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There is a great need to make biological control research applicable to local political and social situations. The objectives of this study were to evaluate the effect of Fusarium oxysporum isolates against opium poppy (Papaver somniferum) and verify their potential as biocontrol agent. For this, a 2-year experiment was carried out under glasshouse (2 trials) and field conditions (2 trials). From infected poppy plants, a total of 16 pathogenic fungal strains were identified as F. oxysporum and used for the experiments. The isolates Ghr18, Ghr5-2, Mr28 and Ghr5-4 caused the highest wilting symptoms on sample plants (P 0.5). Moreover, the results showed a clear host specificity of the selected pathogenic isolates. These results suggest that Fusarium isolates have the potential to be used as biological control agents against poppy plants in the Middle East.

Opium poppy (Papaver somniferum L.) remains the only commercial source for several pharmaceutical alkaloids. In present study, opium poppy plants were genetically manipulated via VIGS technique using pTRV2-BBE (to silencing of BBE1 gene), pTRV2-COM (to simultaneous silencing of T6ODM and CODM genes) and pTRV2-BBE-COM (to simultaneous silencing of T6ODM, CODM and BBE1 genes) constructs. Also, via Agrobacterium-mediated transient expression technique, poppy plants were genetically manipulated using ACS (to simultaneous silencing of T6ODM, CODM and BBE1) and ACS-4ꞌOMT2 (to simultaneous silencing of T6ODM, CODM and BBE1 and over-expression of 4ꞌOMT2) constructs. HPLC analysis showed lower sanguinarine and noscapine levels and elevated levels of morphine in plants infiltrated with pTRV2-BBE construct. In plants infiltrated with pTRV2-COM construct, reduction in the morphine and codeine content and a substantial increase in accumulation of thebaine and papaverine were observed. A substantial increase in the accumulation of thebaine and papaverine and lower levels of morphine, codeine, sanguinarine and noscapine were observed in plants infiltrated with pTRV2-BBE-COM and ACS constructs. Plants infiltrated with ACS-4ꞌOMT2 showed lower levels of codeine, sanguinarine and noscapine and elevated levels of morphine, thebaine and papaverine. Although, previous studies reported the role of our selected genes in benzylisoquinoline alkaloids (BIAs) biosynthetic pathway, but we simultaneously engineered these genes to alter the levels of specific BIAs and take advantage of all genes at the same time. For the first time, we showed different pattern in BIAs accumulation in simultaneous gene manipulation compared with single gene manipulation in previous reports, especially for higher levels of thebaine (about 1550%) and papaverine (about 155%). We also determined potential role of BBE1 gene in noscapine biosynthesis. This is the first report of simultaneous silencing and over-expression of biosynthetic genes in opium poppy and our finding could establish vast potential for metabolic engineering in opium poppy.

Phenylalkylamines, such as the plant compounds ephedrine and pseudoephedrine and the animal neurotransmitters dopamine and adrenaline, compose a large class of natural and synthetic molecules with important physiological functions and pharmaceutically valuable bioactivities. The final steps of ephedrine and pseudoephedrine biosynthesis in members of the plant genus Ephedra involve N-methylation of norephedrine and norpseudoephedrine, respectively. Here, using a plant transcriptome screen, we report the isolation and characterization of an N-methyltransferase (NMT) from Ephedra sinica able to catalyze the formation of (pseudo)ephedrine and other naturally occurring phenylalkylamines, including N-methylcathinone and N-methyl(pseudo)ephedrine. Phenylalkylamine N-methyltransferase (PaNMT) shares substantial amino acid sequence identity with enzymes of the NMT family involved in benzylisoquinoline alkaloid (BIA) metabolism in members of the higher plant order Ranunculales, which includes opium poppy (Papaver somniferum). PaNMT accepted a broad range of substrates with phenylalkylamine, tryptamine, β-carboline, tetrahydroisoquinoline, and BIA structural scaffolds, which is in contrast to the specificity for BIA substrates of NMT enzymes within the Ranunculales. PaNMT transcript levels were highest in young shoots of E. sinica, which corresponded to the location of NMT activity yielding (pseudo)ephedrine, N-methylcathinone, and N-methyl(pseudo)ephedrine, and with in planta accumulation of phenylalkylamines. Co-expression of recombinant genes encoding PaNMT and an ω-transaminase (PP2799) from Pseudomonas putida in Escherichia coli enabled the conversion of exogenous (R)-phenylacetylcarbinol (PAC) and (S)-PAC to ephedrine and pseudoephedrine, respectively. Our work further demonstrates the utility of plant biochemical genomics for the isolation of key enzymes that facilitate microbial engineering for the production of medicinally important metabolites.

Prevailing diversity among germplasm in a crop assists invaluably in crop improvement program. Genetic divergence in 60 germplasm lines of opium poppy was studied based on 13 morphometric characters by using Mahalonobis D2 statistics. The germplasm lines belong to the different eco-geographical origin and are being maintained at CSIR-CIMAP, Lucknow. The ANOVA revealed significant amount of differentiation among germplasm. The genotypes were grouped in eight diverse clusters. Cluster I is the largest group comprises 40 genotypes followed by Cluster II, consisting 12 genotypes and cluster III and IV has 2 genotypes while rest 4 clusters have only a single individual. The maximum intra-cluster distance was observed in cluster IV (93153.35) and minimum for Cluster I (33360.9). Inter-cluster D2 values range from 115261.4 to 4717153. The minimum was between Cluster I and II (115261.4) followed by IV and V (128735.4) and maximum between Cluster I and VII (4717153) followed by Cluster VI and VII (4655531). Based on the mean performance genotypes, G-1 and G-53 can be further selected for exploitation in hybrid breeding program for poppy crop improvement.

Absract The information on gene interactions is a basic requirement for exploitation of genetic variability and advancement in the development of high yielding lines in any crop. The present study was taken to discover the nature of gene action implicated in the inheritance of yield and allied characters in opium poppy. Three testers and 20 lines of opium poppy were crossed in triple test cross design. The results showed that the additive gene action was significant for most of the characters, though, dominance as well as epistasis also had considerable say in the inheritance of characters. The presence of additive effects for all the traits suggested selection based on the studied traits can enhance yield potential effectively in opium poppy. The delayed selection for the traits governed by epistasis may also deliver promising results in opium poppy.

