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Understanding the adaptability of plants to the environment will help us address the challenges posed by the constantly changing climate to sustainable agriculture and industry. The California poppy (Eschscholzia californica Cham.) of Papaveraceae is an ornamentally and medicinally valuable plant species, not only due to its beautiful flower, but also the strong abiotic stress tolerance and production of pharmacologicallly active alkaloids. However, little is known about the mechanisms of its drought tolerance and association with the alkaloids biosynthesis. In this study, significant morphological and physiological changes of E. californica under early and late drought stress were found. Subsequently, the transcriptomic and metabolomic dynamics between normal and drought conditions were independently and jointly analyzed. Transcriptome analysis identified 24,176 differentially expressed genes (DEGs), which were enriched in alkaloid biosynthesis, phytohormone synthesis and signal transduction pathways, amino acid biosynthesis, and reactive oxygen species scavenging, etc. A large number of DEGs encoding transcription factor represent a reservoir for transcriptional regulation of drought stress responses of E. californica. Metabolomic analysis detected 393 drought responsive differentially expressed metabolites (DEMs), predominant in alkaloids and phenolic acids. Consistency between DEGs and DEMs identified by joint analysis revealed critical pathways in response to drought in E. californica and its close association with alkaloid biosynthesis. This study provides new insights into the drought response mechanisms of E. californica. The contribution of alkaloids to drought tolerance is particularly worthy of further in-depth research. [Display omitted] • The first transcriptomic and metabolomic study of drought response in E. californica. • Essential pathways responding to drought were identified in E. californica. • Drought stress promotes biosynthesis of alkaloid , especially isoquinoline alkaloids. • A large number of drought responsive transcription factors were identified. [ABSTRACT FROM AUTHOR]

The effect of iron oxide nanoparticles on medicinal plants biomass biosynthesis and antioxidant defense compounds should be assessed critically to ensure their safe and sustainable use within the agricultural fields and food chains. Current focus has been given to the effects of hematite nanoparticles (α-Fe 2 O 3 NPs, ∼ 27 nm) and ferric ions (Fe3+) at different concentrations [(0, control), 10, 100 and 1000 mg L−1)] on morpho-physiological and biochemical parameters and benzophenanthridine alkaloids content in Eschscholzia californica plants. Upon exposure to α-Fe 2 O 3 NPs, a significant (P < 0.05) inhibition (by 26.6%) of root biomass was observed at the high concentration (1000 mg L−1) over the control. As for the total dry weight of plants, significant (P < 0.01) increase (by 22.7%) was observed at all exposures, except that of 1000 mg L−1 α-Fe 2 O 3 NPs/and Fe+3 treatment which was reduced by 12.8%. The treatment of 100 mg L−1 α-Fe 2 O 3 NPs and Fe3+ caused significant increase in chlorophyll a (29.2% and 40%,), chlorophyll b (59.8% and 49.3%,) and carotenoids (14.3% and 25.4%%,) content when compared to control, respectively. However, accumulation of the lipid peroxidation product, malondialdehyde (MDA), in root and leaf tissues was similar (73.5% vs control) that was induced by 1000 mg L−1 α-Fe 2 O 3 NPs. No significant change in MDA content was observed in leaves and roots of Fe+3 treated plants at all exposure doses. Furthermore, total phenolics and flavonoids content of the leaves and roots exposed to α-Fe 2 O 3 NPs at 100 mg L−1 were significantly (P < 0.01) increased by 41–45%, 27–29%, and 44–48%, respectively, over the control. All obtained extracts revealed strong and significant scavenging effects on DPPH˙ as compared to control, but only exposure to 100 mg L−1 Fe+3 and α-Fe 2 O 3 NPs treatment revealed the highest free radical scavenging/antioxidant activity by 44.2% and 37.65%, respectively. The alkaloid accumulation in leaves exhibited significant (P < 0.05) differences between the control plants and those treated with α-Fe 2 O 3 NPs and Fe+3 of all concentrations. The eshchschollzine and sanguinarine contents showed no significant change in plants treated with Fe+3 concentrations; however, the exposure of high Fe+3 concentration increased (33.4%) the content of chelerythrine compared to control. Foliar applied α-Fe 2 O 3 NPs enhanced the content of californidine (35.4%), eshchschollzine (22.3%), and eshchschollzine (19.1%) compared to control. The observed toxicity was dose-dependent for both α-Fe 2 O 3 NPs and Fe+3. The synthesized endogenous secondary metabolites (e.g., phenolics, flavonoids and benzophenanthridine alkaloids) induce defense mechanism in exposed plants. These results suggest that specific concentrations (10–100 mg L−1) of α-Fe 2 O 3 NPs have extra benefits both as a nanoregulator and nanoelicitor for plant growth promotion and biosynthesis of alkaloids and cellular antioxidants. • Exposure to various doses of α-Fe 2 O 3 NPs and Fe+3 differentially affect root/shoot biomass. • Foliar applied 100 mg L−1 α-Fe 2 O 3 NPs increased photosynthetic pigments content. • The lipid peroxidation product was similarly accumulated in root and leaf tissues of α-Fe 2 O 3 NPs exposed plants. • Hormetic dose-dependent response was found on accumulation of alkaloids in leaves. • Multivariate analyses showed the significance of specific traits for interpreting the observed variance. [ABSTRACT FROM AUTHOR]

