication policies and speaks more broadly to matters of narcotics control, sentiments of anti-Americanism, and notions of democracy in Turkey and the region, past and present. PMID:22164875

  17. "Poppies are democracy!" A critical geopolitics of opium eradication and reintroduction in Turkey.

    PubMed

    Evered, Kyle T

    2011-01-01

    Historical scholarship in traditional geopolitics often relied on documents authored by states and by other influential actors. Although much work in the subfield of critical geopolitics thus far has addressed imbalances constructed in official, academic, and popular media due to a privileging of such narratives, priority might also be given to unearthing and bringing to light alternative geopolitical perspectives from otherwise marginalized populations. Utilizing the early-1970s case of the United States' first “war on drugs,” this article examines the geopolitics of opium-poppy eradication and its consequences within Turkey. Employing not only archival and secondary sources but also oral histories from now-retired poppy farmers, this study examines the diffusion of U.S. antinarcotics policies into the Anatolian countryside and the enduring impressions that the United States and Turkish government created. In doing so, this research gives voice to those farmers targeted by eradication policies and speaks more broadly to matters of narcotics control, sentiments of anti-Americanism, and notions of democracy in Turkey and the region, past and present.

  18. For peace and pain: the medical legitimisation of Afghanistan's poppy crop.

    PubMed

    Attaran, Amir; Boozary, Andrew

    2011-05-01

    Since the overthrow of the Taliban in 2001, there has been an overall increase in illicit opium production in Afghanistan and mounting human losses. The United Nations has attributed 1 million human deaths to Afghan opiates over the past decade. As the war in Afghanistan nears a crucial mark, the NATO coalition forces and Afghan people can no longer afford the same ineffective counternarcotics strategy. This commentary proposes a strategic revision that reframes Afghanistan's poppy problem as an opportunity for global public health. Specifically, The Afghan poppy crop could be repurposed away from illicit drug production, and towards manufacturing licit opioid analgesics to address unmet needs for pain palliation, particularly for diseases such as HIV/AIDS and cancer in the developing world--that is, illegal opium could be converted into legal pain medicine, solving two problems at once. We present a supply-and-demand that illustrates how this useful exchange could be made, and discuss the political opposition that now stands in the way and perpetuates the unsatisfactory status quo in Afghanistan.

  19. Ferric ions involved in the flower color development of the Himalayan blue poppy, Meconopsis grandis.

    PubMed

    Yoshida, Kumi; Kitahara, Sayoko; Ito, Daisuke; Kondo, Tadao

    2006-05-01

    The Himalayan blue poppy, Meconopsis grandis, has sky blue-colored petals, although the anthocyanidin nucleus of the petal pigment is cyanidin. The blue color development in this blue poppy involving ferric ions was therefore studied. We analyzed the vacuolar pH, and the organic and inorganic components of the colored cells. A direct measurement by a proton-selective microelectrode revealed that the vacuolar pH value was 4.8. The concentrations of the total anthocyanins in the colored cells were around 5mM, and ca. three times more concentrated flavonols were detected. Fe was detected by atomic analysis of the colored cells, and the ratio of Fe to anthocyanins was ca. 0.8 eq. By mixing the anthocyanin, flavonol and metal ion components in a buffered aq. solution at pH 5.0, we were able to reproduce the same blue color; the visible absorption spectrum and CD were identical to those in the petals, with Fe(3+), Mg(2+) and flavonol being essential for the blue color. The blue pigment in Meconopsis should be a new type of metal complex pigment that is different from a stoichiometric supramolecular pigment such as commelinin or protocyanin.

  20. For peace and pain: the medical legitimisation of Afghanistan's poppy crop.

    PubMed

    Attaran, Amir; Boozary, Andrew

    2011-05-01

    Since the overthrow of the Taliban in 2001, there has been an overall increase in illicit opium production in Afghanistan and mounting human losses. The United Nations has attributed 1 million human deaths to Afghan opiates over the past decade. As the war in Afghanistan nears a crucial mark, the NATO coalition forces and Afghan people can no longer afford the same ineffective counternarcotics strategy. This commentary proposes a strategic revision that reframes Afghanistan's poppy problem as an opportunity for global public health. Specifically, The Afghan poppy crop could be repurposed away from illicit drug production, and towards manufacturing licit opioid analgesics to address unmet needs for pain palliation, particularly for diseases such as HIV/AIDS and cancer in the developing world--that is, illegal opium could be converted into legal pain medicine, solving two problems at once. We present a supply-and-demand that illustrates how this useful exchange could be made, and discuss the political opposition that now stands in the way and perpetuates the unsatisfactory status quo in Afghanistan. PMID:21282137

  1. A potential signaling role for profilin in pollen of Papaver rhoeas.

    PubMed Central

    Clarke, S R; Staiger, C J; Gibbon, B C; Franklin-Tong, V E

    1998-01-01

    Regulation of pollen tube growth is known to involve alterations in intracellular calcium levels and phosphoinositide signaling, although the mechanisms involved are unclear. However, it appears likely that pollination events involve a complex interplay between signaling pathways and components of the actin cytoskeleton in pollen. In many eukaryotic cells, actin binding proteins function as stimulus-response modulators, translating signals into alterations in the cytoplasmic architecture. In this study, we examined whether profilin, which is a member of this class of signaling intermediate, might play a similar role in pollen. We have analyzed the functional properties of native profilin from pollen of Papaver rhoeas and have investigated the effects of profilin on the phosphorylation of pollen proteins in vitro by adding a slight excess of profilin to cytosolic pollen extracts. We present clear evidence that profilin interacts with soluble pollen components, resulting in dramatic alterations in the phosphorylation of several proteins. We also show, albeit in vitro, the involvement of profilin in modulating the activity of a signaling component(s) affecting protein phosphorylation. Our data, which suggest that pollen profilin can regulate actin-based cytoskeletal protein assembly and protein kinase or phosphatase activity, indicate a possible role for the involvement of profilin in signaling pathways that may regulate pollen tube growth. PMID:9634585

  2. The Occurrence of Flavonoids and Related Compounds in Flower Sections of Papaver nudicaule.

    PubMed

    Dudek, Bettina; Warskulat, Anne-Christin; Schneider, Bernd

    2016-01-01

    Flavonoids play an important role in the pigmentation of flowers; in addition, they protect petals and other flower parts from UV irradiation and oxidative stress. Nudicaulins, flavonoid-derived indole alkaloids, along with pelargonidin, kaempferol, and gossypetin glycosides, are responsible for the color of white, red, orange, and yellow petals of different Papaver nudicaule cultivars. The color of the petals is essential to attract pollinators. We investigated the occurrence of flavonoids in basal and apical petal areas, stamens, and capsules of four differently colored P. nudicaule cultivars by means of chromatographic and spectroscopic methods. The results reveal the specific occurrence of gossypetin glycosides in the basal spot of all cultivars and demonstrate that kaempferol glycosides are the major secondary metabolites in the capsules. Unlike previous reports, the yellow-colored stamens of all four P. nudicaule cultivars are shown to contain not nudicaulins but carotenoids. In addition, the presence of nudicaulins, pelargonidin, and kaempferol glycosides in the apical petal area was confirmed. The flavonoids and related compounds in the investigated flower parts and cultivars of P. nudicaule are profiled, and their potential ecological role is discussed. PMID:27338493

  3. Effects of Papaver rhoeas (L.) extract on formalin-induced pain and inflammation in mice.

    PubMed

    Saeed-Abadi, S; Ranjbaran, M; Jafari, F; Najafi-Abedi, A; Rahmani, B; Esfandiari, B; Delfan, B; Mojabi, N; Ghahramani, M; Sahraei, H

    2012-11-01

    Stress amelioration can improve its metabolic as well as other side effects. In the present study, the effects of hydro-alcoholic extract of Papver rhoeas (L.) on formalin-induced pain and inflammation were investigated in male Swiss-Webster mice (20-25 g). Formalin injects in the plantar portion of mice hind paw and pain was studied for 60 min. The plant extract and other drugs were administered intraperitoneally 30 min before formalin. Experiments showed that administration of extract (25, 50 and 100 mg kg(-1)) could induced analgesia in a dose-response manner in both phases of formalin test. More over, the extract inhibits inflammation induced by formalin injection. Naloxone (4 mg kg(-1)), dextromethorphan (20 mg kg(-1)) and NG-nitro-L-arginine-methylester (L-NAME; 10 mg kg(-1)) reduced the extract analgesia in first but not late phase. Extract administration also increased plasma corticosterone level in dose-dependent manner. It could be concluded that Papaver rhoeas (L.) extract could inhibits acute phase of formalin test in mice by opioidergic, glutamatergic and nitricergic mechanisms. In addition, the extract can induce corticosterone plasma level which may be responsible for inhibition of inflammation and chronic phase of pain induced by formalin.

  4. The Occurrence of Flavonoids and Related Compounds in Flower Sections of Papaver nudicaule

    PubMed Central

    Dudek, Bettina; Warskulat, Anne-Christin; Schneider, Bernd

    2016-01-01

    Flavonoids play an important role in the pigmentation of flowers; in addition, they protect petals and other flower parts from UV irradiation and oxidative stress. Nudicaulins, flavonoid-derived indole alkaloids, along with pelargonidin, kaempferol, and gossypetin glycosides, are responsible for the color of white, red, orange, and yellow petals of different Papaver nudicaule cultivars. The color of the petals is essential to attract pollinators. We investigated the occurrence of flavonoids in basal and apical petal areas, stamens, and capsules of four differently colored P. nudicaule cultivars by means of chromatographic and spectroscopic methods. The results reveal the specific occurrence of gossypetin glycosides in the basal spot of all cultivars and demonstrate that kaempferol glycosides are the major secondary metabolites in the capsules. Unlike previous reports, the yellow-colored stamens of all four P. nudicaule cultivars are shown to contain not nudicaulins but carotenoids. In addition, the presence of nudicaulins, pelargonidin, and kaempferol glycosides in the apical petal area was confirmed. The flavonoids and related compounds in the investigated flower parts and cultivars of P. nudicaule are profiled, and their potential ecological role is discussed. PMID:27338493

  5. Antioxidant, antimutagenic, and anticarcinogenic effects of Papaver rhoeas L. extract on Saccharomyces cerevisiae.

    PubMed

    Todorova, Teodora; Pesheva, Margarita; Gregan, Fridrich; Chankova, Stephka

    2015-04-01

    The aim of this work was to analyze the antioxidant and antimutagenic/anticarcinogenic capacity of Papaver rhoeas L. water extract against standard mutagen/carcinogen methyl methanesulfonate (MMS) and radiomimetic zeocin (Zeo) on a test system Saccharomyces cerevisiae. The following assays were used: 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, quantitative determination of superoxide anion (antireactive oxygen species [antiROS test]), DNA topology assay, D7ts1 test--for antimutagenic--and Ty1 transposition test--for anticarcinogenic effects. Strong pro-oxidative capacity of Zeo was shown to correlate with its well-expressed mutagenic and carcinogenic properties. The mutagenic and carcinogenic effects of MMS were also confirmed. Our data concerning the antioxidant activity of P. rhoeas L. extract revealed that concentration corresponding to IC(50) in the DPPH assay possessed the highest antioxidant activity in the antiROS biological assay. It was also observed that a concentration with 50% scavenging activity expressed the most pronounced antimutagenic properties decreasing Zeo-induced gene conversion twofold, reverse mutation fivefold, and total aberrations fourfold. The same concentration possessed well-expressed anticarcinogenic properties measured as reduction of MMS-induced Ty1 transposition rate fivefold and fourfold when Zeo was used as an inductor. Based on the well-expressed antioxidant, antimutagenic, and anticarcinogenic properties obtained in this work, the P. rhoeas L. extract could be recommended for further investigations and possible use as a food additive.

  6. The Papaver rhoeas S determinants confer self-incompatibility to Arabidopsis thaliana in planta.

    PubMed

    Lin, Zongcheng; Eaves, Deborah J; Sanchez-Moran, Eugenio; Franklin, F Christopher H; Franklin-Tong, Vernonica E

    2015-11-01

    Self-incompatibility (SI) is a major genetically controlled system used to prevent inbreeding in higher plants. S determinants regulate allele-specific rejection of "self" pollen by the pistil. SI is an important model system for cell-to-cell recognition and signaling and could be potentially useful for first-generation (F1) hybrid breeding. To date, the transfer of S determinants has used the complementation of orthologs to "restore" SI in close relatives. We expressed the Papaver rhoeas S determinants PrsS and PrpS in Arabidopsis thaliana. This enabled pistils to reject pollen expressing cognate PrpS. Moreover, plants coexpressing cognate PrpS and PrsS exhibit robust SI. This demonstrates that PrsS and PrpS are sufficient for a functional synthetic S locus in vivo. This transfer of novel S determinants into a highly divergent species (>140 million years apart) with no orthologs suggests their potential utility in crop production. PMID:26542572

  7. Antioxidant and antimicrobial properties of monofloral bee pollen.

    PubMed

    Fatrcová-Šramková, Katarína; Nôžková, Janka; Kačániová, Miroslava; Máriássyová, Magda; Rovná, Katarína; Stričík, Michal

    2013-01-01

    The main aim of this study was to determine antioxidant properties and antibacterial activity of monofloral bee pollen samples to pathogenic bacteria. These samples were collected in different localities in Slovakia. The antioxidant properties of examined plant species were different and decreasing in the following order: Brassica napus subsp. napus L > Papaver somniferum L. > Helianthus annuus L. The antimicrobial effect of the bee product samples were tested by using the agar well diffusion method. The methanol (99.9% and 70%) and the ethanol (96% and 70%) were used for extraction. In this study, five different strains of bacteria were tested: Listeria monocytogenes CCM 4699; Pseudomonas aeruginosa CCM 1960; Staphylococcus aureus CCM 3953; Salmonella enterica CCM 4420; and Escherichia coli CCM 3988. The most sensitive bacteria of the poppy pollen ethanolic extract was Staphylococcus aureus was (70%) The most sensitive bacteria of rape bee pollen methanolic extract (70%) and sunflower ethanolic extract (70%) was Salmonella enterica. PMID:23305281

  8. Short-chain dehydrogenase/reductase catalyzing the final step of noscapine biosynthesis is localized to laticifers in opium poppy.

    PubMed

    Chen, Xue; Facchini, Peter J

    2014-01-01

    The final step in the biosynthesis of the phthalideisoquinoline alkaloid noscapine involves a purported dehydrogenation of the narcotinehemiacetal keto moiety. A short-chain dehydrogenase/reductase (SDR), designated noscapine synthase (NOS), that catalyzes dehydrogenation of narcotinehemiacetal to noscapine was identified in opium poppy and functionally characterized. The NOS gene was isolated using an integrated transcript and metabolite profiling strategy and subsequently expressed in Escherichia coli. Noscapine synthase is highly divergent from other characterized members of the NADPH-dependent SDR superfamily involved in benzylisoquinoline alkaloid metabolism, and it exhibits exclusive substrate specificity for narcotinehemiacetal. Kinetic analyses showed that NOS exhibits higher catalytic efficiency with NAD+ as the cofactor compared with NADP+. Suppression of NOS transcript levels in opium poppy plants subjected to virus-induced gene silencing resulted in a corresponding reduction in the accumulation of noscapine and an increase in narcotinehemiacetal levels in the latex. Noscapine and NOS transcripts were detected in all opium poppy organs, but both were most abundant in stems. Unlike other putative biosynthetic genes clustered in the opium poppy genome, and their corresponding proteins, NOS transcripts and the cognate enzyme were abundant in latex, indicating that noscapine metabolism is completed in a distinct cell type compared with the rest of the pathway.

  9. Determination of inorganic elements in poppy straw by scanning electron microscopy with energy dispersive spectrometry as a means of ascertaining origin.

    PubMed

    Copur, E; Göger, N G; Orbey, T; Sener, B

    2005-01-01

    Cultivation of poppy as a source of opium alkaloids for legitimate medical purposes has a long tradition in Turkey. The main products are poppy straw and concentrate of poppy straw, obtained from dried poppy capsules. The aims of the study reported in the present article were to establish inorganic element profiles for the poppy-growing provinces of Turkey by means of x-ray analysis by scanning electron microscopy with energy dispersive spectrometry (SEM/EDS) and to explore the potential of the technique for determination of origin. Ten elements (sodium, magnesium, silicon, phosphorus, sulphur, chlorine, potassium, calcium, copper and zinc) were analysed in poppy straw samples from 67 towns in nine provinces. As regards the determination of origin, the most significant finding was the presence of copper and zinc in the poppy straw samples from 8 of the 15 towns in Afyon Province. Since those elements are not normally found in soil, it is assumed that their presence is the result of environmental (industrial) contamination. Differences in the samples from the other eight provinces were less significant, possibly a result of their geographical proximity. Nevertheless, differences in the samples were apparent. Because the findings are relative rather than absolute in terms of presence or absence of individual inorganic elements, further research is required to convert them into operationally usable results. The inorganic element profiles generated in the study have been used to form the basis for the development of a comprehensive database on poppy straw samples, which may be used in comparing samples and determining their origin.

  10. Norcoclaurine Synthase Is a Member of the Pathogenesis-Related 10/Bet v1 Protein Family[W

    PubMed Central

    Lee, Eun-Jeong; Facchini, Peter

    2010-01-01

    Norcoclaurine synthase (NCS) catalyzes the first committed step in the biosynthesis of benzylisoquinoline alkaloids (BIAs). NCS from Thalictrum flavum (Tf NCS), Papaver somniferum (Ps NCS1 and Ps NCS2), and Coptis japonica (Cj PR10A) share substantial identity with pathogen-related 10 (PR10) and Bet v1 proteins, whose functions are not well understood. A distinct enzyme (Cj NCS1) with similarity to 2-oxoglutarate–dependent dioxygenases was suggested as the bona fide NCS in C. japonica. Here, we validate the exclusive role of PR10/Bet v1-type NCS enzymes in BIA metabolism. Immunolocalization of Ps NCS2 revealed its cell type–specific occurrence in phloem sieve elements, which contain all other known BIA biosynthetic enzymes. In opium poppy, NCS transcripts and proteins were abundant in root and stem, but at low levels in leaf and carpel. Silencing of NCS in opium poppy profoundly reduced alkaloid levels compared with controls. Immunoprecipitation of NCS from total protein extracts of T. flavum cells resulted in a nearly complete attenuation of NCS activity. A Ps NCS2–green fluorescent protein fusion introduced by microprojectile bombardment into opium poppy cells initially localized to the endoplasmic reticulum but subsequently sorted to the vacuole. In our hands, Cj NCS1 did not catalyze the formation of (S)-norcoclaurine from dopamine and 4-hydroxyphenylacetaldehyde. PMID:21037103

  11. Noscapine comes of age.

    PubMed

    Chen, Xue; Dang, Thu-Thuy T; Facchini, Peter J

    2015-03-01

    Noscapine is a phthalideisoquinoline alkaloid, which represents a class of plant specialized metabolites within the large and structurally diverse group of benzylisoquinoline alkaloids. Along with the narcotic analgesic morphine, noscapine is a major alkaloid in the latex of opium poppy (Papaver somniferum) that has long been used as a cough suppressant and has undergone extensive investigation as a potential anticancer drug. Cultivated opium poppy plants remain the only commercial source of noscapine. Despite its isolation from opium more than two centuries ago, the almost complete biosynthesis of noscapine has only recently been established based on an impressive combination of molecular genetics, functional genomics, and metabolic biochemistry. In this review, we provide a historical account of noscapine from its discovery through to initial investigations of its formation in opium poppy. We also describe recent breakthroughs that have led to an elucidation of the noscapine biosynthetic pathway, and we discuss the pharmacological properties that have prompted intensive evaluation of the potential pharmaceutical applications of noscapine and several semi-synthetic derivatives. Finally, we speculate on the future potential for the production of noscapine using metabolic engineering and synthetic biology in plants and microbes. PMID:25583437

  12. Noscapine comes of age.

    PubMed

    Chen, Xue; Dang, Thu-Thuy T; Facchini, Peter J

    2015-03-01

    Noscapine is a phthalideisoquinoline alkaloid, which represents a class of plant specialized metabolites within the large and structurally diverse group of benzylisoquinoline alkaloids. Along with the narcotic analgesic morphine, noscapine is a major alkaloid in the latex of opium poppy (Papaver somniferum) that has long been used as a cough suppressant and has undergone extensive investigation as a potential anticancer drug. Cultivated opium poppy plants remain the only commercial source of noscapine. Despite its isolation from opium more than two centuries ago, the almost complete biosynthesis of noscapine has only recently been established based on an impressive combination of molecular genetics, functional genomics, and metabolic biochemistry. In this review, we provide a historical account of noscapine from its discovery through to initial investigations of its formation in opium poppy. We also describe recent breakthroughs that have led to an elucidation of the noscapine biosynthetic pathway, and we discuss the pharmacological properties that have prompted intensive evaluation of the potential pharmaceutical applications of noscapine and several semi-synthetic derivatives. Finally, we speculate on the future potential for the production of noscapine using metabolic engineering and synthetic biology in plants and microbes.

  13. Biochemical genomics for gene discovery in benzylisoquinoline alkaloid biosynthesis in opium poppy and related species.

    PubMed

    Dang, Thu Thuy T; Onoyovwi, Akpevwe; Farrow, Scott C; Facchini, Peter J

    2012-01-01

    Benzylisoquinoline alkaloids (BIAs) are a large, diverse group of ∼2500 specialized plant metabolites. Many BIAs display potent pharmacological activities, including the narcotic analgesics codeine and morphine, the vasodilator papaverine, the cough suppressant and potential anticancer drug noscapine, the antimicrobial agents sanguinarine and berberine, and the muscle relaxant (+)-tubocurarine. Opium poppy remains the sole commercial source for codeine, morphine, and a variety of semisynthetic drugs, including oxycodone and buprenorphine, derived primarily from the biosynthetic pathway intermediate thebaine. Recent advances in transcriptomics, proteomics, and metabolomics have created unprecedented opportunities for isolating and characterizing novel BIA biosynthetic genes. Here, we describe the application of next-generation sequencing and cDNA microarrays for selecting gene candidates based on comparative transcriptome analysis. We outline the basic mass spectrometric techniques to perform deep proteome and targeted metabolite analyses on BIA-producing plant tissues and provide methodologies for functionally characterizing biosynthetic gene candidates through in vitro enzyme assays and transient gene silencing in planta. PMID:22999177

  14. Detection of Verticillium wilt of olive trees and downy mildew of opium poppy using hyperspectral and thermal UAV imagery

    NASA Astrophysics Data System (ADS)

    Calderón Madrid, Rocío; Navas Cortés, Juan Antonio; Montes Borrego, Miguel; Landa del Castillo, Blanca Beatriz; Lucena León, Carlos; Jesús Zarco Tejada, Pablo

    2014-05-01

    The present study explored the use of high-resolution thermal, multispectral and hyperspectral imagery as indicators of the infections caused by Verticillium wilt (VW) in olive trees and downy mildew (DM) in opium poppy fields. VW, caused by the soil-borne fungus Verticillium dahliae, and DM, caused by the biotrophic obligate oomycete Peronospora arborescens, are the most economically limiting diseases of olive trees and opium poppy, respectively, worldwide. V. dahliae infects the plant by the roots and colonizes its vascular system, blocking water flow and eventually inducing water stress. P. arborescens colonizes the mesophyll, appearing the first symptoms as small chlorotic leaf lesions, which can evolve to curled and thickened tissues and systemic infections that become deformed and necrotic as the disease develops. The work conducted to detect VW and DM infection consisted on the acquisition of time series of airborne thermal, multispectral and hyperspectral imagery using 2-m and 5-m wingspan electric Unmanned Aerial Vehicles (UAVs) in spring and summer of three consecutive years (2009 to 2011) for VW detection and on three dates in spring of 2009 for DM detection. Two 7-ha commercial olive orchards naturally infected with V. dahliae and two opium poppy field plots artificially infected by P. arborescens were flown. Concurrently to the airborne campaigns, olive orchards and opium poppy fields were assessed "in situ" to assess actual VW severity and DM incidence. Furthermore, field measurements were conducted at leaf and crown level. The field results related to VW detection showed a significant increase in crown temperature (Tc) minus air temperature (Ta) and a decrease in leaf stomatal conductance (G) as VW severity increased. This reduction in G was associated with a significant increase in the Photochemical Reflectance Index (PRI570) and a decrease in chlorophyll fluorescence. DM asymptomatic leaves showed significantly higher NDVI and lower green/red index

  15. Lipoxygenase activity and sanguinarine production in cell suspension cultures of California poppy (Eschscholtzia californica CHAM.).

    PubMed

    Kollárová, R; Oblozinský, M; Kováciková, V; Holková, I; Balazová, A; Pekárová, M; Hoffman, P; Bezáková, L

    2014-08-01

    In this study we investigated the influence of biotic elicitor (phytopathogenic fungus Botrytis cinerea) and abiotic elicitors (methyljasmonate [MJ] and salicylic acid [SA]) on lipoxygenase (LOX) activity and sanguinarine production in cell suspension cultures of California poppy (Eschscholtzia californica CHAM.). We have observed different time effects of elicitors (10, 24, 48 and 72 h) on LOX activity and production of sanguinarine in in vitro cultures. All elicitors used in the experiments evidently increased the LOX activity and sanguinarine production in contrast to control samples. The highest LOX activities were determined in samples elicitated by MJ after 48 h and 72 h and the lowest LOX activities (in contrast to control samples) were detected after biotic elicitation by Botrytis cinerea. These activities showed about 50% lower level against the activities after MJ elicitation. The maximal amount of sanguinarine was observed after 48 h in MJ treated cultures (429.91 mg/g DCW) in comparision with control samples. Although all elicitors affect the sanguinarine production, effect of SA and biotic elicitor on sanguinarine accumulation in in vitrocultures was not so significant than after MJ elicitation.

  16. CYP82Y1 is N-methylcanadine 1-hydroxylase, a key noscapine biosynthetic enzyme in opium poppy.

    PubMed

    Dang, Thu-Thuy T; Facchini, Peter J

    2014-01-24

    Noscapine is a phthalideisoquinoline alkaloid investigated for its potent pharmacological properties. Although structurally elucidated more than a century ago, the biosynthesis of noscapine has not been established. Radiotracer studies have shown that noscapine is derived from the protoberberine alkaloid (S)-scoulerine and has been proposed to proceed through (S)-N-methylcanadine. However, pathway intermediates involved in the conversion of N-methylcanadine to noscapine have not been identified. We report the isolation and characterization of the cytochrome P-450 CYP82Y1, which catalyzes the 1-hydroxylation of N-methylcanadine to 1-hydroxy-N-methylcanadine. Comparison of transcript and metabolite profiles of eight opium poppy chemotypes revealed four cytochrome P-450s, three from the CYP82 and one from the CYP719 families, that were tightly correlated with noscapine accumulation. Recombinant CYP82Y1 was the only enzyme that accepted (R,S)-N-methylcanadine as a substrate with strict specificity and high affinity. As expected, CYP82Y1 was abundantly expressed in opium poppy stems where noscapine accumulation is highest among plant organs. Suppression of CYP82Y1 using virus-induced gene silencing caused a significant reduction in the levels of noscapine, narcotoline, and a putative downstream secoberbine intermediate and also resulted in increased accumulation of the upstream pathway intermediates scoulerine, tetrahydrocolum-bamine, canadine, and N-methylcanadine. The combined biochemical and physiological data support the 1-hydroxylation of (S)-N-methylcanadine catalyzed by CYP82Y1 as the first committed step in the formation of noscapine in opium poppy.

  17. Nocardioides flava sp. nov., isolated from rhizosphere of poppy plant, Republic of Korea.

    PubMed

    Singh, Hina; Yin, Chang Shik

    2016-04-01

    Phylogenetic and taxonomic characterization was performed for bacterium, designated strain THG-DN5.4(T), isolated from the rhizosphere of poppy plant collected from Gyeryongsan, Republic of Korea. Strain THG-DN5.4(T) consists of Gram-stain-positive, aerobic, non-motile rods. The bacteria grow optimally at 18-30 °C, at pH 7.0 and in the presence of 0.5-1.0 % NaCl. Based on 16S rRNA gene sequence analysis, strain THG-DN5.4(T) was found to be most closely related to Nocardioides nitrophenolicus KCTC 047BP(T), followed by Nocardioides ginsengisoli KCTC 19135(T), Nocardioides kongjuensis KCTC 19054(T), Nocardioides simplex KACC 20620(T), Nocardioides aromaticivorans KACC 20613(T), Nocardioides daeguensis KCTC 19799(T) and Nocardioides caeni KCTC 19600(T). The DNA-DNA relatedness between strain THG-DN5.4(T) and closely related phylogenetic neighbors was below 45.0 %, and the DNA G+C content of strain THG-DN5.4(T) was 70.8 mol%. An isoprenoid quinone was identified as MK-8(H4). Strain THG-DN5.4(T) was characterized chemotaxonomically as having LL-diaminopimelic acid in the cell-wall peptidoglycan. The polar lipids were identified as diphosphatidylglycerol, phosphatidylglycerol, some unidentified aminolipids and some unidentified polar lipids. iso-C16:0 and C18:1 ω9c were identified as the major fatty acids present in THG-DN5.4(T). On the basis of a polyphasic taxonomic study, strain THG-DN5.4(T) is considered to represent a novel species of the genus Nocardioides, for which the name Nocardioides flava sp. nov. is proposed. The type strain is THG-DN5.4(T) (=KCTC 39606(T)=CCTCC AB 2015298(T)).

  18. Nocardioides flava sp. nov., isolated from rhizosphere of poppy plant, Republic of Korea.

    PubMed

    Singh, Hina; Yin, Chang Shik

    2016-04-01

    Phylogenetic and taxonomic characterization was performed for bacterium, designated strain THG-DN5.4(T), isolated from the rhizosphere of poppy plant collected from Gyeryongsan, Republic of Korea. Strain THG-DN5.4(T) consists of Gram-stain-positive, aerobic, non-motile rods. The bacteria grow optimally at 18-30 °C, at pH 7.0 and in the presence of 0.5-1.0 % NaCl. Based on 16S rRNA gene sequence analysis, strain THG-DN5.4(T) was found to be most closely related to Nocardioides nitrophenolicus KCTC 047BP(T), followed by Nocardioides ginsengisoli KCTC 19135(T), Nocardioides kongjuensis KCTC 19054(T), Nocardioides simplex KACC 20620(T), Nocardioides aromaticivorans KACC 20613(T), Nocardioides daeguensis KCTC 19799(T) and Nocardioides caeni KCTC 19600(T). The DNA-DNA relatedness between strain THG-DN5.4(T) and closely related phylogenetic neighbors was below 45.0 %, and the DNA G+C content of strain THG-DN5.4(T) was 70.8 mol%. An isoprenoid quinone was identified as MK-8(H4). Strain THG-DN5.4(T) was characterized chemotaxonomically as having LL-diaminopimelic acid in the cell-wall peptidoglycan. The polar lipids were identified as diphosphatidylglycerol, phosphatidylglycerol, some unidentified aminolipids and some unidentified polar lipids. iso-C16:0 and C18:1 ω9c were identified as the major fatty acids present in THG-DN5.4(T). On the basis of a polyphasic taxonomic study, strain THG-DN5.4(T) is considered to represent a novel species of the genus Nocardioides, for which the name Nocardioides flava sp. nov. is proposed. The type strain is THG-DN5.4(T) (=KCTC 39606(T)=CCTCC AB 2015298(T)). PMID:26802009

  19. Dioxygenases catalyze O-demethylation and O,O-demethylenation with widespread roles in benzylisoquinoline alkaloid metabolism in opium poppy.

    PubMed

    Farrow, Scott C; Facchini, Peter J

    2013-10-01

    In opium poppy, the antepenultimate and final steps in morphine biosynthesis are catalyzed by the 2-oxoglutarate/Fe(II)-dependent dioxygenases, thebaine 6-O-demethylase (T6ODM) and codeine O-demethylase (CODM). Further investigation into the biochemical functions of CODM and T6ODM revealed extensive and unexpected roles for such enzymes in the metabolism of protopine, benzo[c]phenanthridine, and rhoeadine alkaloids. When assayed with a wide range of benzylisoquinoline alkaloids, CODM, T6ODM, and the functionally unassigned paralog DIOX2, renamed protopine O-dealkylase, showed novel and efficient dealkylation activities, including regio- and substrate-specific O-demethylation and O,O-demethylenation. Enzymes catalyzing O,O-demethylenation, which cleave a methylenedioxy bridge leaving two hydroxyl groups, have previously not been reported in plants. Similar cleavage of methylenedioxy bridges on substituted amphetamines is catalyzed by heme-dependent cytochromes P450 in mammals. Preferred substrates for O,O-demethylenation by CODM and protopine O-dealkylase were protopine alkaloids that serve as intermediates in the biosynthesis of benzo[c]phenanthridine and rhoeadine derivatives. Virus-induced gene silencing used to suppress the abundance of CODM and/or T6ODM transcripts indicated a direct physiological role for these enzymes in the metabolism of protopine alkaloids, and they revealed their indirect involvement in the formation of the antimicrobial benzo[c]phenanthridine sanguinarine and certain rhoeadine alkaloids in opium poppy.