Opium poppy, a member of the Papaveraceae family, is an ancient herbaceous plant and well-known medical resource in the pharmaceutical industry. However, opium poppies are grown worldwide for producing illicit drugs, significantly increasing the incidence of narcotic drug abuse. Since the narcotic poppy has not yet been genetically investigated, we characterized a novel variable number tandem repeat (VNTR) marker of forensically important poppy species based on the genetic analysis of 164 samples collected from two locations spanning the Jeolla province and Jeju island of South Korea. Comparing analysis of the chloroplast (cp) genome sequences for four representative species of Papaver (Papaver somniferum, Papaver somniferum subs. setigerum, Papaver orientale, and Papaver rhoeas) revealed a unique region with 1–3 repeats for 16 nucleotide motifs in the genome inverted repeat A (IRA, positions 128,651 to 128,698) region. For 16 nucleotide motifs, 3 repeats were found in P. somniferum, and 2 repeats were found in P. somniferum subs. setigerum. Therefore, 10 known and the 133 unknown, seized Papaver species were compared to determine whether the species could be identified via variations in the repeat units. The sizes of a novel VNTR ranged from 181 to 252 bp between the species. Phylogenetic analysis confirmed that a novel VNTR, which we named Pscp1, could clearly distinguish between the narcotic and non-narcotic types of Papaver species based on the patterns of sequence variation. Interestingly, we found that Pscp1 could also distinguish between P. somniferum and P. somniferum subs. setigerum. The regions of eight non-narcotic species displayed similar patterns and also differences were found due to the nucleotide substitution and deletion events. The structural differences of Pscp1 were observed within the two narcotic species or between the narcotic and non-narcotic species, suggesting that these variations may act as a genetic marker. We, therefore, developed a new Pscp1 PCR-capillary electrophoresis (CE) method that can reliably identify the narcotic type of Papaver species. Taken together, our findings suggest that the newly developed Pscp1 can be used as an identification marker of opium poppy, and establish that the Pscp1 genotyping method by PCR-CE is an effective primary screening tool that can also contribute to species discrimination in the field of forensic diagnosis and applications.

Opium poppy belongs to the family − ‘Papaveraceae’ and is well known mainly for its pharmaceutically important alkaloids and seeds. In order to delineate the gene-effects conditioning the expression of seed yield and straw alkaloids, six crosses viz. IS 14 × Sampada, I 14 × Mtu, Sampada × Sujata, CIM Ajay × Dfr 44, Sapna × Sanchita and SPS 49 × TS 1 involving the eleven parents, were sampled for P1, P2 and F1 (=P1 × P2) non segregating, and F2 (=F1), BCP1 (=F1 × P1), and BCP2 (=F1 × P2) segregating generations. The days to flowering (50%) and plant height showed all the three type of interactions (i, j, l) in three crosses, namely C1 − IS 14 × Sampada and C2 − I 14 × Mtu but the traits peduncle length, codeine and thebaine content showed in other three crosses as C1 − IS 14 × Sampada, C2 − I 14 × Mtu, C3 − Sampada × Sujata and C4 − CIM Ajay × Dfr 44 in order. The two type of interactions additive × additive and additive × dominance interactions (i, j,) exhibited by the C6 − SPS 49 × TS 1 for plant height and thebaine content, C5 − Sapna × Sanchita for peduncle length, codeine and narcotine content. The genetic basis of character expression indicated that the interacting crosses showing non-allelic interactions particularly duplicate epistasis in most of the crosses. While most of those without epistatic interactions exhibited superiority over parental mean. On the basis of study results the two specific cross combinations like Sampada × Sujata and CIMAP Ajay × Dfr 44 are selected to ensure tangible superiority for the most economic traits like high seed yield and poppy straw/capsule husk alkaloids including other contributing traits also like early flowering, capsule index for poppy crop improvement programme.

This article presents and implements a theoretical foundation for exploring Afghan farmers’ decisions to cultivate opium poppy that can be used to develop supply-control policy and guide future empirical research. To our knowledge, we are the first to employ a household production model that allows Afghan farmers to choose between opium-poppy and licit-crop cultivation to maximize their expected utility, while weighing the costs, benefits, and risks of agricultural production and consumption jointly. We derive policy lessons from the theoretical analysis and empirical evidence that reflect the complexity of Afghan farmers’ decision-making and the socio-economic and environmental factors that influence it.

Among the opium producing countries, India is one of the major producer of illegal opium in which includes the demand in nationwide and global peddling of opium. Opium is derived from the dried latex of the opium poppy and it is also called as the Papaver Somniferum. It can be taken intravenously injected, smoked, and in the forms of pills. The derivatives of opium are morphine, Thebaine, codeine, noscapine, Papervine. It is used for the medicinal purposes also. The symptoms of the opium are nausea, slowed breathing, constipation, confusion, dry mouth and lining of nose, anxiety, depression, vomiting, insomnia, itching etc. Due to the overdose of the opium, the person can go in coma and death may also occur. Opium affects the many internal organs like brain, respiratory system, central nervous system, gastrointestinal system, immune system, liver. In this review article, aims about the detailed study of the narcotic drug and the toxic effects caused by the humans.

Turkey is the world’s leading country in legal poppy cultivation and morphine production. As with other producing countries, the poppy is traditionally cultivated in Turkey through the sowing of seeds during the autumn. However, if the seasonal rains are insufficient and irregular, this can lead to problems with germination and emergence. In this research, the option of solving these problems by using seedlings in production, instead of seeds, was investigated. The study compares the yield and quality performance of the two production methods, planting seedlings and direct seed sowing in the poppy cultivar ‘Ofis-4’. Field experiments were conducted on three sowing/planting dates, namely October fifteenth, November first, and November fifteenth, during the growing seasons of 2017/18 and 2018/19 in the semi-arid ecological conditions of the southwestern region of Turkey. The plants produced by seedling transplantation and by sowing were compared in terms of the yields of seeds and capsules along with yield-related traits and the contents of seed-fixed oil and capsule opium alkaloids. The planting seedling method was seen to be an average of one week faster than the sowing method. In terms of the sowing/planting time delays in the autumn season, it was also observed that the ripening difference between both propagation methods started to narrow. If the general averages are considered, the highest capsule and seed yields were obtained from planting seedling on October fifteenth, the first planting date, in both years. Since the planting dates were late, the capsule and seed yield decreased while it was found that the contents of capsule alkaloids, except morphine and codeine, were not significantly affected by the production method and sowing/planting dates. On the other hand, the direct sowing method and first sowing date produced the highest morphine (0.53 and 0.58 %, respectively) and codeine (0.101 and 0.126 %, respectively) contents. It was therefore concluded that the seedling planting method can be recommended as an alternative to the direct sowing method traditionally applied in poppy production for the achieving of high capsule and seed yields.