Transcription factors of the WRKY family play pivotal roles in plant defense responses, including the biosynthesis of specialized metabolites. Based on the previous findings of WRKY proteins regulating benzylisoquinoline alkaloid (BIA) biosynthesis, such as CjWRKY1—a regulator of berberine biosynthesis in Coptis japonica—and PsWRKY1—a regulator of morphine biosynthesis in Papaver somniferum—we performed genome-wide characterization of the WRKY gene family in Eschscholzia californica (California poppy), which produces various BIAs. Fifty WRKY genes were identified by homology search and classified into three groups based on phylogenetic, gene structure, and conserved motif analyses. RNA sequencing showed that several EcWRKY genes transiently responded to methyl jasmonate, a known alkaloid inducer, and the expression patterns of these EcWRKY genes were rather similar to those of BIA biosynthetic enzyme genes. Furthermore, tissue expression profiling suggested the involvement of a few subgroup IIc EcWRKYs in the regulation of BIA biosynthesis. Transactivation analysis using luciferase reporter genes harboring the promoters of biosynthetic enzyme genes indicated little activity of subgroup IIc EcWRKYs, suggesting that the transcriptional network of BIA biosynthesis constitutes multiple members. Finally, we investigated the coexpression patterns of EcWRKYs with some transporter genes and discussed the diversified functions of WRKY genes based on a previous finding that CjWRKY1 overexpression in California poppy cells enhanced BIA secretion into the medium.

Genome characterization of California poppy (Eschscholzia californica cv. "Hitoezaki"), which produces pharmaceutically important benzylisoquinoline alkaloids (BIAs), was carried out using the draft genome sequence. The numbers of tRNA and rRNA genes were close to those of the other plant species tested, whereas the frequency of repetitive sequences was distinct from those species. Comparison of the predicted genes with those of Amborella trichopoda, Nelumbo nucifera, Solanum lycopersicum , and Arabidopsis thaliana , and analyses of gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway indicated that the enzyme genes involved in BIA biosynthesis were highly enriched in the California poppy genome. Further comparative analysis using the genome information of Papaver somniferum and Aquilegia coerulea , both BIA-producing plants, revealed that many genes encoding BIA biosynthetic enzymes, transcription factors, transporters, and candidate proteins, possibly related to BIA biosynthesis, were specifically distributed in these plant species. [ABSTRACT FROM AUTHOR]

The nutrient‐rich organic waste generated by ants may affect plant reproductive success directly by enhancing fruit production but also indirectly, by affecting floral traits related with pollinator attraction. Understanding how these soil‐nutrient hot spots influence floral phenotype is relevant to plant–pollination interactions. We experimentally evaluated whether the addition of organic waste from refuse dumps of the leaf‐cutting ant Acromyrmex lobicornis (Hymenoptera: Formicidae: Attini) alters floral traits associated with pollinator attraction in Eschscholzia californica (Ranunculales: Papaveraceae), an entomophilous herb. We analysed flower shape and size using geometric morphometric techniques in plants with and without the addition of refuse‐dumps soil, under greenhouse conditions. We also measured the duration of flowering season, days with new flowers, flower production and floral display size. Plants growing in refuse‐dumps soil showed higher flower shape diversity than those in control soil. Moreover, plants in refuse‐dumps soil showed bigger flower and floral display size, longer flowering season, higher number of flowering days and flower production. As all these variables may potentially increase pollinator visits, plants in refuse‐dumps soil might increase their fitness through enhanced attraction. Our work describes how organic waste from ant nests may enhance floral traits involved in floral attraction, illustrating a novel way of how ants may indirectly benefit plants. [ABSTRACT FROM AUTHOR]

Key message: Viral-induced gene silencing of selected biosynthetic genes decreased overall carotenoid accumulation in California poppy. Regulation of carotenogenesis was linked with pigment sequestration, not changes in biosynthetic gene expression.Abstract: Genes of carotenogenesis are well described, but understanding how they affect carotenoid accumulation has proven difficult because of plant lethality when the pigments are lacking. Here, we used a Tobacco Rattle Virus-based virus-induced-gene-silencing (VIGS) approach in California poppy (Eschscholzia californica) to investigate how silencing of the carotenoid biosynthetic pathway genes affects carotenoid metabolite accumulation and RNA transcript abundance of the carotenoid biosynthetic pathway genes. VIGS of upstream (PDS and ZDS) and downstream (βOH and ZEP) genes reduced transcript abundance of the targeted genes in the poppy petals while having no effect on abundance of the other carotenogenesis genes. Silencing of PDS, ZDS, βOH and ZEP genes reduced total pigment concentration by 75-90% and altered petal colour. HPLC and LC-MS measurements suggested that petal colour changes were caused by substantially altered pigment profiles and quantity. Carotenoid metabolites were different to those normally detected in wild-type petals accumulated but overall carotenoid concentration was less, suggesting the chemical form of carotenoid was important for whether it could be stored at high amounts. In poppy petals, eschscholtzxanthin and retro-carotene-triol were the predominant carotenoids, present mainly as esters. Specific esterification enzymes for specific carotenoids and/or fatty acids appear key for enabling petal carotenoids to accumulate to high amounts. Our findings argue against a direct role for carotenoid metabolites regulating carotenogenesis genes in the petals of California poppy as transcript abundance of carotenogenesis genes studied was unchanged, while the petal carotenoid metabolite profile changed substantially. [ABSTRACT FROM AUTHOR]