  20. Dioxygenases Catalyze O-Demethylation and O,O-Demethylenation with Widespread Roles in Benzylisoquinoline Alkaloid Metabolism in Opium Poppy*

    PubMed Central

    Farrow, Scott C.; Facchini, Peter J.

    2013-01-01

    In opium poppy, the antepenultimate and final steps in morphine biosynthesis are catalyzed by the 2-oxoglutarate/Fe(II)-dependent dioxygenases, thebaine 6-O-demethylase (T6ODM) and codeine O-demethylase (CODM). Further investigation into the biochemical functions of CODM and T6ODM revealed extensive and unexpected roles for such enzymes in the metabolism of protopine, benzo[c]phenanthridine, and rhoeadine alkaloids. When assayed with a wide range of benzylisoquinoline alkaloids, CODM, T6ODM, and the functionally unassigned paralog DIOX2, renamed protopine O-dealkylase, showed novel and efficient dealkylation activities, including regio- and substrate-specific O-demethylation and O,O-demethylenation. Enzymes catalyzing O,O-demethylenation, which cleave a methylenedioxy bridge leaving two hydroxyl groups, have previously not been reported in plants. Similar cleavage of methylenedioxy bridges on substituted amphetamines is catalyzed by heme-dependent cytochromes P450 in mammals. Preferred substrates for O,O-demethylenation by CODM and protopine O-dealkylase were protopine alkaloids that serve as intermediates in the biosynthesis of benzo[c]phenanthridine and rhoeadine derivatives. Virus-induced gene silencing used to suppress the abundance of CODM and/or T6ODM transcripts indicated a direct physiological role for these enzymes in the metabolism of protopine alkaloids, and they revealed their indirect involvement in the formation of the antimicrobial benzo[c]phenanthridine sanguinarine and certain rhoeadine alkaloids in opium poppy. PMID:23928311

  1. Medicinal aspects of opium as described in Avicenna's Canon of Medicine.

    PubMed

    Heydari, Mojtaba; Hashempur, Mohammad Hashem; Zargaran, Arman

    2013-01-01

    Throughout history, opium has been used as a base for the opioid class of drugs used to suppress the central nervous system. Opium is a substance extracted from the opium poppy (Papaver somniferum L.). Its consumption and medicinal application date back to antiquity. In the medieval period, Avicenna, a famous Persian scholar (980-1037 AD) described poppy under the entry Afion of his medical encyclopedia Canon of Medicine. Various effects of opium consumption, both wanted and unwanted are discussed in the encyclopedia. The text mentions the effects of opioids such as analgesic, hypnotic, antitussive, gastrointestinal, cognitive, respiratory depression, neuromuscular disturbance, and sexual dysfunction. It also refers to its potential as a poison. Avicenna describes several methods of delivery and recommendations for doses of the drug. Most of opioid effects described by Avicenna have subsequently been confirmed by modern research, and other references to opium use in medieval texts call for further investigation. This article highlights an important aspect of the medieval history of medicine.

  2. Role of the phloem in the biochemistry and ecophysiology of benzylisoquinoline alkaloid metabolism.

    PubMed

    Lee, Eun-Jeong; Hagel, Jillian M; Facchini, Peter J

    2013-01-01

    Benzylisoquinoline alkaloids (BIAs) are a diverse group of biologically active specialized metabolites produced mainly in four plant families. BIA metabolism is likely of monophyletic origin and involves multiple enzymes yielding structurally diverse compounds. Several BIAs possess defensive properties against pathogenic microorganisms and herbivores. Opium poppy (Papaver somniferum: Papaveraceae) has emerged as a model system to investigate the cellular localization of BIA biosynthesis. Although alkaloids accumulate in the laticifer cytoplasm (latex) of opium poppy, corresponding biosynthetic enzymes and gene transcripts are localized to proximal sieve elements and companion cells, respectively. In contrast, BIA metabolism in the non-laticiferous meadow rue (Thalictrum flavum; Ranunculaceae) occurs independent of the phloem. Evidence points toward the adoption of diverse strategies for the biosynthesis and accumulation of alkaloids as defensive compounds. Recruitment of cell types involved in BIA metabolism, both within and external to the phloem, was likely driven by selection pressures unique to different taxa. The biochemistry, cell biology, ecophysiology, and evolution of BIA metabolism are considered in this context.

  3. Characterization of Neuraminidase Inhibitors in Korean Papaver rhoeas Bee Pollen Contributing to Anti-Influenza Activities In Vitro.

    PubMed

    Lee, In-Kyoung; Hwang, Byung Soon; Kim, Dae-Won; Kim, Ji-Yul; Woo, E-Eum; Lee, Yoon-Ju; Choi, Hwa Jung; Yun, Bong-Sik

    2016-04-01

    The active constituents of Korean Papaver rhoeas bee pollen conferring neuraminidase inhibitory activities (H1N1, H3N2, and H5N1) were investigated. Six flavonoids and one alkaloid were isolated and characterized by nuclear magnetic resonance and mass spectrometry data. These included kaempferol-3-sophoroside (1), kaempferol-3-neohesperidoside (2), kaempferol-3-sambubioside (3), kaempferol-3-glucoside (4), quercetin-3-sophoroside (5), luteolin (6), and chelianthifoline (7). All compounds showed neuraminidase inhibitory activities with IC50 values ranging from 10.7 to 151.1 µM. The most potent neuraminidase inhibitor was luteolin, which was the dominant content in the ethyl acetate fraction. All tested compounds displayed noncompetitive inhibition of H3N2 neuraminidase. Furthermore, compounds 1-7 all reduced the severity of virally induced cytopathic effects as determined by the Madin-Darby canine kidney cell-based assay showing antiviral activity with IC50 values ranging from 10.7 to 33.4 µM (zanamivir: 58.3 µM). The active compounds were quantified by high-performance liquid chromatography, and the total amount of compounds 1-7 made up about 0.592 g/100 g bee pollen, contributing a rich resource of a natural antiviral product. PMID:26848705

  4. Cloning and expression of a distinctive class of self-incompatibility (S) gene from Papaver rhoeas L.

    PubMed Central

    Foote, H C; Ride, J P; Franklin-Tong, V E; Walker, E A; Lawrence, M J; Franklin, F C

    1994-01-01

    We present the identification, cloning, and characterization of a self-incompatibility (S) gene from Papaver rhoeas that has no significant homology to any previously reported gene sequences, including S genes from other species. This result suggests that a different self-incompatibility mechanism may be operating in this species and has important implications for the evolutionary relationships between the S genes. The S1 cDNA was cloned by using an oligonucleotide based upon N-terminal amino acid sequence data from stigmatic proteins that show complete linkage with the S1 gene. The single-copy gene has been expressed in Escherichia coli to test biological activity. Although the recombinant S1 protein (S1e) is not processed in the same way as the protein produced in the plant, it exhibits, in vitro, the specific pollen inhibitory activity expected of an S gene product; pollen carrying the S1 allele is inhibited, whereas pollen not carrying S1 is not inhibited. These results provide definitive demonstration that the product of a cloned S gene has S-specific pollen inhibitory activity. Images PMID:8134385

  5. Phloem-Specific Expression of Tyrosine/Dopa Decarboxylase Genes and the Biosynthesis of Isoquinoline Alkaloids in Opium Poppy.

    PubMed

    Facchini, P. J.; De Luca, V.

    1995-11-01

    Tyrosine/dopa decarboxylase (TYDC) catalyzes the formation of tyramine and dopamine and represents the first steps in the biosynthesis of the large and diverse group of tetrahydroisoquinoline alkaloids. Opium poppy accumulates morphine in aerial organs and roots, whereas sanguinarine, which is derived from a distinct branch pathway, accumulates only in roots. Expression of the TYDC gene family in opium poppy was investigated in relation to the organ-specific biosynthesis of these different types of alkaloids. Members of the TYDC gene family are classified into two groups (represented by TYDC1 and TYDC2) and are differentially expressed. In the mature plant, TYDC2-like transcripts are predominant in stems and are also present in roots, whereas TYDC1-like transcripts are abundant only in roots. In situ hybridization analysis revealed that the expression of TYDC genes is developmentally regulated. TYDC transcripts are associated with vascular tissue in mature roots and stems but are also expressed in cortical tissues at earlier stages of development. Expression of TYDC genes is restricted to metaphloem and to protoxylem in the vascular bundles of mature aerial organs. Localization of TYDC transcripts in the phloem is consistent with the expected developmental origin of laticifers, which are specialized internal secretory cells that accompany vascular tissues in all organs of select species and that contain the alkaloid-rich latex in aerial organs. The differential expression of TYDC genes and the organ-dependent accumulation of different alkaloids suggest a coordinated regulation of specific alkaloid biosynthetic genes that are ultimately controlled by specific developmental programs.

  6. [Determination of poppy ingredients in chafing dish materials by isotopic internal standard coupled with multiple reaction monitoring and online full scan mass spectrometry].

    PubMed

    Zhu, Weixia; Sun, Zhuanlian; Yuan, Ping; Yang, Jizhou; Liu, Yafeng; Sun, Wuyong

    2014-12-01

    A confirmative method was developed for determining five poppy alkaloids including morphine, codeine, papaverine, tibane, noscapine in chafing dish ingredients by high performance liquid chromatography coupled with triple quadrupole linear ion trap mass spectrometry (HPLC-Q Trap MS). The sample was extracted with dilute HCl solution under heating condition. The removal of lipid procedure was performed with hexane. The purification was carried out on a mixed-cation solid-phase extraction column (MCX) and ethyl acetate-methanol containing 5% aqueous ammonia was used for elution. A PAK ST column was used to separate the analytes, and 5 mmol/L ammonium acetate methanol and 10 mmol/L ammonium acetate (pH 3. 6) were used as mobile phases. The five alkaloids was detected in the positive mode simultaneously by multiple reaction monitoring (MRM) and online enhanced product ion full scan (EPI). The LODs were 0.05-0.5 µg/kg and the LOQs were 0. 2-2 µg/kg for the five poppy alkaloids. The overall recoveries of the method varied from 64. 2% to 110. 6%, and the RSD were between 4. 2% and 12. 5%. The EPI mass spectra of positive samples were searched through standard library for qualitative confirmation. The detection of real hot pot material samples showed this method can be used for the simple and accurate determination of the five poppy alkaloid residues in chafing dish.

  7. Unsupervised multiple endmember spectral mixture analysis-based detection of opium poppy fields from an EO-1 Hyperion image in Helmand, Afghanistan.

    PubMed

    Wang, Jian-Jun; Zhang, Yun; Bussink, Coen

    2014-04-01

    Since 1992, Afghanistan has gradually become the primary illicit opium producer in the entire world. To efficiently eradicate the opium poppy, it is crucial for the United Nations Office on Drugs and Crime (UNODC) and Ministry of Counter Narcotics of Afghanistan to monitor opium poppy cultivation timely. In situ detection of opium fields, however, is often expensive, time-consuming and dangerous in Afghanistan. To overcome the constraints of inaccessibility of opium fields, high-resolution (≤1 m) images, like pan-sharpened IKONOS, have been applied in previous studies. Unfortunately, these high-resolution images are expensive when monitoring a large area. In contrast, EO-1 Hyperion imagery, the only source of spaceborne hyperspectral data, has a coarse resolution (30 m), but it is free of charge. Moreover, Hyperion's large number of channels may increase the detection capability of subpixel size targets. Until now, however, little research has been found that identified opium fields from spaceborne or aerial hyperspectral images. Therefore, this study attempts to detect opium fields from a Hyperion image covering a study area in Southwest Afghanistan in a situation where training samples were not available. A proposed methodology based on unsupervised endmember-selection and multiple-endmember spectral mixture analysis can detect opium fields directly from the Hyperion image. The number of poppy pixels was overestimated by 12%.

  8. [Determination of poppy ingredients in chafing dish materials by isotopic internal standard coupled with multiple reaction monitoring and online full scan mass spectrometry].

    PubMed

    Zhu, Weixia; Sun, Zhuanlian; Yuan, Ping; Yang, Jizhou; Liu, Yafeng; Sun, Wuyong

    2014-12-01

    A confirmative method was developed for determining five poppy alkaloids including morphine, codeine, papaverine, tibane, noscapine in chafing dish ingredients by high performance liquid chromatography coupled with triple quadrupole linear ion trap mass spectrometry (HPLC-Q Trap MS). The sample was extracted with dilute HCl solution under heating condition. The removal of lipid procedure was performed with hexane. The purification was carried out on a mixed-cation solid-phase extraction column (MCX) and ethyl acetate-methanol containing 5% aqueous ammonia was used for elution. A PAK ST column was used to separate the analytes, and 5 mmol/L ammonium acetate methanol and 10 mmol/L ammonium acetate (pH 3. 6) were used as mobile phases. The five alkaloids was detected in the positive mode simultaneously by multiple reaction monitoring (MRM) and online enhanced product ion full scan (EPI). The LODs were 0.05-0.5 µg/kg and the LOQs were 0. 2-2 µg/kg for the five poppy alkaloids. The overall recoveries of the method varied from 64. 2% to 110. 6%, and the RSD were between 4. 2% and 12. 5%. The EPI mass spectra of positive samples were searched through standard library for qualitative confirmation. The detection of real hot pot material samples showed this method can be used for the simple and accurate determination of the five poppy alkaloid residues in chafing dish. PMID:25902640

  9. Plant Reproduction: Self-Incompatibility to Go.

    PubMed

    Tovar-Mendez, Alejandro; McClure, Bruce

    2016-02-01

    In a new study, the Papaver rhoeas (poppy family) self-incompatibility system has been transferred into Arabidopsis thaliana, a distantly related plant with a very different floral structure. The simple poppy self-incompatibility system may finally make it possible to introduce this potentially valuable trait into any plant. PMID:26859267

  10. Evidence for the monophyletic evolution of benzylisoquinoline alkaloid biosynthesis in angiosperms.

    PubMed

    Liscombe, David K; Macleod, Benjamin P; Loukanina, Natalia; Nandi, Owi I; Facchini, Peter J

    2005-06-01

    Benzylisoquinoline alkaloids (BIAs) consist of more than 2500 diverse structures largely restricted to the order Ranunculales and the eumagnoliids. However, BIAs also occur in the Rutaceae, Lauraceae, Cornaceae and Nelumbonaceae, and sporadically throughout the order Piperales. Several of these alkaloids function in the defense of plants against herbivores and pathogens - thus, the capacity for BIA biosynthesis is expected to play an important role in the reproductive fitness of certain plants. Biochemical and molecular phylogenetic approaches were used to investigate the evolution of BIA biosynthesis in basal angiosperms. The occurrence of (S)-norcoclaurine synthase (NCS; EC 4.2.1.78) activity in 90 diverse plant species was compared to the distribution of BIAs superimposed onto a molecular phylogeny. These results support the monophyletic origin of BIA biosynthesis prior to the emergence of the eudicots. Phylogenetic analyses of NCS, berberine bridge enzyme and several O-methyltransferases suggest a latent molecular fingerprint for BIA biosynthesis in angiosperms not known to accumulate such alkaloids. The limited occurrence of BIAs outside the Ranunculales and eumagnoliids suggests the requirement for a highly specialized, yet evolutionarily unstable cellular platform to accommodate or reactivate the pathway in divergent taxa. The molecular cloning and functional characterization of NCS from opium poppy (Papaver somniferum L.) is also reported. Pathogenesis-related (PR)10 and Bet v 1 major allergen proteins share homology with NCS, but recombinant polypeptides were devoid of NCS activity.

  11. Evidence for the monophyletic evolution of benzylisoquinoline alkaloid biosynthesis in angiosperms.

    PubMed

    Liscombe, David K; MacLeod, Benjamin P; Loukanina, Natalia; Nandi, Owi I; Facchini, Peter J

    2005-10-01

    Benzylisoquinoline alkaloids (BIAs) consist of more than 2500 diverse structures largely restricted to the order Ranunculales and the eumagnoliids. However, BIAs also occur in the Rutaceae, Lauraceae, Cornaceae and Nelumbonaceae, and sporadically throughout the order Piperales. Several of these alkaloids function in the defense of plants against herbivores and pathogens--thus the capacity for BIA biosynthesis is expected to play an important role in the reproductive fitness of certain plants. Biochemical and molecular phylogenetic approaches were used to investigate the evolution of BIA biosynthesis in basal angiosperms. The occurrence of (S)-norcoclaurine synthase (NCS; EC 4.2.1.78) activity in 90 diverse plant species was compared to the distribution of BIAs superimposed onto a molecular phylogeny. These results support the monophyletic origin of BIA biosynthesis prior to the emergence of the eudicots. Phylogenetic analysis of NCS, berberine bridge enzyme and several O-methyltransferases suggest a latent molecular fingerprint for BIA biosynthesis in angiosperms not known to accumulate such alkaloids. The limited occurrence of BIAs outside the Ranunculales and eumagnoliids suggests the requirement for a highly specialized, yet evolutionarily unstable cellular platform to accommodate or reactivate the pathway in divergent taxa. The molecular cloning and functional characterization of NCS from opium poppy (Papaver somniferum L.) is also reported. Pathogenesis--related (PR)10 and Bet v 1 major allergen proteins share homology with NCS, but recombinant polypeptides were devoid of NCS activity.

  12. Blind column selection protocol for two-dimensional high performance liquid chromatography.

    PubMed

    Burns, Niki K; Andrighetto, Luke M; Conlan, Xavier A; Purcell, Stuart D; Barnett, Neil W; Denning, Jacquie; Francis, Paul S; Stevenson, Paul G

    2016-07-01

    The selection of two orthogonal columns for two-dimensional high performance liquid chromatography (LC×LC) separation of natural product extracts can be a labour intensive and time consuming process and in many cases is an entirely trial-and-error approach. This paper introduces a blind optimisation method for column selection of a black box of constituent components. A data processing pipeline, created in the open source application OpenMS®, was developed to map the components within the mixture of equal mass across a library of HPLC columns; LC×LC separation space utilisation was compared by measuring the fractional surface coverage, fcoverage. It was found that for a test mixture from an opium poppy (Papaver somniferum) extract, the combination of diphenyl and C18 stationary phases provided a predicted fcoverage of 0.48 and was matched with an actual usage of 0.43. OpenMS®, in conjunction with algorithms designed in house, have allowed for a significantly quicker selection of two orthogonal columns, which have been optimised for a LC×LC separation of crude extractions of plant material. PMID:27154652

  13. Growth of Pollen Tubes of Papaver rhoeas Is Regulated by a Slow-Moving Calcium Wave Propagated by Inositol 1,4,5-Trisphosphate.

    PubMed Central

    Franklin-Tong, V. E.; Drobak, B. K.; Allan, A. C.; Watkins, PAC.; Trewavas, A. J.

    1996-01-01

    A signaling role for cytosolic free Ca2+ ([Ca2+]i) in regulating Papaver rhoeas pollen tube growth during the self-incompatibility response has been demonstrated previously. In this article, we investigate the involvement of the phosphoinositide signal transduction pathway in Ca2+-mediated pollen tube inhibition. We demonstrate that P. rhoeas pollen tubes have a Ca2+-dependent polyphosphoinositide-specific phospholipase C activity that is inhibited by neomycin. [Ca2+]i imaging after photolysis of caged inositol (1,4,5)-trisphosphate (Ins[1,4,5]P3) in pollen tubes demonstrated that Ins(1,4,5)P3 could induce Ca2+ release, which was inhibited by heparin and neomycin. Mastoparan, which stimulated Ins(1,4,5)P3 production, also induced a rapid increase in Ca2+, which was inhibited by neomycin. These data provide direct evidence for the involvement of a functional phosphoinositide signal-transducing system in the regulation of pollen tube growth. We suggest that the observed Ca2+ increases are mediated, at least in part, by Ins(1,4,5)P3-induced Ca2+ release. Furthermore, we provide data suggesting that Ca2+ waves, which have not previously been reported in plant cells, can be induced in pollen tubes. PMID:12239415

  14. Cytochrome P450 3A Enzymes Catalyze the O6-Demethylation of Thebaine, a Key Step in Endogenous Mammalian Morphine Biosynthesis*

    PubMed Central

    Kramlinger, Valerie M.; Alvarado Rojas, Mónica; Kanamori, Tatsuyuki; Guengerich, F. Peter

    2015-01-01

    Morphine, first characterized in opium from the poppy Papaver somniferum, is one of the strongest known analgesics. Endogenous morphine has been identified in several mammalian cells and tissues. The synthetic pathway of morphine in the opium poppy has been elucidated. The presence of common intermediates in plants and mammals suggests that biosynthesis occurs through similar pathways (beginning with the amino acid l-tyrosine), and the pathway has been completely delineated in plants. Some of the enzymes in the mammalian pathway have been identified and characterized. Two of the latter steps in the morphine biosynthesis pathway are demethylation of thebaine at the O3- and the O6-positions, the latter of which has been difficult to demonstrate. The plant enzymes responsible for both the O3-demethylation and the O6-demethylation are members of the FeII/α-ketoglutarate-dependent dioxygenase family. Previous studies showed that human cytochrome P450 (P450) 2D6 can catalyze thebaine O3-demethylation. We report that demethylation of thebaine at the O6-position is selectively catalyzed by human P450s 3A4 and 3A5, with the latter being more efficient, and rat P450 3A2. Our results do not support O6-demethylation of thebaine by an FeII/α-ketoglutarate-dependent dioxygenase. In rat brain microsomes, O6-demethylation was inhibited by ketoconazole, but not sulfaphenazole, suggesting that P450 3A enzymes are responsible for this activity in the brain. An alternate pathway to morphine, oripavine O6-demethylation, was not detected. The major enzymatic steps in mammalian morphine synthesis have now been identified. PMID:26157146

  15. Cytochrome P450 3A Enzymes Catalyze the O6-Demethylation of Thebaine, a Key Step in Endogenous Mammalian Morphine Biosynthesis.

    PubMed

    Kramlinger, Valerie M; Alvarado Rojas, Mónica; Kanamori, Tatsuyuki; Guengerich, F Peter

    2015-08-14

    Morphine, first characterized in opium from the poppy Papaver somniferum, is one of the strongest known analgesics. Endogenous morphine has been identified in several mammalian cells and tissues. The synthetic pathway of morphine in the opium poppy has been elucidated. The presence of common intermediates in plants and mammals suggests that biosynthesis occurs through similar pathways (beginning with the amino acid L-tyrosine), and the pathway has been completely delineated in plants. Some of the enzymes in the mammalian pathway have been identified and characterized. Two of the latter steps in the morphine biosynthesis pathway are demethylation of thebaine at the O(3)- and the O(6)-positions, the latter of which has been difficult to demonstrate. The plant enzymes responsible for both the O(3)-demethylation and the O(6)-demethylation are members of the Fe(II)/α-ketoglutarate-dependent dioxygenase family. Previous studies showed that human cytochrome P450 (P450) 2D6 can catalyze thebaine O(3)-demethylation. We report that demethylation of thebaine at the O(6)-position is selectively catalyzed by human P450s 3A4 and 3A5, with the latter being more efficient, and rat P450 3A2. Our results do not support O(6)-demethylation of thebaine by an Fe(II)/α-ketoglutarate-dependent dioxygenase. In rat brain microsomes, O(6)-demethylation was inhibited by ketoconazole, but not sulfaphenazole, suggesting that P450 3A enzymes are responsible for this activity in the brain. An alternate pathway to morphine, oripavine O(6)-demethylation, was not detected. The major enzymatic steps in mammalian morphine synthesis have now been identified.

  16. Cytochrome P450 3A Enzymes Catalyze the O6-Demethylation of Thebaine, a Key Step in Endogenous Mammalian Morphine Biosynthesis.

    PubMed

    Kramlinger, Valerie M; Alvarado Rojas, Mónica; Kanamori, Tatsuyuki; Guengerich, F Peter

    2015-08-14

    Morphine, first characterized in opium from the poppy Papaver somniferum, is one of the strongest known analgesics. Endogenous morphine has been identified in several mammalian cells and tissues. The synthetic pathway of morphine in the opium poppy has been elucidated. The presence of common intermediates in plants and mammals suggests that biosynthesis occurs through similar pathways (beginning with the amino acid L-tyrosine), and the pathway has been completely delineated in plants. Some of the enzymes in the mammalian pathway have been identified and characterized. Two of the latter steps in the morphine biosynthesis pathway are demethylation of thebaine at the O(3)- and the O(6)-positions, the latter of which has been difficult to demonstrate. The plant enzymes responsible for both the O(3)-demethylation and the O(6)-demethylation are members of the Fe(II)/α-ketoglutarate-dependent dioxygenase family. Previous studies showed that human cytochrome P450 (P450) 2D6 can catalyze thebaine O(3)-demethylation. We report that demethylation of thebaine at the O(6)-position is selectively catalyzed by human P450s 3A4 and 3A5, with the latter being more efficient, and rat P450 3A2. Our results do not support O(6)-demethylation of thebaine by an Fe(II)/α-ketoglutarate-dependent dioxygenase. In rat brain microsomes, O(6)-demethylation was inhibited by ketoconazole, but not sulfaphenazole, suggesting that P450 3A enzymes are responsible for this activity in the brain. An alternate pathway to morphine, oripavine O(6)-demethylation, was not detected. The major enzymatic steps in mammalian morphine synthesis have now been identified. PMID:26157146

  17. Plug-and-Play Benzylisoquinoline Alkaloid Biosynthetic Gene Discovery in Engineered Yeast.

    PubMed

    Morris, J S; Dastmalchi, M; Li, J; Chang, L; Chen, X; Hagel, J M; Facchini, P J

    2016-01-01

    Benzylisoquinoline alkaloid (BIA) metabolism has been the focus of a considerable research effort over the past half-century, primarily because of the pharmaceutical importance of several compounds produced by opium poppy (Papaver somniferum). Advancements in genomics technologies have substantially accelerated the rate of gene discovery over the past decade, such that most biosynthetic enzymes involved in the formation of the major alkaloids of opium poppy have now been isolated and partially characterized. Not unexpectedly, the availability of all perceived biosynthetic genes has facilitated the reconstitution of several BIA pathways in microbial hosts, including yeast (Saccharomyces cerevisiae). Product yields are currently insufficient to consider the commercial production of high-value BIAs, such as morphine. However, the rudimentary success demonstrated by the uncomplicated and routine assembly of a multitude of characterized BIA biosynthetic genes provides a valuable gene discovery tool for the rapid functional identification of the plethora of gene candidates available through increasingly accessible genomic, transcriptomic, and proteomic databases. BIA biosynthetic gene discovery represents a substantial research opportunity largely owing to the wealth of existing enzyme data mostly obtained from a single plant species. Functionally novel enzymes and variants with potential metabolic engineering applications can be considered the primary targets. Selection of candidates from sequence repositories is facilitated by the monophyletic relationship among biosynthetic genes belonging to a wide range of enzyme families, such as the numerous cytochromes P450 and AdoMet-dependent O- and N-methyltransferases that operate in BIA metabolism. We describe methods for the rapid functional screening of uncharacterized gene candidates encoding potential BIA biosynthetic enzymes using yeast strains engineered to perform selected metabolic conversions. As an initial

  18. Draft Genome Sequence of Agrobacterium sp. Strain R89-1, a Morphine Alkaloid-Biotransforming Bacterium

    PubMed Central

    Kyslíková, Eva

    2016-01-01

    Agrobacterium sp. strain R89-1 isolated from composted wastes of Papaver somniferum can effectively biotransform codeine/morphine into 14-OH-derivatives. Here, we present a 4.7-Mb assembly of the R89-1 strain genome. The draft shows that the strain R89-1 represents a distinct phylogenetic lineage within the genus Agrobacterium. PMID:27056219

  19. Characterization of two methylenedioxy bridge-forming cytochrome P450-dependent enzymes of alkaloid formation in the Mexican prickly poppy Argemone mexicana.

    PubMed

    Díaz Chávez, Maria Luisa; Rolf, Megan; Gesell, Andreas; Kutchan, Toni M

    2011-03-01

    Formation of the methylenedioxy bridge is an integral step in the biosynthesis of benzo[c]phenanthridine and protoberberine alkaloids in the Papaveraceae family of plants. This reaction in plants is catalyzed by cytochrome P450-dependent enzymes. Two cDNAs that encode cytochrome P450 enzymes belonging to the CYP719 family were identified upon interrogation of an EST dataset prepared from 2-month-old plantlets of the Mexican prickly poppy Argemone mexicana that accumulated the benzo[c]phenanthridine alkaloid sanguinarine and the protoberberine alkaloid berberine. CYP719A13 and CYP719A14 are 58% identical to each other and 77% and 60% identical, respectively, to stylopine synthase CYP719A2 of benzo[c]phenanthridine biosynthesis in Eschscholzia californica. Functional heterologous expression of CYP719A14 and CYP719A13 in Spodoptera frugiperda Sf9 cells produced recombinant enzymes that catalyzed the formation of the methylenedioxy bridge of (S)-cheilanthifoline from (S)-scoulerine and of (S)-stylopine from (S)-cheilanthifoline, respectively. Twenty-seven potential substrates were tested with each enzyme. Whereas CYP719A14 transformed only (S)-scoulerine to (S)-cheilanthifoline (K(m) 1.9±0.3; k(cat)/K(m) 1.7), CYP719A13 converted (S)-tetrahydrocolumbamine to (S)-canadine (K(m) 2.7±1.3; k(cat)/K(m) 12.8), (S)-cheilanthifoline to (S)-stylopine (K(m) 5.2±3.0; k(cat)/K(m) 2.6) and (S)-scoulerine to (S)-nandinine (K(m) 8.1±1.9; k(cat)/K(m) 0.7). These results indicate that although CYP719A14 participates in only sanguinarine biosynthesis, CYP719A13 can be involved in both sanguinarine and berberine formation in A. mexicana.

  20. The seirena B Class Floral Homeotic Mutant of California Poppy (Eschscholzia californica) Reveals a Function of the Enigmatic PI Motif in the Formation of Specific Multimeric MADS Domain Protein Complexes[C][W][OA

    PubMed Central

    Lange, Matthias; Orashakova, Svetlana; Lange, Sabrina; Melzer, Rainer; Theißen, Günter; Smyth, David R.; Becker, Annette

    2013-01-01

    The products of B class floral homeotic genes specify petal and stamen identity, and loss of B function results in homeotic conversions of petals into sepals and stamens into carpels. Here, we describe the molecular characterization of seirena-1 (sei-1), a mutant from the basal eudicot California poppy (Eschscholzia californica) that shows homeotic changes characteristic of floral homeotic B class mutants. SEI has been previously described as EScaGLO, one of four B class–related MADS box genes in California poppy. The C terminus of SEI, including the highly conserved PI motif, is truncated in sei-1 proteins. Nevertheless, like the wild-type SEI protein, the sei-1 mutant protein is able to bind CArG-boxes and can form homodimers, heterodimers, and several higher order complexes with other MADS domain proteins. However, unlike the wild type, the mutant protein is not able to mediate higher order complexes consisting of specific B, C, and putative E class related proteins likely involved in specifying stamen identity. Within the PI motif, five highly conserved N-terminal amino acids are specifically required for this interaction. Several families lack this short conserved sequence, including the Brassicaceae, and we propose an evolutionary scenario to explain these functional differences. PMID:23444328

  1. Benzylisoquinoline alkaloid metabolism: a century of discovery and a brave new world.

    PubMed

    Hagel, Jillian M; Facchini, Peter J

    2013-05-01

    Benzylisoquinoline alkaloids (BIAs) are a structurally diverse group of plant specialized metabolites with a long history of investigation. Although the ecophysiological functions of most BIAs are unknown, the medicinal properties of many compounds have been exploited for centuries. These include the narcotic analgesics codeine and morphine, the antimicrobial agents sanguinarine and berberine, and the antitussive and anticancer drug noscapine. BIA biosynthesis involves a restricted number of enzyme types that catalyze landmark coupling reactions and subsequent functional group modifications. A pathogenesis-related (PR)10/Bet v1 'Pictet-Spenglerase', several O-methyl-, N-methyl- and O-acetyltransferases, cytochromes P450, FAD-dependent oxidases, non-heme dioxygenases and NADPH-dependent reductases have been implicated in the multistep pathways leading to structurally diverse alkaloids. A small number of plant species, including opium poppy (Papaver somniferum) and other members of the Ranunculales, have emerged as model systems to study BIA metabolism. The expansion of resources to include a wider range of plant species is creating an opportunity to investigate previously uncharacterized BIA pathways. Contemporary knowledge of BIA metabolism reflects over a century of research coupled with the development of key innovations such as radioactive tracing, enzyme isolation and molecular cloning, and functional genomics approaches such as virus-induced gene silencing. Recently, the emergence of transcriptomics, proteomics and metabolomics has expedited the discovery of new BIA biosynthetic genes. The growing repository of BIA biosynthetic genes is providing the parts required to apply emerging synthetic biology platforms to the development of production systems in microbes as an alternative to plants as a commecial source of valuable BIAs.

