Laticifers in stamens of Papaver somniferum L

Summary Articulated anastomosing laticifers were identified at both light and electron microscopic levels in the stamens of Papaver somniferum L. They were observed associated with the phloem forming a continuous system from the filament into the anther of the stamen. Laticifers, which were comparable in structure to laticifers found elsewhere in the plant, possessed numerous vesicles of different sizes within the protoplast.     

Oxidation of tyrosine by Papaver somniferum latex

The polyphenolase complex isolated from the organelles which sedimented at 1000 g from the latex of Papaver somniferum was found to be composed of soluble and mernbrane-bound fractions. Partial purification resolved two polyphenolases, only one of which utilized tyrosine, a probable precursor of the alkaloid morphine. Activity of these two polyphenolase fractions was shown to change during the development of the capsule.     

Mineral Nutrition and Morphine Production in Papaver somniferum

Mineral nutrition of poppy (Paparer somniferum L.) was studied in its effects on morphine production. Hydroponic cultures were carried out with nutritive solutions percolating over sand. The anion NO_3- is the most efficient form of nitrogen for the production of fresh matter, dry matter or total morphine; in the latter respect, it rates higher than the NH₄NO₃ form which, on the other hand, gives higher alkaloid contents, while both cation NH₄⁺ and urea have depressive effects. Phosphates have apparently little effect on the growth of the poppy, but solutions enriched with assimilable phosphate do stimulate flower proliferation and fruit development, without increasing markedly the total morphine output. Mg²⁺ and Ca²⁺ are important factors; Mg deficiencies will bring about a marked elongation of stems and early flowering, without any notable decrease in morphine outputs; conversely, Ca deficiencies will cause a drop in alkaloid production, while more calcium in the solution will give stronger elongation, a larger number of capsules and a marked increase in the weight of dry matter and morphine outputs, without any marked change in content. Sodium will favour poppy development (flowers and capsules) and will increase both the content and output of morphine. Na⁺ should therefore be introduced whenever possible in the fertilizing of poppy crops.     

Genomic organization of the glutathione S-transferase family in insects

Cytosolic glutathione S-transferases (GSTs) are a large and diverse gene family in insects. They are classified into six major subclasses. Sigma, Omega, Zeta, and Theta have representatives across Metazoa while Delta and Epsilon are specific to Insecta and Holometabola, respectively. In this study, GSTs are assigned to a subclass by a combination of literature, phylogenetic, and genomic evidence. Moreover, it is confirmed that GSTs frequently cluster by genomic position as a result of recent gene expansions. These expansions are largely explained by the number of protein-coding genes in the genome, although life history is another contributing factor.     

Somatic embryogenesis in the opium poppy, Papaver somniferum

A procedure is described for the induction of somatic embryos in the opium poppy. Papaver somniferum L. Callus was obtained from seedling hypocotyls on an agar solidified medium [Murashige and Skoog (1962) Physiol. Plant. 15: 473–497] containing 0.25 mg/l (1.2 μ M ) kinetin and 2.0 mg/l (10.7 μ M ) naphtalene acetic acid (NAA). Suspension cultures were initiated from callus using a liquid medium in which 2.0 mg/l (9.0 μ M ) 2,4-dichlorophenoxyacetic acid was substituted for NAA. Meristemoids, spheres of closely packed cells, developed in suspensions and on the surface of a few callus cultures. Differentiation of meristemoids into somatic embryos was accomplished by removing growth regulators from the liquid medium. Embryoids appeared morphologically normal and similar to torpedo stage embryos, however, they possessed mature tracheary elements and laticifers in areas that should have contained only procambium. Whole plants have been obtained by placing embryos in the light on solid medium that also lacked growth regulators.     

Intracellular delivery of glutathione S-transferase into mammalian cells

Protein transduction domains (PTDs) derived from human immunodeficiency virus Tat protein and herpes simplex virus VP22 protein are useful for the delivery of non-membrane-permeating polar or large molecules into living cells. In the course of our study aiming at evaluating PTD, we unexpectedly found that the fluorescent-dye-labeled glutathione S-transferase (GST) from Schistosoma japonicum without known PTDs was delivered into COS7 cells. The intracellular transduction of GST was also observed in HeLa, NIH3T3, and PC12 cells, as well as in hippocampal primary neurons, indicating that a wide range of cell types is permissive for GST transduction. Furthermore, we showed that the immunosuppressive peptide VIVIT fused with GST successfully inhibits NFAT activation. These results suggest that GST is a novel PTD which may be useful in the intracellular delivery of biologically active molecules, such as small-molecule drugs, bioactive peptides, or proteins.     

Bound morphine and codeine in the capsule of Papaver somniferum

The polysaccharide fraction of the capsule of Papaver somniferum contained bound morphine and codeine. The alkaloids appear to be bound to the polymer by two different types of linkage.     