Summary: The petals of Eschscholzia californica (California poppy) are robust, pliable and typically coloured intensely orange or yellow owing to the presence of carotenoid pigments; they are also highly reflective at certain angles, producing a silky effect. To understand the mechanisms behind colour enhancement and reflectivity in California poppy, which represents a model species among early‐divergent eudicots, we explored the development, ultrastructure, pigment composition and optical properties of the petals using light microscopy and electron microscopy combined with both spectrophotometry and goniometry. The elongated petal epidermal cells each possess a densely thickened prism‐like ridge that is composed primarily of cell wall. The surface ridges strongly focus incident light onto the pigments, which are located in plastids at the cell base. Our results indicate that this highly unusual, deeply ridged surface structure not only enhances the deep colour response in this desert species, but also results in strongly angle‐dependent ‘silky’ reflectivity that is anisotropic and mostly directional. [ABSTRACT FROM AUTHOR]

Severe infection by powdery mildew was observed on California poppy (Eschscholzia californica) plants in Texcoco, State of Mexico, during spring of 2014 to 2017. Symptoms included colonies of white to greyish mildew-like growth on the abaxial and adaxial surfaces of the leaves. The identification of the fungal species was performed by examination of morphological structures using light microscopy and scanning electron microscope (SEM), as well as sequence analysis of the 5ʹ-end of 28S rRNA gene and the internal transcribed spacer (ITS) region of rDNA. Using the combination of morphological characterization and a phylogenetic analysis using Bayesian inference, the fungal agent was identified as Erysiphe eschscholziae. Pathogenicity tests were conducted on leaves of California poppy plants, and Koch’s postulates were fulfilled. This is the first report of E. eschscholziae causing powdery mildew on Eschscholzia californica in Mexico. [ABSTRACT FROM AUTHOR]

The berberine bridge enzyme from the California poppy Eschscholzia californica (EcBBE) catalyzes the oxidative cyclization of (S)‐reticuline to (S)‐scoulerine, that is, the formation of the berberine bridge in the biosynthesis of benzylisoquinoline alkaloids. Interestingly, a large number of BBE‐like genes have been identified in plants that lack alkaloid biosynthesis. This finding raised the question of the primordial role of BBE in the plant kingdom, which prompted us to investigate the closest relative of EcBBE in Physcomitrella patens (PpBBE1), the most basal plant harboring a BBE‐like gene. Here, we report the biochemical, structural, and in vivo characterization of PpBBE1. Our studies revealed that PpBBE1 is structurally and biochemically very similar to EcBBE. In contrast to EcBBE, we found that PpBBE1 catalyzes the oxidation of the disaccharide cellobiose to the corresponding lactone, that is, PpBBE1 is a cellobiose oxidase. The enzymatic reaction mechanism was characterized by a structure‐guided mutagenesis approach that enabled us to assign a catalytic role to amino acid residues in the active site of PpBBE1. In vivo experiments revealed the highest level of PpBBE1 expression in chloronema, the earliest stage of the plant's life cycle, where carbon metabolism is strongly upregulated. It was also shown that the enzyme is secreted to the extracellular space, where it may be involved in later steps of cellulose degradation, thereby allowing the moss to make use of cellulose for energy production. Overall, our results suggest that the primordial role of BBE‐like enzymes in plants revolved around primary metabolic reactions in carbohydrate utilization. Database: Structural data are available in the PDB under the accession numbers 6EO4 and 6EO5. [ABSTRACT FROM AUTHOR]

Land plants produce specialized low molecular weight metabolites to adapt to various environmental stressors, such as UV radiation, pathogen infection, wounding and animal feeding damage. Due to the large variety of stresses, plants produce various chemicals, particularly plant species-specific alkaloids, through specialized biosynthetic pathways. In this study, using a draft genome sequence and querying known biosynthetic cytochrome P450 (P450) enzymeencoding genes, we characterized the P450 genes involved in benzylisoquinoline alkaloid (BIA) biosynthesis in California poppy (Eschscholzia californica), as P450s are key enzymes involved in the diversification of specialized metabolism. Our in silico studies showed that all identified enzyme-encoding genes involved in BIA biosynthesis were found in the draft genome sequence of approximately 489 Mb, which covered approximately 97% of the whole genome (502 Mb). Further analyses showed that some P450 families involved in BIA biosynthesis, i.e. the CYP80, CYP82 and CYP719 families, were more enriched in the genome of E. californica than in the genome of Arabidopsis thaliana, a plant that does not produce BIAs. CYP82 family genes were highly abundant, so we measured the expression of CYP82 genes with respect to alkaloid accumulation in different plant tissues and two cell lines whose BIA production differs to estimate the functions of the genes. Further characterization revealed two highly homologous P450s (CYP82P2 and CYP82P3) that exhibited 10-hydroxylase activities with different substrate specificities. Here, we discuss the evolution of the P450 genes and the potential for further genome mining of the genes encoding the enzymes involved in BIA biosynthesis. [ABSTRACT FROM AUTHOR]