  2. Functional Characterization of 4′OMT and 7OMT Genes in BIA Biosynthesis

    PubMed Central

    Gurkok, Tugba; Ozhuner, Esma; Parmaksiz, Iskender; Özcan, Sebahattin; Turktas, Mine; İpek, Arif; Demirtas, Ibrahim; Okay, Sezer; Unver, Turgay

    2016-01-01

    Alkaloids are diverse group of secondary metabolites generally found in plants. Opium poppy (Papaver somniferum L.), the only commercial source of morphinan alkaloids, has been used as a medicinal plant since ancient times. It produces benzylisoquinoline alkaloids (BIA) including the narcotic analgesic morphine, the muscle relaxant papaverine, and the anti-cancer agent noscapine. Though BIAs play crucial roles in many biological mechanisms their steps in biosynthesis and the responsible genes remain to be revealed. In this study, expressions of 3-hydroxy-N-methylcoclaurine 4′–methyltransferase (4′OMT) and reticuline 7-O-methyltransferase (7OMT) genes were subjected to manipulation to functionally characterize their roles in BIA biosynthesis. Measurements of alkaloid accumulation were performed in leaf, stem, and capsule tissues accordingly. Suppression of 4′OMT expression caused reduction in the total alkaloid content in stem tissue whereas total alkaloid content was significantly induced in the capsule. Silencing of the 7OMT gene also caused repression in total alkaloid content in the stem. On the other hand, over-expression of 4′OMT and 7OMT resulted in higher morphine accumulation in the stem but suppressed amount in the capsule. Moreover, differential expression in several BIA synthesis genes (CNMT, TYDC, 6OMT, SAT, COR, 4′OMT, and 7OMT) were observed upon manipulation of 4′OMT and 7OMT expression. Upon silencing and overexpression applications, tissue specific effects of these genes were identified. Manipulation of 4′OMT and 7OMT genes caused differentiated accumulation of BIAs including morphine and noscapine in capsule and stem tissues. PMID:26909086

  3. Functional Characterization of 4'OMT and 7OMT Genes in BIA Biosynthesis.

    PubMed

    Gurkok, Tugba; Ozhuner, Esma; Parmaksiz, Iskender; Özcan, Sebahattin; Turktas, Mine; İpek, Arif; Demirtas, Ibrahim; Okay, Sezer; Unver, Turgay

    2016-01-01

    Alkaloids are diverse group of secondary metabolites generally found in plants. Opium poppy (Papaver somniferum L.), the only commercial source of morphinan alkaloids, has been used as a medicinal plant since ancient times. It produces benzylisoquinoline alkaloids (BIA) including the narcotic analgesic morphine, the muscle relaxant papaverine, and the anti-cancer agent noscapine. Though BIAs play crucial roles in many biological mechanisms their steps in biosynthesis and the responsible genes remain to be revealed. In this study, expressions of 3-hydroxy-N-methylcoclaurine 4'-methyltransferase (4'OMT) and reticuline 7-O-methyltransferase (7OMT) genes were subjected to manipulation to functionally characterize their roles in BIA biosynthesis. Measurements of alkaloid accumulation were performed in leaf, stem, and capsule tissues accordingly. Suppression of 4'OMT expression caused reduction in the total alkaloid content in stem tissue whereas total alkaloid content was significantly induced in the capsule. Silencing of the 7OMT gene also caused repression in total alkaloid content in the stem. On the other hand, over-expression of 4'OMT and 7OMT resulted in higher morphine accumulation in the stem but suppressed amount in the capsule. Moreover, differential expression in several BIA synthesis genes (CNMT, TYDC, 6OMT, SAT, COR, 4'OMT, and 7OMT) were observed upon manipulation of 4'OMT and 7OMT expression. Upon silencing and overexpression applications, tissue specific effects of these genes were identified. Manipulation of 4'OMT and 7OMT genes caused differentiated accumulation of BIAs including morphine and noscapine in capsule and stem tissues. PMID:26909086

  4. Light microscopy can reveal the consumption of a mixture of psychotropic plant and fungal material in suspicious death.

    PubMed

    Wiltshire, Patricia E J; Hawksworth, David L; Edwards, Kevin J

    2015-08-01

    Light microscopical examination of plant and fungal remains in the post mortem gut may be capable of demonstrating the ingestion of unexpected natural psychotropic materials. This is demonstrated here in a case in which a 'shaman' was accused of causing the death of a young man. The deceased had participated in a ceremony which involved the drinking of ayahuasca in order to induce a psychotropic experience. Ayahuasca is an infusion of Banisteriopsis caapi (ayahuasca vine), which produces a monoamine oxidase inhibitor, and one or more additional tropical plants, generally Psychotria viridis (chacruna) which produces dimethyltryptamine (DMT). The monoamine oxidase inhibitor prevents DMT from being broken down in the gut, so enabling its passage into the bloodstream and across the blood/brain barrier. Toxicological tests for DMT demonstrated the presence of this compound in the body. The deceased was reported to be in the habit of using Psilocybe semilanceata (liberty cap). This fungus (popularly called magic mushroom) contains psilocybin which is hydrolysed in the gut to psilocin; this compound mimics a serotonin uptake inhibitor, and also invokes psychotropic experiences. Microscopical examination established that the ileum and colon contained spores of Psilocybe and, in addition, pollen of Cannabis sativa and seeds of Papaver cf. somniferum (opium poppy). Both the plant species yield psychotropic substances. Palynological and mycological analysis of containers from the deceased person's dwelling also yielded abundant trace evidence of pertinent pollen and spores. The police had requested analysis for DMT but there was no screening for other psychotropic substances. Investigators were surprised that a mixture of hallucinogenic materials had been consumed by the deceased. The charge was modified from manslaughter to possession of a 'Class A' drug as the deceased had been consuming psychotropic substances not administered by the 'shaman'. Where death involving drugs

  5. Assortment of the plants in the Medieval diet in Czech countries (based on archaeobotanical finds).

    PubMed

    Culíková, V

    2000-01-01

    The finds of remains of useful plants in archaeological objects were drawing attention already in the last century. However, it was not until after the World War II, in relation with renovation of historical cores of the towns, when a separate branch of the botany--archaeobotany constituted. In the Czech Republic it was E. Opravil who was the first to become engaged in a systematic archaeobotanical research at the beginning of the 60's. All the vegetal remains--fruits and seeds on the first place as well as pollen grains from prehistoric times to the post-medieval period, are the subjects of archaeobotany. The medieval waste pits are the richest sources of the vegetal remains. They contain a lot of diasporas of cultural as well as wild plants. The remains of useful plants are an authentic document of the vegetal component of diet. Even though some vegetal species, such as for example garlic, onion or stalk-plants, about whose cultivation we are informed from literature, occur only seldom, there appeared also the species not mentioned in literature. All the basic cereals have been known in our country already since the early Middle Ages. Wheat (Triticum aestivum) and rye (Secale cereale), as well as millet (Panicum miliaceum) that, along with buckwheat (Fagopyrum esculentum), were replaced by potatoes in the modern times were the most important. Also leguminous plants were of great importance in the diet even though their charred seeds seldom preserved. Remains of oil plants such as hemp (Cannabis sativa), flax (Linum usitatissimum) and poppy (Papaver somniferum) preserved well. It was successfully proved that cucumber (Cucumis sativus), among other numerous kinds of vegetables, was introduced by the Slavs already in the 8th century in Moravia and in the 9th century in Bohemia. The assortment of fruit was rich--especially the assortment of cultivated fruit. Among the imported kinds of fruit the first place occupied figs (Ficus carica), from the numerous finds it can

  6. Assortment of the plants in the Medieval diet in Czech countries (based on archaeobotanical finds).

    PubMed

    Culíková, V

    2000-01-01

    The finds of remains of useful plants in archaeological objects were drawing attention already in the last century. However, it was not until after the World War II, in relation with renovation of historical cores of the towns, when a separate branch of the botany--archaeobotany constituted. In the Czech Republic it was E. Opravil who was the first to become engaged in a systematic archaeobotanical research at the beginning of the 60's. All the vegetal remains--fruits and seeds on the first place as well as pollen grains from prehistoric times to the post-medieval period, are the subjects of archaeobotany. The medieval waste pits are the richest sources of the vegetal remains. They contain a lot of diasporas of cultural as well as wild plants. The remains of useful plants are an authentic document of the vegetal component of diet. Even though some vegetal species, such as for example garlic, onion or stalk-plants, about whose cultivation we are informed from literature, occur only seldom, there appeared also the species not mentioned in literature. All the basic cereals have been known in our country already since the early Middle Ages. Wheat (Triticum aestivum) and rye (Secale cereale), as well as millet (Panicum miliaceum) that, along with buckwheat (Fagopyrum esculentum), were replaced by potatoes in the modern times were the most important. Also leguminous plants were of great importance in the diet even though their charred seeds seldom preserved. Remains of oil plants such as hemp (Cannabis sativa), flax (Linum usitatissimum) and poppy (Papaver somniferum) preserved well. It was successfully proved that cucumber (Cucumis sativus), among other numerous kinds of vegetables, was introduced by the Slavs already in the 8th century in Moravia and in the 9th century in Bohemia. The assortment of fruit was rich--especially the assortment of cultivated fruit. Among the imported kinds of fruit the first place occupied figs (Ficus carica), from the numerous finds it can

  7. Light microscopy can reveal the consumption of a mixture of psychotropic plant and fungal material in suspicious death.

    PubMed

    Wiltshire, Patricia E J; Hawksworth, David L; Edwards, Kevin J

    2015-08-01

    Light microscopical examination of plant and fungal remains in the post mortem gut may be capable of demonstrating the ingestion of unexpected natural psychotropic materials. This is demonstrated here in a case in which a 'shaman' was accused of causing the death of a young man. The deceased had participated in a ceremony which involved the drinking of ayahuasca in order to induce a psychotropic experience. Ayahuasca is an infusion of Banisteriopsis caapi (ayahuasca vine), which produces a monoamine oxidase inhibitor, and one or more additional tropical plants, generally Psychotria viridis (chacruna) which produces dimethyltryptamine (DMT). The monoamine oxidase inhibitor prevents DMT from being broken down in the gut, so enabling its passage into the bloodstream and across the blood/brain barrier. Toxicological tests for DMT demonstrated the presence of this compound in the body. The deceased was reported to be in the habit of using Psilocybe semilanceata (liberty cap). This fungus (popularly called magic mushroom) contains psilocybin which is hydrolysed in the gut to psilocin; this compound mimics a serotonin uptake inhibitor, and also invokes psychotropic experiences. Microscopical examination established that the ileum and colon contained spores of Psilocybe and, in addition, pollen of Cannabis sativa and seeds of Papaver cf. somniferum (opium poppy). Both the plant species yield psychotropic substances. Palynological and mycological analysis of containers from the deceased person's dwelling also yielded abundant trace evidence of pertinent pollen and spores. The police had requested analysis for DMT but there was no screening for other psychotropic substances. Investigators were surprised that a mixture of hallucinogenic materials had been consumed by the deceased. The charge was modified from manslaughter to possession of a 'Class A' drug as the deceased had been consuming psychotropic substances not administered by the 'shaman'. Where death involving drugs

  8. Cutting Down the Tall Poppies: Horizontal Violence.

    ERIC Educational Resources Information Center

    Funk, Carole

    Many women in educational leadership positions experience negative treatment from female teachers and female superintendents. This phenomenon is known as horizontal violence, "the curious behavior of members of oppressed groups who often lash out at their peers in response to oppression instead of attacking their oppressors." This paper explores…

  9. Small Poppies: Highly Gifted Children in the Early Years.

    ERIC Educational Resources Information Center

    Gross, Miraca U. M.

    1999-01-01

    Discusses the underidentification of young highly-gifted children and describes the developmental differences in highly-gifted children. Issues in the identification of young highly-gifted children are addressed, along with suggestions for teacher nomination, parent nomination, ability and achievement testing, and appropriate placements of…

  10. Plant remains from an Early Iron Age well at Hajndl, Slovenia.

    PubMed

    Sostarić, Renata; Alegro, Antun; Hrsak, Vladimir; Stancić, Zvjezdana; Küster, Hansjörg

    2009-12-01

    The archaeobotanical samples analysed derive from a well-preserved well dated to the Early Iron Age (Hallstatt) according to pottery found in the sampled layers and on a radiocarbon dating (720-520 cal B.C.) of the wooden construction of the well. Cultivated plants (Panicum miliaceum, Linum usitatissimum, Papaver somniferum and Camelina sativa) were recorded in relatively small numbers whereas primary cereals are lacking. Together with quite a large number of accompanied weeds they suggest agriculture activities, but a mixture of weeds and ruderal plants was probably also growing inside the relatively large settlement complex and could have been included in the assemblages just by chance. The plant species composition indicates local vegetation developed under strong anthropogenic influence and on mainly moist and nitrogen-rich soils. Grassland plants and an almost complete lacking of tree and shrub species characteristic for the climax vegetation (deciduous mesophilous mixed forests) of the region also indicate at least local human activity.

  11. Unprecedented Utilization of Pelargonidin and Indole for the Biosynthesis of Plant Indole Alkaloids

    PubMed Central

    Warskulat, Anne‐Christin; Tatsis, Evangelos C.; Dudek, Bettina; Kai, Marco; Lorenz, Sybille

    2016-01-01

    Abstract Nudicaulins are a group of indole alkaloid glycosides responsible for the color of yellow petals of Papaver nudicaule (Iceland poppy). The unique aglycone scaffold of these alkaloids attracted our interest as one of the most unusual flavonoid‐indole hybrid structures that occur in nature. Stable isotope labeling experiments with sliced petals identified free indole, but not tryptamine or l‐tryptophan, as one of the two key biosynthetic precursors of the nudicaulin aglycone. Pelargonidin was identified as the second key precursor, contributing the polyphenolic unit to the nudicaulin molecule. This finding was inferred from the temporary accumulation of pelargonidin glycosides in the petals during flower bud development and a drop at the point in time when nudicaulin levels start to increase. The precursor‐directed incorporation of cyanidin into a new 3′‐hydroxynudicaulin strongly supports the hypothesis that anthocyanins are involved in the biosynthesis of nudicaulins. PMID:26670055

  12. Unprecedented Utilization of Pelargonidin and Indole for the Biosynthesis of Plant Indole Alkaloids.

    PubMed

    Warskulat, Anne-Christin; Tatsis, Evangelos C; Dudek, Bettina; Kai, Marco; Lorenz, Sybille; Schneider, Bernd

    2016-02-15

    Nudicaulins are a group of indole alkaloid glycosides responsible for the color of yellow petals of Papaver nudicaule (Iceland poppy). The unique aglycone scaffold of these alkaloids attracted our interest as one of the most unusual flavonoid-indole hybrid structures that occur in nature. Stable isotope labeling experiments with sliced petals identified free indole, but not tryptamine or l-tryptophan, as one of the two key biosynthetic precursors of the nudicaulin aglycone. Pelargonidin was identified as the second key precursor, contributing the polyphenolic unit to the nudicaulin molecule. This finding was inferred from the temporary accumulation of pelargonidin glycosides in the petals during flower bud development and a drop at the point in time when nudicaulin levels start to increase. The precursor-directed incorporation of cyanidin into a new 3'-hydroxynudicaulin strongly supports the hypothesis that anthocyanins are involved in the biosynthesis of nudicaulins. PMID:26670055

  13. The In vitro anti-acne activity of two unani drugs

    PubMed Central

    Chaudhary, Shahid Shah; Tariq, Mohd.; Zaman, Roohi; Imtiyaz, Shaikh

    2013-01-01

    Background: Acne is the most common disorder treated by dermatologists. As many as 80-90% of all adolescents have some type of acne and 30% of them require medical treatment. It is an inflammatory disease of the pilosebaceous unit characterized by the formation of open and closed comedones, papules, pustules, nodules, and cysts. Aims: The present study was conducted to investigate the in vitro anti-acne activity of two Unani single drugs Darchini (Cinnamomum zeylanicum Bl.) and Tukhm Khashkhash (Papaver somniferum L. seeds). Materials and Methods: The antibacterial activity of aqueous, ethanolic and hydroalcoholic extracts of both drugs were investigated against two acne causing bacteria, i.e., Propionibacterium acne and Staphylococcus epidermidis using well diffusion method. Results: The result showed that both drugs were active against the two bacteria. Against P. acne aqueous and ethanolic extract of Darchini and Tukhm Khashkhash showed the zone of inhibition of 18 ± 1.02 mm and 18 ± 1.6 mm and 13 ± 1.04 mm and 14 ± 1.8 mm, respectively. Against S. epidermidis aqueous, hydroalcoholic and ethanolic extracts of Darchini showed 22 ± 1.7 mm, 22 ± 1.2 mm and 15 ± 1.8 mm zone of inhibition respectively. Hydroalcoholic and ethanolic extracts of Tukhm Khashkhash showed 15 ± 1.09 mm and 13 ± 1.6 mm zone of inhibition respectively. Conclusion: This suggests that C. zeylanicum and P. somniferum have potential against acne causing bacteria and hence they can be used in topical anti-acne preparations and may address the antibiotic resistance of the bacteria. PMID:25161328

  14. Identification of a unique 2-oxoglutarate-dependent flavone 7-O-demethylase completes the elucidation of the lipophilic flavone network in basil.

    PubMed

    Berim, Anna; Kim, Min-Jeong; Gang, David R

    2015-01-01

    Small molecule demethylation is considered unusual in plants. Of the studied instances, the N-demethylation of nicotine is catalyzed by a Cyt P450 monooxygenase, while the O-dealkylation of alkaloids in Papaver somniferum is mediated by 2-oxoglutarate-dependent dioxygenases (2-ODDs). This report describes a 2-ODD regiospecifically catalyzing the 7-O-demethylation of methoxylated flavones in peltate trichomes of sweet basil (Ocimum basilicum L.). Three candidate 2-ODDs were identified in the basil trichome transcriptome database. Only the candidate designated ObF7ODM1 was found to be active with and highly specific for the proposed natural substrates, gardenin B and 8-hydroxysalvigenin. Of the characterized 2-ODDs, ObF7ODM1 is most closely related to O-demethylases from Papaver. The demethylase activity in trichomes from four basil chemotypes matches well with the abundance of ObF7ODM1 peptides and transcripts in the same trichome preparations. Treatment of basil plants with a 2-ODD inhibitor prohexadione-calcium significantly reduced the accumulation of 7-O-demethylated flavone nevadensin, confirming the involvement of a 2-ODD in its formation. Notably, the full-length open reading frame of ObF7ODM1 contains a second in-frame AUG codon 57 nucleotides downstream of the first translation initiation codon. Both AUG codons are recognized by bacterial translation machinery during heterologous gene expression. The N-truncated ObF7ODM1 is nearly inactive. The N-terminus essential for activity is unique to ObF7ODM1 and does not align with the sequences of other 2-ODDs. Further studies will reveal whether alternative translation initiation plays a role in regulating the O-demethylase activity in planta. Molecular identification of the flavone 7-O-demethylase completes the biochemical elucidation of the lipophilic flavone network in basil.

  15. Identification of a unique 2-oxoglutarate-dependent flavone 7-O-demethylase completes the elucidation of the lipophilic flavone network in basil.

    PubMed

    Berim, Anna; Kim, Min-Jeong; Gang, David R

    2015-01-01

    Small molecule demethylation is considered unusual in plants. Of the studied instances, the N-demethylation of nicotine is catalyzed by a Cyt P450 monooxygenase, while the O-dealkylation of alkaloids in Papaver somniferum is mediated by 2-oxoglutarate-dependent dioxygenases (2-ODDs). This report describes a 2-ODD regiospecifically catalyzing the 7-O-demethylation of methoxylated flavones in peltate trichomes of sweet basil (Ocimum basilicum L.). Three candidate 2-ODDs were identified in the basil trichome transcriptome database. Only the candidate designated ObF7ODM1 was found to be active with and highly specific for the proposed natural substrates, gardenin B and 8-hydroxysalvigenin. Of the characterized 2-ODDs, ObF7ODM1 is most closely related to O-demethylases from Papaver. The demethylase activity in trichomes from four basil chemotypes matches well with the abundance of ObF7ODM1 peptides and transcripts in the same trichome preparations. Treatment of basil plants with a 2-ODD inhibitor prohexadione-calcium significantly reduced the accumulation of 7-O-demethylated flavone nevadensin, confirming the involvement of a 2-ODD in its formation. Notably, the full-length open reading frame of ObF7ODM1 contains a second in-frame AUG codon 57 nucleotides downstream of the first translation initiation codon. Both AUG codons are recognized by bacterial translation machinery during heterologous gene expression. The N-truncated ObF7ODM1 is nearly inactive. The N-terminus essential for activity is unique to ObF7ODM1 and does not align with the sequences of other 2-ODDs. Further studies will reveal whether alternative translation initiation plays a role in regulating the O-demethylase activity in planta. Molecular identification of the flavone 7-O-demethylase completes the biochemical elucidation of the lipophilic flavone network in basil. PMID:25378691

  16. Pain and Poppies: The Good, the Bad, and the Ugly of Opioid Analgesics

    PubMed Central

    Al-Hasani, Ream; Salvemini, Daniela; Salter, Michael W.; Gutstein, Howard

    2015-01-01

    Treating pain is one of the most difficult challenges in medicine and a key facet of disease management. The isolation of morphine by Friedrich Sertürner in 1804 added an essential pharmacological tool in the treatment of pain and spawned the discovery of a new class of drugs known collectively as opioid analgesics. Revered for their potent pain-relieving effects, even Morpheus the god of dreams could not have dreamt that his opium tincture would be both a gift and a burden to humankind. To date, morphine and other opioids remain essential analgesics for alleviating pain. However, their use is plagued by major side effects, such as analgesic tolerance (diminished pain-relieving effects), hyperalgesia (increased pain sensitivity), and drug dependence. This review highlights recent advances in understanding the key causes of these adverse effects and explores the effect of chronic pain on opioid reward. SIGNIFICANCE STATEMENT Chronic pain is pervasive and afflicts >100 million Americans. Treating pain in these individuals is notoriously difficult and often requires opioids, one of the most powerful and effective classes of drugs used for controlling pain. However, their use is plagued by major side effects, such as a loss of pain-relieving effects (analgesic tolerance), paradoxical pain (hyperalgesia), and addiction. Despite the potential side effects, opioids remain the pharmacological cornerstone of modern pain therapy. This review highlights recent breakthroughs in understanding the key causes of these adverse effects and explores the cellular control of opioid systems in reward and aversion. The findings will challenge traditional views of the good, the bad, and the ugly of opioids. PMID:26468188

  17. Modulation of CYPs, P-gp, and PXR by Eschscholzia californica (California poppy) and its alkaloids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Eschscholzia californica Cham., a native US plant, is traditionally used as a sedative, analgesic and anxiolytic herb. With the rapid rise in the use of herbal supplements together with over the counter (OTC) and prescription drugs, the risk for potential herb-drug interactions is also increasing. M...

  18. Modulation of CYPs, P-gp, and PXR by Eschscholzia californica (California Poppy) and Its Alkaloids.

    PubMed

    Manda, Vamshi K; Ibrahim, Mohamed A; Dale, Olivia R; Kumarihamy, Mallika; Cutler, Stephen J; Khan, Ikhlas A; Walker, Larry A; Muhammad, Ilias; Khan, Shabana I

    2016-04-01

    Eschscholzia californica, a native US plant, is traditionally used as a sedative, analgesic, and anxiolytic herb. With the rapid rise in the use of herbal supplements together with over-the-counter and prescription drugs, the risk for potential herb-drug interactions is also increasing. Most of the clinically relevant pharmacokinetic drug interactions occur due to modulation of cytochrome P450 enzymes (CYPs), P-glycoprotein, and the pregnane X receptor by concomitantly used herbs. This study aimed to determine the effects of an EtOH extract, aqueous extract (tea), basic CHCl3 fractions, and isolated major alkaloids, namely protopine (1), escholtzine (2), allocryptopine (3), and californidine (4), of E. californica on the activity of cytochrome P450s, P-glycoprotein and the pregnane X receptor. The EtOH extract and fractions showed strong time-dependent inhibition of CYP 3A4, CYP 2C9, and CYP 2C19, and reversible inhibition of CYP 2D6. Among the alkaloids, escholtzine (2) and allocryptopine (3) exhibited time-dependent inhibition of CYP 3A4, CYP 2C9, and CYP 2C19 (IC50 shift ratio > 2), while protopine (1) and allocryptopine (3) showed reversible inhibition of CYP 2D6 enzyme. A significant activation of the pregnane X receptor (> 2-fold) was observed with the EtOH extract, basic CHCl3 fraction, and alkaloids (except protopine), which resulted into an increased expression of mRNA and the activity of CYP 3A4 and CYP 1A2. The expression of P-glycoprotein was unaffected. However, aqueous extract (tea) and its main alkaloid californidine (4) did not affect cytochrome P450s, P-glycoprotein, or the pregnane X receptor. This data suggests that EtOH extract of E. californica and its major alkaloids have a potential of causing interactions with drugs that are metabolized by cytochrome P450s, while the tea seems to be safer. PMID:27054913

  19. Laūq: A Sustained-Release Dosage Form for Respiratory Disorders in Traditional Persian Medicine.

    PubMed

    Karegar-Borzi, Hossein; Salehi, Mehdi; Rahimi, Roja

    2016-01-01

    Laūq is a pharmaceutical dosage form that had been mainly used for the treatment of various respiratory disorders in traditional Persian medicine. It is important from 2 aspects: a dosage form with efficient and optimum delivery of drugs to the respiratory tract and biological effects of its ingredients. Natural medicine in laūq has been demonstrated to act in respiratory disorders by their antitussive, antiallergic, anti-inflammatory, antioxidant, spasmolytic, and antibacterial activities. Some of these natural remedies act by most of the mentioned mechanisms such as Cydonia oblonga, Glycyrrhiza glabra, Crocus sativus, Hyssopus officinalis, Foeniculum vulgare, and honey. However, the evidence is limited including Cassia fistula, Papaver somniferum, and Drimia maritima. According to positive pharmacokinetic and pharmacodynamic aspects of laūqs, they may be considered as efficient dosage forms for delivery of drugs to the respiratory tract. For better compatibility of patients, it could be substituted laūqs with newer drug delivery systems like lozenges.

  20. The Noscapine Chronicle: A Pharmaco-Historic Biography of the Opiate Alkaloid Family and its Clinical Applications

    PubMed Central

    Rida, Padmashree C. G.; LiVecche, Dillon; Ogden, Angela; Zhou, Jun; Aneja, Ritu

    2016-01-01

    Given its manifold potential therapeutic applications and amenability to modification, noscapine is a veritable “Renaissance drug” worthy of commemoration. Perhaps the only facet of noscapine’s profile more astounding than its versatility is its virtual lack of side effects and addictive properties, which distinguishes it from other denizens of Papaver somniferum. This review intimately chronicles the rich intellectual and pharmacological history behind the noscapine family of compounds, the length of whose arms was revealed over decades of patient scholarship and experimentation. We discuss the intriguing story of this family of nontoxic alkaloids, from noscapine’s purification from opium at the turn of the 19th century in Paris to the recent torrent of rationally designed analogs with tremendous anticancer potential. In between, noscapine’s unique pharmacology; impact on cellular signaling pathways, the mitotic spindle, and centrosome clustering; use as an antimalarial drug and cough suppressant; and exceptional potential as a treatment for polycystic ovarian syndrome, strokes, and diverse malignancies are catalogued. Seminal experiments involving some of its more promising analogs, such as amino-noscapine, 9-nitronoscapine, 9-bromonoscapine, and reduced bromonoscapine, are also detailed. Finally, the bright future of these oftentimes even more exceptional derivatives is described, rounding out a portrait of a truly remarkable family of compounds. PMID:26179481

  1. Progress Toward the Development of Noscapine and Derivatives as Anticancer Agents.

    PubMed

    DeBono, Aaron; Capuano, Ben; Scammells, Peter J

    2015-08-13

    Many nitrogen-moiety containing alkaloids derived from plant origins are bioactive and play a significant role in human health and emerging medicine. Noscapine, a phthalideisoquinoline alkaloid derived from Papaver somniferum, has been used as a cough suppressant since the mid 1950s, illustrating a good safety profile. Noscapine has since been discovered to arrest cells at mitosis, albeit with moderately weak activity. Immunofluorescence staining of microtubules after 24 h of noscapine exposure at 20 μM elucidated chromosomal abnormalities and the inability of chromosomes to complete congression to the equatorial plane for proper mitotic separation ( Proc. Natl. Acad. Sci. U. S. A. 1998 , 95 , 1601 - 1606 ). A number of noscapine analogues possessing various modifications have been described within the literature and have shown significantly improved antiprolific profiles for a large variety of cancer cell lines. Several semisynthetic antimitotic alkaloids are emerging as possible candidates as novel anticancer therapies. This perspective discusses the advancing understanding of noscapine and related analogues in the fight against malignant disease. PMID:25811651

  2. Laūq: A Sustained-Release Dosage Form for Respiratory Disorders in Traditional Persian Medicine.

    PubMed

    Karegar-Borzi, Hossein; Salehi, Mehdi; Rahimi, Roja

    2016-01-01

    Laūq is a pharmaceutical dosage form that had been mainly used for the treatment of various respiratory disorders in traditional Persian medicine. It is important from 2 aspects: a dosage form with efficient and optimum delivery of drugs to the respiratory tract and biological effects of its ingredients. Natural medicine in laūq has been demonstrated to act in respiratory disorders by their antitussive, antiallergic, anti-inflammatory, antioxidant, spasmolytic, and antibacterial activities. Some of these natural remedies act by most of the mentioned mechanisms such as Cydonia oblonga, Glycyrrhiza glabra, Crocus sativus, Hyssopus officinalis, Foeniculum vulgare, and honey. However, the evidence is limited including Cassia fistula, Papaver somniferum, and Drimia maritima. According to positive pharmacokinetic and pharmacodynamic aspects of laūqs, they may be considered as efficient dosage forms for delivery of drugs to the respiratory tract. For better compatibility of patients, it could be substituted laūqs with newer drug delivery systems like lozenges. PMID:26008751

  3. The Noscapine Chronicle: A Pharmaco-Historic Biography of the Opiate Alkaloid Family and its Clinical Applications.

    PubMed

    Rida, Padmashree C G; LiVecche, Dillon; Ogden, Angela; Zhou, Jun; Aneja, Ritu

    2015-09-01

    Given its manifold potential therapeutic applications and amenability to modification, noscapine is a veritable "Renaissance drug" worthy of commemoration. Perhaps the only facet of noscapine's profile more astounding than its versatility is its virtual lack of side effects and addictive properties, which distinguishes it from other denizens of Papaver somniferum. This review intimately chronicles the rich intellectual and pharmacological history behind the noscapine family of compounds, the length of whose arms was revealed over decades of patient scholarship and experimentation. We discuss the intriguing story of this family of nontoxic alkaloids, from noscapine's purification from opium at the turn of the 19th century in Paris to the recent torrent of rationally designed analogs with tremendous anticancer potential. In between, noscapine's unique pharmacology; impact on cellular signaling pathways, the mitotic spindle, and centrosome clustering; use as an antimalarial drug and cough suppressant; and exceptional potential as a treatment for polycystic ovarian syndrome, strokes, and diverse malignancies are catalogued. Seminal experiments involving some of its more promising analogs, such as amino-noscapine, 9-nitronoscapine, 9-bromonoscapine, and reduced bromonoscapine, are also detailed. Finally, the bright future of these oftentimes even more exceptional derivatives is described, rounding out a portrait of a truly remarkable family of compounds. PMID:26179481

  4. Synthesis of Morphinan Alkaloids in Saccharomyces cerevisiae

    PubMed Central

    Fossati, Elena; Narcross, Lauren; Ekins, Andrew; Falgueyret, Jean-Pierre; Martin, Vincent J. J.