The importance of alkaloids of opium poppy is increasing constantly worldwide to fulfill the demand of pharmaceutical industries. India is one of the countries to produce gum opium and contain many potential genotypes to attend the recent demand of alkaloids. So, for proper utilization of the available potential genotypes, the present investigation was carried out to study the genetic diversity and genetic differentiation along with marker-trait association based on quantitative traits as well as AFLP marker in a large number of germplasm lines of opium poppy which were collected from different parts of India. Various genetic parameters and correlation among different traits were also worked out to find out the particular traits enhancing yield potential. Eight AFLP primer pairs were deployed that generated 140 polymorphic bands with fragment ranging in size from 50 to 498 bp in size. The maximum gene diversity was found in the germplasm lines of UP. Most of the variability (96%) was partitioned into within populations implying that collection strategies for conservation should focus on a few populations with many individuals across the ecological amplitude of the population. Genetic differentiation was FST = 0.0413, that implies more or less complete panmixis. The morphine was found associated with seven AFLP loci, such as E-AAG/M-CAG_76, E-AAC/M-CAG_102, E-ACT/M-CTA_181, E-ACG/M-CTC_167, E-ACG/M-CTC_176, E-ACG/M-CTC_194, and E-ACG/M-CTC_102 which showed stability by both models (GLM, MLM) of analysis while three markers (E-ACT/M-CAA_53,E-ACT/M-CAA_243,E-AGG/M-CTG_241) for codeine, four markers (E-ACT/M-CAA_193,E-AAC/M-CAG_151, E-AAC/M-CAG_91, and E-AGC/M-CTA_75) for thebaine, six for narcotine (E-ACT/M-CAA_85, E-AGG/M-CTG_75, E-AAC/M-CAG_116, E-ACT/M-CTA_122, E-ACT/M-CTA_182, and E-ACT/M-CTA_209), two markers (E-AAG/M-CAG_303, and E-AAC/M-CAG_210) for papaverine and five markers (E-ACT/M-CAA_76, E-AAG/M-CAG_122, E-AAG/M-CAG_245, E-AGG/M-CTG_84, and E-AGG/M-CTG_219) for opium yield were found associated (P

Noted as a rich source of thebaine and a potential alternative to Papaver somniferum for production of codeine and some semi-synthetic antagonist drugs (namely naloxone and naltrexone), Papaver bracteatum is a perennial species belongs to the section oxytona. Benzylisoquinoline alkaloids (BIAs) as a diverse class of alkaloids comprises about 2500 various structures contain medically important pharmaceuticals. Developmental and inducible factors control the alkaloid biosynthesis in opium poppy. To make better understanding on comparative developmental accumulation of alkaloid content and tyrosine/dopa decarboxilase (TYDC), berberine bridge enzyme (BBE), salutardinol acetyl transferase (SAT) and codeinone reductase (COR) gene transcripts in different organs of the species, current study was conducted. The accumulation of thebaine was higher in the capsule at pendulous bud (10.66 mg/gr dry weight) following in the root at bud initiation (10.25 mg/gr dry weight). The minimum amount of thebaine was observed in the petal of flowering stage (4.57 mg/gr dry weight). Although in some cases the fluctuation of thebaine content could be coordinated to the up or down-regulation of gene transcripts, however, the presence of considerable exceptions complicate making of a comprehensive conclusion.

The current study was undertaken to estimate the morphological and molecular diversity present among the 51 accessions of Indian opium poppy germplasm using Mahalanobis D2 and SCoT (Start Codon Targeted Polymorphism) marker analyses, respectively. A good range of morphological variations were observed among the accessions. The accessions were clumped into nine clusters and the morphological diversity recorded was 69%. Clusters VII and IX showed the maximum inter-cluster distance (117.97) whereas it was found to be minimum (21.53) in case of clusters II and III. The morphological trait, seed yield per capsule, contributed maximum (17.30%) towards the genetic divergence followed by thebaine content (14.56%) and papaverine content (14.06%). In contrast to the morphological diversity, genetic diversity at molecular level was found to be limited (33%), although successfully detected by utilizing DNA markers targeting coding regions of genome (SCoT markers). The outcome of the study has prospects in identification of lines with desirable traits to be utilized in future breeding programmes.

Poppy has been a plant of much importance and interest since time immemorial. In our 196 accessions, we observed, identified and selected distinct plants containing specific nature of capsules. These capsules if properly grouped according to the number of stigmatic rays – ranging from five to sixteen can become a useful morphological marker for farmers which will assist them instantaneously for selection of yield parameters related to their interests (seeds and or opiate alkaloids). In this investigation we explored the relation between notches in capsules with the agro-economical traits – seed yield and major opiate alkaloid content using plant breeding, bio-statistical and analytical tools and techniques for the first time in this crop. Briefly our study indicated that the rate of grain filling in poppy seeds is indirectly related and the intensity of colour and density of latex is directly related to the population of notches on their respective capsules.

Successful development of stable and adaptable crop varieties solely depend on the positive results obtained from the interaction between genotype and environment that consequently has significant impact on breeding strategies. GEI is a finest breeding approach that helps in the development of varieties for specific or wide adaptation, depends on the expression of stability. The present study was aimed to find out the stable thebaine rich line out of the seven advanced breeding high thebaine lines which have been developed for the first time through rigrous selection from advance generations of interspecific population derived from cross between Papaver somniferum L. and Pa paver setigerum DC. based on the estimate of genotype × environment interaction (GEI). The results of ANOVA showed wide variability in the genotypes under testing which suggested that the genotypes interacted differentially with respect to yield performance, so further stability analysis across environments should be followed before being released for commercial cultivation. The overall stability analysis following different stability models (including regression and graphical models) concluded that the lines NBIHT-1, NBIHT-3 and NBIHT-4 were highly stable and adaptable for seed, opium and thebaine content in different agro-climatic conditions Lines. NBIHT-5 and NBIHT-6 can also perform drastically well in specific climatic conditions to obtain maximum gain. The study concluded that the GGE Biplot model proposed by Yan and Hunt is best suited for stability analysis than other different stability models due to ease in visualization of stable genotypes from the graphical representation. Commercialization of such developed high thebaine lines having more than 10% thebaine in comparison to the existing genotypes having 1–2% thebaine could revolutionize pharmaceutical industries worldwide.