Transcription factors control many processes in plants and have high potentials to manipulate specialized metabolic pathways. Transcriptional regulation of the biosynthesis of monoterpenoid indole alkaloids (MIAs), nicotine alkaloids, and benzylisoquinoline alkaloids (BIAs) has been characterized using Catharanthus roseus, Nicotiana and Coptis plants. However, metabolic engineering in which specific transcription factors are used in alkaloid biosynthesis is limited. In this study, we characterized the effects of ectopic expression of CjWRKY1, which is a transcriptional activator with many targets in BIA biosynthesis in Coptis japonica (Ranunculaceae) and Eschscholzia californica (California poppy, Papaveraceae). Heterologous expression of CjWRKY1 in cultured California poppy cells induced increases in transcripts of several genes encoding BIA biosynthetic enzymes. Metabolite analyses indicated that the overexpression of the CjWRKY1 gene also induced increases in the accumulation of BIAs such as sanguinarine, chelerythrine, chelirubine, protopine, allocryptopine, and 10-hydroxychelerythrine in the culture medium. Previous characterization of EcbHLH1 and current results indicated that both transcription factors, WRKY1 and bHLH1, are substantially involved in the regulation of BIA biosynthesis. We discuss the function of CjWRKY1 in E. californica cells and its potential for metabolic engineering in BIA biosynthesis. [ABSTRACT FROM AUTHOR]

Background: Reductions of genetic diversity and phenotypic changes in invasive plants are often observed to occur at high elevations. Genetic/phenotypic changes of invasive plants along elevation help to understand mechanisms of the presumed resistance of mountain ecosystems to invasion. Aims: To assess genetic variability and phenotypic plasticity along an elevation gradient of Eschscholzia californica in the Andes, central Chile. Methods: Eleven microsatellites were used to describe the genetic structure and the allelic diversity individuals, distributed at three elevations and two sites. We assessed the number of flowers per plant, floral biomass, leaf area, number of leaves, vegetative biomass and plant height of plants at each elevation. Results: Genetic diversity as genetic structure did not decrease with elevations. Plant height and flower numbers decreased while leaf number and vegetative biomass increased with elevation. The ratio of the number of flowers to vegetative biomass, decreased significantly with elevation. Conclusions: Strong genetic differences among elevations and similar genetic diversity along elevation do not suggest dispersal limitation to higher elevation. Reduction of reproductive and vegetative traits concomitantly with an increase of the reproductive cost suggests reproductive stress with increasing elevation, reducing the invasiveness of this species to higher elevation. [ABSTRACT FROM AUTHOR]

California poppy (Eschscholzia californica), a member of the Papaveraceae family, produces many biologically active benzylisoquinoline alkaloids (BIAs), such as sanguinarine, macarpine and chelerythrine. Sanguinarine biosynthesis has been elucidated at the molecular level, and its biosynthetic genes have been isolated and used in synthetic biology approaches to produce BIAs in vitro. However, several genes involved in the biosynthesis of macarpine and chelerythrine have not yet been characterized. In this study, we report the isolation and characterization of a novel O-methyltransferase (OMT) involved in the biosynthesis of partially characterized BIAs, especially chelerythrine. A search of the RNA sequence database from NCBI and PhytoMetaSyn for the conserved OMT domain identified 68 new OMT-like sequences, of which the longest 22 sequences were selected based on sequence similarity. Based on their expression in cell lines with different macarpine/chelerythrine profiles, we selected three OMTs (G2, G3 and G11) for further characterization. G3 expression in Escherichia coli indicated O-methylation activity of the simple benzylisoquinolines, including reticuline and norreticuline, and the protoberberine scoulerine with dual regio-reactivities. G3 produced 7-O-methylated, 30-O-methylated and dual O-methylated products from reticuline and norreticuline, and 9-O-methylated tetrahydrocolumbamine, 2-O-methylscoulerine and tetrahydropalmatine from scoulerine. Further enzymatic analyses suggested that G3 is a scoulerine-9-O-methyltransferase for the biosynthesis of chelerythrine in California poppy. In the present study, we discuss the physiological role of G3 in BIA biosynthesis. [ABSTRACT FROM AUTHOR]

Background: Seed mass is a life history trait that is related to invasiveness. Under limiting conditions, a trade-off is observed whereby an increase in seed mass occurs at the expense of seed numbers; if the above trade-off holds across climatic gradients it can provide an opportunity to assess the fitness/invasive potential of exotic plants. Aims: To examine the variation in the life history traits ofEschscholzia californicapopulations across climatic gradients and to relate these traits to observed invasiveness. Methods: We examined 19 populations in Chile. For each population we related seed mass, seed number, the slope of trade-off between seed mass and number and plant density with annual precipitation and mean annual temperature. Results: Seed number and the coefficient of variation in seed mass were positively correlated with climatic variables. Trade-off was detected in 26% of the populations and no relationship was detected with climatic gradient. Plant density was negatively associated with precipitation. Conclusions: The results suggest that forE. californicaproducing seeds with a variety of sizes is an optimal strategy to face geographic heterogeneity and hence to increase its invasiveness. Increased production of seeds at the cool and wet southern limit of the current range of the species does not contribute to an increase in its invasiveness. [ABSTRACT FROM PUBLISHER]

The article presents abstracts on topics of medicinal agents derived from plants which include the changes in expression of selected genes in California poppy seedlings, genetic resources of medicinal plants and phytochemical analysis of amorpha fruticosa.