    2015-01-01

    Morphinan alkaloids are the most powerful narcotic analgesics currently used to treat moderate to severe and chronic pain. The feasibility of morphinan synthesis in recombinant Saccharomyces cerevisiae starting from the precursor (R,S)-norlaudanosoline was investigated. Chiral analysis of the reticuline produced by the expression of opium poppy methyltransferases showed strict enantioselectivity for (S)-reticuline starting from (R,S)-norlaudanosoline. In addition, the P. somniferum enzymes salutaridine synthase (PsSAS), salutaridine reductase (PsSAR) and salutaridinol acetyltransferase (PsSAT) were functionally co-expressed in S. cerevisiae and optimization of the pH conditions allowed for productive spontaneous rearrangement of salutaridinol-7-O-acetate and synthesis of thebaine from (R)-reticuline. Finally, we reconstituted a 7-gene pathway for the production of codeine and morphine from (R)-reticuline. Yeast cell feeding assays using (R)-reticuline, salutaridine or codeine as substrates showed that all enzymes were functionally co-expressed in yeast and that activity of salutaridine reductase and codeine-O-demethylase likely limit flux to morphine synthesis. The results of this study describe a significant advance for the synthesis of morphinans in S. cerevisiae and pave the way for their complete synthesis in recombinant microbes. PMID:25905794

  5. PreBotzinger complex neurokinin-1 receptor-expressing neurons mediate opioid-induced respiratory depression.

    PubMed

    Montandon, Gaspard; Qin, Wuxuan; Liu, Hattie; Ren, Jun; Greer, John J; Horner, Richard L

    2011-01-26

    The analgesic properties of the opium poppy Papever somniferum were first mentioned by Hippocrates around 400 BC, and opioid analgesics remain the mainstay of pain management today. These drugs can cause the serious side-effect of respiratory depression that can be lethal with overdose, however the critical brain sites and neurochemical identity of the neurons mediating this depression are unknown. By locally manipulating neurotransmission in the adult rat, we identify the critical site of the medulla, the preBötzinger complex, that mediates opioid-induced respiratory depression in vivo. Here we show that opioids at the preBötzinger complex cause respiratory depression or fatal apnea, with anesthesia and deep-sleep being particularly vulnerable states for opioid-induced respiratory depression. Importantly, we establish that the preBötzinger complex is fully responsible for respiratory rate suppression following systemic administration of opioid analgesics. The site in the medulla most sensitive to opioids corresponds to a region expressing neurokinin-1 receptors, and we show in rhythmically active brainstem section in vitro that neurokinin-1 receptor-expressing preBötzinger complex neurons are selectively inhibited by opioids. In summary, neurokinin-1 receptor-expressing preBötzinger complex neurons constitute the critical site mediating opioid-induced respiratory rate depression, and the key therapeutic target for its prevention or reversal.

  6. Synthesis of Morphinan Alkaloids in Saccharomyces cerevisiae.

    PubMed

    Fossati, Elena; Narcross, Lauren; Ekins, Andrew; Falgueyret, Jean-Pierre; Martin, Vincent J J

    2015-01-01

    Morphinan alkaloids are the most powerful narcotic analgesics currently used to treat moderate to severe and chronic pain. The feasibility of morphinan synthesis in recombinant Saccharomyces cerevisiae starting from the precursor (R,S)-norlaudanosoline was investigated. Chiral analysis of the reticuline produced by the expression of opium poppy methyltransferases showed strict enantioselectivity for (S)-reticuline starting from (R,S)-norlaudanosoline. In addition, the P. somniferum enzymes salutaridine synthase (PsSAS), salutaridine reductase (PsSAR) and salutaridinol acetyltransferase (PsSAT) were functionally co-expressed in S. cerevisiae and optimization of the pH conditions allowed for productive spontaneous rearrangement of salutaridinol-7-O-acetate and synthesis of thebaine from (R)-reticuline. Finally, we reconstituted a 7-gene pathway for the production of codeine and morphine from (R)-reticuline. Yeast cell feeding assays using (R)-reticuline, salutaridine or codeine as substrates showed that all enzymes were functionally co-expressed in yeast and that activity of salutaridine reductase and codeine-O-demethylase likely limit flux to morphine synthesis. The results of this study describe a significant advance for the synthesis of morphinans in S. cerevisiae and pave the way for their complete synthesis in recombinant microbes. PMID:25905794

  7. 75 FR 34114 - Pesticide Products; Registration Applications

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-16

    ..., oil radish, poppy seed, rapeseed, sesame, sweet rocket, calendula, castor oil plant, Chinese... pleasure, hare's ear mustard, lesquerella, lunaria, meadowfoam, milkweed, mustard seed, oil radish, poppy... mustard, lesquerella, lunaria, meadowfoam, milkweed, mustard seed, oil radish, poppy seed,...

  8. Pollen foraging: learning a complex motor skill by bumblebees (Bombus terrestris)

    NASA Astrophysics Data System (ADS)

    Raine, Nigel E.; Chittka, Lars

    2007-06-01

    To investigate how bumblebees (Bombus terrestris) learn the complex motor skills involved in pollen foraging, we observed naïve workers foraging on arrays of nectarless poppy flowers (Papaver rhoeas) in a greenhouse. Foraging skills were quantified by measuring the pollen load collected during each foraging bout and relating this to the number of flowers visited and bout duration on two consecutive days. The pollen standing crop (PSC) in each flower decreased drastically from 0530 to 0900 hours. Therefore, we related foraging performance to the changing levels of pollen available (per flower) and found that collection rate increased over the course of four consecutive foraging bouts (comprising between 277 and 354 individual flower visits), suggesting that learning to forage for pollen represents a substantial time investment for individual foragers. The pollen collection rate and size of pollen loads collected at the start of day 2 were markedly lower than at the end of day 1, suggesting that components of pollen foraging behaviour could be subject to imperfect overnight retention. Our results suggest that learning the necessary motor skills to collect pollen effectively from morphologically simple flowers takes three times as many visits as learning how to handle the most morphologically complex flowers to extract nectar, potentially explaining why bees are more specialised in their choice of pollen flowers.

  9. The rules of drug taking: wine and poppy derivatives in the ancient world. V. Sobriety or postponement of drunkenness?

    PubMed

    Nencini, P

    1997-04-01

    In spite of declared intentions to maintain control of drinking, we have cues that during the symposium enough wine was ingested to get intoxicated. Hence, it was widely debated how much wine could be safety drunk, and the conclusion was that it was a matter of individual endurance. Apparently, elders were less prone to wine-induced loss of rationality and control. In conclusion, the Ancients were aware of potential dangers of wine and that its consumption needed rules, but apparently these rules existed for being broken.

  10. The rules of drug taking: wine and poppy derivatives in the ancient world. IV. The rules of temperance.

    PubMed

    Nencini, P

    1997-03-01

    Drinking in moderation was considered fully appropriate for Greek citizens and, accordingly, wine played a central role in the symposium, where adult men met to enjoy poems, music, conversation, and sex. To reach the goal of remaining "euphoric" without getting intoxicated, wine was drunk diluted. Usually, three parts of water were added to one part of wine. Since drinking diluted wine produces a slow rise of alcoholemia, the pharmacological rationale of this habit seems to stand on two factors: 1) feeling relaxed and euphoric are mainly perceived during the rise of alcoholemia: 2) alcohol intake can be titrated as long as intoxication is not reached.

  11. Late Neolithic vegetation history at the pile-dwelling site of Palù di Livenza (northeastern Italy

    NASA Astrophysics Data System (ADS)

    Pini, Roberta

    2004-12-01

    The Late Neolithic pile-dwelling of Palù di Livenza yielded archaeological remains typical of the Square Mouth Pottery and Lagozza Cultures. A palynological investigation reveals important changes in the vegetation due to anthropogenic pressure. Between ca. 6590 and 5960 cal. yr BP, dense oak wood forests with deciduous Quercus, Fagus and Corylus extended around the mire, with no signs of human impact. The establishment of the pile-dwelling, dated to ca. 5960 cal. yr BP, led to a strong reduction of forests, reclamation of wetlands, and expansion of herbaceous communities, with cultivated species, infestant weeds, nitrophilous and ruderal herbs, pastures and meadows. According to AMS dates and previous archaeological chronologies, the pile-dwelling persisted for about 700 years (from ca. 5960 to 5260 cal. yr BP). The history of the pile-dwelling after ca. 5260 cal. yr BP cannot be reconstructed because of recent contamination of the top part of the section. Rarefaction analysis was applied to estimate changes of palynological richness through time: the highest E(Tn) (between 56 and 69 taxa) are contemporaneous with the local development of the pile-dwelling. The comparison of pollen data with archaeobotanical evidence indicates that Fragaria vesca, Malus sylvestris, Papaver somniferum and Physalis alkekengi were gathered at some distance from the site and that Linum usitatissimum is strongly under-represented in pollen samples. Crop cultivation can be estimated for a radius of several hundred metres around the mire. Palù di Livenza is significant in the context of Neolithic archaeobotany of northern Italy and neighbouring countries. Copyright

  12. Virus-induced gene silencing (VIGS) in Cysticapnos vesicaria, a zygomorphic-flowered Papaveraceae (Ranunculales, basal eudicots)

    PubMed Central

    Hidalgo, Oriane; Bartholmes, Conny; Gleissberg, Stefan

    2012-01-01

    Background and Aims Studies of evolutionary diversification in the basal eudicot family Papaveraceae, such as the transition from actinomorphy to zygomorphy, are hampered by the lack of comparative functional studies. So far, gene silencing methods are only available in the actinomorphic species Eschscholzia californica and Papaver somniferum. This study addresses the amenability of Cysticapnos vesicaria, a derived fumitory with zygomorphic flowers, to virus-induced gene silencing (VIGS), and describes vegetative and reproductive traits in this species. Methods VIGS-mediated downregulation of the C. vesicaria PHYTOENE DESATURASE gene (CvPDS) and of the FLORICAULA gene CvFLO was carried out using Agrobacterium tumefaciens transfer of Tobacco rattle virus (TRV)-based vectors. Wild-type and vector-treated plants were characterized using reverse transcription–PCR (RT–PCR), in situ hybridization, and macroscopic and scanning electron microscopic imaging. Key Results Cysticapnos vesicaria germinates rapidly, can be grown at high density, has a short life cycle and is self-compatible. Inoculation of C. vesicaria with a CvPDS-VIGS vector resulted in strong photobleaching of green parts and reduction of endogenous CvPDS transcript levels. Gene silencing persisted during inflorescence development until fruit set. Inoculation of plants with CvFLO-VIGS affected floral phyllotaxis, symmetry and floral organ identities. Conclusions The high penetrance, severity and stability of pTRV-mediated silencing, including the induction of meristem-related phenotypes, make C. vesicaria a very promising new focus species for evolutionary–developmental (evo–devo) studies in the Papaveraceae. This now enables comparative studies of flower symmetry, inflorescence determinacy and other traits that diversified in the Papaveraceae. PMID:22307568

  13. Phylogenomic and structural analyses of 18 complete plastomes across nearly all families of early-diverging eudicots, including an angiosperm-wide analysis of IR gene content evolution.

    PubMed

    Sun, Yanxia; Moore, Michael J; Zhang, Shoujun; Soltis, Pamela S; Soltis, Douglas E; Zhao, Tingting; Meng, Aiping; Li, Xiaodong; Li, Jianqiang; Wang, Hengchang

    2016-03-01

    The grade of early-diverging eudicots includes five major lineages: Ranunculales, Trochodendrales, Buxales, Proteales and Sabiaceae. To examine the evolution of plastome structure in early-diverging eudicots, we determined the complete plastome sequences of eight previously unsequenced early-diverging eudicot taxa, Pachysandra terminalis (Buxaceae), Meliosma aff. cuneifolia (Sabiaceae), Sabia yunnanensis (Sabiaceae), Epimedium sagittatum (Berberidaceae), Euptelea pleiosperma (Eupteleaceae), Akebia trifoliata (Lardizabalaceae), Stephania japonica (Menispermaceae) and Papaver somniferum (Papaveraceae), and compared them to previously published plastomes of the early-diverging eudicots Buxus, Tetracentron, Trochodendron, Nelumbo, Platanus, Nandina, Megaleranthis, Ranunculus, Mahonia and Macadamia. All of the newly sequenced plastomes share the same 79 protein-coding genes, 4 rRNA genes, and 30 tRNA genes, except for that of Epimedium, in which infA is pseudogenized and clpP is highly divergent and possibly a pseudogene. The boundaries of the plastid Inverted Repeat (IR) were found to vary significantly across early-diverging eudicots; IRs ranged from 24.3 to 36.4kb in length and contained from 18 to 33 genes. Based on gene content, the IR was classified into six types, with shifts among types characterized by high levels of homoplasy. Reconstruction of ancestral IR gene content suggested that 18 genes were likely present in the IR region of the ancestor of eudicots. Maximum likelihood phylogenetic analysis of a 79-gene, 97-taxon data set that included all available early-diverging eudicots and representative sampling of remaining angiosperm diversity largely agreed with previous estimates of early-diverging eudicot relationships, but resolved Trochodendrales rather than Buxales as sister to Gunneridae, albeit with relatively weak bootstrap support, conflicting with what has been found for these three clades in most previous analyses. In addition, Proteales was

  14. Reproduction and pollination of the endangered dwarf bear-poppy Arctomecon humilis (Papaveraceae)across a quarter century: unraveling of a pollination web?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    ABSTRACT.—Arctomecon humilis, a rare gypsophile of the extreme northeastern Mojave Desert, is restricted to a few isolated populations in Washington County, Utah, USA. At several times in the past quarter century, we have studied the breeding system and reproductive success of this endangered specie...

  15. 21 CFR 1301.34 - Application for importation of Schedule I and II substances.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... will be permitted to import only: (i) Such amounts of crude opium, poppy straw, concentrate of poppy... as may be relevant to and consistent with the public health and safety. (c) In determining...

  16. 21 CFR 1301.34 - Application for importation of Schedule I and II substances.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... will be permitted to import only: (i) Such amounts of crude opium, poppy straw, concentrate of poppy... as may be relevant to and consistent with the public health and safety. (c) In determining...

  17. 21 CFR 1301.34 - Application for importation of Schedule I and II substances.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... will be permitted to import only: (i) Such amounts of crude opium, poppy straw, concentrate of poppy... as may be relevant to and consistent with the public health and safety. (c) In determining...

  18. 21 CFR 1301.34 - Application for importation of Schedule I and II substances.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... will be permitted to import only: (i) Such amounts of crude opium, poppy straw, concentrate of poppy... as may be relevant to and consistent with the public health and safety. (c) In determining...

  19. 21 CFR 1312.13 - Issuance of import permit.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ..., poppy straw and concentrate of poppy straw) having as its source: (1) Turkey, (2) India, (3) Spain, (4... narcotic raw material imported into the United States annually shall have as its source Spain,...

  20. 77 FR 30026 - Manufacturer of Controlled Substances Notice of Application, Ampac Fine Chemicals LLC.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-21

    ... Thebaine (9333) II Poppy Straw Concentrate (9670) II The company is a contract manufacturer. In reference to Poppy Straw Concentrate the company will manufacture Thebaine intermediates for sale to...

  1. 78 FR 46372 - Manufacturer of Controlled Substances; Notice of Application; Ampac Fine Chemicals, LLC.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-07-31

    ... Methylphenidate (1724) II Thebaine (9333) II Poppy Straw Concentrate (9670) II Tapentadol (9780) II The company is a contract manufacturer. In reference to Poppy Straw Concentrate the company will...

  2. 76 FR 32225 - Manufacturer Of Controlled Substances; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-03

    ... basic classes of controlled substances: Drug Schedule Thebaine (9333) II Poppy Straw Concentrate (9670) II The company is a contract manufacturer. In reference to Poppy Straw Concentrate the company...

  3. 75 FR 32506 - Importer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-08

    ... 20, 2009 and published in the Federal Register on October 28, 2009 (74 FR 55583), Cody Laboratories... listed in schedule II: Drug Schedule Raw Opium (9600) II Concentrate of Poppy Straw (9670) II The company... from raw opium, poppy straw, and concentrate of poppy straw. No comments or objections have...

  4. 77 FR 31389 - Importer of Controlled Substances; Notice of Registration; Penick Corporation

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-25

    ... Notice dated April 17, 2012 and published in the Federal Register on April 26, 2012, 77 FR 24983, Penick... applications to import narcotic raw material are not appropriate. 72 FR 3417 (2007) DEA has considered the... substances: Drug Schedule Coca Leaves (9040) II Raw Opium (9600) II Poppy Straw (9650) II Poppy...

  5. 78 FR 64014 - Importer of Controlled Substances; Notice of Application; Penick Corporation

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-25

    ... material are not appropriate. 72 FR 3417 (2007). As noted in a previous notice published in the Federal Register on September 23, 1975, 40 FR 43745, all applicants for registration to import a basic class of any... (9040) II Opium, raw (9600) II Poppy Straw (9650) II Poppy Straw Concentrate (9670) II The company...

  6. 75 FR 22844 - Importer of Controlled Substances; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-30

    ... Technologies, 72 FR 3417 (2007), comments and requests for hearings on applications to import narcotic raw... 23, 1975, (40 FR 43745), all applicants for registration to import a basic class of any controlled... Opium (9600) II Poppy Straw (9650) II Concentrate of Poppy Straw (9670) II The company plans to...

  7. 77 FR 24983 - Importer of Controlled Substances; Notice of Application Penick Corporation

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-26

    ... not appropriate. 72 FR 3417(2007). As noted in a previous notice published in the Federal Register on September 23, 1975, 40 FR 43745, all applicants for registration to import a basic class of any controlled... Opium (9600) II Poppy Straw (9650) II Concentrate of Poppy Straw (9670) II The company plans to...

  8. 75 FR 9613 - Importer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-03

    ... 20, 2009, and published in the Federal Register on November 30, 2009 (74 FR 62598), Aptuit, 10245... (DEA) to be registered as an importer of Poppy Straw Concentrate (9670), a basic class of controlled... contain trace amounts of the controlled substance normally found in poppy straw concentrate for...

  9. 76 FR 81979 - Manufacturer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-29

    ... 25, 2011, and published in the Federal Register on June 3, 2011, 76 FR 32225, AMPAC Fine Chemicals... controlled substances: Drug Schedule Thebaine (9333) II Poppy Straw Concentrate (9670) II The company is a contract manufacturer. In reference to Poppy Straw Concentrate the company will manufacture...

  10. 75 FR 9614 - Importer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-03

    ... 16, 2009, and published in the Federal Register on October 28, 2009 (74 FR 55584), Clinical Supplies... Administration (DEA) to be registered as an importer of Poppy Straw Concentrate (9670), a basic class of... wounds which contains trace amounts of controlled substances normally found in poppy straw...

  11. 78 FR 19016 - Importer of Controlled Substances; Notice of Registration; Johnson Matthey, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-28

    ... Notice dated November 19, 2012, and published in the Federal Register on November 27, 2012, 77 FR 70824... Thebaine (9333) II Opium, raw (9600) II Noroxymorphone (9668) II Poppy Straw Concentrate (9670) II...

  12. 75 FR 32505 - Importer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-08

    ... 21, 2009, and published in the Federal Register on October 29, 2009 (74 FR 55858), Johnson Matthey... Leaves (9040) II Thebaine (9333) II Opium, raw (9600) II Noroxymorphone (9668) II Poppy Straw...

  13. 75 FR 44285 - Importer of Controlled Substances: Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-28

    ..., 2010 and published in the Federal Register on April 30, 2010, (75 FR 22844), Penick Corporation, 33... schedule II: Drug Schedule Coca Leaves (9040) II Opium, raw (9600) II Poppy Straw (9650) II Concentrate...

  14. 76 FR 30969 - Importer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-27

    ... 18, 2011, and published in the Federal Register on February 2, 2011, 76 FR 5827, Mallinckrodt Inc... substances: Drug Schedule Phenylacetone (8501) II Coca Leaves (9040) II Opium, raw (9600) II Poppy...

  15. 76 FR 7234 - Importer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-09

    ..., 2010, and published in the Federal Register on October 20, 2010, 75 FR 64743, Johnson Matthey, Inc... (9333) II Opium, raw (9600) II Noroxymorphone (9668) II Concentrate of Poppy Straw (9670) II The...

  16. 75 FR 10312 - Importer of Controlled Substances Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-05

    ... 23, 2009, and published in the Federal Register on December 2, 2009 (74 FR 63155), Cambrex Charles... listed in schedule II: Drug Schedule Opium, raw (9600) II Poppy Straw Concentrate (9670) II The...

  17. 49 CFR 40.293 - What is the SAP's function in conducting the initial evaluation of an employee?

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... violation of a DOT drug or alcohol regulation (e.g., related to assertions of use of hemp oil, “medical marijuana” use, “contact positives,” poppy seed ingestion, job stress); or (3) Personal opinions you...

  18. 49 CFR 40.293 - What is the SAP's function in conducting the initial evaluation of an employee?

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... violation of a DOT drug or alcohol regulation (e.g., related to assertions of use of hemp oil, “medical marijuana” use, “contact positives,” poppy seed ingestion, job stress); or (3) Personal opinions you...

  19. 49 CFR 40.293 - What is the SAP's function in conducting the initial evaluation of an employee?

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... violation of a DOT drug or alcohol regulation (e.g., related to assertions of use of hemp oil, “medical marijuana” use, “contact positives,” poppy seed ingestion, job stress); or (3) Personal opinions you...

  20. 49 CFR 40.293 - What is the SAP's function in conducting the initial evaluation of an employee?

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... violation of a DOT drug or alcohol regulation (e.g., related to assertions of use of hemp oil, “medical marijuana” use, “contact positives,” poppy seed ingestion, job stress); or (3) Personal opinions you...

  1. 49 CFR 40.293 - What is the SAP's function in conducting the initial evaluation of an employee?

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... violation of a DOT drug or alcohol regulation (e.g., related to assertions of use of hemp oil, “medical marijuana” use, “contact positives,” poppy seed ingestion, job stress); or (3) Personal opinions you...

  2. 75 FR 69462 - Importer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-12

    ..., 2010, and published in the Federal Register on September 1, 2010 (75 FR 53718), Cambrex Charles City... schedule II: Drug Schedule Opium, raw (9600) II Poppy Straw Concentrate (9670) II The company plans...

  3. 75 FR 69462 - Importer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-12

    ..., 2010, and published in the Federal Register on June 28, 2010 (75 FR 36680), Rhodes Technologies, 498... schedule II: Drug Schedule Raw Opium (9600) II Concentrate of Poppy Straw (9670) II The company plans...

  4. 75 FR 69461 - Importer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-12

    ..., 2010, and published in the Federal Register on ] September 1, 2010 (75 FR 53719), Noramco Inc., 1440... (DEA) to be registered as an importer of Poppy Straw Concentrate (9670), a basic class of...

  5. Draft Genome Sequence of Agrobacterium sp. Strain R89-1, a Morphine Alkaloid-Biotransforming Bacterium.

    PubMed

    Zahradník, Jiří; Kyslíková, Eva; Kyslík, Pavel

    2016-01-01

    Agrobacteriumsp. strain R89-1 isolated from composted wastes ofPapaver somniferumcan effectively biotransform codeine/morphine into 14-OH-derivatives. Here, we present a 4.7-Mb assembly of the R89-1 strain genome. The draft shows that the strain R89-1 represents a distinct phylogenetic lineage within the genusAgrobacterium. PMID:27056219

  6. 76 FR 16413 - Pesticide Products; Registration Applications

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-23

    ... treatment on alfalfa, potato, and rice; seed treatment on alfalfa, barley, dry bean and peas (crop subgroup 6C), potato, rice, soybeans, triticale, and wheat; and nursery seed and seedlings of conifers and..., lunaria, meadowfoam, milkweed, mustard seed, oil radish, poppy seed, rapeseed (canola), sesame, and...

  7. 76 FR 77253 - Importer of Controlled Substances; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-12

    ... applications to import narcotic raw material are not appropriate, in accordance with 72 FR 3417 (2007). In... a previous notice published in the Federal Register on September 23, 1975, 40 FR 43745, all...) II Opium, raw (9600) II Noroxymorphone (9668) II Poppy Straw Concentrate (9670) II The company...

  8. 21 CFR 1312.13 - Issuance of import permit.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... drug listed in Schedule III, IV, or V if he finds: (1) That the substance is crude opium, poppy straw... the Act (21 U.S.C. 952(a)(1) or (a)(2)(A)), the importation of approved narcotic raw material...

  9. 75 FR 1812 - Importer of Controlled Substances; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-01-13

    ... Application pertaining to Rhodes Technologies, 72 FR 3417 (2007), comments and requests for hearings on... a previous notice published in the Federal Register on September 23, 1975, (40 FR 43745), all... schedule II: Drug Schedule Phenylacetone (8501) II Coca Leaves (9040) II Opium, raw (9600) II Poppy...

  10. 77 FR 70824 - Importer of Controlled Substances; Notice of Application; Johnson Matthey, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-11-27

    ... hearings on applications to import narcotic raw material are not appropriate, in accordance with 72 FR 3417...). As noted in a previous notice published in the Federal Register on September 23, 1975, 40 FR 43745... Thebaine (9333) II Opium, raw (9600) II Noroxymorphone (9668) II Poppy Straw Concentrate (9670) II...

  11. 77 FR 64142 - Importer of Controlled Substances, Notice of Application, Noramco, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-18

    ... requests for hearings on applications to import narcotic raw material are not appropriate. 72 FR 3417 (2007...). As noted in a previous notice published in the Federal Register on September 23, 1975, 40 FR 43745-46... controlled substances: Drug Schedule Phenylacetone (8501) II Opium, raw (9600) II Poppy Straw...

  12. 21 CFR 1312.13 - Issuance of import permit.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... drug listed in Schedule III, IV, or V if he finds: (1) That the substance is crude opium, poppy straw... the Act (21 U.S.C. 952(a)(1) or (a)(2)(A)), the importation of approved narcotic raw material...

  13. 76 FR 586 - Importer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-05

    ... 19, 2010, and published in the Federal Register on October 26, 2010, 75 FR 65658, Noramco, Inc... basic classes of controlled substances: Drug Schedule Opium, raw (9600) II Concentrate of Poppy Straw (9670) II Tapentadol (9780) II The company plans to import the Raw Opium (9600) and Concentrate of...

  14. 21 CFR 1312.13 - Issuance of import permit.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... drug listed in Schedule III, IV, or V if he finds: (1) That the substance is crude opium, poppy straw... the Act (21 U.S.C. 952(a)(1) or (a)(2)(A)), the importation of approved narcotic raw material...

  15. 75 FR 64743 - Importer of Controlled Substances; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-20

    ... Notice of Application pertaining to Rhodes Technologies, 72 FR 3417 (2007), comments and requests for..., (40 FR 43745), all applicants for registration to import a basic classes of any controlled substances... Thebaine (9333) II Opium, raw (9600) II Noroxymorphone (9668) II Poppy Straw Concentrate (9670) II...

  16. 76 FR 51400 - Importer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-18

    ... 28, 2011, and published in the Federal Register on May 4, 2011 76 FR 25374, Rhodes Technologies, 498... explained in the Correction to Notice of Application pertaining to Rhodes Technologies, 72 FR 3417 (2007...: Drug Schedule Raw Opium (9600) II Concentrate of Poppy Straw II (9670) The company plans to import...

  17. 76 FR 587 - Manufacturer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-05

    ... August 2, 2010, and published in the Federal Register on September 1, 2010, (75 FR 53721), Cambrex... Dextropropoxyphene, bulk II (non-dosage forms) (9273) Morphine (9300) II Thebaine (9333) II Raw Opium (9600) II Opium extracts (9610) II Opium, powdered (9639) II Opium, granulated (9640) II Poppy Straw (9650) II...

  18. 77 FR 43862 - Importer of Controlled Substances; Notice of Registration; Noramco, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-07-26

    ... dated May 15, 2012, and published in the Federal Register on May 25, 2012, 77 FR 31388, Noramco, Inc... requests for hearings on applications to import narcotic raw material are not appropriate. 72 FR 3417 (2007...: Drug Schedule Phenylacetone (8501) II Opium, raw (9600) II Poppy Straw Concentrate (9670) II...

  19. 77 FR 55503 - Importer of Controlled Substances; Notice of Registration; Cambrex Charles City, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-10

    ..., Inc. By Notice dated June 18, 2012, and published in the Federal Register on June 26, 2012, 77 FR... to import narcotic raw material are not appropriate. 72 FR 3417 (2007). DEA has considered the... Phenylacetone (8501) II Opium, raw (9600) II Poppy Straw Concentrate (9670) II The company plans to import...

  20. 77 FR 16262 - Importer of Controlled Substances, Notice of Registration; Johnson Matthey, Inc., Pharmaceutical...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-20

    ..., 2011, 76 FR 77253, Johnson Matthey, Inc., Pharmaceutical Materials, 2003 Nolte Drive, West Deptford... not appropriate, 72 FR 3417 (2007). Regarding all other basic classes of controlled substances, no... (9040) II Thebaine (9333) II Opium, raw (9600) II Noroxymorphone (9668) II Poppy Straw Concentrate...

  1. 21 CFR 1312.13 - Issuance of import permit.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... drug listed in Schedule III, IV, or V if he finds: (1) That the substance is crude opium, poppy straw... the Act (21 U.S.C. 952(a)(1) or (a)(2)(A)), the importation of approved narcotic raw material...

  2. 77 FR 40086 - Importer of Controlled Substances, Notice of Application, Chattem Chemicals Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-07-06

    ... to import narcotic raw material are not appropriate. 72 FR 3417 (2007). Any bulk manufacturer who is...). As noted in a previous notice published in the Federal Register on September 23, 1975, 40 FR 43745-46... Phenylacetone (8501) II Opium, raw (9600) II Poppy Straw Concentrate (9670) II Tapentadol (9780) II The...

  3. 76 FR 5827 - Importer of Controlled Substances; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-02

    ... Application pertaining to Rhodes Technologies, 72 FR 3417 (2007), comments and requests for hearings on... notice published in the Federal Register on September 23, 1975, (40 FR 43745), all applicants for...: Drug Schedule Phenylacetone (8501) II Coca Leaves (9040) II Opium, raw (9600) II Poppy...

  4. 76 FR 51398 - Importer of Controlled Substances; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-18

    ... Application pertaining to Rhodes Technologies, 72 FR 3417 (2007), comments and requests for hearings on...). As noted in a previous notice published in the Federal Register on September 23, 1975, 40 FR 43745...: Drug Schedule Phenylacetone (8501) II Opium, raw (9600) II Poppy Straw Concentrate (9670) II...

  5. 78 FR 33440 - Importer of Controlled Substances, Notice of Registration; Rhodes Technologies

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-04

    ... Notice dated April 10, 2013, and published in the Federal Register on April 19, 2013, 78 FR 23594, Rhodes... FR 3417 (2007). DEA has considered the factors in 21 U.S.C. 823(a) and 952(a), and determined that... substances: Drug Schedule Opium Raw (9600) II Poppy Straw Concentrate (9670) II The company plans to...

  6. 77 FR 5846 - Importer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-06

    ... 28, 2011, and published in the Federal Register on October 7, 2011, 76 FR 62447, Cody Laboratories... material are not appropriate, 72 FR 3417 (2007). DEA has considered the factors in 21 U.S.C. 823(a) and 952... substances: Drug Schedule Opium, raw (9600) II Poppy Straw Concentrate (9670) II The company plans to...

  7. 78 FR 64020 - Manufacturer of Controlled Substances, Notice of Registration, Siegfried USA, LLC

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-25

    ... The Notice dated July 23, 2013, and published in the Federal Register on August 1, 2013, 78 FR 46613... registration as a Bulk Manufacturer of drug codes Opium, raw (9600), and Poppy Straw Concentrate (9670). On August 6, 2013, Siegfried (USA), LLC., subsequently withdrew this request for drug code Opium, raw...

  8. 77 FR 35057 - Importer of Controlled Substances; Notice of Registration; Rhodes Technologies

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-12

    ... Notice dated April 17, 2012, and published in the Federal Register on April 26, 2012, 77 FR 24984, Rhodes... and requests for hearings on applications to import narcotic raw material are not appropriate. 72 FR...: Drug Schedule Opium, raw (9600) II Poppy Straw Concentrate (9670) II The company plans to import...

  9. 77 FR 60143 - Importer of Controlled Substances; Notice of Registration; Cody Laboratories, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-02

    .... By Notice dated July 17, 2012, and published in the Federal Register on July 26, 2012, 77 FR 43861... FR 3417 (2007). Regarding Tapentadol, no comments or objections have been received. DEA has... classes of controlled substances: Drug Schedule Opium, raw (9600) II Concentrate Poppy Straw (9670)...

  10. 77 FR 5846 - Importer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-06

    ..., 2011, and published in the Federal Register on August 18, 2011, 76 FR 51398, Cambrex Charles City, Inc... are not appropriate, in accordance with 72 FR 3417 (2007). DEA has considered the factors in 21 U.S.C...: Drug Schedule Phenylacetone (8501) II Opium, raw (9600) II Poppy Straw Concentrate (9670) II...

  11. 77 FR 43861 - Importer of Controlled Substances; Notice Of Application; Cody Laboratories, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-07-26

    ... and requests for hearings on applications to import narcotic raw material are not appropriate. 72 FR... previous notice published in the Federal Register on September 23, 1975, 40 FR 43745, all applicants for... substances: Drug Schedule Opium, Raw (9600) II Concentrate Poppy Straw (9670) II Tapentadol (9780) II...

  12. 75 FR 65658 - Importer of Controlled Substances; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-26

    .... As explained in the Correction to Notice of Application pertaining to Rhodes Technologies, 72 FR 3417... Register on September 23, 1975, (40 FR 43745-46), all applicants for registration to import a basic class... controlled substances listed in schedule II: Drug Schedule Raw Opium (9600) II Concentrate of Poppy...