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.

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. (C) 2015 Elsevier B.V. All rights reserved., 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 for 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.

Inhabitants of the Northwest European Plain were rather late in adopting crop cultivation. Most of their crop plants were originally domesticated in the Near East, but on the way to northwestern Europe three of the original founder crops were lost: chickpea, bitter vetch, and lentil. Surprisingly, one plant was added: opium poppy. Another new plant, rye-brome, just missed out becoming established as a true crop. The final adoption of crop cultivation may have been facilitated by the appearance of summer varieties of the cereals grown. Whether the fields were the short-lived fields connected with a slash-and-burn technique or longer-lived is open to debate, but the effect of agriculture was that the landscape became more open, sufficiently open to allow the use of the ard and the wagon.

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.

Cytochrome P450 enzymes are a promising tool for the late-stage diversification of lead drug candidates and can provide an alternative route to structural modifications that are difficult to achieve with synthetic chemistry. In this study, a library of P450BM3 mutants was produced using site-directed mutagenesis and the enzymes screened for metabolism of the opium poppy alkaloid noscapine, a drug with anticancer activity. Of the 18 enzyme mutants screened, 12 showed an ability to metabolise noscapine that was not present in the wild-type enzyme. Five noscapine metabolites were detected by LC-MS/MS, with the major metabolite for all mutants being N-demethylated noscapine. The highest observed regioselectivity for N-demethylation was 88%. Two hydroxylated metabolites, a catechol and two C-C cleavage products were also detected. P450-mediated production of hydroxylated and N-demethylated noscapine structures may be useful for the development of noscapine analogues with improved biological activity. The variation in substrate turnover, coupling efficiency and product distribution between the active mutants was considered alongside in silico docking experiments to gain insight into structural and functional effects of the introduced mutations. Selected mutants were identified as targets for further mutagenesis to improve activity and when coupled with an optimised process may provide a route for the preparative-scale production of noscapine metabolites. Keywords: Cytochrome P450, N-demethylation, Hydroxylation, Site-directed mutagenesis, Biocatalysis

In present study, the energetic and exergetic performances of a diesel engine fueled with conventional diesel and various biodiesel fuels were investigated experimentally. Energetic and exergetic performance parameters of the engine fueled with neat diesel, neat opium poppy and neat palm biodiesel fuels were calculated and compared with each other. Moreover, the sustainability analysis was carried out in the study. The findings of this study indicated that the energetic and exergetic performance parameters were profoundly affected by the engine speed and the fuel type. Both the thermal and exergetic efficiencies were the highest at the engine speed of 1600 rpm for all tested fuels. At this engine speed, the lowest thermal efficiency of the engine occurred with palm biodiesel as 34.92%, while the highest thermal efficiency value was 35.85% with diesel fuel. The exergy efficiencies of corresponding fuel types ranged from 32.50% to 33.64% for palm biodiesel and diesel fuels, respectively. In addition, the difference between the exergy efficiencies of diesel fuel and opium poppy biodiesel fuel was about 0.90%. Both the energy and exergy efficiencies diminished with the increment of engine speed higher than 1600 rpm for all fuel types. The maximum values of sustainability index for all fuel types were obtained at the engine speed of 1600 rpm. The best exergetically-sustainable fuel was the diesel fuel followed by the opium poppy biodiesel fuel. According to the results, opium poppy biodiesel might replace with the conventional diesel fuel due to the similarity of energetic and exergetic performance parameters.

The present research work was undertaken to discern the character associations and the direct as well as indirect effect of component traits on opium yield in biparental inbreeding populations (BIPs) derived from four segregating base populations of the crosses NB-1Kr40-3/3×NB-1Kr30 + 0.2–2/1, NB-5Kr40-7/2 × 58/1, NB-1Kr30 + 0.2–2/1 × 58/1, and NB-1Kr40-3/3×NB-5Kr40-7/2 through North Carolina Design-I (NCD-I). Ample amount of inherent genetic association among the traits was witnessed as the magnitudes of genotypic correlations (rg) were larger than phenotypic correlations (rp) in BIPs developed from all the four populations. Opium yield showed negative correlation with morphine content in all the BIPs derived from four different base populations. It was concluded that selection based on branches/plant, leaves/plant, stem diameter, capsule size, capsule weight/plant, seed yield/plant, and husk yield/plant or concurrent selection for more than one trait may augment the opium yield in opium poppy.

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.

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.

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.

Opium poppy ( Papaver somniferum L.) is an important medicinal plant produces more than 80 alkaloids belonging to various tetrahydrobenzylisoquinoline derived classes. These alkaloids are obtained from the capsules and straw of the plant. Information on the nature and magnitude of gene effects are required for genetic improvement. Therefore, the continuous assessment of newer breeding materials is mandatory on part of breeders. The objective of this study was undertaken to understand the particular gene action involved in the inheritance of yield and component traits. Two families (VG26 × VG20 and SG35II × VE01) of opium poppy were analyzed to study the gene actions involved in the inheritance of yield and component traits (plant height, leaves per plant, capsules per plant, peduncle length, capsule index, seed and straw yield per plant and morphine content). Simple additive, dominance, and epistatic genetic components were found to be significant. Dominance effect (‘ h ’) was higher than additive effect (‘ d ’) for capsule index and morphine content. Digenic interaction indicated the prevalence of dominance × dominance (‘ l ’) followed by additive × dominance (‘ j ’) type epistasis. The opposite sign of dominance (‘ h ’) and dominance × dominance (‘ l ’) indicated duplicate epistasis for all the traits. Biparental mating followed by recurrent selection involving desired recombinants may be utilized to improve the component traits.

Opium poppy, Papaver somniferum L ., is one of the ancient herbal medicines. In addition to this medical use of latex, opium that is extracted from the immature seed capsule, it is also used illegally for pleasure. It is being produced in great quantities in Turkey especially in Afyonkarahisar city. The seeds of opium poppy plant have high ratio oil content. The opium poppy seeds and oil of these seeds are purely used as an ingredient in production of bakery products. In this study, biodiesel evaluation of the opium poppy seeds that have a high oil ratio is aimed. Alkali catalyzed (NaOH) single-phase reaction was preferred to produce biodiesel from opium poppy oil. The parameters like catalyst concentration, methanol ratio, reaction temperature were optimized and biodiesel production was obtained with high yield in reaction time of 75 min. The methyl ester content in the opium poppy oil biodiesel was determined with Gas Chromatography–Frame Ionized Detector (GC–FID). In optimum conditions, methanol ratio and catalyst concentration was determined as 20 wt% and 0.5 wt%, respectively. The reaction temperature was optimized as 60 °C. Biodiesel was obtained from the opium poppy oil under optimum conditions. Some basic features of the produced methyl esters were determined.