Cell cultures of Eschscholzia californica react to a fungal elicitor by the overproduction of antimicrobial benzophenanthridine alkaloids. The signal cascade toward the expression of biosynthetic enzymes includes (1) the activation of phospholipase A2 at the plasma membrane, resulting in a peak of lysophosphatidylcholine, and (2) a subsequent, transient efflux of vacuolar protons, resulting in a peak of cytosolic H+. This study demonstrates that one of the Na+/H+ antiporters acting at the tonoplast of E. californica cells mediates this proton flux. Four antiporter-encoding genes were isolated and cloned from complementary DNA (EcNHX1-EcNHX4). RNA interference-based, simultaneous silencing of EcNHX1, EcNHX3, and EcNHX4 resulted in stable cell lines with largely diminished capacities of (1) sodium-dependent efflux of vacuolar protons and (2) elicitor-triggered overproduction of alkaloids. Each of the four EcNHX genes of E. californica reconstituted the lack of Na+-dependent H+ efflux in a βnhx null mutant of Saccharomyces cerevisiae. Only the yeast strain transformed with and expressing the EcNHX1 gene displayed Na+-dependent proton fluxes that were stimulated by lysophosphatidylcholine, thus giving rise to a net efflux of vacuolar H+. This finding was supported by three-dimensional protein homology models that predict a plausible recognition site for lysophosphatidylcholine only in EcNHX1. We conclude that the EcNHX1 antiporter functions in the elicitor-initiated expression of alkaloid biosynthetic genes by recruiting the vacuolar proton pool for the signaling process. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The component Allee effect has been defined as 'a positive relationship between any measure of individual fitness and the number or density of conspecifics'. Larger plant populations or large patches have shown a higher pollinator visitation rate, which may give rise to an Allee effect in reproduction of the plants. We experimentally tested the effect of number of conspecifics on reproduction and pollinator visitation in Eschscholzia californica Cham., an invasive plant in Chile. We then built patches with two, eight and 16 flowering individuals of E. californica (11 replicates per treatment) in an area characterised by dominance of the study species. We found that E. californica exhibits a component Allee effect, as the number of individuals of this species has a positive effect on individual seed set. However, individual fruit production was not affected by the number of plants examined. Pollinator visitation rate was also independent of the number of plants, so this factor would not explain the Allee effect. This rate was positively correlated with the total number of flowers in the patches. We also found that the number of plants did not affect the seed mass or proportion of germinated seeds in the patches. Higher pollen availability in patches with 16 plants and pollination by wind could explain the Allee effect. The component Allee effect identified could lead to a weak demographic Allee effect that might reduce the rate of spread of E. californica. Knowledge of this would be useful for management of this invasive plant in Chile. [ABSTRACT FROM AUTHOR]

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, CjbHLHI, comprehensively regulates berberine biosynthesis in Coptis japonica. Interestingly, CjbHLH1 homologous genes were found in many IQAproducing plant species, which suggests that non-MYC2- type CjbHLH homologs are specifically associated with IQA biosynthesis. To test whether CjbHLHI 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 EcbHLHIs regulate IQA biosynthesis in California poppy like CjbHLHI in C. japonica. [ABSTRACT FROM AUTHOR]

Species climate requirements are useful for predicting their geographic distribution. It is often assumed that the niche requirements for invasive plants are conserved during invasion, especially when the invaded regions share similar climate conditions. California and central Chile have a remarkable degree of convergence in their vegetation structure, and a similar Mediterranean climate. Such similarities make these geographic areas an interesting natural experiment for testing climatic niche dynamics and the equilibrium of invasive species in a new environment. We tested to see if the climatic niche of Eschscholzia californica is conserved in the invaded range (central Chile), and we assessed whether the invasion process has reached a biogeographical equilibrium, i.e., occupy all the suitable geographic locations that have suitable conditions under native niche requirements. We compared the climatic niche in the native and invaded ranges as well as the projected potential geographic distribution in the invaded range. In order to compare climatic niches, we conducted a Principal Component Analysis (PCA) and Species Distribution Models (SDMs), to estimate E. californica's potential geographic distribution. We also used SDMs to predict altitudinal distribution limits in central Chile. Our results indicated that the climatic niche occupied by E. californica in the invaded range is firmly conserved, occupying a subset of the native climatic niche but leaving a substantial fraction of it unfilled. Comparisons of projected SDMs for central Chile indicate a similarity, yet the projection from native range predicted a larger geographic distribution in central Chile compared to the prediction of the model constructed for central Chile. The projected niche occupancy profile from California predicted a higher mean elevation than that projected from central Chile. We concluded that the invasion process of E. californica in central Chile is consistent with climatic niche conservatism but there is potential for further expansion in Chile. [ABSTRACT FROM AUTHOR]

In compound leaves, leaflet primordia are initiated directionally along the lateral sides. Our understanding of the molecular basis of leaflet initiation has improved, but the regulatory mechanisms underlying spatio-temporal patterns remain unclear. In this study, we investigated the mechanisms of acropetal (from the base to the tip) progression of leaflet initiation in Eschscholzia californica. We established an ultraviolet-laser ablation system to manipulate compound-leaf development. Local ablation at the leaflet incipient site generated leaves with asymmetric morphology. In the majority of cases, leaflets that were initiated on the ablated sides shifted apically. Finite time-course observation revealed that the timing of leaflet initiation was delayed, but the distance from the leaf tip did not decrease. These results were suggestive of the local spacing mechanism in leaflet initiation, whereby the distance from the leaf tip and adjacent pre-existing leaflet determines the position of leaflet initiation. To understand how such a local patterning mechanism generates a global pattern of successive leaflet initiation, we assessed the growth rate gradient along the apical-basal axis. Our time-course analysis revealed differential growth rates along the apical-basal axis of the leaf, which can explain the acropetal progression of leaflet initiation. We propose that a leaflet is initiated at a site where the distances from pre-existing leaflets and the leaf tip are sufficient. Furthermore, the differential growth rate may be a developmental factor underlying the directionality of leaflet initiation. [ABSTRACT FROM AUTHOR]