  13. 77 FR 31388 - Importer of Controlled Substances; Notice of Application; Noramco, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-25

    ... appropriate. 72 FR 3417 (2007). In regard to the non-narcotic raw material, any bulk manufacturer who is... September 23, 1975, 40 FR 43745-46, all applicants for registration to import a basic class of any... controlled substances: Drug Schedule Phenylacetone (8501) II Opium, raw (9600) II Poppy Straw...

  14. 76 FR 35241 - Importer of Controlled Substances; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-16

    ... explained in the Correction to Notice of Application pertaining to Rhodes Technologies, 72 FR 3417 (2007... (f). As noted in a previous notice published in the Federal Register on September 23, 1975, 40 FR... (8501) II Raw Opium (9600) II Concentrate of Poppy Straw (9670) II The company plans to import...

  15. 78 FR 17230 - Importer of Controlled Substances, Notice of Registration, Noramco, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-20

    ... dated October 9, 2012, and published in the Federal Register on October 18, 2012, 77 FR 64142, Noramco... requests for hearings on applications to import narcotic raw ] material are not appropriate. 72 FR 3417... controlled substances: Drug Schedule Phenylacetone (8501) II Opium, raw (9600) II Poppy Straw...

  16. 78 FR 64015 - Importer of Controlled Substances; Notice of Application; Noramco, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-25

    ... requests for hearings on applications to import narcotic raw material are not appropriate. 72 FR 3417(2007...). As noted in a previous notice published in the Federal Register on September 23, 1975, 40 FR 43745-46... controlled substances: Drug Schedule Phenylacetone (8501) II Opium, raw (9600) II Poppy Straw...

  17. 76 FR 62447 - Importer of Controlled Substances; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-07

    ... the Correction to Notice of Application pertaining to Rhodes Technologies, 72 FR 3417 (2007), comments... noted in a previous notice published in the Federal Register on September 23, 1975, 40 FR 43745, all... following basic classes of controlled substances: Drug Schedule Opium, raw (9600) II Poppy Straw...

  18. 78 FR 64015 - Importer of Controlled Substances, Notice of Registration, Siegfried (USA), LLC

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-25

    ... Notice dated August 2, 2013, and published in the Federal Register on ] August 14, 2013, 78 FR 49546... appropriate. 72 FR 3417 (2007). DEA has considered the factors in 21 U.S.C. 823(a) and 952(a), and determined... classes of controlled substances: Drug Schedule Opium, raw (9600) II Poppy Straw Concentrate (9670) II...

  19. 76 FR 42732 - Importer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-19

    ..., 2011 and published in the Federal Register on June 16, 2011, 76 FR 35239, Penick Corporation, 33... Application pertaining to Rhodes Technologies, 72 FR 2417 (2007), comments and requests for hearings on...: Drug Schedule Coca Leaves (9040) II Opium, raw (9600) II Poppy Straw (9650) II Concentrate of...

  20. 76 FR 17967 - Importer of Controlled Substances; Notice of Registration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-31

    ... 19, 2010, and published in the Federal Register on October 26, 2010, 75 FR 65658, Cody Laboratories... substances: Drug Schedule Raw Opium (9600) II Concentrate of Poppy Straw (9670) II The company plans to... registered with DEA as a manufacturer of several controlled substances that are manufactured from raw...

  1. La Artesania Mexicana (Mexican Handicrafts).

    ERIC Educational Resources Information Center

    Steele, Bettina

    This booklet contains instructions in English and Spanish for making eleven typical Mexican craft articles. The instructions are accompanied by pen-and-ink drawings. The objects are (1) "La Rosa" (The Rose); (2) "El Crisantemo" (The Chrysanthemum); (3) "La Amapola" (The Poppy); (4) "Ojos de Dios" (God's Eyes); (5) "Ojitos con dos caras" (Two-Sided…

  2. 40 CFR 180.582 - Pyraclostrobin; tolerances for residues.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... seed, seed 0.45 Nut, tree, group 14 0.04 Oat, grain 1.2 Oat, hay 18 Oat, straw 15 Oil radish, seed 0.45... Peanut, refined oil 0.1 Peppermint, tops 8.0 Pistachio 0.7 Poppy, seed 0.45 Radish, tops 16...

  3. 40 CFR 180.582 - Pyraclostrobin; tolerances for residues.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... seed, seed 0.45 Nut, tree, group 14 0.04 Oat, grain 1.2 Oat, hay 18 Oat, straw 15 Oil radish, seed 0.45... Peanut, refined oil 0.1 Peppermint, tops 8.0 Pistachio 0.7 Poppy, seed 0.45 Radish, tops 16...

  4. 40 CFR 180.582 - Pyraclostrobin; tolerances for residues.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 0.45 Nut, tree, group 14 0.04 Oat, grain 1.2 Oat, hay 18 Oat, straw 15 Oil radish, seed 0.45 Papaya... Peanut, refined oil 0.1 Peppermint, tops 8.0 Pistachio 0.7 Poppy, seed 0.45 Radish, tops 16...

  5. 40 CFR 180.582 - Pyraclostrobin; tolerances for residues.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... seed, seed 0.45 Nut, tree, group 14 0.04 Oat, grain 1.2 Oat, hay 18 Oat, straw 15 Oil radish, seed 0.45... Peanut, refined oil 0.1 Peppermint, tops 8.0 Pistachio 0.7 Poppy, seed 0.45 Radish, tops 16...

  6. 78 FR 64013 - Importer of Controlled Substances; Notice of Application; Cambrex Charles City, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-25

    ... appropriate. 72 FR 3417 (2007). Any bulk manufacturer who is presently, or is applying to be, registered with... published in the Federal Register on September 23, 1975, 40 FR 43745, all applicants for registration to... Cocaine (9041) II Opium, raw (9600) II Poppy Straw Concentrate (9670) II The company plans to import...

  7. 77 FR 19717 - Importer of Controlled Substances; Notice of Registration Mallinckrodt LLC

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-02

    ...). Comments and requests for hearings on applications to import narcotic raw material are not appropriate, 72... Notice dated January 23, 2012, and published in the Federal Register on January 31, 2012, 77 FR 4831... substances: Drug Schedule Phenylacetone (8501) II Coca Leaves (9040) II Opium, raw (9600) II Poppy...

  8. 77 FR 38085 - Importer of Controlled Substances; Notice of Application; Cambrex Charles City, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-26

    ... requests for hearings on applications to import narcotic raw material are not appropriate. 72 FR 3417 (2007... Opium, raw (9600) II Poppy Straw Concentrate (9670) II The company plans to import the listed controlled... September 23, 1975, 40 FR 43745, all applicants for registration to import a basic class of any...

  9. My General Education: "'Discovering" Amitav Ghosh

    ERIC Educational Resources Information Center

    Richards, Sandra L.

    2015-01-01

    Asked to recommend a faculty reading that might counteract our habits of overspecialization, I nominate Amitav Ghosh's "Sea of Poppies" (2008), the first novel in his "Ibis" trilogy. Reading Ghosh has offered me not only a fascinating introduction to the Indian Ocean world but also an occasion to reflect upon the dynamics of a…

  10. Simulating the Afghanistan-Pakistan opium supply chain

    SciTech Connect

    Watkins, Jennifer H; MacKerrow, Edward P; Merritt, Terence M

    2010-04-08

    This paper outlines an opium supply chain using the Hilmand province of Afghanistan as exemplar. The opium supply chain model follows the transformation of opium poppy seed through cultivation and chemical alteration to brown heroin base. The purpose of modeling and simulating the Afghanistan-Pakistan opium supply chain is to discover and test strategies that will disrupt this criminal enterprise.

  11. 78 FR 17230 - Importer of Controlled Substances, Notice of Application, Noramco Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-20

    ... request for hearings on applications to import narcotic raw material are not appropriate. 72 FR 3417 (2007... Federal Register on September 23, 1975, 40 FR 43745-46, all applicants for registration to import a basic... schedule II: Drug Schedule Phenylacetone (8501) II Thebaine (9333) II Noroxymorphone (9668) II Poppy...

  12. 77 FR 55505 - Manufacturer of Controlled Substances; Notice of Registration; AMPAC Fine Chemicals LLC

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-10

    ... LLC By Notice dated May 11, 2012, and published in the Federal Register on May 21, 2012, 77 FR 30026... FR 3417 (2007). DEA has considered the factors in 21 USC Sec. 823(a) and determined that the... following basic classes of controlled substances: Drug Schedule Thebaine (9333) II Poppy Straw...

  13. 75 FR 53719 - Importer of Controlled Substances; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-01

    ... Technologies, 72 FR 3417 (2007), comments and requests for hearings on applications to import narcotic raw... 23, 1975, (40 FR 43745), all applicants for registration to import a basic class of any controlled... Drug Enforcement Administration (DEA) for registration as an importer of Poppy Straw Concentrate...

  14. 77 FR 24984 - Importer of Controlled Substances; Notice of Application; Rhodes Technologies

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-26

    ... hearings on applications to import narcotic raw material are not appropriate. 72 FR 3417 (2007). As noted in a previous notice published in the Federal Register on September 23, 1975, 40 FR 43745, all... Opium, Raw (9600) II Poppy Straw Concentrate (9670) II The company plans to import the listed...

  15. Wet solids flow enhancement

    SciTech Connect

    Caram, H.S.; Agrawal, D.K.; Foster, N.

    1997-07-01

    The objective was to visualize the flow of granular materials in the silo using Nuclear Magnetic Resonance. This was done by introducing traces. Mustard seeds and poppy seeds were used as trace particles. The region sampled was a cylinder 25 mm in diameter and 40 mm in length. Eight slices containing 128 by 128 to 256 by 256 pixels were generated for each image.

  16. 78 FR 69134 - Manufacturer of Controlled Substances; Notice of Registration: AMPAC Fine Chemicals, LLC.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-18

    ..., LLC. By Notice dated July 23, 2013, and published in the Federal Register on July 31, 2013, 78 FR... to import narcotic raw material are not appropriate. 72 FR 3417 (2007). DEA has considered the... Thebaine (9333) II Poppy Straw Concentrate (9670) II Tapentadol (9780) II The company is a...

  17. Instructional Resources. The Significance of Form: Ceramics.

    ERIC Educational Resources Information Center

    Zawatsky, Carole; And Others

    1989-01-01

    Presents four lesson plans designed to teach K-12 students about ceramics and the artists using the medium. Each lesson is centered around one ceramic piece: (1) "Wall Clock," by the Chantilly Porcelain Factory; (2) "Poppy Vase," by Adelaide Robineau; (3) "Laughing Eyes," by Pablo Picasso; and (4) "Ceramic Drum Jar," by Tsayutitsa. (GEA)

  18. 78 FR 35052 - Importer of Controlled Substances; Notice of Registration; Noramco, Inc.

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-11

    ... dated March 12, 2013, and published in the Federal Register on March 20, 2013, 78 FR 17230, Noramco, Inc... raw material are not appropriate. 72 FR 3417 (2007). DEA has considered the factors in 21 U.S.C. 823(a... Phenylacetone (8501) II Thebaine (9333) II Poppy Straw Concentrate (9670) II Tapentadol (9780) II ] The...

  19. Mexican Folkart for Children.

    ERIC Educational Resources Information Center

    Dominguez, Graciela; And Others

    Directions, suggested materials, and illustrations are given for making paper mache pinatas and masks, cascarones, Ojos de Dios, maracas, dresser scarf embroidery, burlap murals, yarn designs, paper plate trays, paper cut designs, the poppy, sarape aprons, and paper Mexican dolls. Filled with candy and broken, the pinata is used on most Mexican…

  20. Heroin

    MedlinePlus

    ... Search Share Print Home » Drugs of Abuse » Heroin Heroin Email Facebook Twitter Brief Description Heroin is an opioid drug that is synthesized from ... seed pod of the Asian opium poppy plant. Heroin usually appears as a white or brown powder ...

  1. Experience in the use of hyperspectral data for the detection of vegetation containing narcotic substances

    NASA Astrophysics Data System (ADS)

    Sedelnikov, V. P.; Lukashevich, E. L.; Karpukhina, O. A.

    2014-12-01

    This paper provides the characteristics of an experimental sample of a hyperspectral videospectrometer Sokol-SCP and presents examples of the hyperspectral data received as a result of flight tests. The results of the detection of vegetation containing narcotic substances by spectral attributes using the obtained hyperspectral information are considered. The opportunity for using the hyperspectral data for detection of cannabis and papaver sites, including those in mixed crops with masking vegetation, is confirmed.

  2. Evolutionary Genetics of an S-Like Polymorphism in Papaveraceae with Putative Function in Self-Incompatibility

    PubMed Central

    Paape, Timothy; Miyake, Takashi; Takebayashi, Naoki; Wolf, Diana; Kohn, Joshua R.

    2011-01-01

    Background Papaver rhoeas possesses a gametophytic self-incompatibility (SI) system not homologous to any other SI mechanism characterized at the molecular level. Four previously published full length stigmatic S-alleles from the genus Papaver exhibited remarkable sequence divergence, but these studies failed to amplify additional S-alleles despite crossing evidence for more than 60 S-alleles in Papaver rhoeas alone. Methodology/Principal Findings Using RT-PCR we identified 87 unique putative stigmatic S-allele sequences from the Papaveraceae Argemone munita, Papaver mcconnellii, P. nudicuale, Platystemon californicus and Romneya coulteri. Hand pollinations among two full-sib families of both A. munita and P. californicus indicate a strong correlation between the putative S-genotype and observed incompatibility phenotype. However, we also found more than two S-like sequences in some individuals of A. munita and P. californicus, with two products co-segregating in both full-sib families of P. californicus. Pairwise sequence divergence estimates within and among taxa show Papaver stigmatic S-alleles to be the most variable with lower divergence among putative S-alleles from other Papaveraceae. Genealogical analysis indicates little shared ancestral polymorphism among S-like sequences from different genera. Lack of shared ancestral polymorphism could be due to long divergence times among genera studied, reduced levels of balancing selection if some or all S-like sequences do not function in incompatibility, population bottlenecks, or different levels of recombination among taxa. Preliminary estimates of positive selection find many sites under selective constraint with a few undergoing positive selection, suggesting that self-recognition may depend on amino acid substitutions at only a few sites. Conclusions/Significance Because of the strong correlation between genotype and SI phenotype, sequences reported here represent either functional stylar S-alleles, tightly

  3. Mycobiota and mycotoxins in bee pollen collected from different areas of Slovakia.

    PubMed

    Kačániová, Miroslava; Juráček, Miroslav; Chlebo, Róbert; Kňazovická, Vladimíra; Kadasi-Horáková, Miriam; Kunová, Simona; Lejková, Jadža; Haščík, Peter; Mareček, Ján; Simko, Milan

    2011-01-01

    Contamination by microscopic fungi and mycotoxins in different bee pollen samples, which were stored under three different ways of storing as freezing, drying and UV radiation, was investigated. During spring 2009, 45 samples of bee-collected pollen were gathered from beekeepers who placed their bee colonies on monocultures of sunflower, rape and poppy fields within their flying distance. Bee pollen was collected from bees' legs by special devices placed at the entrance to hives. Samples were examined for the concentration and identification of microscopic fungi able to grow on Malt and Czapek-Dox agar and mycotoxins content [deoxynivalenol (DON), T-2 toxin (T-2), zearalenone (ZON) and total aflatoxins (AFL), fumonisins (FUM), ochratoxins (OTA)] by direct competitive enzyme-linked immunosorbent assays (ELISA). The total number of microscopic fungi in this study ranged from 2.98 ± 0.02 in frozen sunflower bee pollen to 4.06 ± 0.10 log cfu.g(-1) in sunflower bee pollen after UV radiation. In this study, 449 isolates belonging to 21 fungal species representing 9 genera were found in 45 samples of bee pollen. The total isolates were detected in frozen poppy pollen 29, rape pollen 40, sunflower pollen 80, in dried poppy pollen 12, rape pollen 36, sunflower 78, in poppy pollen after UV radiation treatment 54, rape 59 and sunflower 58. The most frequent isolates of microscopic fungi found in bee pollen samples of all prevalent species were Mucor mucedo (49 isolates), Alternaria alternata (40 isolates), Mucor hiemalis (40 isolates), Aspergillus fumigatus (33 isolates) and Cladosporium cladosporioides (31 isolates). The most frequently found isolates were detected in sunflower bee pollen frozen (80 isolates) and the lowest number of isolates was observed in poppy bee pollen dried (12 isolates). The most prevalent mycotoxin of poppy bee pollen was ZON (361.55 ± 0.26 μg.kg(-1)), in rape bee pollen T-2 toxin (265.40 ± 0.18 μg.kg(-1)) and in sunflower bee pollen T-2 toxin

  4. In-planta detection and monitorization of endophytic colonization by a Beauveria bassiana strain using a new-developed nested and quantitative PCR-based assay and confocal laser scanning microscopy.

    PubMed

    Landa, B B; López-Díaz, C; Jiménez-Fernández, D; Montes-Borrego, M; Muñoz-Ledesma, F J; Ortiz-Urquiza, A; Quesada-Moraga, E

    2013-10-01

    Beauveria bassiana strain 04/01-Tip obtained from larvae of the opium poppy stem gall Iraella luteipes endophytically colonizes opium poppy plants and protect it against this pest. Development of a specific, rapid and sensitive technique that allows accurately determining the process and factors leading to the establishment of this strain in opium poppy plants would be essential to achieve its efficient control in a large field scale. For that purpose in the present study, species-specific primers that can be used in conventional or quantitative PCR protocols were developed for specifically identification and detection of B. bassiana in plant tissues. The combination of the designed BB.fw/BB.rv primer set with the universal ITS1-F/ITS4 primer set in a two-step nested-PCR approach, has allowed the amplification of up to 10fg of B. bassiana. This represented an increase in sensitivity of 10000- and 1000-fold of detection than when using the BB.fw/BB.rv primers in a single or single-tube semi-nested PCR approaches, respectively. The BB.fw and BB.rv primer set were subsequently optimized to be used in real time quantitative PCR assays and allowed to accurately quantify B. bassiana DNA in different plant DNA backgrounds (leaves and seeds) without losing accuracy and efficiency. The qPCR protocol was used to monitor the endophytic colonization of opium poppy leaves byB. bassiana after inoculation with the strain EABb 04/01-Tip, detecting as low as 26fg of target DNA in leaves and a decrease in fungal biomass over time. PCR quantification data were supported in parallel with CLMS by the monitoring of spatial and temporal patterns of leaf and stem colonization using a GFP-tagged transformant of the B. bassiana EABb 04/01-Tip strain, which enabled to demonstrate that B. bassiana effectively colonizes aerial tissues of opium poppy plants mainly through intercellular spaces and even leaf trichomes. A decline in endophytic colonization was also observed by the last sampling

  5. Effects of gluten-free breads, with varying functional supplements, on the biochemical parameters and antioxidant status of rat serum.

    PubMed

    Świeca, Michał; Reguła, Julita; Suliburska, Joanna; Złotek, Urszula; Gawlik-Dziki, Urszula

    2015-09-01

    This paper examines the effects of gluten-free bread enriched with functional ingredients (milk powder, poppy, sunflower and pumpkin seeds, egg yolk, carum, hazel nuts and amaranth) on the morphological and biochemical parameters and antioxidant status of rats serum. Rats were provided test diets--gluten-free breads and water ad libitum. After 14 days, the animals were weighed and killed. A hazel nut-amaranth bread diet significantly increased the level of thrombocytes when compared to control bread. A mixed bread diet significantly decreased cholesterol levels in rats. All fortified breads decreased triglyceride levels and alanine transaminase activity and caused an increase in antiradical activity of the serum. In rats fed with poppy-milk bread, milk-seed bread and mixed bread, a marked decrease in superoxide dismutase activity was found. Enriched breads reduced the levels of triglyceride and improved the antiradical properties of serum, although the physiological relevance of this needs to be confirmed by human studies.

  6. Nuclear magnetic resonance studies of granular flows. Technical progress report, January--March 1994

    SciTech Connect

    Not Available

    1994-07-01

    The authors have completed additional NMR imaging experiments for larger mustard seeds with average diameter 2.4 mm and poppy seeds having a flat, circular shape with a cusp. Overall flow profiles of the larger mustard seeds are quantitatively similar to those from smaller mustard seeds reported previously, i.e., the size increase by a factor of 1.6 did not affect the flow profiles. Poppy seeds, on the other hand, were easy to break when they collided and were sheared. Therefore, fines due to breakage formed a small core which always persisted in the experiment. For the first time it is possible to obtain velocity and diffusion for both constituents in a multiphase flow during the segregation process. This will certainly lead to a much better understanding of granular flows.

  7. Effects of gluten-free breads, with varying functional supplements, on the biochemical parameters and antioxidant status of rat serum.

    PubMed

    Świeca, Michał; Reguła, Julita; Suliburska, Joanna; Złotek, Urszula; Gawlik-Dziki, Urszula

    2015-09-01

    This paper examines the effects of gluten-free bread enriched with functional ingredients (milk powder, poppy, sunflower and pumpkin seeds, egg yolk, carum, hazel nuts and amaranth) on the morphological and biochemical parameters and antioxidant status of rats serum. Rats were provided test diets--gluten-free breads and water ad libitum. After 14 days, the animals were weighed and killed. A hazel nut-amaranth bread diet significantly increased the level of thrombocytes when compared to control bread. A mixed bread diet significantly decreased cholesterol levels in rats. All fortified breads decreased triglyceride levels and alanine transaminase activity and caused an increase in antiradical activity of the serum. In rats fed with poppy-milk bread, milk-seed bread and mixed bread, a marked decrease in superoxide dismutase activity was found. Enriched breads reduced the levels of triglyceride and improved the antiradical properties of serum, although the physiological relevance of this needs to be confirmed by human studies. PMID:25842337

  8. [Latex allergy].

    PubMed

    Richter, J; Susický, P

    2000-04-01

    The authors describe a case of an allergic affection in a patient with occupational exposure to latex allergens with a history of anaphylactic reaction to poppy seed and reaction to the antigens of apples, oranges, tangerines, peanuts and bananas, revealed by the method CAP Phadiatop. A marked reaction was initiated after the use of a shampoo containing volatile banana oil. The authors emphasize the high incidence of latex allergy, the manifestations of which may be encountered also in clinical ophthalmology. PMID:10874793

  9. Antioxidant Potential of the Extracts, Fractions and Oils Derived from Oilseeds.

    PubMed

    Ishtiaque, Shagufta; Khan, Nasir; Siddiqui, Muhammad A; Siddiqi, Rahmanullah; Naz, Shahina

    2013-10-15

    The polyphenolic extracts and oils were obtained from ajwain, mustard, fenugreek and poppy seeds. The extracts were partitioned into acidic and neutral polyphenolic fractions and following estimation of total phenolics in the crude extract, acidic and neutral fractions and oil, all were analyzed for their DPPH (2,2-diphenyl-1-picrylhydrazyl) scavenging potential, ferric reducing ability and chelating power. The highest amount of polyphenols was found in ajwain (8330 ± 107), then in mustard seeds (2844 ± 56.00) and in fenugreek (1130 ± 29.00), and least in poppy seeds (937 ± 18.52). The higher amounts of polyphenols were estimated in neutral fraction compared to acidic (p < 0.05). % Inhibition of DPPH by the crude extract and fractions of all oilseeds was quite significant, being higher for acidic than neutral. The highest % DPPH inhibition was shown by ajwain extract than mustard > fenugreek and least by poppy seed extracts (p < 0.05). The reducing power and the chelating effect of the oilseeds followed the same order as DPPH, but higher % chelation was shown by neutral than acidic fraction (p < 0.05). Though low in polyphenols, the oil fractions were as strong antioxidants as the acidic one. Though oilseeds are used in very small quantity in food, they are potential sources of natural antioxidants and may replace synthetic ones.

  10. Segregation in horizontal rotating cylinders: radial and axial band formation, band traveling and merging studied by Magnetic Resonance Imaging.

    NASA Astrophysics Data System (ADS)

    Nguyen, Thoa; Sederman, Andrew; Gladden, Lynn

    2007-03-01

    Radial and axial segregations are investigated by Magnetic Resonance Imaging (MRI). For the first time, full 3D structures and real-time 2D MRI movies showing the progress of segregation over many hours are reported. Data were acquired with high temporal (74 ms) and in-plane spatial resolutions (1 mm x 1 mm), giving new insights into the underlying mechanisms. The mixture composition can be quantified throughout segregation. The cylinder to be considered is 48 mm in diameter, up to 50 cm long and filled to 50 -- 82% by volume with millet and poppy seeds at a 3:1 ratio. In particular, the effects of filling fraction, cylinder length and rotational speed on segregation are addressed. Radial segregation is found to be driven by both core diffusion and the free surface. The former is dominant in the cylindrical core buried under the avalanche layer in systems over 75% full while the latter is significant at lower filling levels. Axial segregation is characterized by band formation, traveling, and merging. In all cases studied, the formation of poppy-rich bands is observed, after which individual bands start to travel at ˜3 μm s-1 until they are within ˜3 cm of a stationary band. Adjacent bands then merge into a single, enlarged poppy band as millet seeds move out of the merging region.

  11. Quantitative wear and wear damage analysis of composite resins in vitro.

    PubMed

    Koottathape, Natthavoot; Takahashi, Hidekazu; Iwasaki, Naohiko; Kanehira, Masafumi; Finger, Werner J

    2014-01-01

    The aim of this study was to investigate volume loss and worn surfaces' morphologies of eight composite resins: Durafill VS (DUR), Clearfil AP-X (APX), Filtek Z250 (Z250), Filtek Supreme XT (FIL), Kalore (KAL), MI Flow (MFL), Venus Diamond (VED) and Venus Pearl (VEP). Disc-shaped specimens were fabricated and mounted in a ball-on-disc wear testing machine and abraded in water or with the third-body media, poppy seed slurry and polymethyl methacrylate (PMMA) slurry. Volume loss (n=5) was determined after 50k sliding cycles, and analyzed using two-way ANOVA (α=0.05). The worn surfaces were examined with SEM. Two-way ANOVA suggested significant interaction between composite and wear condition. DUR, KAL and MFL showed low wear in water. DUR, Z250 and FIL showed moderate wear with PMMA slurry, whereas APX, KAL and MFL were deeper abraded. Under the action of poppy seed slurry DUR proved high volume loss. SEM showed that Z250, FIL and MFL were uniformly abraded in water. KAL and MFL with poppy seed were heavily destructed, whereas VED and VEP appeared very smooth. KAL and MFL abraded with PMMA slurry showed many cracks, but VEP remained crack-free and smooth. Volume loss and worn surfaces' morphologies varied with type of composite and third-body media used. PMID:24219861

  12. Essential Oil Composition and Antigermination Activity of Artemisia dracunculus (Tarragon).

    PubMed

    Fraternale, Daniele; Flamini, Guido; Ricci, Donata

    2015-08-01

    The chemical composition of an Italian oil of tarragon (Artemisia dracunculus L.) was analyzed by GC/EIMS. The major compound of the oil was estragole (73.3%), followed by limonene (5.4%), (E)-β-ocimene (5.3%), β-pinene (3.4%) and (Z)-β-ocimene (3.0%). The essential oil was tested "in vitro" for its antigermination activity against Raphanus sativus L., Lepidium sativum L, Papaver rhoeas L. and Avena fatua L. seeds and demonstrating a good inhibitory activity in a dose-dependent way. PMID:26434144

  13. Essential Oil Composition and Antigermination Activity of Artemisia dracunculus (Tarragon).

    PubMed

    Fraternale, Daniele; Flamini, Guido; Ricci, Donata

    2015-08-01

    The chemical composition of an Italian oil of tarragon (Artemisia dracunculus L.) was analyzed by GC/EIMS. The major compound of the oil was estragole (73.3%), followed by limonene (5.4%), (E)-β-ocimene (5.3%), β-pinene (3.4%) and (Z)-β-ocimene (3.0%). The essential oil was tested "in vitro" for its antigermination activity against Raphanus sativus L., Lepidium sativum L, Papaver rhoeas L. and Avena fatua L. seeds and demonstrating a good inhibitory activity in a dose-dependent way.

  14. Deceive, and be deceived!

    PubMed

    Schuiling, G A

    2004-06-01

    In their struggle for life, many species, for example prokaryotes, plants and animals, defend themselves by producing substances which may poison or disorientate (potential) enemies (grazers/predators etc), or signal information which puts (potential) enemies on the wrong track. Examples of poisonous/ disorientating substances include prussic acid (e.g. almonds) or morphine (papaver), examples of 'false' visual cues include the appearance of harmless hoverflies which strongly resemble 'dangerous' wasps or certain orchid flowers which resemble the female sex organs of certain insects ('mimicry'). Misleading is an essential part of communication systems. PMID:15715040

  15. Diagnosing the inter-model spread in snow water equivalent for CMIP5 over Southwest Asia

    NASA Astrophysics Data System (ADS)

    Mankin, J. S.; Scherer, M.; Diffenbaugh, N. S.

    2012-12-01

    Recent analysis of the CMIP5 set of integrations has highlighted a wide divergence in the models' ability to resolve historical observations of snow water equivalent (SWE) throughout the Northern Hemisphere. However, despite the difficulty of resolving SWE in hindcasts, there exists a consistent signal in the magnitude of SWE decline under the RCP8.5 forcing scenario among the CMIP5. Separately, our work has established that lower yields in irrigated wheat induce Afghan farmers to plant more opium poppy, a more drought resistant crop planted at the same time. In Southwest and Central Asia, subsistence and industrial agriculture rely on irrigation supplied by runoff from upland snowmelt, and crop yields, including those of wheat and poppy, are influenced by this water availability. If water availability attenuates driving yield declines in staple crops like wheat, farmers in Afghanistan can be driven to cultivate more opium poppy in response—a crop that has a complex influence on stability and conflict there. Bounding the ensemble uncertainty in model simulations of SWE is an important step in assessing the ways in which farmer decisions have and will be constrained. Therefore, diagnosing the sources of model divergence in this important metric for subsistence and large-scale agriculture in Southwest and Central Asia is a first step for improving model resolution of such processes in projections of climate change. We present initial results that quantify the extent to which snow albedo feedback (SAF) parameterizations among the models in CMIP5 influence SWE simulation uncertainties over Southwest and Central Asia.

  16. Hector Berlioz and other famous artists with opium abuse.

    PubMed

    Wolf, Paul L

    2010-01-01

    The effect of opium on the creativity and productivity of a famous composer of classical music, an essayist, and poets including Hector Berlioz, Thomas De Quincy, Samuel Taylor Coleridge, John Keats, and Jean Cocteau, is described. Opium is a narcotic drug prepared from the juice of the unripe seed capsules of the opium poppy. It contains alkaloids such as morphine, codeine, and papaverine. Medically it is used to relieve pain and produce sleep. It is used as an intoxicant. Alcohol and opium were commonly relied on in the 19th century, especially by artists, to stimulate creativity and relieve stress. These artists described the effect of opium on their creativity and productivity.

  17. Recent Advances in the Synthesis of Morphine and Related Alkaloids

    NASA Astrophysics Data System (ADS)

    Chida, Noritaka

    Morphine, an alkaloid isolated from the opium poppy, has been widely used as an analgesic, and has been a fascinating synthetic target of organic chemists. After the first total synthesis reported in 1952, a number of synthetic studies toward morphine have been reported, and findings obtained in such studies have greatly contributed to the progress of synthetic organic chemistry as well as medicinal chemistry. This review provides an overview of recent studies toward the total synthesis of morphine and related alkaloids. Work reported in the literature since 2004 will be reviewed.

  18. WET SOLIDS FLOW ENHANCEMENT

    SciTech Connect

    Hugo S. Caram; Natalie Foster

    1997-03-31

    The objective was to visualize the flow of granular materials in flat bottomed silo. This was done by for dry materials introducing mustard seeds and poppy seeds as tracer particles and imaging them using Nuclear Magnetic Resonance. The region sampled was a cylinder 25 mm in diameter and 40 mm in length. Eight slices containing 128*128 to 256*256 pixels were generated for each image. The size of the silo was limited by the size of the high resolution NMR imager available. Cross-sections of 150mm flat bottomed silos, with the tracer layers immobilized by a gel, showed similar qualitative patterns for both dry and wet granular solids.

  19. Granular convection observed by magnetic resonance imaging

    SciTech Connect

    Ehrichs, E.E.; Jaeger, H.M.; Knight, J.B.; Nagel, S.R.; Karczmar, G.S.; Kuperman, V.Yu.

    1995-03-17

    Vibrations in a granular material can spontaneously produce convection rolls reminiscent of those seen in fluids. Magnetic resonance imaging provides a sensitive and noninvasive probe for the detection of these convection currents, which have otherwise been difficult to observe. A magnetic resonance imaging study of convection in a column of poppy seeds yielded data about the detailed shape of the convection rolls and the depth dependence of the convection velocity. The velocity was found to decrease exponentially with depth; a simple model for this behavior is presented here. 31 refs., 4 figs.

  20. Granular convection observed by magnetic resonance imaging

    NASA Astrophysics Data System (ADS)

    Ehrichs, E. E.; Jaeger, H. M.; Karczmar, Greg S.; Knight, James B.; Kuperman, Vadim Yu.; Nagel, Sidney R.