Archaeobotanical analyses were carried out on layers from the site of Piazza Garibaldi in Parma, a town located in the plain of Emilia Romagna, in northern Italy. The layers dated to the 3rd–2nd centuries BC, around the time of the foundation of the Roman town, and to the 10th–11th centuries AD. According to archaeological data, the site was a sacred area in Roman times, while it was a market square in Mediaeval times. Data from pollen and seeds/fruits were useful for both palaeoenvironmental and palaeoethnobotanical reconstructions, and together with NPPs including parasite remains contributed to add details on the function of the site in the different chronological phases. Since Roman times, woods have grown far from the site, and human activities highly influenced the landscape. Cereals, legumes and hemp were cultivated together with figs, grapevines, and a number of medicinal, vegetables and spice plants. Altogether, data confirmed the presence of votive offerings, and particularly the association of opium poppy and cereals suggested that they were mainly offered to Ceres, the goddess of crops and soil fertility. In the Middle Ages, cereal fields, together with legumes, grapevines and fruit trees continued to be grown in the area, but olive trees and Prunoideae revealed a different arboricultural economy. The analyses of plant and parasite remains in four pits and one latrine suggested that their infillings consisted of waste, human and animal excrements, deteriorated vegetable food and marcs. Human parasite eggs of Ascaris and Trichuris were found in the latrine, while parasites of animals were found also in the pits. The interpretation of archaeobotanical data was so tightly linked to the archaeological context that similar spectra must be differently interpreted in the two chronological phases.

Opium is a narcotic obtained from opium poppy and raw materials of heroin for illegal drug trade. Information on opium production is important for governments and international communities to understand the scale of illegal drug trade and to implement drug fighting policies. Opium production can be estimated from opium yield and poppy cultivation acreage. Poppy cultivation acreage in turn can be estimated using remote sensing technique. This study explores the feasibility of assessing opium yield using remote sensing technique. The objective of this study is to find an effective biophysical indicator that can be related to both opium yield estimation and remote sensing measurements. Data are obtained from three years ground experiments conducted in an official farm in North-Western China. Opium yield and main plant and environmental parameters of opium poppy are measured. A linear regression analysis measures the relationship between opium yield and main plant parameters. A strong correlation between opium yield and leaf area index (LAI) in flowering and harvesting periods of opium poppy is observed (R^2=0.826 and 0.862 respectively). LAI is a good indicator during flowering and harvesting periods and can be used to build a model for opium yield estimation. Furthermore, LAI can be retrieved using remote sensing at a region scale. The study shows that remote sensing could be used to estimate opium yield in opium poppy concentrated regions, and that a more comprehensive and precise opium production information could be obtained for further analysis of illegal drug circulating trend and policy.

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.

Autopolyploidy is advantageous for plant metabolism in terms of elevated rates of synthesis or a higher variability of metabolically relevant compounds. In the present study, successful induction of polyploidy was achieved through applying colchicine soaked cotton on shoot meristem. The ploidy level of the developed tetraploids was confirmed through microscopic observations of stomata and chromosomal studies. Chromosome number in the developed tetraploids were 2 x = 2 n = 44 as compared to the control having 2 x = 2 n = 22. Alkaloid profile of both treated and control plants showed a significant enhancement from 25% to 50% in morphine content. Expression analysis through semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) of various known genes involved in the biosynthesis of morphinanes showed increased expression in tetraploids. Gene expression analysis of different polyploidy series can supplement our understanding related to molecular mechanism involved in increased alkaloid biosynthesis during polyploidization.

Papaverine is one of the earliest opium alkaloids for which a biosynthetic hypothesis was developed on theoretical grounds. Norlaudanosoline (=tetrahydropapaveroline) was claimed as the immediate precursor alkaloid for a multitude of nitrogen containing plant metabolites. This tetrahydroxylated compound was proposed to be fully O-methylated. The resulting tetrahydropapaverine should then aromatize to papaverine. In view of experimental data, this pathway has to be revised. Precursor administration to 8-day-old seedlings of Papaver followed by direct examination of the metabolic fate of the stable-isotope-labeled precursors in the total plant extract, without further purification of the metabolites, led to elucidation of the papaverine pathway in vivo. The central and earliest benzylisoquinoline alkaloid is not the tetraoxygenated norlaudanosoline, but instead the trihydroxylated norcoclaurine that is further converted into (S)-reticuline, the established precursor for poppy alkaloids. The papaverine pathway is opened by the methylation of (S)-reticuline to generate (S)-laudanine. A second methylation at the 3' position of laudanine leads to laudanosine, both known alkaloids from the opium poppy. Subsequent N-demethylation of laudanosine yields the known precursor of papaverine: tetrahydropapaverine. Inspection of the subsequent aromatization reaction established the presence of an intermediate, 1,2-dihydropapaverine, which has been characterized. The final step to papaverine is dehydrogenation of the 1,2-bond, yielding the target compound papaverine. We conclusively show herein that the previously claimed norreticuline does not play a role in the biosynthesis of papaverine.

Morphine biosynthesis was genetically engineered in an industrial elite line of the opium poppy (Papaver somniferum L.), to modify the production of alkaloids in plants. The cytochrome P-450-dependent monooxygenase (S)-N-methylcoclaurine 3′-hydroxylase (CYP80B3) lies on the pathway to the benzylisoquinoline alkaloid branch point intermediate (S)-reticuline. Overexpression of cyp80b3 cDNA resulted in an up to 450% increase in the amount of total alkaloid in latex. This increase occurred either without changing the ratio of the individual alkaloids, or together with an overall increase in the ratio of morphine. Correspondingly, antisense-cyp80b3 cDNA expressed in opium poppy caused a reduction of total alkaloid in latex up to 84%, suggesting that the observed phenotypes were dependent on the presence of the transgene. This study found compelling evidence, that cyp80b3 is a key regulation step in morphine biosynthesis and provides practical means to genetically engineer valuable secondary metabolites in this important medicinal plant.