Carpel development and evolution are central issues for plant biology. The conservation of genetic functions conferring carpel identity has been widely studied in higher plants. However, although genetic networks directing the development of characteristic features of angiosperm carpels such as stigma and style are increasingly known in Arabidopsis thaliana, little information is available on the conservation and diversification of these networks in other species. Here, we have studied the functional conservation of NGATHA transcription factors in widely divergent species within the eudicots., We determined by in situ hybridization the expression patterns of NGATHA orthologs in Eschscholzia californica and Nicotiana benthamiana. Virus-induced gene silencing ( VIGS)-mediated inactivation of NGATHA genes in both species was performed and different microscopy techniques were used for phenotypic characterization., We found the expression patterns of Ec NGA and Nb NGA genes during flower development to be highly similar to each other, as well as to those reported for Arabidopsis NGATHA genes. Inactivation of Ec NGA and Nb NGA also caused severe defects in style and stigma development in both species., These results demonstrate the widely conserved essential role of NGATHA genes in style and stigma specification and suggest that the angiosperm-specific NGATHA genes were likely recruited to direct a carpel-specific developmental program. [ABSTRACT FROM AUTHOR]

In angiosperms, the shoot apical meristem is at the origin of leaves and stems and is eventually transformed into the floral meristem. Class I knotted-like homeobox ( KNOX I) genes are known as crucial regulators of shoot meristem formation and maintenance. KNOX I genes maintain the undifferentiated state of the apical meristem and are locally downregulated upon leaf initiation. In Arabidopsis, KNOX I genes, especially SHOOTMERISTEMLESS ( STM), have been shown to regulate flower development and the formation of carpels. We investigated the role of STM-like genes in the reproductive development of Eschscholzia californica, to learn more about the evolution of KNOX I gene function in basal eudicots. We identified two orthologs of STM in Eschscholzia, EcSTM1 and EcSTM2, which are predominantly expressed in floral tissues. In contrast, a KNAT1/ BP-like and a KNAT2/ 6-like KNOX I gene are mainly expressed in vegetative organs. Virus-induced gene silencing (VIGS) was used to knockdown gene expression, revealing that both EcSTM genes are required for the formation of reproductive organs. Silencing of EcSTM1 resulted in the loss of the gynoecium and a reduced number of stamens. EcSTM2-VIGS flowers had reduced and defective gynoecia and a stronger reduction in the number of stamen than observed in EcSTM1-VIGS. Co-silencing of both genes led to more pronounced phenotypes. In addition, silencing of EcSTM2 alone or together with EcSTM1 resulted in altered patterns of internodal elongation and sometimes in other floral defects. Our data suggest that some aspects of STM function present in Arabidopsis evolved already before the basal eudicots diverged from core eudicots. [ABSTRACT FROM AUTHOR]

Abstract: Benzophenanthridine alkaloids, such as sanguinarine, are produced from reticuline, a common intermediate in benzylisoquinoline alkaloid biosynthesis, via protopine. Four cytochrome P450s are involved in the biosynthesis of sanguinarine from reticuline; i.e. cheilanthifoline synthase (CYP719A5; EC 1.14.21.2.), stylopine synthase (CYP719A2/A3; EC 1.14.21.1.), N-methylstylopine hydroxylase (MSH) and protopine 6-hydroxylase (P6H; EC 1.14.13.55.). In this study, a cDNA of P6H was isolated from cultured Eschscholzia californica cells, based on an integrated analysis of metabolites and transcript expression profiles of transgenic cells with Coptis japonica scoulerine-9-O-methyltransferase. Using the full-length candidate cDNA for P6H (CYP82N2v2), recombinant protein was produced in Saccharomyces cerevisiae for characterization. The microsomal fraction containing recombinant CYP82N2v2 showed typical reduced CO-difference spectra of P450, and production of dihydrosanguinarine and dihydrochelerythrine from protopine and allocryptopine, respectively. Further characterization of the substrate-specificity of CYP82N2v2 indicated that 6-hydroxylation played a role in the reaction. [Copyright &y& Elsevier]

* Premise of the study: On a compound leaf, leaflet primordia are repetitively formed along the apical--basal axis, with the direction varying among taxa. Why and how the directions vary among species is yet to be solved, although a change in a single factor was proposed to cause the variation. In this study, we compared two species in the Papaveraceae with different directions of leaflet initiation, Chelidonium majus subsp. asiaticum (basipetal) and Eschscholzia californica (acropetal). Because E. californica has been studied in some detail, we focused on C. majus and asked how basipetal pattern is achieved. * Methods: Since only immature leaf primordial tissue has leaflet-generating competency, we performed histological and gene expression analyses on markers of the tissue maturation state. In addition, we performed a time-course analysis of leaf primordial growth. * Key results: Quantitative reverse transcription-PCR analysis demonstrated that a putative regulator of tissue maturation in C. majus, the CINCINNATA homolog, had higher expression in apical parts than in basal parts during the organogenetic phase. In contrast, expression of the CIN homolog was not elevated in either the apical or basal parts in E. californica during the organogenetic phase. * Conclusions: In C. majus, apical parts of leaf primordia have already lost leaflet-generating competency during the organogenetic phase. We propose that precocious progression of the maturation process instructs basipetal progression of leaflet initiation in C. majus. This is not the mirror image of data on E. californica, which shows the opposite direction in leaflet formation, indicating that variation in direction is not attributable to a change in a single factor. [ABSTRACT FROM AUTHOR]