    1995-03-01

    Vibrations in a granular material can spontaneously produce convection rolls reminiscent of those seen in fluids. Magnetic resonance imaging provides a sensitive and noninvasive probe for the detection of these convection currents, which have otherwise been difficult to observe. A magnetic resonance imaging study of convection in a column of poppy seeds yielded data about the detailed shape of the convection rolls and the depth dependence of the convection velocity. The velocity was found to decrease exponentially with depth; a simple model for this behavior is presented here.

  1. Granular Segregation with Anisotropic Particles

    NASA Astrophysics Data System (ADS)

    Sykes, Tim

    2005-11-01

    The results from experimental investigations of horizontally vibrated mixtures of anisotropic poppy seeds and long chains of linked spheres will be presented. A critical packing fraction was observed to be required to initiate a transition to segregation. The average size of the resulting patterns was measured and the concentration ratio of the mixtures was varied by changing the number of chains present in the mixtures. A change in the order of the transition, from second to first order with associated hysteresis, was observed as the chain number was reduced. This gave rise to three distinct regions of behaviour: segregated, mixed and a bi-stable state.

  2. LHC II system sensitivity to magnetic fluids

    NASA Astrophysics Data System (ADS)

    Cotae, Vlad; Creanga, Ioan

    2005-03-01

    Experiments have been designed to reveal the influences of ferrofluid treatment and static magnetic field exposure on the photosynthetic system II, where the light harvesting complex (LHC II) controls the ratio chlorophyll a/ chlorophyll b (revealing, indirectly, the photosynthesis rate). Spectrophotometric measurement of chlorophyll content revealed different influences for relatively low ferrofluid concentrations (10-30 μl/l) in comparison to higher concentrations (70-100 μl/l). The overlapped effect of the static magnetic field shaped better the stimulatory ferrofluid action on LHC II system in young poppy plantlets.

  3. Self-organized patterns in collections of chains

    NASA Astrophysics Data System (ADS)

    Sykes, T.; Mullin, T.

    2009-11-01

    Results are presented of an experimental investigation into patterned segregation in thin layers of poppy seeds and short lengths of metal chains subjected to vibration. Critical phenomena are uncovered and both continuous and discontinuous transitions are observed. A phase diagram for the behavior is mapped out and a tricritical point that separates hysteretic from continuous segregation is identified. Remarkable similarities are found between the observed behavior in this driven granular system and phase separation phenomena in mixtures where the dynamics of the constituent components are markedly different.

  4. Elemental characterization of wild edible plants from countryside and urban areas.

    PubMed

    Renna, Massimiliano; Cocozza, Claudio; Gonnella, Maria; Abdelrahman, Hamada; Santamaria, Pietro

    2015-06-15

    Thirteen elements (Na, K, Ca, Mg, Fe, Mn, Cu, Zn, Cr, Co, Cd, Ni and Pb) in 11 different wild edible plants (WEP) (Amaranthus retroflexus, Foeniculum vulgare, Cichorium intybus, Glebionis coronaria, Sonchus spp., Borago officinalis, Diplotaxis tenuifolia, Sinapis arvensis, Papaver rhoeas, Plantago lagopus and Portulaca oleracea) collected from countryside and urban areas of Bari (Italy) were determined. B.officinalis and P.rhoeas could represent good nutritional sources of Mn and Fe, respectively, as well as A.retroflexus and S.arvensis for Ca. High intake of Pb and Cd could come from P.lagopus and A.retroflexus (1.40 and 0.13 mg kg(-1) FW, respectively). WEP may give a substantial contribution to the elements intake for consumers, but in some cases they may supply high level of elements potentially toxic for human health. Anyway, both ANOVA and PCA analyses have highlighted the low influence of the harvesting site on the elements content. PMID:25660854

  5. [The War on "Red Drugs": Anticommunism and Drug Policy in Republic of Korea, 1945-1960].

    PubMed

    Park, Ji-Young

    2016-04-01

    This paper investigates the discourses and policies on narcotics in Republic of Korea from 1945 to 1960. Since the Liberation the narcotic problem was regarded as the vestige of Japanese imperialism. which was expected to be cleaned up. The image of narcotic crimes as the legacy of the colonial past was turned into as the result of the Red Army's tactics to attack on the liberalist camp around the Korean war. The government of ROK represented the source of the illegal drugs as the Red army and the spy from North Korea. The anticommunist discourse about narcotics described the spies, who introduced the enormous amount of poppies into ROK and brought about the addicts, as the social evil. Through this discourse on poppies from North Korea, the government of ROK emphasized the immorality of the communists reinforcing the anticommunist regime, which was inevitable for the government of ROK to legitimize the division of Korea and the establishment of the government alone. This paper examines how the discourses and policies on narcotics in ROK was shaped and transformed from 1945 to 1960 focusing the relationship between the them and the political context such as anticommunism, Korean war, the division of Korea, and etc. This approach would be helpful to reveal the effect of the ROK's own political situation to the public health system involving the management for drugs. PMID:27301856

  6. Opium Field Detection in South Oxfordshire Using SAR Polarimetry

    NASA Astrophysics Data System (ADS)

    Walker, Nick; Marino, Armando

    2011-03-01

    To-date the use of satellite imagery to monitor the growth of illicit crops such as marijuana, opium and coca has mostly been conducted using optical frequencies. However, it is well known that while optical imagery can be hampered by localised aerosols such as thin clouds, cirrus, haze and smoke, these do not present a problem for Synthetic Aperture Radar (SAR). In recent years a new generation of satellite borne sensors have also been equipped with enhanced polarimetric capabilities, which can potentially help with detecting and classifying different terrain types. For these reasons we believe it is useful to consider whether high resolution polarimetric SAR data can be applied to illicit crop detection.In this paper we present the results of an experiment whereby opium poppy fields were successfully detected in the south Oxfordshire region in the UK using RadarSat-2 quad-polarisation imagery. It should be noted that these crops are not being grown illicitly but instead are being cultivated for medicinal reasons in parts of the UK. It is interesting to note that the poppies cultivated for opium in the UK have white flowers rather than the more familiar red as can be seen from the photograph in Figure 1, which was taken 11 days earlier in the season compared to Figure 4 and Figure 5.

  7. Synthesis of the skeleton of the morphine molecule by mammalian liver.

    PubMed

    Weitz, C J; Faull, K F; Goldstein, A

    The possibility that morphine could be synthesized in animals has long been considered and a pathway in mammalian brain analogous to that in the opium poppy has been proposed. Substances have been detected in mammalian brain that are recognized by antisera raised against morphine. Recently we reported the presence of three such immunoreactive substances in bovine hypothalamus and adrenal, and in rat brain, and the definitive identification of two of them by gas chromatography-mass spectrometry as morphine and codeine. Incorporation of a labelled precursor has demonstrated the biosynthesis of morphine in the opium poppy from tyrosine-derived units (see Fig. 1). Intramolecular coupling of reticuline to form salutaridine is the critical step that generates the morphine skeleton (morphinan) and the stereochemistry of the morphinan series. We now report the conversion in vivo and in vitro of reticuline to salutaridine by rat liver, but this conversion is not detectable in rat brain and bovine adrenal. This is the first direct demonstration of the synthesis of a morphinan in an animal tissue and also supports the hypothesis that morphine and codeine in brain and adrenal are of endogenous origin.

  8. Historical trends in the production and consumption of illicit drugs in Mexico: implications for the prevention of blood borne infections.

    PubMed

    Bucardo, Jesus; Brouwer, Kimberly C; Magis-Rodríguez, Carlos; Ramos, Rebeca; Fraga, Miguel; Perez, Saida G; Patterson, Thomas L; Strathdee, Steffanie A

    2005-09-01

    Mexico has cultivated opium poppy since before the 1900's and has been an important transit route for South American cocaine for decades. However, only recently has drug use, particularly injection drug use, been documented as an important problem. Heroin is the most common drug used by Mexican injection drug users (IDUs). Increased cultivation of opium poppy in some Mexican states, lower prices for black tar heroin and increased security at U.S.-Mexican border crossings may be contributing factors to heroin use, especially in border cities. Risky practices among IDUs, including needle sharing and shooting gallery attendance are common, whereas perceived risk for acquiring blood borne infections is low. Although reported AIDS cases attributed to IDU in Mexico have been low, data from sentinel populations, such as pregnant women in the Mexican-U.S. border city of Tijuana, suggest an increase in HIV prevalence associated with drug use. Given widespread risk behaviors and rising numbers of blood borne infections among IDUs in Mexican-U.S. border cities, there is an urgent need for increased disease surveillance and culturally appropriate interventions to prevent potential epidemics of blood borne infections. We review available literature on the history of opium production in Mexico, recent trends in drug use and its implications, and the Mexican response, with special emphasis on the border cities of Ciudad Juarez and Tijuana.

  9. Factors affecting the oil agglomeration of Sivas-Divrigi Ulucayir lignite

    SciTech Connect

    Unal, I.; Gorgun Ersan, M.

    2007-07-01

    In the coal industry, the coal particles need to be decreased to a very fine size because of the need of removing inorganic materials from coal. Oil agglomeration is a kind of coal cleaning technique that is used for separation of organic and inorganic parts of fine sized coal. In this study, the oil agglomeration of Sivas-Divrigi (S-D) Ulucayir lignite was carried out by using kerosene, diesel oil, fuel oil, poppy oil, and sunflower oil. The amount of bridging oil was varied from 5% to 25% of the amount of lignite. The effect of oil amount, oil type, solid content, agitation rate and time, pH on agglomeration performance was investigated. Maximum recovery value of 98.18% was observed by using poppy oil. In order to investigate the effect of pH on agglomeration NaOH and HCl is added to the slurry in various amounts. It is decided that the best agglomeration condition is obtained at low pH values. The effect of nonionic surface active agent (Igepal-CA 630) on agglomeration is investigated by adding to the slurry and it is observed that the grade is increased with the amount of surface active agent.

  10. Conservation and canalization of gene expression during angiosperm diversification accompany the origin and evolution of the flower.

    PubMed

    Chanderbali, André S; Yoo, Mi-Jeong; Zahn, Laura M; Brockington, Samuel F; Wall, P Kerr; Gitzendanner, Matthew A; Albert, Victor A; Leebens-Mack, James; Altman, Naomi S; Ma, Hong; dePamphilis, Claude W; Soltis, Douglas E; Soltis, Pamela S

    2010-12-28

    The origin and rapid diversification of the angiosperms (Darwin's "Abominable Mystery") has engaged generations of researchers. Here, we examine the floral genetic programs of phylogenetically pivotal angiosperms (water lily, avocado, California poppy, and Arabidopsis) and a nonflowering seed plant (a cycad) to obtain insight into the origin and subsequent evolution of the flower. Transcriptional cascades with broadly overlapping spatial domains, resembling the hypothesized ancestral gymnosperm program, are deployed across morphologically intergrading organs in water lily and avocado flowers. In contrast, spatially discrete transcriptional programs in distinct floral organs characterize the more recently derived angiosperm lineages represented by California poppy and Arabidopsis. Deep evolutionary conservation in the genetic programs of putatively homologous floral organs traces to those operating in gymnosperm reproductive cones. Female gymnosperm cones and angiosperm carpels share conserved genetic features, which may be associated with the ovule developmental program common to both organs. However, male gymnosperm cones share genetic features with both perianth (sterile attractive and protective) organs and stamens, supporting the evolutionary origin of the floral perianth from the male genetic program of seed plants. PMID:21149731

  11. Antimicrobial polyphenols from small tropical fruits, tea and spice oilseeds.

    PubMed

    Aman, Sahar; Naim, Asma; Siddiqi, Rahmanullah; Naz, Shahina

    2014-06-01

    The polyphenolic fractions of fruits: Terminalia catappa, Carissa carandas, Ziziphus nummularia; spice oilseeds: thymol, mustard, fenugreek and poppy seeds; and herb: green and black teas were analyzed for their total phenolics, flavonoids and antimicrobial potential. All fractions from fruits, except anthocyanin of C. carandas, displayed substantial antibacterial activity in accordance to their phenolic contents, the difference in activity being quite significant (p < 0.05), highest for T. catappa (minimum inhibitory concentration, MIC: 7.8-1000 microg/mL) and lowest for C. carandas (MIC: 62.5-1000 microg/mL). With few exceptions, both green and black teas' fractions inhibited the tested strains, however, green tea fractions (MIC: 15.63-125 microg/mL) were more active than black (MIC: 31.25-1000 microg/mL) and neutral were more active than their corresponding acidic fractions. Oil fractions of all oilseeds were found to be more active than their polyphenolic fractions, their antibacterial action decreased in the order thymol > mustard > fenugreek > poppy seeds (p < 0.05). Though the fruits used for the study are underutilized and have been emphasized for processed products, they may potentially be important to fight against pathogenic bacteria in view of their MICs. The teas and oilseeds, though a small part of total food intake, are more functional and active against the tested bacterial species and may find potential applications in therapeutics and food preservation.

  12. Anaphylaxis to buckwheat in an atopic child: a risk factor for severe allergy to nuts and seeds?

    PubMed

    Varga, Eva-Maria; Kollmann, Dagmar; Zach, Maximilian; Bohle, Barbara

    2011-01-01

    Common buckwheat (Fagopyrum esculentum) is known to cause severe anaphylactic reactions in adult individuals. However, type I allergy to buckwheat is rarely seen in children. We report on a 7-year-old boy who developed a grade III anaphylactic reaction after consumption of a cake containing buckwheat flour. Prior to this incident, the boy had developed severe allergic reactions to hazelnuts and suffered from an oral allergy syndrome to poppy seed. Analysis of the patient's IgE reactivity by immunoblotting experiments revealed that he was sensitized to members of the 2S albumin and 11S globulin protein families in buckwheat. Additionally, cross-reactivity was found between the 11S globulins in buckwheat, poppy and hazelnut. IgE inhibition experiments indicated that the 11S globulin in buckwheat was the initial sensitizing protein. We conclude that 11S globulins in buckwheat have the potential to induce IgE antibodies cross-reactive with 11S globulins in other, botanically unrelated foods and may induce anaphylactic reactions.

  13. Lethal Lullabies: A History of Opium Use in Infants.

    PubMed

    Obladen, Michael

    2016-02-01

    Poppy extract accompanied the human infant for more than 3 millenia. Motives for its use included excessive crying, suspected pain, and diarrhea. In antiquity, infantile sleeplessness was regarded as a disease. When treatment with opium was recommended by Galen, Rhazes, and Avicenna, baby sedation made its way into early medical treatises and pediatric instructions. Dabbing maternal nipples with bitter substances and drugging the infant with opium were used to hasten weaning. A freerider of gum lancing, opiates joined the treatment of difficult teething in the 17th century. Foundling hospitals and wet-nurses used them extensively. With industrialization, private use was rampant among the working class. In German-speaking countries, poppy extracts were administered in soups and pacifiers. In English-speaking countries, proprietary drugs containing opium were marketed under names such as soothers, nostrums, anodynes, cordials, preservatives, and specifics and sold at the doorstep or in grocery stores. Opium's toxicity for infants was common knowledge; thousands of cases of lethal intoxication had been reported from antiquity. What is remarkable is that the willingness to use it in infants persisted and that physicians continued to prescribe it for babies. Unregulated trade, and even that protected by governments, led to greatly increased private use of opiates during the 19th century. Intoxication became a significant factor in infant mortality. As late as 1912, the International Hague Convention forced governments to implement legislation that effectively curtailed access to opium and broke the dangerous habit of sedating infants.

  14. Conservation and canalization of gene expression during angiosperm diversification accompany the origin and evolution of the flower.

    PubMed

    Chanderbali, André S; Yoo, Mi-Jeong; Zahn, Laura M; Brockington, Samuel F; Wall, P Kerr; Gitzendanner, Matthew A; Albert, Victor A; Leebens-Mack, James; Altman, Naomi S; Ma, Hong; dePamphilis, Claude W; Soltis, Douglas E; Soltis, Pamela S

    2010-12-28

    The origin and rapid diversification of the angiosperms (Darwin's "Abominable Mystery") has engaged generations of researchers. Here, we examine the floral genetic programs of phylogenetically pivotal angiosperms (water lily, avocado, California poppy, and Arabidopsis) and a nonflowering seed plant (a cycad) to obtain insight into the origin and subsequent evolution of the flower. Transcriptional cascades with broadly overlapping spatial domains, resembling the hypothesized ancestral gymnosperm program, are deployed across morphologically intergrading organs in water lily and avocado flowers. In contrast, spatially discrete transcriptional programs in distinct floral organs characterize the more recently derived angiosperm lineages represented by California poppy and Arabidopsis. Deep evolutionary conservation in the genetic programs of putatively homologous floral organs traces to those operating in gymnosperm reproductive cones. Female gymnosperm cones and angiosperm carpels share conserved genetic features, which may be associated with the ovule developmental program common to both organs. However, male gymnosperm cones share genetic features with both perianth (sterile attractive and protective) organs and stamens, supporting the evolutionary origin of the floral perianth from the male genetic program of seed plants.

  15. Historical trends in the production and consumption of illicit drugs in Mexico: Implications for the prevention of blood borne infections

    PubMed Central

    Bucardo, Jesus; Brouwer, Kimberly C.; Magis-Rodríguez, Carlos; Ramos, Rebeca; Fraga, Miguel; Perez, Saida G.; Patterson, Thomas L.; Strathdee, Steffanie A.

    2007-01-01

    Mexico has cultivated opium poppy since before the 1900’s and has been an important transit route for South American cocaine for decades. However, only recently has drug use, particularly injection drug use, been documented as an important problem. Heroin is the most common drug used by Mexican injection drug users (IDUs). Increased cultivation of opium poppy in some Mexican states, lower prices for black tar heroin and increased security at U.S.-Mexican border crossings may be contributing factors to heroin use, especially in border cities. Risky practices among IDUs, including needle sharing and shooting gallery attendance are common, whereas perceived risk for acquiring blood borne infections is low. Although reported AIDS cases attributed to IDU in Mexico have been low, data from sentinel populations, such as pregnant women in the Mexican-U.S. border city of Tijuana, suggest an increase in HIV prevalence associated with drug use. Given widespread risk behaviors and rising numbers of blood borne infections among IDUs in Mexican-U.S. border cities, there is an urgent need for increased disease surveillance and culturally appropriate interventions to prevent potential epidemics of blood borne infections. We review available literature on the history of opium production in Mexico, recent trends in drug use and its implications, and the Mexican response, with special emphasis on the border cities of Ciudad Juarez and Tijuana. PMID:16102372

  16. Lethal Lullabies: A History of Opium Use in Infants.

    PubMed

    Obladen, Michael

    2016-02-01

    Poppy extract accompanied the human infant for more than 3 millenia. Motives for its use included excessive crying, suspected pain, and diarrhea. In antiquity, infantile sleeplessness was regarded as a disease. When treatment with opium was recommended by Galen, Rhazes, and Avicenna, baby sedation made its way into early medical treatises and pediatric instructions. Dabbing maternal nipples with bitter substances and drugging the infant with opium were used to hasten weaning. A freerider of gum lancing, opiates joined the treatment of difficult teething in the 17th century. Foundling hospitals and wet-nurses used them extensively. With industrialization, private use was rampant among the working class. In German-speaking countries, poppy extracts were administered in soups and pacifiers. In English-speaking countries, proprietary drugs containing opium were marketed under names such as soothers, nostrums, anodynes, cordials, preservatives, and specifics and sold at the doorstep or in grocery stores. Opium's toxicity for infants was common knowledge; thousands of cases of lethal intoxication had been reported from antiquity. What is remarkable is that the willingness to use it in infants persisted and that physicians continued to prescribe it for babies. Unregulated trade, and even that protected by governments, led to greatly increased private use of opiates during the 19th century. Intoxication became a significant factor in infant mortality. As late as 1912, the International Hague Convention forced governments to implement legislation that effectively curtailed access to opium and broke the dangerous habit of sedating infants. PMID:26163533

  17. [The War on "Red Drugs": Anticommunism and Drug Policy in Republic of Korea, 1945-1960].

    PubMed

    Park, Ji-Young

    2016-04-01

    This paper investigates the discourses and policies on narcotics in Republic of Korea from 1945 to 1960. Since the Liberation the narcotic problem was regarded as the vestige of Japanese imperialism. which was expected to be cleaned up. The image of narcotic crimes as the legacy of the colonial past was turned into as the result of the Red Army's tactics to attack on the liberalist camp around the Korean war. The government of ROK represented the source of the illegal drugs as the Red army and the spy from North Korea. The anticommunist discourse about narcotics described the spies, who introduced the enormous amount of poppies into ROK and brought about the addicts, as the social evil. Through this discourse on poppies from North Korea, the government of ROK emphasized the immorality of the communists reinforcing the anticommunist regime, which was inevitable for the government of ROK to legitimize the division of Korea and the establishment of the government alone. This paper examines how the discourses and policies on narcotics in ROK was shaped and transformed from 1945 to 1960 focusing the relationship between the them and the political context such as anticommunism, Korean war, the division of Korea, and etc. This approach would be helpful to reveal the effect of the ROK's own political situation to the public health system involving the management for drugs.

  18. Biochemistry and Occurrence of O-Demethylation in Plant Metabolism

    PubMed Central

    Hagel, Jillian M.; Facchini, Peter J.

    2010-01-01

    Demethylases play a pivitol role in numerous biological processes from covalent histone modification and DNA repair to specialized metabolism in plants and microorganisms. Enzymes that catalyze O- and N-demethylation include 2-oxoglutarate (2OG)/Fe(II)-dependent dioxygenases, cytochromes P450, Rieske-domain proteins and flavin adenine dinucleotide (FAD)-dependent oxidases. Proposed mechanisms for demethylation by 2OG/Fe(II)-dependent enzymes involve hydroxylation at the O- or N-linked methyl group followed by formaldehyde elimination. Members of this enzyme family catalyze a wide variety of reactions in diverse plant metabolic pathways. Recently, we showed that 2OG/Fe(II)-dependent dioxygenases catalyze the unique O-demethylation steps of morphine biosynthesis in opium poppy, which provides a rational basis for the widespread occurrence of demethylases in benzylisoquinoline alkaloid metabolism. PMID:21423357

  19. Isolation and Partial Characterization of a New Lectin from Seeds of the Greater Celandine (Chelidonium majus) 1

    PubMed Central

    Peumans, Willy J.; De Ley, Marc; Stinissen, Hetty M.; Broekaert, Willem F.

    1985-01-01

    Seeds of the greater celandine (Chelidonium majus L.) contain a lectin which could be isolated using a combination of affinity chromatography on chitin and ion exchange chromatography on sulphopropyl-Sephadex. The purified lectin was partially characterized with respect to its biochemical and physicochemical properties. It is a small dimeric protein composed of two different subunits of Mr 9,500 and 11,500, respectively. Its amino acid composition is typified by high contents of glycine and cysteine. No covalently bound carbohydrate could be detected. Hapten inhibition experiments indicated that the lectin exhibits specificity towards oligomers of N-acetylglucosamine, the potency of inhibition increasing with chain length up to four residues. The greater celandine lectin is the first lectin to be isolated from a species belonging to the plant family Papaveraceae (poppy family). Although it represents a new type of plant lectin, resemblances to phytohemaglutinins from diverse taxonomic origin are obvious. Images Fig. 2 PMID:16664249

  20. [Milk of paradise? Opium and opiates in nineteenth and twentieth century literature].

    PubMed

    Schäfer, D

    2007-08-01

    One cannot have an idea of this multifaceted theme without its medical and cultural-historical background. After a history of several thousand years as a remedy and consumer good, around 1800 this poppy drug was in the focus of public attention due to Brownianism, at first as an often self-prescribed unspecific remedy against physical and mental pain. Many representatives of the early Romanticism knew it from personal experience. However, it was the publication of Thomas de Quincey's Confessions of an English Opium-Eater (1821/1822) which made it a subject of international debate in accordance with the programmatic statements of writers of that epoque and corresponding to the antibourgeois attitude of these men. It became a motif of a counter-world experience and a subject and cause of lyric-subjective reflection as well as a possible premise of poetic creativity. PMID:17593402

  1. Detection of sesame seed DNA in foods using real-time PCR.

    PubMed

    Brzezinski, Jennifer L

    2007-04-01

    The detection of potentially allergenic foods, such as sesame seeds, in food products is a major concern for the food-processing industry. A real-time PCR method was designed to determine if sesame seed DNA is present in food products. The PCR reaction amplifies a 66-bp fragment of the sesame seed 2S albumin gene, which is detected with a sesame-specific, dual-labeled TaqMan probe. This reaction will not amplify DNA derived from other seeds present in baked goods, such as pumpkin, poppy, and sunflower seeds. Additionally, this assay will not cross-react with DNA from several tree nut species, such as almond, Brazil nut, cashew, hazelnut, and walnut, as well as four varieties of peanut. This assay is sensitive enough to detect 5 pg of purified sesame seed DNA, as well as sesame seed DNA in a spiked wheat cracker sample.

  2. Papaverine intoxication in a newborn: an unusual case report.

    PubMed

    Kahramaner, Zelal; Erdemir, Aydın; Türkoğlu, Ebru; Coşar, Hese; Sütçüoğlu, Sümer; Özer, Esra Arun

    2014-01-01

    Herbal agents are increasingly used for medicinal purposes, but there is a lack of knowledge about the content of these agents. Indiscriminate use of herbal agents may cause severe side effects and also death. We report a newborn who developed convulsions and respiratory arrest after oral intake of an opium poppy preparation containing papaverine for its antitussive effect. The infant experienced a good outcome with supportive treatment. To the best of our knowledge, this is the first time a newborn with papaverine intoxication has been described. Parents should avoid self-medication of their children, and the possibility of exposure to foreign products should be kept in mind in any seizure of a newborn with unexplained origin.

  3. Mammalian opiate alkaloid synthesis: lessons derived from plant biochemistry.

    PubMed

    Meijerink, W J; Molina, P E; Abumrad, N N

    1999-09-01

    The presence of opiate receptors in mammalian tissues has stimulated the search for endogenous ligands to these receptors and has led to the discovery and characterization of endogenous opioid peptides. However, recent studies have provided evidence for the presence of opiate alkaloids in mammalian tissues and for their endogenous synthesis. The study of their origin and synthetic pathway has been significantly influenced by the early classical biochemical studies performed in plants. This review is a historical account of the use and abuse of opiates, the elucidation of morphine's synthetic pathway in the poppy plant, and the subsequent characterization of its presence in mammalian tissues. Clearly, our understanding of its synthetic pathway and regulation is a reflection of observations originally made in plant biochemistry.

  4. Total biosynthesis of opiates by stepwise fermentation using engineered Escherichia coli.

    PubMed

    Nakagawa, Akira; Matsumura, Eitaro; Koyanagi, Takashi; Katayama, Takane; Kawano, Noriaki; Yoshimatsu, Kayo; Yamamoto, Kenji; Kumagai, Hidehiko; Sato, Fumihiko; Minami, Hiromichi

    2016-02-05

    Opiates such as morphine and codeine are mainly obtained by extraction from opium poppies. Fermentative opiate production in microbes has also been investigated, and complete biosynthesis of opiates from a simple carbon source has recently been accomplished in yeast. Here we demonstrate that Escherichia coli serves as an efficient, robust and flexible platform for total opiate synthesis. Thebaine, the most important raw material in opioid preparations, is produced by stepwise culture of four engineered strains at yields of 2.1 mg l(-1) from glycerol, corresponding to a 300-fold increase from recently developed yeast systems. This improvement is presumably due to strong activity of enzymes related to thebaine synthesis from (R)-reticuline in E. coli. Furthermore, by adding two genes to the thebaine production system, we demonstrate the biosynthesis of hydrocodone, a clinically important opioid. Improvements in opiate production in this E. coli system represent a major step towards the development of alternative opiate production systems.

  5. UN/Thai programme for drug abuse control in Thailand-A report on phase I:February 1972-June 1979.

    PubMed

    Williams, I M

    1979-01-01

    The paper describes the principal achievements of the UN/Thai programme for drug abuse control which was carried out during the years 1972-1979. The programme was of a pilot nature and covered five selected key villages and 25 satellite villages in the north of Thailand. The major components of the programme were crop replacement and community development, treatment and rehabilitation, and drug information and education. The Royal Thai Government has accepted the achievements of this project as models for greatly expanded programmes for the eradication of the opium poppy cultivation. Two key villages-Ban Mae Tho and Hui Tund Jaw which were included in a pilot programme have provided to be opium free.

  6. Treatment of depression.

    PubMed

    Payk, T R

    1994-10-01

    Depressions are the most common psychiatric diseases. For treatment, plant extracts have been used for thousands of years: examples are extracts from the (sleeping) poppy (opium), deadly nightshade (Atropa belladonna), Indian hemp (hashish), henbane (hyoscyamine), thorn apple (scopolamine), and St. John's wort (hypericum oil). In addition, psychotherapeutic measures, like playing music, dancing, playing theatre, and also the temple sleep, were used. In the 19th century, the introduction of brome (1826), codeine (1832), chloral hydrate (1869), and paraldehyde (1882), as well as the barbiturates (at the turn of the century) introduced significant improvements in pharmacotherapy. The modern thymoleptica therapy started in 1957 with the introduction of imipramine. Now about 40 active antidepressants are marketed. New drug developments should be characterized mainly by an improvement in tolerance.

  7. Studying Ancient Anthropogenic Impacts on Current Floral Biodiversity in the Southern Levant as reflected by the Philistine Migration

    PubMed Central

    Frumin, Suembikya; Maeir, Aren M.; Kolska Horwitz, Liora; Weiss, Ehud

    2015-01-01

    Human migrations across geographic boundaries can facilitate the introduction of new husbandry practices and dispersal of plants and animals, resulting in changes in biodiversity. As previously demonstrated, the 12th century BCE Philistine migration–to the southern Levantine littoral, involved the transportation of pigs from Europe, engendering long term genetic displacement of local Near Eastern haplotypes. Building on this, and combining biogeographical methods of Floral List comparisons with archaeological data, we have elucidated the Philistine impact on Southern Levantine floral ecosystems. We demonstrate that previously unexploited local plants were incorporated into the Philistine milieu, and new species were introduced–from Europe, the Aegean, Egypt and Mesopotamia –resulting in the earliest locally cultivated sycamore, cumin, coriander, bay tree and opium poppy. This research has highlighted the impact of past cultures on the formation of floral ecosystems and their long-term effects on contemporary local biological diversity. PMID:26304818

  8. Total biosynthesis of opiates by stepwise fermentation using engineered Escherichia coli

    PubMed Central

    Nakagawa, Akira; Matsumura, Eitaro; Koyanagi, Takashi; Katayama, Takane; Kawano, Noriaki; Yoshimatsu, Kayo; Yamamoto, Kenji; Kumagai, Hidehiko; Sato, Fumihiko; Minami, Hiromichi

    2016-01-01

    Opiates such as morphine and codeine are mainly obtained by extraction from opium poppies. Fermentative opiate production in microbes has also been investigated, and complete biosynthesis of opiates from a simple carbon source has recently been accomplished in yeast. Here we demonstrate that Escherichia coli serves as an efficient, robust and flexible platform for total opiate synthesis. Thebaine, the most important raw material in opioid preparations, is produced by stepwise culture of four engineered strains at yields of 2.1 mg l−1 from glycerol, corresponding to a 300-fold increase from recently developed yeast systems. This improvement is presumably due to strong activity of enzymes related to thebaine synthesis from (R)-reticuline in E. coli. Furthermore, by adding two genes to the thebaine production system, we demonstrate the biosynthesis of hydrocodone, a clinically important opioid. Improvements in opiate production in this E. coli system represent a major step towards the development of alternative opiate production systems. PMID:26847395

  9. Complete biosynthesis of opioids in yeast

    PubMed Central

    Galanie, Stephanie; Thodey, Kate; Trenchard, Isis J.; Interrante, Maria Filsinger; Smolke, Christina D.

    2016-01-01

    Opioids are the primary drugs used in Western medicine for pain management and palliative care. Farming of opium poppies remains the sole source of these essential medicines despite diverse market demands and uncertainty in crop yields due to weather, climate change, and pests. Here, we engineered yeast to produce the selected opioid compounds thebaine and hydrocodone starting from sugar. All work was conducted in a laboratory that is permitted and secured for work with controlled substances. We combined enzyme discovery, enzyme engineering, and pathway and strain optimization to realize full opiate biosynthesis in yeast. The resulting opioid biosynthesis strains required expression of 21 (thebaine) and 23 (hydrocodone) enzyme activities from plants, mammals, bacteria, and yeast itself. This is a proof-of-principle, and major hurdles remain before optimization and scale up could be achieved. Open discussions of options for governing this technology are also needed in order to responsibly realize alternative supplies for these medically relevant compounds. PMID:26272907

  10. Drying of solids in fluidized beds

    SciTech Connect

    Kannan, C.S.; Thomas, P.P.; Varma, Y.B.G.