This study presents what we believe to be the first formal evaluation of the Taliban crackdown against opium poppy cultivation in Afghanistan. Afghanistan was the main source of the world’s illicit heroin supply for most of the 1990s. From late 2000 and the year that followed, the Taliban enforced a ban on poppy farming via threats, forced eradication, and public punishment of transgressors. The result was a 99% reduction in the area of opium poppy farming in Taliban-controlled areas. The evaluation uses multiple comparison areas: the non-Taliban area of Afghanistan, neighbouring countries, the non-contiguous comparison area of Myanmar (Burma), and, the rest of the world. Alternative possible causes of the reduction such as drought, migration or changes in global opium markets are reviewed and excluded. It is concluded that the reduction in Afghan poppy cultivation was due to the enforcement action by the Taliban. Globally, the net result of the intervention produced an estimated 35% reduction in poppy cultivation and a 65% reduction in the potential illicit heroin supply from harvests in 2001. Though Afghan poppy growing returned to previous levels after the fall of the Taliban government, this may have been the most effective drug control action of modern times. © 2004 Published by Elsevier B.V.

0 d aded the country, it was claimed that generally statistics on fghanistan were “wild guesses based on inadequate data” Dupree, 1980). A report published in 2002 suggested that Afghanistan is one of the least researched countries in the orld. A dearth of pre-war data, the inherent difficulties f conducting research in a war zone and the low priority ccorded to Afghanistan before 9/11 by the international ommunity have all contributed to this problem” (Bhatia, oodhand, Atmar, Pain, & Suleman, 2002). It is a country hat is only now in the process of being popularly discovered y the international community. Even measuring a socially visible, if sometimes not easly accessible (at least from the ground), phenomenon like ultivated fields of opium poppy, remarkable for their bright olours while in bloom, is problematic. According to UNDC, in 2002 there were 74,000 ha planted with poppy opium cultivation and production in Afghanistan serve as crude indicators of the ground realities. One continuing problem is the tendency for politicians to seek ‘quick fix’ solutions to complex social problems, for journalists to seek fast and easy soundbites to sell stories, and for commentators to focus on the “success” of the Taliban ban in eradicating poppy cultivation in 2001 while ignoring the complex historical, political and socio-economic motivations and dynamics that led up to it in the first place. Not to mention the steady increase in opium cultivation and production that characterised the first 5 years of Taliban rule. Such unidimensional views run the risk of encouraging comparisons between Taliban methods then and their potential replication for current poppy reduction strategies in Afghanistan or elsewhere. To call the ban “the most effective drug control enforcement action of modern times” (Farrell & Thorne, UNODC, 2002) while for the same year the US claimed here were only 30,750 ha (US Department of State, 2003), ess than 50% of the UNODC figure. This also meant, accordng to US figures, cultivation had doubled by 2003 as their stimate for that year was 61,000 ha. For the same year UN2005) is disingenuous, as effectiveness must be premised on the use of methods and tactics acceptable within a framework of international law and human rights. Certainly focusing on whether opium cultivation and production figures for 1 year have risen or fallen makes little sense unless they are context o c p t t i v c DC estimated 80,000 ha, only an 8% increase from 2002. hile such discrepancies may be explained away by the use f different methodologies, the UN basing its figures on a ombination of comprehensive ground surveys and analysis f images from commercial satellites, the US on a sample

Opium poppy (Papaver somniferum) produces a large number of benzylisoquinoline alkaloids, including morphine and sanguinarine, derived from tyrosine via the branch-point intermediate (S)-reticuline. Molecular clones for the three methlytransferases involved in (S)-reticuline biosynthesis, (S)-norcoclaurine-6-O-methyltransferase (6OMT), (S)-3'-hydroxy-N-methylcoclaurine-4'-O-methyltransferase (4'OMT), and (S)-coclaurine N-methyltransferase (CNMT), were isolated from opium poppy and shown to share extensive homology with the corresponding cDNAs from Japanese goldthread (Coptis japonica). These cDNAs were used together with previously isolated clones for tyrosine/dopa decarboxylase (TYDC), (S)-N-methylcoclaurine-3'-hydroxylase (CYP80B1), berberine bridge enzyme, (BBE), (7S)-salutaridinol 7-O-acetyltransferase (SAT), and codeinone reductase (COR), to compare the accumulation of gene transcripts encoding eight alkaloid biosynthetic enzymes in opium poppy. Transcript levels generally increased in developing seedlings and were consistently high in stems and flower buds, but were more variable in roots and leaves of mature plants. The accumulation of each transcript, with the exception of COR, showed a marked induction in response to elicitor treatment or wounding of cultured cells. Specific gene transcript levels often correlated with the accumulation of morphine or sanguinarine, with notable exceptions. Our data suggest some degree of coordination in the developmental and inducible regulation of alkaloid biosynthetic genes in opium poppy.

A wealth of prehistoric archaeological findings unearthed by archaeologists has brought forward a rich harvest of information about the use of opium poppy in the Minoan civilization. They reveal the widespread use of opium for cult rituals or healing purposes in the east Mediterranean and Crete at least since 5th century BC, when no written references existed. Poppy capsules ornamenting figurines, bas-reliefs, vases, pins and jewelry imply the multiple symbolic meanings of opium—healing, fertility, wealth and immortality. The Minoan “goddess of poppies, patronage of healing,” dated from 1300 BC, wearing in her head three hairpins of poppy capsules, is a distinct example of the knowledge in the Minoan world of the healing and soporific properties of opium and also of the methods of extracting it from the poppy capsule. The presence of opium poppy in different objects of everyday life like lekythia, rings, pyxis from Crete and Mycenae also suggests a long tradition of the use of opium byproducts in the ancient East Mediterranean.

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).

A variant plant of opium poppy ( Papaver somniferum L.) having high thebaine was obtained from the dose of 10 kR (kilo rad) + 0.4% EMS during an extensive mutation breeding experiment. The M 2 seeds of variant plant were subjected to in vitro studies to investigate the prospects of thebaine production through tissue culture. Alkaloid profile of variant showed higher thebaine in stem followed by leaf callus, stem callus and cotyledons. Such a high content of thebaine from tissue raised plant material opens a new vista for the extraction of thebaine at commercial level. A comparison was also made between in vivo and in vitro cost of production and extraction of thebaine.