The plasma membrane of the California poppy is known to harbour a PLA2 (phospholipase A2) that is associated with the Gɑ protein which facilitates its activation by a yeast glycoprotein, thereby eliciting the biosynthesis of phytoalexins. To understand the functional architecture of the protein complex, we titrated purified plasma membranes with the Gɑ protein (native or recombinant) and found that critical amounts of this subunit keep PLA2 in a low-activity state from which it is released either by elicitor plus GTP or by raising the Gɑ concentration, which probably causes oligomerization of Gɑ, as supported by FRET (fluorescence resonance energy transfer)-orientated fluorescence imaging and a semiquantitative split-ubiquitin assay. All effects of Gɑ were blocked by specific antibodies. A low-Gɑ mutant showed elevated PLA2 activity and lacked the GTP-dependent stimulation by elicitor, but regained this capability after pre-incubation with Gɑ. The inhibition by Gɑ and the GTP-dependent stimulation of PLA2 were diminished by inhibitors of peptidylprolyl cis-trans isomerases. A cyclophilin was identified by sequence in the plasma membrane and in immunoprecipitates with anti-Gɑ antibodies. We conclude that soluble and target-associated Gɑ interact at the plasma membrane to build complexes of varying architecture and signal amplification. Protein-folding activity is probably required to convey conformational transitions from Gɑ to its target PLA2. [ABSTRACT FROM AUTHOR]

Flowers are reproductive shoots produced by determinate floral meristems. The role of FLORICAULA/ LEAFY-like genes in the specification of flowers varies between lineages and has not been characterized in a basal eudicot species. Here we report the phenotypic effects of virus-induced silencing of EcFLO, a FLORICAULA/LEAFY homologue in the California poppy Eschscholzia californica. EcFLO silencing resulted in repeated sepal and petal whorl formation and internodes between calyxes. A subset of silenced flowers showed reduced petal and stamen numbers and mosaic identities of floral organs. We also provide expression data of other floral regulator genes and conclude that EcFLO contributes to certain aspects of flower development, reflecting both functional conservation and differentiation among basal eudicots, core eudicots, and monocot grasses. EcFLO is involved in stamen and petal initiation, floral organ identity delimitation, suppression of internodes, and limitation of the number of perianth whorls. Further, silencing of EcFLO allows insights into perigynous flower development in Eschscholzia. We demonstrate that receptacle cup formation, by which the perianth and the androecium are elevated above the gynoecium late in flower development, responds to the initiation of the synsepalous calyx. [ABSTRACT FROM AUTHOR]

The article examines the phylogenetic relationships and patterns of character evolution among the California poppies and their relatives (Papaveraceae, Eschscholtzieae). Morphological comparisons and parsimony were used in the study, along with likelihood-based phylogenetic analyses of two chloroplast and two nuclear DNA regions. The subspecies of both Eschscholzia californica and Eschscholzia lemmonii has been promoted to specific status based on the results of the study.

The Crabs Claw (CRC) YABBY gene is required for regulating carpel development in angiosperms and has played an important role in nectary evolution during core eudicot speciation. The function or expression of CRC-like genes has been explored in two basal eudicots, Eschscholzia californica and Aquilegia formosa. To further investigate the function of CRC orthologous genes related to evolution of carpel and nectary development in basal eudicots, a CRC ortholog, EsCRC, was isolated and characterized from Epimedium sagittatum (Sieb. and Zucc.) Maxim. A phylogenetic analysis of EsCRC and previously identified CRC-like genes placed EsCRC within the basal eudicot lineage. Gene expression results suggest that EsCRC is involved in the development of sepals and carpels, but not nectaries. Phenotypic complementation of the Arabidopsis mutant crc-1 was achieved by constitutive expression of EsCRC. In addition, over-expression of EsCRC in Arabidopsis and tobacco gave rise to abaxially curled leaves. Transgenic results together with the gene expression analysis suggest that EsCRC may maintain a conserved function in carpel development and also play a novel role related to sepal formation. Absence of EsCRC and ElCRC expression in nectaries further indicates that nectary development in non-core eudicots is unrelated to expression of CRC-like genes. [ABSTRACT FROM AUTHOR]

The revegetation of soils affected by historic depositions of an industrial complex in Central Chile was studied. The plant re-colonization from the existing soil seed bank and changes in the physico-chemical properties of the soil were evaluated in field plots amended with lime and/or compost. We found that the application of lime and/or compost decreased the Cu2+ ion activity in the soil solution and the exchangeable Cu in the soil, showing an effective Cu immobilization in the topsoil. Whereas lime application had no effect on plant productivity in comparison with the unamended control, the application of compost and lime+compost increased the plant cover and aboveground biomass due to the higher nutrient availability and water-holding capacity of the compost-amended soils. Although the Cu2+ activity and the exchangeable Cu were markedly lower in the amended soils than in the unamended control, the shoot Cu concentrations of Lolium spp. and Eschscholzia californica did not differ between the treatments. [ABSTRACT FROM AUTHOR]