    1995-09-01

    Fluidized bed drying is advantageously adopted in industrial practice for drying of granular solids such as grains, fertilizers, chemicals, and minerals either for long shelf life or to facilitate further processing or handling. Solids are dried in batch and in continuous fluidized beds corresponding to cross-flow and countercurrent flow of phases covering a wide range in drying conditions. Materials that essentially dry with constant drying rate and then give a falling drying rate approximately linear with respect to solids moisture content (sand) as well as those with an extensive falling rate period with the subsequent falling rate being a curve with respect to the moisture content (mustard, ragi, poppy seeds) are chosen for the study. The performance of the continuous fluidized bed driers is compared with that of batch fluidized bed driers; the performance is predicted using batch kinetics, the residence time distribution of solids, and the contact efficiency between the phases.

  11. Natural products for chronic cough: Text mining the East Asian historical literature for future therapeutics.

    PubMed

    Shergis, Johannah Linda; Wu, Lei; May, Brian H; Zhang, Anthony Lin; Guo, Xinfeng; Lu, Chuanjian; Xue, Charlie Changli

    2015-08-01

    Chronic cough is a significant health burden. Patients experience variable benefits from over the counter and prescribed products, but there is an unmet need to provide more effective treatments. Natural products have been used to treat cough and some plant compounds such as pseudoephedrine from ephedra and codeine from opium poppy have been developed into drugs. Text mining historical literature may offer new insight for future therapeutic development. We identified natural products used in the East Asian historical literature to treat chronic cough. Evaluation of the historical literature revealed 331 natural products used to treat chronic cough. Products included plants, minerals and animal substances. These natural products were found in 75 different books published between AD 363 and 1911. Of the 331 products, the 10 most frequently and continually used products were examined, taking into consideration findings from contemporary experimental studies. The natural products identified are promising and offer new directions in therapeutic development for treating chronic cough.

  12. Studying Ancient Anthropogenic Impacts on Current Floral Biodiversity in the Southern Levant as reflected by the Philistine Migration.

    PubMed

    Frumin, Suembikya; Maeir, Aren M; Kolska Horwitz, Liora; Weiss, Ehud

    2015-08-25

    Human migrations across geographic boundaries can facilitate the introduction of new husbandry practices and dispersal of plants and animals, resulting in changes in biodiversity. As previously demonstrated, the 12th century BCE Philistine migration-to the southern Levantine littoral, involved the transportation of pigs from Europe, engendering long term genetic displacement of local Near Eastern haplotypes. Building on this, and combining biogeographical methods of Floral List comparisons with archaeological data, we have elucidated the Philistine impact on Southern Levantine floral ecosystems. We demonstrate that previously unexploited local plants were incorporated into the Philistine milieu, and new species were introduced-from Europe, the Aegean, Egypt and Mesopotamia -resulting in the earliest locally cultivated sycamore, cumin, coriander, bay tree and opium poppy. This research has highlighted the impact of past cultures on the formation of floral ecosystems and their long-term effects on contemporary local biological diversity.

  13. Evolution of the local packing density in a sheared granular material

    NASA Astrophysics Data System (ADS)

    Umbanhowar, Paul; van Hecke, Martin; Sakaie, Ken

    2005-11-01

    The local particle packing density, η, in a bed of poppy seeds sheared in a vertical, split-bottom cylindrical container is measured using magnetic resonance imaging. Wide shear zones away from the wall are generated when a thin concentric disk at the bottom of the cylinder, and with radius less than the cylinder radius, is rotated slowly. Experiments reveal that, initially, a shear band forms at the outer edge of the disk which decreases in radius with decreasing depth. Material near the shear zone dilates with a corresponding decrease in η of about 15%. The radial extent of the dilated region increases approximately logarithmically with increasing rotation in shallow layers. In relatively deep layers, the shear zone is at first entirely below the free surface, but with further rotation grows vertically creating a low η core. We discuss the relation between the previously reported universal velocity profiles across the shear zone and the spatial and temporal evolution of the packing density.

  14. Application of k- and q-space encoding NMR techniques on granular media in a 3D model fluidized bed reactor

    NASA Astrophysics Data System (ADS)

    Harms, Silke; Stapf, Siegfried; Blümich, Bernhard

    2006-02-01

    A combination of PFG-NMR imaging and velocity encoding methods was applied to investigate the dynamic behavior of a bed of poppy seeds subjected to air flow, representing a model setup for fluidized bed reactors. The particle motion is described both from a statistical point of view, by determining propagators and dispersion coefficients representing an average over the whole bed volume, as well as combined with spatial resolution by generating velocity maps. Velocity images of different horizontal slices in the bed confirm the notion of a toroidal particle flow pattern inside the shallow granular bed. Despite the need of considerable averaging due to the random motion of the relatively few particles in the bed, quantitative velocity images and statistical information about the random particle motion can be obtained from monitoring the fluid component in the seeds by conventional spin-echo techniques.

  15. [The use of opium in Roman society and the dependence of Princeps Marcus Aurelius].

    PubMed

    Trancas, Bruno; Borja Santos, Nuno; Patrício, Luís D

    2008-01-01

    Opium was known and frequently used in Roman society. Medical practice recognized its usefulness as an analgesic, soporific, anti-tussic or anti-diarrheic agent, as well as other currently unsupported uses with quasi-magical properties. It was additionally used as an ingredient in antidotes, panaceas and poisons. The authors present a non-exhaustive compilation of opium use according to medical doctors, writers and encyclopaedists of the time. Mythological and literary representations of the opium poppy reflected its diverse roles, being associated with prosperity and fertility, sleep, death and the underworld and with the art of medicine. Despite its free and routine use, there is no solid evidence of addiction, except the putative case of emperor Marcus Aurelius, consistently reported as one of the most likely cases of addiction to opium.

  16. Complete biosynthesis of opioids in yeast.

    PubMed

    Galanie, Stephanie; Thodey, Kate; Trenchard, Isis J; Filsinger Interrante, Maria; Smolke, Christina D

    2015-09-01

    Opioids are the primary drugs used in Western medicine for pain management and palliative care. Farming of opium poppies remains the sole source of these essential medicines, despite diverse market demands and uncertainty in crop yields due to weather, climate change, and pests. We engineered yeast to produce the selected opioid compounds thebaine and hydrocodone starting from sugar. All work was conducted in a laboratory that is permitted and secured for work with controlled substances. We combined enzyme discovery, enzyme engineering, and pathway and strain optimization to realize full opiate biosynthesis in yeast. The resulting opioid biosynthesis strains required the expression of 21 (thebaine) and 23 (hydrocodone) enzyme activities from plants, mammals, bacteria, and yeast itself. This is a proof of principle, and major hurdles remain before optimization and scale-up could be achieved. Open discussions of options for governing this technology are also needed in order to responsibly realize alternative supplies for these medically relevant compounds. PMID:26272907

  17. Total biosynthesis of opiates by stepwise fermentation using engineered Escherichia coli.

    PubMed

    Nakagawa, Akira; Matsumura, Eitaro; Koyanagi, Takashi; Katayama, Takane; Kawano, Noriaki; Yoshimatsu, Kayo; Yamamoto, Kenji; Kumagai, Hidehiko; Sato, Fumihiko; Minami, Hiromichi

    2016-01-01

    Opiates such as morphine and codeine are mainly obtained by extraction from opium poppies. Fermentative opiate production in microbes has also been investigated, and complete biosynthesis of opiates from a simple carbon source has recently been accomplished in yeast. Here we demonstrate that Escherichia coli serves as an efficient, robust and flexible platform for total opiate synthesis. Thebaine, the most important raw material in opioid preparations, is produced by stepwise culture of four engineered strains at yields of 2.1 mg l(-1) from glycerol, corresponding to a 300-fold increase from recently developed yeast systems. This improvement is presumably due to strong activity of enzymes related to thebaine synthesis from (R)-reticuline in E. coli. Furthermore, by adding two genes to the thebaine production system, we demonstrate the biosynthesis of hydrocodone, a clinically important opioid. Improvements in opiate production in this E. coli system represent a major step towards the development of alternative opiate production systems. PMID:26847395

  18. Strategies to Improve Bioavailability and In Vivo Efficacy of the Endogenous Opioid Peptides Endomorphin-1 and Endomorphin-2.

    PubMed

    De Marco, Rossella; Janecka, Anna

    2015-01-01

    Morphine and the other alkaloids found in the opium poppy plant still represent the preferred therapeutic tools to treat severe pain in first aid protocols, as well as chronic pain. The use of the opiate alkaloids is accompanied by several unwanted side effects; additionally, some forms of pain are resistant to standard treatments (e.g. neuropathic pain from cancer). For these reasons, there is currently renewed interest in the design and assay of modified versions of the potent endogenous opioid peptides endomorphin-1 and endomorphin-2. This review presents a selection of the strategies directed at preparing highly stable peptidomimetics of the endomorphins, and of the strategies aimed at improving central nervous system bioavailability, for which increased in vivo antinociceptive efficacy was clearly demonstrated. PMID:26279081

  19. Studying Ancient Anthropogenic Impacts on Current Floral Biodiversity in the Southern Levant as reflected by the Philistine Migration.

    PubMed

    Frumin, Suembikya; Maeir, Aren M; Kolska Horwitz, Liora; Weiss, Ehud

    2015-01-01

    Human migrations across geographic boundaries can facilitate the introduction of new husbandry practices and dispersal of plants and animals, resulting in changes in biodiversity. As previously demonstrated, the 12th century BCE Philistine migration-to the southern Levantine littoral, involved the transportation of pigs from Europe, engendering long term genetic displacement of local Near Eastern haplotypes. Building on this, and combining biogeographical methods of Floral List comparisons with archaeological data, we have elucidated the Philistine impact on Southern Levantine floral ecosystems. We demonstrate that previously unexploited local plants were incorporated into the Philistine milieu, and new species were introduced-from Europe, the Aegean, Egypt and Mesopotamia -resulting in the earliest locally cultivated sycamore, cumin, coriander, bay tree and opium poppy. This research has highlighted the impact of past cultures on the formation of floral ecosystems and their long-term effects on contemporary local biological diversity. PMID:26304818

  20. Natural products for chronic cough: Text mining the East Asian historical literature for future therapeutics.

    PubMed

    Shergis, Johannah Linda; Wu, Lei; May, Brian H; Zhang, Anthony Lin; Guo, Xinfeng; Lu, Chuanjian; Xue, Charlie Changli

    2015-08-01

    Chronic cough is a significant health burden. Patients experience variable benefits from over the counter and prescribed products, but there is an unmet need to provide more effective treatments. Natural products have been used to treat cough and some plant compounds such as pseudoephedrine from ephedra and codeine from opium poppy have been developed into drugs. Text mining historical literature may offer new insight for future therapeutic development. We identified natural products used in the East Asian historical literature to treat chronic cough. Evaluation of the historical literature revealed 331 natural products used to treat chronic cough. Products included plants, minerals and animal substances. These natural products were found in 75 different books published between AD 363 and 1911. Of the 331 products, the 10 most frequently and continually used products were examined, taking into consideration findings from contemporary experimental studies. The natural products identified are promising and offer new directions in therapeutic development for treating chronic cough. PMID:25901012

  1. Papaverine intoxication in a newborn: an unusual case report.

    PubMed

    Kahramaner, Zelal; Erdemir, Aydın; Türkoğlu, Ebru; Coşar, Hese; Sütçüoğlu, Sümer; Özer, Esra Arun

    2014-01-01

    Herbal agents are increasingly used for medicinal purposes, but there is a lack of knowledge about the content of these agents. Indiscriminate use of herbal agents may cause severe side effects and also death. We report a newborn who developed convulsions and respiratory arrest after oral intake of an opium poppy preparation containing papaverine for its antitussive effect. The infant experienced a good outcome with supportive treatment. To the best of our knowledge, this is the first time a newborn with papaverine intoxication has been described. Parents should avoid self-medication of their children, and the possibility of exposure to foreign products should be kept in mind in any seizure of a newborn with unexplained origin. PMID:26022590

  2. CjbHLH1 homologs regulate sanguinarine biosynthesis in Eschscholzia californica cells.

    PubMed

    Yamada, Yasuyuki; Motomura, Yukiya; Sato, Fumihiko

    2015-05-01

    Isoquinoline alkaloids (IQAs), terpenoid indole alkaloid and nicotine are some of the most studied alkaloids. Recently, several groups have reported that the biosynthesis of these alkaloids is regulated by basic helix-loop-helix (bHLH) transcription factors. Whereas the biosyntheses of nicotine and terpenoid indole alkaloid in Nicotiana plants and Catharanthus roseus are directly or indirectly regulated by Arabidopsis thaliana MYC2 homologs, a non-MYC2-type bHLH transcription factor, CjbHLH1, comprehensively regulates berberine biosynthesis in Coptis japonica. Interestingly, CjbHLH1 homologous genes were found in many IQA-producing plant species, which suggests that non-MYC2-type CjbHLH homologs are specifically associated with IQA biosynthesis. To test whether CjbHLH1 homologs are involved in the biosynthesis of IQA in a plant other than C. japonica, we isolated two genes homologous to CjbHLH1, i.e. EcbHLH1-1 and EcbHLH1-2, from Eschscholzia californica (California poppy). Stable transformants in which the expression levels of EcbHLH1 genes were constitutively suppressed by RNA interference (RNAi) showed a reduced expression of some IQA biosynthetic enzyme genes. A metabolite analysis confirmed that the suppression of EcbHLH1, particularly EcbHLH1-2, caused a decrease in sanguinarine accumulation in transgenic cultured cells. These results indicate that non-MYC2-type EcbHLH1s regulate IQA biosynthesis in California poppy like CjbHLH1 in C. japonica.

  3. Characterization of rapeseed (Brassica napus) oils by bulk C, O, H, and fatty acid C stable isotope analyses.

    PubMed

    Richter, Eva Katharina; Spangenberg, Jorge E; Kreuzer, Michael; Leiber, Florian

    2010-07-14

    Rapeseed ( Brassica napus ) oils differing in cultivar, sites of growth, and harvest year were characterized by fatty acid concentrations and carbon, hydrogen, and oxygen stable isotope analyses of bulk oils (delta(13)C(bulk), delta(2)H(bulk), delta(18)O(bulk) values) and individual fatty acids (delta(13)C(FA)). The delta(13)C(bulk), delta(2)H(bulk), and delta(18)O(bulk) values were determined by continuous flow combustion and high-temperature conversion elemental analyzer-isotope ratio mass spectrometry (EA/IRMS, TC-EA/IRMS). The delta(13)C(FA) values were determined using gas chromatography--combustion-isotope ratio mass spectrometry (GC/C/IRMS). For comparison, other C(3) vegetable oils rich in linolenic acid (flax and false flax oils) and rich in linoleic acid (poppy, sunflower, and safflower oils) were submitted to the same chemical and isotopic analyses. The bulk and molecular delta(13)C values were typical for C(3) plants. The delta(13)C value of palmitic acid (delta(13)C(16:0)) and n-3 alpha-linolenic acid (delta(13)C(18:3n-3)) differed (p < 0.001) between rape, flax, and poppy oils. Also within species, significant differences of delta(13)C(FA) were observed (p < 0.01). The hydrogen and oxygen isotope compositions of rape oil differed between cultivars (p < 0.05). Major differences in the individual delta(13)C(FA) values were found. A plant-specific carbon isotope fractionation occurs during the biosynthesis of the fatty acids and particularly during desaturation of C(18) acids in rape and flax. Bulk oil and specific fatty acid stable isotope analysis might be useful in tracing dietary lipids differing in their origin.

  4. Reclamation and groundwater remediation at a hydrocarbon site in Alaska

    SciTech Connect

    Ririe, G.T.; Drake, L.D.; Olson, S.S.

    1997-12-31

    As part of a joint hydrocarbon cleanup project between Unocal and Marathon, we have initiated the use of constructed wetlands for restoration of the 40-acre Poppy Lane gravel pit located near Kenai, Alaska. Gravel excavated from this site was used to construct roads and drilling pads in the 1960`-70`s. During this period it was also used as a refuse dump for waste from the Kenai gas field and from local residents. The bulk wastes were removed and pockets of oily sand were removed, treated and returned to a stockpile on the site. This left the site with residual pockets of hydrocarbon-impacted sand (<1000 TPH) plus traces of hydrocarbon contamination in the uppermost shallow groundwater flowing through the outwash gravels. The final part of the cleanup will be land restoration and bioremediation of the final traces of hydrocarbons, which are predominantly diesel-range. High resolution gas chromatography analysis indicated that common plants already growing on the site (willow, cottonwood, and alder) did not concentrate diesel-range petroleum hydrocarbons in their foliage when growing in soils containing these contaminants. As part of the planned restoration and shallow groundwater remediation, two 1/3 acre test plots were constructed to promote in-situ biodegradation processes. In spring 1995, the first test, a tree root-barrier plot, was planted with dormant cuttings of four native wetland tree and shrub species, which were planted to depths up to five feet. Alder and elderberry did not succeed under any conditions, nor did any species planted in standing water. For cottonwood and willow species, approximately one half of each rooted and survived. When the water table dropped the second year, the willow cuttings rooted deeper in the vadose zone, while cottonwood did not. As a result of these findings, a tree root-barrier wetland is not considered to be a viable option for groundwater treatment at Poppy Lane.

  5. Influence of volume fraction on the dynamics of granular impact

    NASA Astrophysics Data System (ADS)

    Umbanhowar, Paul; Yang, Ding; Goldman, Daniel

    2008-11-01

    Variation of the volume fraction φ of non-cohesive granular media causes disproportionate changes in the forces exerted on impacting objects and, consequently, the impact kinematics. In our experiments, a computer controlled air fluidized granular bed is used to vary φ from 0.58 (low) to 0.62 (high) for 0.3 mm diameter glass spheres and 1̃ mm poppy seeds. An accelerometer attached to a 4.0 cm diameter steel sphere measures collision forces for initial impact velocities ranging from 0.5 to 3.5 m/s. As an example of the dramatic changes produced by varying φ, time series of the force during impact with poppy seeds at an impact velocity of 1 m/s change from monotonically increasing with slope 100 N/s at φ=0.59 to monotonically decreasing with slope -100 N/s at φ=0.62; glass beads show similar behavior. Increasing φ from low to high at fixed collision velocity causes the penetration depth to decrease monotonically by approximately 50%. However, for the same parameters, the collision duration changes little, decreasing by 10% as φ is increased from 0.58 to 0.6 and then increasing by about 3% as φ is increased to 0.63. Our impact simulations exhibit the same collision dynamics vs. φ and reveal qualitative differences in grain velocity fields and local volume fraction changes between low and high φ states. Support by the Burroughs Wellcome Fund and the Army Research Lab MAST CTA.

  6. Late snowmelt delays plant development and results in lower reproductive success in the High Arctic.

    PubMed

    Cooper, Elisabeth J; Dullinger, Stefan; Semenchuk, Philipp

    2011-01-01

    In tundra areas where the growing season is short, any delay in the start of summer may have a considerable effect on plant development, growth and reproductive success. Climate models suggest long-term changes in winter precipitation in the Arctic, which may lead to deeper snow cover and a resultant delay in date of snow melt. In this paper, we investigated the role of snow depth and melt out date on the phenological development and reproductive success of vascular plants in Adventdalen, Svalbard (78° 10'N, 16° 06'E). Effects of natural variations in snow accumulation were demonstrated using two vegetation types (snow depth: meadow 21 cm, heath 32 cm), and fences were used to experimentally increase snow depth by over 1m. Phenological delay was greatest directly after snowmelt in the earlier phenological phases, and had the largest effect on the early development of those species which normally green-up early (i.e. Dryas, Papaver, Salix, Saxifraga). Compressed growing seasons and length of the reproductive period led to a reduced reproductive success in some of the study species. There were fewer flowers, fewer plots with dispersing seeds, and lower germination rates. This can have consequences for plant establishment and community composition in the long-term.

  7. Study on the weediness of winter wheat in a long-term fertilization field experiment.

    PubMed

    Lehoczky, E; Kismányoky, A; Kismányoky, T

    2006-01-01

    The study was carried out in Keszthely, in the long-term fertilization field experiment in April of 2005. In the experiment we had opportunity to compare the weediness in NPK and NPK + FYM* treatments, and we could study the effect of increasing N dosis on the weeds and winter wheat. The weed survey was made on the 20th of April at the end of tillering. For the weed survey used the Balázs-Ujvárosi method. After that we collected all the weeds from the plots per 1 m2. We counted, measured the fresh and dry matter weight of aerial parts. Winter wheat sampels were taken also from all plots (1 running meter per plot). In the experiment 10 weed species were found, 9 annual: Ambrosia artemisiifolia, Consolida regalis, Galium aparine, Lamiunt amplexicaule, Matricaria inodora, Papaver rhoeas, Stellaria media, Veronica hederifolia, Veronica triphyllos, and 1 perennial: Cirsium arvense. Veronica hederifolia was the dominant species in both fertilized plots, Stellaria media has the second highest weed coverage. The manuring treatments, and the N-dosis has important and significantly effect to the weedeness and the biomass production of winter wheat. On the control plots was the relation of biomass weight of weeds the highest. This relation reduced to the effect of N treatments, wich had an favorable effect on the winter wheat. PMID:17390822

  8. Differential enhancement of benzophenanthridine alkaloid content in cell suspension cultures of Sanguinaria canadensis under conditions of combined hormonal deprivation and fungal elicitation.

    PubMed

    Cline, S D; McHale, R J; Coscia, C J

    1993-08-01

    An elicitation protocol, resulting in the accumulation of sanguinarine in suspension cultures of Papaver bracteatum, was assessed for induction of the same alkaloid in Sanguinaria canadensis. Although only a trace constituent of P. bracteatum plants, sanguinarine is a major alkaloid (1-3% dry wt) of S. canadensis rhizomes. By combining hormonal deprivation for various intervals and a 3-day fungal (Verticillium dahliae) elicitation, benzophenanthridine alkaloid accumulation was induced in S. canadensis cell suspensions. Chelirubine content increased (0.1-1.3% dry wt) consistently in elicited cell cultures while chelerythrine (0.01-0.10% dry wt) and sanguinarine (0-0.02% dry wt) levels were considerably less. Alkaloid accumulation always occurred upon removal of hormone but only at certain time intervals in the log phase upon fungal elicitation. Levels of dopamine, a precursor of the alkaloids, fluctuated over the incubation period, but displayed a 2- to 6-fold increase in cell suspensions grown without hormone. In some experiments dopamine accumulated to levels > 20% dry wt, and these increases were enhanced by the addition of fungal elicitor. Although the same fungal elicitor induces benzophenanthridines in taxonomically related S. canadensis and P. bracteatum, it did not elicit the accumulation of the same alkaloid in the two different plant cultures.

  9. [Psychoactive plant species--actual list of plants prohibited in Poland].

    PubMed

    Simonienko, Katarzyna; Waszkiewicz, Napoleon; Szulc, Agata

    2013-01-01

    According to the Act on Counteracting Drug Addiction (20-th of March, 2009, Dz. U. Nr 63 poz. 520.) the list of federally prohibited plants in Poland was expanded to include 16 new species. Until that time the only illegal plant materials were cannabis, papaver, coca and most of their products. The actual list of herbal narcotics includes species which significantly influence on the central nervous system work but which are rarely described in the national literature. The plants usually come from distant places, where--among primeval cultures--are used for ritual purposes. In our civilization the plants are usually used experimentally, recreationally or to gain particular narcotic effects. The results of the consumption vary: they can be specific or less typical, imitate other substances intake, mental disorders or different pathological states. The plant active substances can interact with other medicaments, be toxic to internal organs, cause serious threat to health or even death. This article describes the sixteen plant species, which are now prohibited in Poland, their biochemical ingredients and their influence on the human organism.

  10. Spray droplet size, drift potential, and risks to nontarget organisms from aerially applied glyphosate for coca control in Colombia.

    PubMed

    Hewitt, Andrew J; Solomon, Keith R; Marshall, E J P

    2009-01-01

    A wind tunnel atomization study was conducted to measure the emission droplet size spectra for water and Glyphos (a glyphosate formulation sold in Colombia) + Cosmo-flux sprays for aerial application to control coca and poppy crops in Colombia. The droplet size spectra were measured in a wind tunnel for an Accu-Flo nozzle (with 16 size 0.085 [2.16 mm] orifices), under appropriate simulated aircraft speeds (up to 333 km/h), using a laser diffraction instrument covering a dynamic size range for droplets of 0.5 to 3,500 microm. The spray drift potential of the glyphosate was modeled using the AGDISP spray application and drift model, using input parameters representative of those occurring in Colombia for typical aerial application operations. The droplet size spectra for tank mixes containing glyphosate and Cosmo-Flux were considerably finer than water and became finer with higher aircraft speeds. The tank mix with 44% glyphosate had a D(v0.5) of 128 microm, while the value at the 4.9% glyphosate rate was 140 microm. These are classified as very fine to fine sprays. Despite being relatively fine, modeling showed that the droplets would not evaporate as rapidly as most similarly sized agricultural sprays because the nonvolatile proportion of the tank mix (active and inert adjuvant ingredients) was large. Thus, longer range drift is small and most drift that does occur will deposit relatively close to the application area. Drift will only occur downwind and, with winds of velocity less than the modeled maximum of 9 km/h, the drift distance would be substantially reduced. Spray drift potential might be additionally reduced through various practices such as the selection of nozzles, tank mix adjuvants, aircraft speeds, and spray pressures that would produce coarser sprays. Species sensitivity distributions to glyphosate were constructed for plants and amphibians. Based on modeled drift and 5th centile concentrations, appropriate no-spray buffer zones (distance from the

  11. Perceptions of policy and political leadership in nursing in New Zealand.

    PubMed

    Donovan, Donna J; Diers, Donna; Carryer, Jenny

    2012-07-01

    This qualitative study was focused on the landscape of nursing policy and political leadership in New Zealand. A volunteer sample (N = 18) of nurse leaders (Fellows of the College of Nurses Aotearoa (NZ) Inc) drawn from across the country was interviewed with respect to issues that affect their interest in participating in political action and policy work. The framework of stages of nursing's political development published by Cohen and colleagues (1996) was used as an interview guide. Respondents were asked to describe their own stage of political development, their perception of the political development of New Zealand nurses and nursing organisations at large, and also their thoughts on what could be done to better position nursing in healthcare policy development. In general, respondents agreed that the major nursing organisations in New Zealand (the College of Nurses-- Aotearoa and New Zealand Nurses Organisation [NZNO]) were moving toward increasing policy sophistication. Qualitative content analysis suggested five themes which, taken together, describe nursing's policy/political development in New Zealand: languaging; succession/legacy planning; Tall Poppies and Queen Bees; "it's a small country"; and speaking with one voice. Although limited by sample size, the information collected provides a beginning focus for discussion that can steer New Zealand nursing activities toward the wider involvement of nurse leaders in healthcare policy work on behalf of the discipline.

  12. Urinary excretion of morphine and biosynthetic precursors in mice

    PubMed Central

    Grobe, Nadja; Lamshöft, Marc; Orth, Robert G.; Dräger, Birgit; Kutchan, Toni M.; Zenk, Meinhart H.; Spiteller, Michael

    2010-01-01

    It has been firmly established that humans excrete a small but steady amount of the isoquinoline alkaloid morphine in their urine. It is unclear whether it is of dietary or endogenous origin. There is no doubt that a simple isoquinoline alkaloid, tetrahydropapaveroline (THP), is found in human and rodent brain as well as in human urine. This suggests a potential biogenetic relationship between both alkaloids. Unlabeled THP or [1,3,4-D3]-THP was injected intraperitoneally into mice and the urine was analyzed. This potential precursor was extensively metabolized (96%). Among the metabolites found was the phenol-coupled product salutaridine, the known morphine precursor in the opium poppy plant. Synthetic [7D]-salutaridinol, the biosynthetic reduction product of salutaridine, injected intraperitoneally into live animals led to the formation of [7D]-thebaine, which was excreted in urine. [N-CD3]-thebaine was also administered and yielded [N-CD3]-morphine and the congeners [N-CD3]-codeine and [N-CD3]-oripavine in urine. These results show for the first time that live animals have the biosynthetic capability to convert a normal constituent of rodents, THP, to morphine. Morphine and its precursors are normally not found in tissues or organs, presumably due to metabolic breakdown. Hence, only that portion of the isoquinoline alkaloids excreted in urine unmetabolized can be detected. Analysis of urine by high resolution-mass spectrometry proved to be a powerful method for tracking endogenous morphine and its biosynthetic precursors. PMID:20421505

  13. A review of characterization of tocotrienols from plant oils and foods.

    PubMed

    Ahsan, Haseeb; Ahad, Amjid; Siddiqui, Waseem A

    2015-04-01

    Tocotrienols, members of the vitamin E family, are natural compounds found in a number of vegetable oils, wheat germ, barley and certain types of nuts and grains. Vegetable oils provide the best sources of these vitamin E forms, particularly palm oil and rice bran oil contain higher amounts of tocotrienols. Other sources of tocotrienols include grape fruit seed oil, oats, hazelnuts, maize, olive oil, buckthorn berry, rye, flax seed oil, poppy seed oil and sunflower oil. Tocotrienols are of four types, viz. alpha (α), beta (β), gamma (γ) and delta (δ). Unlike tocopherols, tocotrienols are unsaturated and possess an isoprenoid side chain. A number of researchers have developed methods for the extraction, analysis, identification and quantification of different types of vitamin E compounds. This article constitutes an in-depth review of the chemistry and extraction of the unsaturated vitamin E derivatives, tocotrienols, from various sources using different methods. This review article lists the different techniques that are used in the characterization and purification of tocotrienols such as soxhlet and solid-liquid extractions, saponification method, chromatography (thin layer, column chromatography, gas chromatography, supercritical fluid, high performance), capillary electrochromatography and mass spectrometry. Some of the methods described were able to identify one form or type while others could analyse all the analogues of tocotrienol molecules. Hence, this article will be helpful in understanding the various methods used in the characterization of this lesser known vitamin E variant.

  14. Forest plunder in Southeast Asia: an environmental security nexus in Burma and Cambodia.

    PubMed

    Talbott, K; Brown, M

    1998-01-01

    This article discusses the cycle of conversion, consumption, and corruption that undermines the environment and civil society in Cambodia and Burma (Myanmar). In these countries, forests are declining in patterns similar to other Southeast Asian deforestation. Illegal logging, prostitution, and heroin trafficking constitute the bulk of Cambodia's shadow economy. Revenues are used to provide financial support for political causes and build the private wealth of the elite. Major political and guerilla groups and the Cambodian military have been major beneficiaries of logging revenue, supported private sector forestry in many military zones, and facilitated logging and trade. About 40% of land goes to forest concessions granted to Southeast Asian companies, and revenues bypass the regular state budget. In Burma, the cease fire agreements in the early 1990s, led to remote border area forests being opened up to large, nonsustainable commercial timber mining. Land was divided into ethnic and government controlled areas. Timber profits were funneled into a business owned by members of the new ruling force, the SLORC, and used to launder drug exports and profits. Trading partners include Thailand, and most recently, China. It is speculated that deforested areas are replanted with opium poppies, and trade routes carry timber and heroin. The unregulated logging industry and the lack of financial accounting of the timber trade undermine the structures of civil society and good governance. Forest policies appear progressive but are in reality unenforced. Politics and agreements in both countries are closely tied to deforestation issues. PMID:12321720

  15. Hypersensitivities to sesame and other common edible seeds.

    PubMed

    Patel, A; Bahna, S L

    2016-10-01

    Several seeds have been increasingly incorporated in various food items, with consequent risk of hypersensitivity reactions that are often severe. Identification of the specific seed as the culprit is often not explored or is difficult to verify. In this article, we reviewed the English literature from January 1930 to March 2016 using PubMed and Google Scholar searching for publications relevant to hypersensitivity to common edible seeds, namely sesame, sunflower seed, poppy seed, pumpkin seed, flaxseed, and mustard seed. Considering the worldwide consumption of those seeds, the number of published articles on the subject was relatively small and was mainly as case reports rather than large series. Allergy to sesame was more reported than to other seeds, with an estimated prevalence of 0.1-0.2%. In this review, we summarize the information relevant to each of the six seeds and their oils regarding the manifestations, routes of exposure, identified major allergens, and cross-reactivity with other seeds or other foods. We also addressed the important role of a thorough history taking in suspecting seed allergy, the limited reliability of routine diagnostic procedures, and the importance of verification by appropriate challenge tests. At present, management is basically dietary avoidance and the use of symptomatic medications that may include epinephrine auto-injectors. We did not encounter any well-designed studies on immunotherapy for seed allergy, but it is hoped that such a gap be filled by the development of safe effective protocols in the near future. PMID:27332789

  16. Multivariate Analysis of Combined Fourier Transform Near-Infrared Spectrometry (FT-NIR) and Raman Datasets for Improved Discrimination of Drying Oils.