New insights into the evolution of metazoan tyrosinase gene family

Tyrosinases, widely distributed among animals, plants and fungi, are involved in the biosynthesis of melanin, a pigment that has been exploited, in the course of evolution, to serve different functions. We conducted a deep evolutionary analysis of tyrosinase family amongst metazoa, thanks to the availability of new sequenced genomes, assessing that tyrosinases (tyr) represent a distinctive feature of all the organisms included in our study and, interestingly, they show an independent expansion in most of the analyzed phyla. Tyrosinase-related proteins (tyrp), which derive from tyr but show distinct key residues in the catalytic domain, constitute an invention of chordate lineage. In addition we here reported a detailed study of the expression territories of the ascidian Ciona intestinalis tyr and tyrps. Furthermore, we put efforts in the identification of the regulatory sequences responsible for their expression in pigment cell lineage. Collectively, the results reported here enlarge our knowledge about the tyrosinase gene family as valuable resource for understanding the genetic components involved in pigment cells evolution and development.     

Variability of flavonol contents during floral morphogenesis in Papaver somniferum L

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.     

Some enzymes of general metabolism in the latex of Papaver somniferum

Enzymes of general metabolism have been determined in the latex of Papaver somniferum in an attempt to elucidate further the nature of the 1000 g130 min organelles and their role in alkaloid biogenesis. A number of enzymes involved in the glyoxylic acid and tricarboxylic acid cycles have been found, namely, aconitase, isocitrate dehydrogenase, succinate dehydrogenase, fumarase, malate dehydrogenase and isocitrate lyase. Two enzymes of glycolysis, namely, pyruvate kinase and lactate dehydrogenase, as well as enzymes associated with peroxisomes (glyoxylate reductase, catalase) and lysosomes (arylesterase, acid phosphatase) have been studied. Finally, some enzymes previously reported as occurring in poppy seedlings have been investigated, namely peroxidase, glutamate—oxaloacetate and glutamate-pyruvate transaminases, together with phenylalanine, tyrosine, DOPA and glutamic acid decarboxylases.     

A genomics approach to the comprehensive analysis of the glutathione S-transferase gene family in soybean and maize

By BLAST searching a large expressed sequence tag database for glutathione S-transferase (GST) sequences we have identified 25 soybean (Glycine max) and 42 maize (Zea mays) clones and obtained accurate full-length GST sequences. These clones probably represent the majority of members of the GST multigene family in these species. Plant GSTs are divided according to sequence similarity into three categories: types I, II, and III. Among these GSTs only the active site serine, as well as another serine and arginine in or near the "G-site" are conserved throughout. Type III GSTs have four conserved sequence patches mapping to distinct structural features. Expression analysis reveals the distribution of GSTs in different tissues and treatments: Maize GSTI is overall the most highly expressed in maize, whereas the previously unknown GmGST 8 is most abundant in soybean. Using DNA microarray analysis we observed increased expression among the type III GSTs after inducer treatment of maize shoots, with different genes responding to different treatments. Protein activity for a subset of GSTs varied widely with seven substrates, and any GST exhibiting greater than marginal activity with chloro-2,4 dinitrobenzene activity also exhibited significant activity with all other substrates, suggesting broad individual enzyme substrate specificity.     

罂粟正名考

十世纪前罂粟传入中国后,随着岁月的迁移出现了大量的中文名称。其中不少的名称曾作为正名载于古籍。本文讨论了古代罂粟用名的混乱和正名的选用。     

The Ultrastructure of the Alkaloidal Vesicles of Papaver somniferum latex

Papaver somniferum latex contains abundant small vesicles. Theirultrastructure was studied in tissue sections from adult plantsand in sections of sequential fractions of centrifuged latex.The vesicles were found to exist in two forms, the first witha smooth but progressively granulated outer membrane and thesecond, probably derived from the first, with adherent ‘cap-like’structures which in the heavier centrifuged fractions possesseda zonally-ordered interior. These vesicle fractions were active in synthesizing morphineand the name ‘alkaloidal vesicle’ is proposed forthem. Papaver somniferum latex also contains an organelle whichwas found to resemble a complex organelle present in the latexof Hevea brasiliensis. Its function is not yet known.     

Molecular characterization of an anthocyanin-related glutathione S-transferase gene in cyclamen

Anthocyanins are a subclass of flavonoids and are a major contributor to flower colors ranging from red to blue and purple. Previous studies in model and ornamental plants indicate a member of the glutathione S-transferase (GST) gene family is involved in vacuolar accumulation of anthocyanins. In order to identify the anthocyanin-related GST in cyclamen, degenerate PCR was performed using total RNA from immature young petals. Four candidates of GSTs (CkmGST1 to CkmGST4) were isolated. Phylogenetic analysis indicated that CkmGST3 was closely related to PhAN9, an anthocyanin-related GST of petunia, and this clade was clustered with other known anthocyanin-related GSTs. Expression analysis at different developmental stages of petals revealed that CkmGST3 was strongly expressed in paler pigmented petals than in fully pigmented petals, in contrast to the constitutive expression of the other three candidates during petal development. This expression pattern of CkmGST3 was correlated with those of other anthocyanin biosynthetic genes such as CkmF3'5'H and CkmDFR2. Molecular complementation of Arabidopsis tt19, a knockout mutant of an anthocyanin-related GST gene, demonstrated that CkmGST3 could complement the anthocyanin-less phenotype of tt19. Transgenic plants that expressed the other three CkmGSTs did not show anthocyanin accumulation. These results indicate CkmGST3 functions in anthocyanin accumulation in cyclamen.