Summary An efficient Agrobacterium -mediated protocol has been developed for the stable genetic transformation of intact opium poppy, Papaver somniferum L., plants via shoot organogenesis. Excised cotyledons were cocultivated with the disarmed A. tumefaciens strain GV3101 carrying the pBI121 binary vector, and incubated on an optimized shoot induction medium consisting of B5 salts and vitamins, 30 g L −1 sucrose, 2 mg L −1 6-benzylaminopurine, 5 mg L −1 AgNO 3 , and 3 g L −1 Gelrite. Except for the cocultivation medium, all formulations included 30 mg L −1 paromomycin as the selective agent, and 200 mg L −1 timentin to eliminate the Agrobacterium . Eight-week-old, paromomycin-resistant shoots were transferred to an optimized root induction medium consisting of B5 salts and vitamins, 0.5 mg L −1 indole-3-acetic acid, 0.5 mg L −1 indole-3-butyric acid, and either 5 mg L −1 AgNO 3 or 40 mg L −1 putrescine. About 15 % of the regenerated shoots developed roots within eight weeks. Regenerated plants were transferred to soil, where they grew normally and set seed. Detection of the neomycin phosphotransferase gene, the high levels of β-glucuronidase (GUS) mRNA and enzyme activity, and the cytohistochemical localization of GUS activity in all organs, confirmed the genetic transformation of the regenerated plants. The transformation process did not alter the normal alkaloid content of opium poppy; thus, the reported protocol could serve as a valuable tool to investigate the molecular and metabolic regulation of benzylisoquinoline alkaloid biosynthesis.

In this paper we show the diurnal variation in latex water content in the laticifer system of the opium poppy and relate this variation to the observed variation in alkaloid concentration in the latex. A pronounced, opposed variation to the diurnal water fluctuations are shown for the major opium alkaloids, morphine, codeine and noscapine in the poppy cultivar Parmo under dry weather conditions in Denmark in 1994. Diurnal fluctuations of the alkaloid concentration in the opium poppy latex have previously been explained as a result of rapid metabolism in the biosynthetic sequence leading to morphine. Our results, however, disagree with this explanation as a significant correlation between the diurnal fluctuation in the latex water content and that of the alkaloids morphine, codeine and noscapine can be demonstrated. It is concluded that the observed diurnal fluctuations in the concentration of the major alkaloids are not reflections of enzymatic processes but the result of water transport between the laticifers and the surrounding vascular tissue.

Biomass of formae speciales of Fusarium oxysporum pathogenic to the narcotic plants coca ( Erythroxylum coca ) and opium poppy ( Papaver somniferum ) was incorporated into wheat flour–kaolin granules (Pesta). Shelf life (inoculum viability) of samples with the coca pathogen (strain EN4-S; 2.6 × 10 7 colony-forming units (cfu)/g, mostly chlamydospores) was at least 1 year at 35°C when stored at a water activity ( a W ) of 0.12 (equal to 12% relative humidity) and 2 years when stored at 25°C. Shelf life of samples with the poppy pathogen (strain CP3A; 3.4 × 10 7 cfu/g, mostly microconidia) was about 10 months at 33°C and 0.33 a W . Shelf life was maximized by optimizing water activity (near 0.12 a W for F. oxysporum ) and incorporating at least 5 × 10 6 cfu/g of chlamydospore inoculum. Multikilogram quantities of granular formulations were produced by scaling up the production process by using twin-screw extrusion and fluid-bed drying.

The opium poppy, Papaver somniferum L., and its narcotic and analgesic alkaloids, have an ancient history of use (and abuse) by humankind. A recent article by Allen and co-workers describes the metabolic engineering of morphine biosynthesis to block morphine formation and accumulate a potentially valuable pathway intermediate, (S)-reticuline. This work highlights the potential for modifying the production of pharmaceuticals in plants, but also raises questions about the complex regulation of biosynthetic pathways.

Two early and potential rate-limiting steps in the biosynthesis of isoquinoline alkaloids, such as morphine and codeine, in opium poppy (Papaver somniferum) involve decarboxylation of L-tyrosine and L-dihydroxyphenylalanine (L-dopa) to yield tyramine and dopamine, respectively. A DNA fragment was amplified by polymerase chain reaction (PCR) using degenerate primers designed to two highly conserved domains found in other aromatic amino acid decarboxylases. A poppy seedling cDNA library was screened with this PCR product and a cDNA (cTYDC1) for tyrosine/dopa decarboxylase (TYDC/DODC) was isolated. Two other independent cDNAs (cTYDC2 and cTYDC3) encoding TYDC/DODC were isolated by heterologous screening with a plant tryptophan decarboxylase (TDC) cDNA as probe. A poppy genomic library was screened with cTYDC1 and two intronless genomic clones (gTYDC1 and gTYDC4) were isolated. The deduced amino acid sequences of all poppy clones share extensive identity with other reported pyridoxal phosphate-dependent decarboxylases from both plants and animals. Based on sequence homology, members of the gene family were divided into two subsets (cTYDC1 and gTYDC4; cTYDC2 and cTYDC3) of proteins with predicted M(r) = 56,983 and 59,323, respectively. Within each subset the clones exhibit greater than 90& identity, whereas clones between subsets share less than 75& identity. Expression of gTYDC1 and cTYDC2 as beta-galactosidase fusion proteins in Escherichia coli resulted in catalytically active enzymes immunodetectable with TDC-specific polyclonal antibodies. Each enzyme showed marginally higher substrate specificity for L-dopa over L-tyrosine, but did not accept L-tryptophan and L-phenylalanine as substrates. Genomic DNA blot-hybridization analysis revealed 6 to 8 genes homologous to cTYDC1 and 4 to 6 genes homologous to cTYDC2 in the tetraploid poppy genome. A premature translation stop codon was found in the gTYDC4 clone suggesting that it may not encode a functional protein. RNA blot-hybridization with probes specific to the gTYDC1- or cTYDC2-like subsets showed that members of the TYDC gene family are differentially expressed in various plant tissues.