    PubMed

    Carlesi, Serena; Ricci, Marilena; Cucci, Costanza; La Nasa, Jacopo; Lofrumento, Cristiana; Picollo, Marcello; Becucci, Maurizio

    2015-07-01

    This work explores the application of chemometric techniques to the analysis of lipidic paint binders (i.e., drying oils) by means of Raman and near-infrared spectroscopy. These binders have been widely used by artists throughout history, both individually and in mixtures. We prepared various model samples of the pure binders (linseed, poppy seed, and walnut oils) obtained from different manufacturers. These model samples were left to dry and then characterized by Raman and reflectance near-infrared spectroscopy. Multivariate analysis was performed by applying principal component analysis (PCA) on the first derivative of the corresponding Raman spectra (1800-750 cm(-1)), near-infrared spectra (6000-3900 cm(-1)), and their combination to test whether spectral differences could enable samples to be distinguished on the basis of their composition. The vibrational bands we found most useful to discriminate between the different products we studied are the fundamental ν(C=C) stretching and methylenic stretching and bending combination bands. The results of the multivariate analysis demonstrated the potential of chemometric approaches for characterizing and identifying drying oils, and also for gaining a deeper insight into the aging process. Comparison with high-performance liquid chromatography data was conducted to check the PCA results. PMID:26036244

  17. Evaluation of mycotoxins, mycobiota, and toxigenic fungi in selected medicinal plants of Khyber Pakhtunkhwa, Pakistan.

    PubMed

    Ahmad, Bashir; Ashiq, Samina; Hussain, Arshad; Bashir, Shumaila; Hussain, Mubbashir

    2014-01-01

    Medicinal plants are used worldwide to treat a variety of ailments. Due to the provenance of medicinal plants, they are subjected to contamination by moulds, which may be responsible for spoilage and production of mycotoxins. The investigation was designed to throw light on mycological and mycotoxicological status of some medicinal plants from Pakistan and the result showed 30 % and 26.7 % samples were contaminated with aflatoxins and ochratoxin A, respectively. Mould contamination was present in 90 % samples, of which 70 % exceeded the permissible limits. Opium poppy, licorice root, and Indian rennet were most contaminated samples. The predominant moulds found were Aspergillus flavus, Aspergillus niger, Aspergillus parasiticus, and Penicillium spp. and 31 % of the 47 isolates tested were found to be toxigenic. The findings indicate that the contamination in the medicinal plants may contribute to adverse human health problems. This information would prove helpful for regulatory agencies to establish limits for these contaminants in medicinal plants and will explore ways for export of herbal products to countries where more stringent permissible limits of mycotoxins exist. The study is first of its kind in the country reporting natural occurrence of mycotoxins in medicinal plants in Pakistan.

  18. Environmentally Clean Mitigation of Undesirable Plant Life Using Lasers

    SciTech Connect

    Rubenchik, A M; McGrann, T J; Yamamoto, R M; Parker, J M

    2009-07-01

    This concept comprises a method for environmentally clean destruction of undesirable plant life using visible or infrared radiation. We believe that during the blossom stage, plant life is very sensitive to electromagnetic radiation, with an enhanced sensitivity to specific spectral ranges. Small doses of irradiation can arrest further plant growth, cause flower destruction or promote plant death. Surrounding plants, which are not in the blossoming stage, should not be affected. Our proposed mechanism to initiate this effect is radiation produced by a laser. Tender parts of the blossom possess enhanced absorptivity in some spectral ranges. This absorption can increase the local tissue temperature by several degrees, which is sufficient to induce bio-tissue damage. In some instances, the radiation may actually stimulate plant growth, as an alternative for use in increased crop production. This would be dependent on factors such as plant type, the wavelength of the laser radiation being used and the amount of the radiation dose. Practical, economically viable realization of this concept is possible today with the advent of high efficiency, compact and powerful laser diodes. The laser diodes provide an efficient, environmentally clean source of radiation at a variety of power levels and radiation wavelengths. Figure 1 shows the overall concept, with the laser diodes mounted on a movable platform, traversing and directing the laser radiation over a field of opium poppies.

  19. Illegal drugs, anti-drug policy failure, and the need for institutional reforms in Colombia.

    PubMed

    Thoumi, Francisco E

    2012-01-01

    This paper is inspired by two anomalies encountered in the study of the illegal drugs industry. First, despite the very high profits of coca/cocaine and poppy/opium/heroin production, most countries that can produce do not. Why, for example, does Colombia face much greater competition in the international coffee, banana, and other legal product markets than in cocaine? And second, though illegal drugs are clearly associated with violence, why is it that illegal drug trafficking organizations have been so much more violent in Colombia and Mexico than in the rest of the world? The answers to these questions cannot be found in factors external to Colombia (and Mexico). They require identifying the societal weaknesses of each country. To do so, the history of the illegal drugs industry is surveyed, a simple model of human behavior that stresses the conflict between formal (legal) and informal (socially accepted) norms as a source of the weaknesses that make societies vulnerable is formulated. The reasons why there is a wide gap between formal and informal norms in Colombia are explored and the effectiveness of anti-drug policies is considered to explain why they fail to achieve their posited goals. The essay ends with reflections and conclusion on the need for institutional change.

  20. Analysis of methylsterol fractions from twenty vegetable oils.

    PubMed

    Jeong, T M; Itoh, T; Tamura, T; Matsumoto, T

    1975-10-01

    The 4-monomethylsterol and 4,4-dimethylsterol fractions were separated from the unsaponifiables of 20 vegetable oils by preparative thin layer chromatography, and their compositions were determined by gas liquid chromatography. Tentative identification of the individual components of these fractions was carried out by gas liquid chromatography and combined gas liquid chromatography-mass spectrometry. Among 4-monomethylsterols, obtusifoliol, gramisterol, and citrostadienol occur abundantly in most of the oils. Cycloeucalenol also occurs in some of the oils as a major component of 4-monomethylsterols. Other 4-monomethylsterols tentatively identified are: lophenol, 31-norlanosterol, 31-norcycloartenol, and 31-norlanostenol and/or 4alpha-methylzymostenol. Among 4,4-dimethylsterols, cycloartenol and 24-methylenecycloartanol followed by beta-amyrin and cycloartanol are common to most of the oils. Butyrospermol, alpha-amyrin, lupeol, and cyclobranol together with a 4,4-dimethylsterol, presumably lanostenol, occur in some of the oils. Cyclolaudenol is present in poppy seed oil. Besides these compounds, each of the oils contains some unidentified members of 4-monomethylsterols and 4,4-dimethylsterols. The methylsterol fraction of capsicum seed oil as compared with that of the other oils is characterized by its very high content of lophenol and cycloartanol together with three other members, presumably 31-norlanostenol, 4alpha-methylzymostenol, and lanostenol.

  1. Forest Conflict in Thailand: Northern Minorities in Focus

    NASA Astrophysics Data System (ADS)

    Hares, Minna

    2009-03-01

    This paper aims at exploring the local background of and solutions to the forest conflict in upland areas inhabited by ethnic minorities, who are called hill tribes, in northern Thailand. A so-called hill tribe problem has been officially identified as a result of the slash-and-burn cultivation and other perceived problems, such as opium poppy cultivation, illegal immigration, and the suspicion of disloyalty to the state. This has created distrust and tension between the groups and authorities. The local conflict has recently been related to the dilemma of conserving the forest from all human interference, while many people live and make their livelihood within and adjacent to the protected areas. Furthermore, as the results imply, strictly protected areas and reforestation have also increased the competition over land and natural resources and, thereby, the likelihood of local conflicts. The scarcity and pollution of water, illegal logging, and poor fire control have contributed to the conflicts between local communities. The conflicts between the local communities and officials have been nourished by political and public discussions. Using definitions and terms with negative connotations and ignoring the heterogeneity between the groups or labeling some groups as malevolent have increased distrust and strengthened existing stereotypical images. Conflict resolution starts with efforts toward better mutual understanding, and changes in structures and attitudes are necessary. Local cooperation, utilization of traditional methods, and local institutions are central to conflict solving.

  2. Urinary excretion of morphine and biosynthetic precursors in mice.

    PubMed

    Grobe, Nadja; Lamshöft, Marc; Orth, Robert G; Dräger, Birgit; Kutchan, Toni M; Zenk, Meinhart H; Spiteller, Michael

    2010-05-01

    It has been firmly established that humans excrete a small but steady amount of the isoquinoline alkaloid morphine in their urine. It is unclear whether it is of dietary or endogenous origin. There is no doubt that a simple isoquinoline alkaloid, tetrahydropapaveroline (THP), is found in human and rodent brain as well as in human urine. This suggests a potential biogenetic relationship between both alkaloids. Unlabeled THP or [1,3,4-D(3)]-THP was injected intraperitoneally into mice and the urine was analyzed. This potential precursor was extensively metabolized (96%). Among the metabolites found was the phenol-coupled product salutaridine, the known morphine precursor in the opium poppy plant. Synthetic [7D]-salutaridinol, the biosynthetic reduction product of salutaridine, injected intraperitoneally into live animals led to the formation of [7D]-thebaine, which was excreted in urine. [N-CD(3)]-thebaine was also administered and yielded [N-CD(3)]-morphine and the congeners [N-CD(3)]-codeine and [N-CD(3)]-oripavine in urine. These results show for the first time that live animals have the biosynthetic capability to convert a normal constituent of rodents, THP, to morphine. Morphine and its precursors are normally not found in tissues or organs, presumably due to metabolic breakdown. Hence, only that portion of the isoquinoline alkaloids excreted in urine unmetabolized can be detected. Analysis of urine by high resolution-mass spectrometry proved to be a powerful method for tracking endogenous morphine and its biosynthetic precursors.

  3. Simulating PSFs for WFIRST and JWST with WebbPSF

    NASA Astrophysics Data System (ADS)

    Long, Joseph D.; Perrin, Marshall D.; Van Der Marel, Roeland P.

    2016-01-01

    Accurate models of a telescope's point spread function are key to predicting its performance and extracting information from observations. Developed at STScI since 2010, WebbPSF is a flexible Python-based PSF simulation tool initially developed for JWST's imaging, spectroscopy, and coronagraphic instruments. We present improvements that allow this tool to simulate PSFs for the WFIRST wide-field imaging mode, as well as additional spectroscopy modes for the NIRSpec, MIRI, and NIRISS instruments on JWST. The WFIRST wide field imaging mode is also the first WebbPSF model to simulate PSF variation across the entire field of view. These variations are included in the Fraunhofer-domain PSF calculation as Zernike polynomial terms up to Z22. As WFIRST is still early in its development, high-spatial-frequency wavefront errors (beyond Z22) are incorporated using an optical path difference map from another notable 2.4 meter space telescope. Common infrastructure to build simulated optical instruments has been made available as POPPY (Physical Optics Propagation in Python), an open-source library that has seen contributions from users in astronomy and beyond.

  4. Assessment of the microbiological safety of edible dried seeds from retail premises in the United Kingdom with a focus on Salmonella spp.

    PubMed

    Willis, Caroline; Little, Christine L; Sagoo, Satnam; de Pinna, Elizabeth; Threlfall, John

    2009-12-01

    Sesame seed products have recently been associated with a number of Salmonella outbreaks in the UK and elsewhere. Aside from sesame seeds, there is little published information on the prevalence of Salmonella spp. in edible seeds. A study of 3735 samples of retail edible dried seeds in the UK was therefore carried out between October 2007 and March 2008 to assess their microbiological safety in relation to Salmonella contamination and levels of Escherichia coli, an indicator of faecal contamination. Overall, Salmonella was detected in 23 samples (0.6%), of which over half (57%) were sesame seeds. Other seeds contaminated with Salmonella were linseed (1 sample), sunflower (1 sample), alfalfa (1 sample), melon (4 samples) and mixed seeds (3 samples). E. coli was detected in 9% of samples, with 1.5% containing unsatisfactory levels (> or = 10(2)/g). These included melon, pumpkin, sesame, hemp, poppy, linseed, sunflower and mixed seeds. The UK retailers affected by the detection of Salmonella in their products recalled the contaminated batches, and Food Standards Agency food alerts were issued to advise against the consumption of affected seed products. This study highlights the importance of good hygiene practices and effective decontamination procedures during the production of these products.

  5. The effects of adulterants and selected ingested compounds on drugs-of-abuse testing in urine.

    PubMed

    Dasgupta, Amitava

    2007-09-01

    Household chemicals such as bleach, table salt, laundry detergent, toilet bowl cleaner, vinegar, lemon juice, and eyedrops are used for adulterating urine specimens. Most of these adulterants except eyedrops can be detected by routine specimen integrity tests (creatinine, pH, temperature, and specific gravity); however, certain adulterants such as Klear, Whizzies, Urine Luck, and Stealth cannot. These adulterants can successfully mask drug testing if the concentrations of certain abused drugs are moderate. Several spot tests have been described to detect the presence of such adulterants in urine. Urine dipsticks are commercially available for detecting the presence of such adulterants, along with performance of tests for creatinine, pH, and specific gravity. Certain hair shampoo and saliva-cleaning mouthwashes are available to escape detection in hair or saliva samples, but the effectiveness of such products in masking drugs-of-abuse testing has not been demonstrated. Ingestion of poppy seed cake may result in positive screening test results for opiates, and hemp oil exposure can cause positive results for marijuana. These would be identified as true-positive results in drugs-of-abuse testing even though they do not represent the actual drug of abuse.

  6. Feeding outside the forest: the importance of crop raiding and an invasive weed in the diet of gallery forest ring-tailed lemurs (Lemur catta) following a cyclone at the Beza Mahafaly special reserve, Madagascar.

    PubMed

    LaFleur, M; Gould, L

    2009-01-01

    In January 2005, a cyclone hit southern Madagascar, including the Beza Mahafaly Special Reserve, disrupting the flowering/fruiting cycle of Tamarindus indica, leaving Lemur catta without its major food resource during reproductive periods. We studied two adjacent groups of L. catta during the late gestation period, and both groups ventured outside the reserve to feed. The Red group (RG) fed daily on cultivated sweet potato (Ipomoea batatas) leaves in a nearby field, and both groups consumed leaves and stems of the invasive terrestrial flowering herb Mexican prickly poppy (Argemone mexicana), growing outside the reserve. The Green group (GG) spent significantly more time feeding than did RG, and more time feeding inside the forest compared to outside. The members of RG spent half of their time feeding in the crops, and nearly half of their diet consisted of easy-to-process sweet potato leaves. Additionally, RG defended and restricted GG's access to the crop territory. Of the two non-forest foods, A. mexicana leaves were higher in protein and most minerals (P, Mg, K and Na, but not Ca) and lower in fiber than sweet potato leaves, but sweet potato leaves were preferred by RG. L. catta is a markedly flexible primate with respect to diet, and switches to fallback foods from outside the forest during periods of low food availability. In the highly seasonal and unpredictable climate of southern Madagascar, such behavioral adaptations are important to the survival of this species. PMID:19776607

  7. Sensitive dependence of the motion of a legged robot on granular media

    PubMed Central

    Li, Chen; Umbanhowar, Paul B.; Komsuoglu, Haldun; Koditschek, Daniel E.; Goldman, Daniel I.

    2009-01-01

    Legged locomotion on flowing ground (e.g., granular media) is unlike locomotion on hard ground because feet experience both solid- and fluid-like forces during surface penetration. Recent bioinspired legged robots display speed relative to body size on hard ground comparable with high-performing organisms like cockroaches but suffer significant performance loss on flowing materials like sand. In laboratory experiments, we study the performance (speed) of a small (2.3 kg) 6-legged robot, SandBot, as it runs on a bed of granular media (1-mm poppy seeds). For an alternating tripod gait on the granular bed, standard gait control parameters achieve speeds at best 2 orders of magnitude smaller than the 2 body lengths/s (≈60 cm/s) for motion on hard ground. However, empirical adjustment of these control parameters away from the hard ground settings restores good performance, yielding top speeds of 30 cm/s. Robot speed depends sensitively on the packing fraction φ and the limb frequency ω, and a dramatic transition from rotary walking to slow swimming occurs when φ becomes small enough and/or ω large enough. We propose a kinematic model of the rotary walking mode based on generic features of penetration and slip of a curved limb in granular media. The model captures the dependence of robot speed on limb frequency and the transition between walking and swimming modes but highlights the need for a deeper understanding of the physics of granular media. PMID:19204285

  8. PAHs, PCBs, PBDEs and Pesticides in Cold-Pressed Vegetable Oils.

    PubMed

    Roszko, M; Szterk, A; Szymczyk, K; Waszkiewicz-Robak, B

    2012-03-01

    The aim of this study was to investigate levels of polychlorinated biphenyls (marker and dioxin-like congeners), polycyclic aromatic hydrocarbons (EPA 15 + 1), polybrominated diphenyl ethers (14 predominant congeners) and pesticides (74 compounds) in various cold-pressed vegetable oils. Poppy seed oil, rapeseed oil, sesame seed oil, pumpkinseed oil, hempseed oil, linaire oil, borage oil and evening star oil were investigated. Results of this study revealed that concentrations of PCBs, PBDEs and PAHs were low in majority of the investigated samples. However, high concentrations of organophosphorus insecticides were found. Chlorpyrifos methyl and pirimiphos methyl were the pesticide residues most commonly found in the studied oils. Concentration of 15 + 1 EPA PAHs was within the 17.85-37.16 μg kg(-1) range, concentration of (marker) PCBs varied from 127 to 24,882 pg g(-1), dioxin-like TEQ values were below 0.1 pg TEQ g(-1). Concentration of PBDEs was below LOQ in most cases. PMID:22389518

  9. Structures of Two Arabidopsis thaliana Major Latex Proteins Represent Novel Helix-Grip Folds

    PubMed Central

    Lytle, Betsy L.; Song, Jikui; de la Cruz, Norberto B.; Peterson, Francis C.; Johnson, Kenneth A.; Bingman, Craig A.; Phillips, George N.; Volkman, Brian F.

    2010-01-01

    The major latex proteins (MLP) are a protein family first identified in the latex of opium poppy. They are found only in plants and have 24 identified members in Arabidopsis alone as well as in other plants such as peach, strawberry, melon, cucumber, and soybean. While the function of the MLPs is unknown, they have been associated with fruit and flower development and in pathogen defense responses. Based on modest sequence similarity, they have been characterized as members of the Bet v 1 protein superfamily; however, no structures have yet been reported. As part of an ongoing structural genomics effort, we determined the structures of two Arabidopsis thaliana MLPs: the solution structure of MLP28 (gene product of At1g70830.1) and the crystal structure of At1g24000.1. The structures revealed distinct differences when compared to one another and to the typical Bet v 1 fold. Nevertheless, NMR titration experiments demonstrated that the characteristic Bet v 1 hydrophobic binding pocket of At1g24000.1 is able to bind a ligand, suggesting that it plays a role in the function of the MLPs. A structure-based sequence analysis identified conserved hydrophobic residues in the long alpha helix that contribute to the binding cavity and may specify preferred ligands for the MLP family. PMID:19326460

  10. Modulatory Effects of Eschscholzia californica Alkaloids on Recombinant GABAA Receptors

    PubMed Central

    Fedurco, Milan; Gregorová, Jana; Šebrlová, Kristýna; Kantorová, Jana; Peš, Ondřej; Baur, Roland; Sigel, Erwin; Táborská, Eva

    2015-01-01

    The California poppy (Eschscholzia californica Cham.) contains a variety of natural compounds including several alkaloids found exclusively in this plant. Because of the sedative, anxiolytic, and analgesic effects, this herb is currently sold in pharmacies in many countries. However, our understanding of these biological effects at the molecular level is still lacking. Alkaloids detected in E. californica could be hypothesized to act at GABAA receptors, which are widely expressed in the brain mainly at the inhibitory interneurons. Electrophysiological studies on a recombinant α1β2γ2 GABAA receptor showed no effect of N-methyllaurotetanine at concentrations lower than 30 μM. However, (S)-reticuline behaved as positive allosteric modulator at the α3, α5, and α6 isoforms of GABAA receptors. The depressant properties of aerial parts of E. californica are assigned to chloride-current modulation by (S)-reticuline at the α3β2γ2 and α5β2γ2 GABAA receptors. Interestingly, α1, α3, and α5 were not significantly affected by (R)-reticuline, 1,2-tetrahydroreticuline, codeine, and morphine—suspected (S)-reticuline metabolites in the rodent brain. PMID:26509084

  11. Antitumor effects of the benzophenanthridine alkaloid sanguinarine: Evidence and perspectives

    PubMed Central

    Gaziano, Roberta; Moroni, Gabriella; Buè, Cristina; Miele, Martino Tony; Sinibaldi-Vallebona, Paola; Pica, Francesca

    2016-01-01

    Historically, natural products have represented a significant source of anticancer agents, with plant-derived drugs becoming increasingly explored. In particular, sanguinarine is a benzophenanthridine alkaloid obtained from the root of Sanguinaria canadensis, and from other poppy Fumaria species, with recognized anti-microbial, anti-oxidant and anti-inflammatory properties. Recently, increasing evidence that sanguinarine exibits anticancer potential through its capability of inducing apoptosis and/or antiproliferative effects on tumor cells, has been proved. Moreover, its antitumor seems to be due not only to its pro-apoptotic and inhibitory effects on tumor growth, but also to its antiangiogenic and anti-invasive properties. Although the precise mechanisms underlying the antitumor activity of this compound remain not fully understood, in this review we will focus on the most recent findings about the cellular and molecular pathways affected by sanguinarine, together with the rationale of its potential application in clinic. The complex of data currently available suggest the potential application of sanguinarine as an adjuvant in the therapy of cancer, but further pre-clinical studies are needed before such an antitumor strategy can be effectively translated in the clinical practice. PMID:26798435

  12. Comparison of oilseed yields: a preliminary review

    SciTech Connect

    Duke, J.A.; Bagby, M.O.

    1982-01-01

    It was assumed that for most oilseed crops, 90% of the oil yield might be considered as profit. To compare oil seeds, pertinent portions of the yield and energy paragraphs from a summary published by Dr. Duke for DOE Grant No. 59-2246-1-6-054-0 with Dr. Bagby as ADODR were reproduced. The seed yields ranged from 200 to 14,000 kg/ha, the low one too low to consider and the high one suspiciously high. The yield of 14,000 kg oil per hectare is equivalent to more than 30 barrels of oil per hectare. The energy species included ambrette, tung-oil tree, cashew, wood-oil tree, mu-oil tree, peanut, mustard greens; rape, colza; black mustard, turnip, safflower, colocynth, coconut, crambe, African oil palm, soybean, cotton, sunflower, Eastern black walnut, Engligh walnut, meadow foam, flax, macadamia nuts, opium poppy, perilla, almond, castorbean, Chinese tallow tree, sesame, jojoba, yellow mustard, stokes' aster, and Zanzibar oilvine. 1 table. (DP)

  13. Effect of seeds on bile-enzymatic-gravimetric analysis of total dietary fiber.

    PubMed

    Cranker, K J; Phillips, K M; Gonzales, M C; Stewart, K K

    1997-01-01

    Dietary fiber sometimes is defined chemically as nonstarch polysaccharides plus lignin or as other specific chemical entities. Analysis of dietary fiber according to a chemical definition typically involves gas chromatography, which allows separation and quantitation of chemical constituents that are added to arrive at a dietary fiber value. Other definitions of fiber are broader, defining it to be whatever is not digested in the alimentary tract. Analytically, this definition translates into the gravimetric sum of the material remaining after a series of enzymatic and chemical treatments that simulate in vivo digestion. Various methods reflect the gravimetric definition, which might include as dietary fiber some protein, resistant starch, and even lipids that are not digested by particular assay conditions. We used a recently proposed bile-enzymatic-gravimetric assay for total dietary fiber on commonly consumed seeds (hulled and unhulled sesame, caraway, and poppy) and visually found these seeds to be undigested. We then determined the impact of the undigested seeds on measured dietary fiber content by spiking homogenized daily menus with 5% by weight of these seeds and calculating recoveries with 2 assumptions: seeds are 100% fiber because they are not digested, and the fiber content of seeds is as determined by assay. Calculated recoveries were very different depending on which assumption was made (71-90% or 99-109%, respectively), and the difference was closely related to the seed's protein content.

  14. Investigation of damping liquids for aircraft instruments

    NASA Technical Reports Server (NTRS)

    Keulegan, G H

    1929-01-01

    This report covers the results of an investigation carried on at the Bureau of Standards under a research authorization from, and with the financial assistance of, the National Advisory Committee for Aeronautics. The choice of a damping liquid for aircraft instruments is difficult owing to the range of temperature at which aircraft operate. Temperature changes affect the viscosity tremendously. The investigation was undertaken with the object of finding liquids of various viscosities otherwise suitable which had a minimum change in viscosity with temperature. The new data relate largely to solutions. The effect of temperature on the kinematic viscosity of the following liquids and solutions was determined in the temperature interval -18 degrees to +30 degrees C. (1) solutions of animal and vegetable oils in xylene. These were poppy-seed oil, two samples of neat's-foot oils, castor oil, and linseed oil. (2) solutions of mineral oil in xylene. These were Squibb's petrolatum of naphthene base and transformer oil. (3) glycerine solutions in ethyl alcohol and in mixture of 50-50 ethyl alcohol and water. (4) mixtures of normal butyl alcohol with methyl alcohol. (5) individual liquids, kerosene, mineral spirits, xylene, recoil oil. The apparatus consisted of four capillary-tube viscometers, which were immersed in a liquid bath in order to secure temperature control. The method of calibration and the related experimental data are presented.

  15. Antitumor effects of the benzophenanthridine alkaloid sanguinarine: Evidence and perspectives.

    PubMed

    Gaziano, Roberta; Moroni, Gabriella; Buè, Cristina; Miele, Martino Tony; Sinibaldi-Vallebona, Paola; Pica, Francesca

    2016-01-15

    Historically, natural products have represented a significant source of anticancer agents, with plant-derived drugs becoming increasingly explored. In particular, sanguinarine is a benzophenanthridine alkaloid obtained from the root of Sanguinaria canadensis, and from other poppy Fumaria species, with recognized anti-microbial, anti-oxidant and anti-inflammatory properties. Recently, increasing evidence that sanguinarine exibits anticancer potential through its capability of inducing apoptosis and/or antiproliferative effects on tumor cells, has been proved. Moreover, its antitumor seems to be due not only to its pro-apoptotic and inhibitory effects on tumor growth, but also to its antiangiogenic and anti-invasive properties. Although the precise mechanisms underlying the antitumor activity of this compound remain not fully understood, in this review we will focus on the most recent findings about the cellular and molecular pathways affected by sanguinarine, together with the rationale of its potential application in clinic. The complex of data currently available suggest the potential application of sanguinarine as an adjuvant in the therapy of cancer, but further pre-clinical studies are needed before such an antitumor strategy can be effectively translated in the clinical practice.

  16. California Rare Endemics and Climate Change

    NASA Astrophysics Data System (ADS)

    Espinoza, M.

    2010-12-01

    California is known for its wide variety of endemic flora, from its annuals such as the Eschscholzia californica (California poppy) to the perennials like the Arctostaphylos pallida (Alameda manzanita), which happens to be a rare species. Each species plays an important role in the biodiversity of California, yet there are species that are threatened, not only by human interaction and urbanization, but by climate change. Species that we seldom see are now on the verge of becoming eradicated; rare endemics similar to Arctostaphylos pallida are now facing a new challenge that may severely impair their survival. The climate has changed significantly over the twentieth century and it has affected the distribution of rare endemics in California, both geographically as well as within their climatic and edaphic niches. Lilaeopsis masonii is just one rare endemic, however it serves as a representative of the other 23 species that were studied. Using Maxent, a climate-modeling program, it was viable to construct two climate envelopes of the masonii species: the early century envelope (1930-1959) and the later century envelope (1990-2009). When these two climate envelopes were compared, it became clear that the later century climate envelope had contracted radically, reshaping the climate niche of all rare endemics in California due to an increase in temperature. It is possible to conclude that the future of rare endemics hangs in the balance, where one degree higher in temperature is enough to topple the scale.

  17. Microencapsulation of anti-tumor, antibiotic and thrombolytic drugs in microgravity

    NASA Technical Reports Server (NTRS)

    Morrison, Dennis R.; Mosier, Benjamin; Cassanto, John

    1994-01-01

    Encapsulation of cytotoxic or labile drugs enables targeted delivery and sustained release kinetics that are not available with intravenous injection. A new liquid-liquid diffusion process has been developed for forming unique microcapsules that contain both aqueous and hydrocarbon soluble drugs. Microgravity experiments, on sounding rockets (1989-92) and Shuttle missions STS-52 (1992) and STS-56 (1993) using an automated Materials Dispersion Apparatus, produced multi-lamellar microcapsules containing both Cis-platinum (anti-tumor drug) and iodinated poppy seed oil (a radiocontrast medium), surrounded by a polyglyceride skin. Microcapsules formed with amoxicillin (antibiotic) or urokinase (a clot dissolving enzyme), co-encapsulated with IPO, are still intact after two years. Microcapsules were formed with the drug so concentrated that crystals formed inside. Multi-layered microspheres, with both hydrophobic drug compartments, can enable diffusion of complementary drugs from the same microcapsule, e.g. antibiotics and immuno-stimulants to treat resistant infections or multiple fibrinolytic drugs to dissolve emboli. Co-encapsulation of enough radio-contrast medium enables oncologists to monitor the delivery of anti-tumor microcapsules to target tumors using computerized tomography and radiography that would track the distribution of microcapsules after release from the intra-arterial catheter. These microcapsules could have important applications in chemotheraphy of certain liver, kidney, brain and other tumors.

  18. Evolutionary and neuropsychological perspectives on addictive behaviors and addictive substances: relevance to the “food addiction” construct

    PubMed Central

    Davis, Caroline

    2014-01-01

    It has been argued that food cannot be “addictive”, unlike conventional drugs of abuse, because it is an essential part of life. In this paper, evidence is reviewed, largely from an evolutionary psychobiological perspective, that plant-based psychoactive drugs (such as those derived from the opium poppy and the coca leaf) and gambling-related behaviors were once adaptive for human health and survival in a similar manner as energy-based foods were for nourishment. “Evolutionary mismatch” viewpoints contend that certain behaviors were enhanced during the hunter-gatherer lifestyle – from which our genetic endowment had its origins – because they bestowed both survival and reproductive advantages to the species. However, in the context of advanced technology and other rapid environmental changes, these same behaviors have tended to become maladaptive and greatly overexpressed. Similar to the manufactured purification of psychotropic plant-based substances, the reward impact of processed and hyperpalatable foods, with their high levels of sugar, fat, and salt, is much increased from foods produced in nature. It is concluded therefore that what was once beneficial and necessary for our survival has been altered and ultraprocessed into edible products that may be disadvantageous and potentially addictive. PMID:25540603

  19. Forest conflict in Thailand: northern minorities in focus.

    PubMed

    Hares, Minna

    2009-03-01

    This paper aims at exploring the local background of and solutions to the forest conflict in upland areas inhabited by ethnic minorities, who are called hill tribes, in northern Thailand. A so-called hill tribe problem has been officially identified as a result of the slash-and-burn cultivation and other perceived problems, such as opium poppy cultivation, illegal immigration, and the suspicion of disloyalty to the state. This has created distrust and tension between the groups and authorities. The local conflict has recently been related to the dilemma of conserving the forest from all human interference, while many people live and make their livelihood within and adjacent to the protected areas. Furthermore, as the results imply, strictly protected areas and reforestation have also increased the competition over land and natural resources and, thereby, the likelihood of local conflicts. The scarcity and pollution of water, illegal logging, and poor fire control have contributed to the conflicts between local communities. The conflicts between the local communities and officials have been nourished by political and public discussions. Using definitions and terms with negative connotations and ignoring the heterogeneity between the groups or labeling some groups as malevolent have increased distrust and strengthened existing stereotypical images. Conflict resolution starts with efforts toward better mutual understanding, and changes in structures and attitudes are necessary. Local cooperation, utilization of traditional methods, and local institutions are central to conflict solving. PMID:19067036

  20. Forest conflict in Thailand: northern minorities in focus.

    PubMed

    Hares, Minna

    2009-03-01

    This paper aims at exploring the local background of and solutions to the forest conflict in upland areas inhabited by ethnic minorities, who are called hill tribes, in northern Thailand. A so-called hill tribe problem has been officially identified as a result of the slash-and-burn cultivation and other perceived problems, such as opium poppy cultivation, illegal immigration, and the suspicion of disloyalty to the state. This has created distrust and tension between the groups and authorities. The local conflict has recently been related to the dilemma of conserving the forest from all human interference, while many people live and make their livelihood within and adjacent to the protected areas. Furthermore, as the results imply, strictly protected areas and reforestation have also increased the competition over land and natural resources and, thereby, the likelihood of local conflicts. The scarcity and pollution of water, illegal logging, and poor fire control have contributed to the conflicts between local communities. The conflicts between the local communities and officials have been nourished by political and public discussions. Using definitions and terms with negative connotations and ignoring the heterogeneity between the groups or labeling some groups as malevolent have increased distrust and strengthened existing stereotypical images. Conflict resolution starts with efforts toward better mutual understanding, and changes in structures and attitudes are necessary. Local cooperation, utilization of traditional methods, and local institutions are central to conflict solving.