1,2-Dehydroreticuline reductase, the NADPH-dependent enzyme which reduces stereospecifically 1,2-dehydroreticuline to ( R )-reticuline has been discovered in seedlings of the opium poppy ( Papaver somniferum ). The enzyme has been purified to apparent electrophoretic homogeneity by ammonium sulphate precipitation and five subsequent column chromatography steps. The isolated enzyme is a single polypeptide with M r 30 000 and has a pH optimum at 8.5 and a temperature optimum at 30°. The apparent K m values for 1,2-dehydroreticuline and NADPH are 10 and 7 μM, respectively. The enzyme mediates the transfer of the pro- S -hydride of NADPH to C-1 of 1,2-dehydroreticuline with high substrate specificity; neither 1,2-dehydronorreticuline nor 1,2-dehydrococlaurine are utilized by the enzyme. The enzyme activity is inhibited by ( S )- and ( R )-reticuline with I 50 values of 0.05 and 0.10 mM, respectively. The reductase is a cytosolic enzyme and present only in morphinan alkaloid-containing plants. This highly species-, substrate- and stereospecific enzyme catalyses the provision of ( R )-reticuline for the formation of morphinan alkaloids that possess also ( R )-configuration at that chiral centre.

Tetrahydropapaveroline is an endogenous complex alkaloid derived from dopamine through the oxidation by monoamine oxidase. This alkaloid is considered to be involved in the pathogenesis of alcoholism and to act as a false neurotransmitter. Recently the (S) enantiomer was proposed to be a precursor of morphine biosynthesis in the opium poppy. In this paper stereo-chemical characteristic of tetrahydropapaveroline in human brains was examined. In all four control human brains examined, only the (S)-tetrahydropapaveroline was detected. The concentrations were 0.12-0.22 pmol/g wet weight of brain tissue, and the presence of alcohol in blood did not affect the concentration. The results suggest that (S)-tetrahydropapaveroline may be enantio-selectively synthesized in human brain and it may be an intermediate of the de novo synthesis of morphine analogues.

A cytosolic NADPH 2 dependent enzyme which reduces stereospecifically 1,2-dehydroreticuline to yield (R)-reticuline, the biosynthetic precursor of the morphinan alkaloids, has been discovered, purified to homogeneity, and characterized from Papaver somniferum seedlings.

Introduction Papaver somniferum L. or opium poppy also known in Morocco as “KHERCHACHA” is one of the most used plant in traditional medicine for ritual, prophylactic and therapeutic purposes. we report two cas of poisoning by Papaver somniferum L. Cases reports Twin male infants, aged two months, coming from healthy pregnancy and delivery, were admitted to the emergency for breathing difficulty with cyanosis of the extremities. Clinical exam in admission found respiratory distress with respiratory pause in two infants. One of them had in addition an obnubilation and pupils in bilateral myosis. The interrogation had found the concept of administration of opium poppy for the two children to calm them down and make them sleep. Biological and radiological tests were normal. The toxicological analysis of urine was positive for opiates. Both brothers received symptomatic treatment with favorable outcome. Conclusion Poisoning poppy poisoning remains one of the most dangerous plants. In our context, its use in infants remains an aggravating aspect that requires information and education of mothers. The physicians should be aware of this type of poisoning.

Professors Farrell and Thorne (2005) have written an interesting paper that shows that the Taliban’s ban on opium poppy cultivation in 2000 resulted in dramatic declines of 90.8% in the number of hectares cultivated with poppy in Afghanistan 2002 to levels close to those of 2000. Farrell and Thorne do not pretend to draw conclusions about the possible sustainability of the poppy ban had the Taliban remained in power. There is no question that the Taliban’s ban on poppy was and 35% in the size of illegal poppy plantings in the world. Since the Taliban controlled area in Afghanistan had the highest opium yield per hectare, the declines in opium production were even larger: 94.3% in Afghanistan and 65.3% in the world! Farrell and Thorne, without praising or defending the Taliban, argue that “this may have been the most effective drug control action of modern times” achieved though a combination of “three principal techniques: the threat of punishment, the close local monitoring and eradication of continued poppy farming, plus the public punishment of transgressors”. The authors explore and reject alternative explanations to the poppy crop decline such as “a change in the global heroin market” and bad weather. They also present reasons why the Taliban decided to implement a ban on poppy plantings that emphasize the isolation that the Taliban regime was experiextraordinarily successful in 2001 and confirms that an extremely authoritarian government that has control over the territory can eliminate illicit crops. The authors’ arguments about the events leading to the ban are however based on a partial picture of what occurred in Afghanistan and present an overly positive view of what UNODC achieved. Following the convention used by Farrell and Thorne I use UNODC any time I refer to the United Nations office charged with drug control programmes, even though their acronym and functions have changed through time. I would like to argue that UNODC had less leverage on the Taliban than Farrell and Thorne believe, that a significant change in the world’s heroin market was an important event leading to the poppy ban, and that there are strong indications that the Taliban would not have been able to sustain the poppy ban. First, it is true that UNODC had established contacts with encing from the international community in 1998, the Muslim fundamentalist rejection of mind altering drugs and the Taliban’s assertion that the three year drought experienced by Afghanistan was God’s punishment for the peasants’ cultivation of poppy. Besides, the Taliban had been in contact w O h f c

Rationale The opium poppy, Papaver somniferum L., is the source of both poppy seeds and opium. The commercially available seeds are widely used as ingredients for various kinds of food. IgE-mediated sensitization to poppy seeds is rare but if present, clinical symptoms are usually severe. Cross-reactivity between poppy seeds and other food allergens have been described with sesame, hazelnut, rye grain and kiwi fruit. We report a novel cross-reactivity with buckwheat in a 17-year-old female with poppy seed anaphylaxis. Methods Skin prick test, CAP-FEIA, CAP-inhibition tests, immunoblotting, immunoblot-inhibition and open oral food challenge tests were performed. Results Prick tests with native poppy seeds were strongly positive. Total serum IgE was raised to 646 kU/l, and allergen specific IgE against poppy seed (287 kU/l), hazelnut, sesame, buckwheat, brasil nut, pistachio, cashew nut and almond was detected by CAP-FEIA. CAP inhibition tests demonstrated a cross-reactivity from poppy seed to buckwheat but not to natural rubber latex. Immunoblotting of poppy seed extracts with our patients serum revealed 4 distinct allergen bands, which were inhibited after preincubation with buckwheat extract. The immunoblot for opium latex or raw opium was negative. Oral food challenge with up to 10 g of unground poppy seeds were negative, whereas 1 g of ground poppy seeds led to urticaria, heat sensation, pruritus, conjunctivitis and dyspnoe followed by a threefold raise of mast cell tryptase in serum (5.4 to 17.6 μg/l) within 3 hours. Conclusions Buckwheat must be considered as a novel cross-reactive allergen in at least some poppy seed allergic patients.