Bees are important flower-visiting insects that display differential occurrences at food resources throughout urban and wildland landscapes. This study examined the visitation rates and foraging patterns of eight taxonomic groups of bees that are common to California poppies, Eschscholzia californica, in both landscape types. Bee occurrence was documented in relation to floral resource characteristics (patch area, poppy coverage, and poppy density), local landscape characteristics (distance to the wildland-urban interface, distance to riparian areas, distance to green space, and land use), and regional landscape context (urban versus wildland). Similar abundance and richness measures were recorded at both urban and wildland poppy patches, but community composition varied in each landscape. Bumble bees were more abundant at poppies in the wildland whereas species in the family Halictidae (sweat bees) were more abundant at poppies within the urban landscape. Resource patch size and density consistently correlated with increased bee presence for all bee types foraging in the wildland. Individual patterns of occurrence in the urban landscape were somewhat divergent; the foraging dynamics of larger bodied-bees ( Bombus vosnesenskii and Megachile species) correlated significantly with resource patch size and density, whereas smaller-bodied bees (family Halictidae and Andrena species) were influenced by landscape characteristics such as distance to the wildland-urban interface and distance to riparian areas. In summary, the surrounding landscape had an influence on community composition, but the magnitude of the floral resource present at a site and factors relating to foraging energetics were dominant drivers of local occurrence. These results suggest that management strategies that provide dense and abundant floral resources should be successful in attracting bees, irrespective of their location within the urban matrix. [ABSTRACT FROM AUTHOR]

YABBY genes are seed plant-specific transcriptional regulators that are involved in diverse aspects of leaf, shoot and flower development. A series of duplications gave rise to five gene groups found throughout flowering plants. In Arabidopsis and other species, expression of two gene groups, CRABS CLAW and INNER NO OUTER, is restricted to floral organs. In contrast, members of the FILAMENTOUS FLOWER, YABBY2 and YABBY5 gene groups are also expressed in leaves and have been termed 'vegetative YABBYs'. How the five paralogue groups evolved and how their expression and function diversified have remained largely unresolved, precluding a reconstruction of the natural history of this gene family. Here, we report new genes from Eschscholzia californica (Ranunculales, Papaveraceae) that we use together with currently available database sequences in a comprehensive phylogenetic re-evaluation of the YABBY gene family. Multilayered Bayesian analysis covering seed plants allowed us to locate Eschscholzia YABBY sequences within the gene family phylogeny. We established that vegetative YABBYs do not form a monophyletic clade, and that CRABS CLAW and FILAMENTOUS FLOWER arose from a common ancestor gene. INNER NO OUTER genes are sister to that ancestral gene. We identified several conserved motifs outside of known amino acid domains that define all five angiosperm YABBY gene clades. Further, we inferred the evolution of gene expression and provide evidence for release of purifying constraint in certain branches of the gene family tree. Finally, we report expression patterns for five Eschscholzia YABBY genes consistent with functional conservation between early-diverged and core eudicots. [ABSTRACT FROM AUTHOR]

Background: There is conclusive evidence that there are fitness costs of plant defense and that herbivores can drive selection for defense. However, most work has focused on above-ground interactions, even though belowground herbivory may have greater impacts on individual plants than above-ground herbivory. Given the role of belowground plant structures in resource acquisition and storage, research on belowground herbivores has much to contribute to theories on the evolution of plant defense. Pocket gophers (Geomyidae) provide an excellent opportunity to study root herbivory. These subterranean rodents spend their entire lives belowground and specialize on consuming belowground plant parts. Methodology and Principal Findings: We compared the root defenses of native forbs from mainland populations (with a history of gopher herbivory) to island populations (free from gophers for up to 500,000 years). Defense includes both resistance against herbivores and tolerance of herbivore damage. We used three approaches to compare these traits in island and mainland populations of two native California forbs: 1) Eschscholzia californica populations were assayed to compare alkaloid deterrents, 2) captive gophers were used to test the palatability of E. californica roots and 3) simulated root herbivory assessed tolerance to root damage in Deinandra fasciculata and E. californica. Mainland forms of E. californica contained 2.5 times greater concentration of alkaloids and were less palatable to gophers than island forms. Mainland forms of D. fasciculata and, to a lesser extent, E. californica were also more tolerant of root damage than island conspecifics. Interestingly, undamaged island individuals of D. fasciculata produced significantly more fruit than either damaged or undamaged mainland individuals. Conclusions and Significance: These results suggest that mainland plants are effective at deterring and tolerating pocket gopher herbivory. Results also suggest that both forms of defense are costly to fitness and thus reduced in the absence of the putative target herbivore. [ABSTRACT FROM AUTHOR]

The model basal eudicot plant California poppy (Eschscholzia californica Cham.) typically has intense yellow to orange petals and orange pollen due to pigmentation by carotenoids. Flower color variants ranging from white to yellow and orange are common. We analyzed flower color inheritance in a diverse range of white and yellow color variants with reduced carotenoid content. The inheritance of the petal–pollen color of 24 variant flowers was investigated through complementation analysis by hybridization between different color variants and screening F1, F2, and BC1 populations for segregation of petal–pollen color. All white and yellow flower color variants exhibited the pleiotropic effect with each mutation influencing both petal and pollen color, with both petal and pollen color phenotypes coinherited. A total of 5 complementation groups were identified with the color variants behaving as single recessive loci. Epistatic interactions among the loci were also identified. The white/yellow California poppy color variants described in this paper represent a unique genetic resource for analysis of carotenoid biosynthesis in this basal eudicot species. [ABSTRACT FROM PUBLISHER]

Many plant cells respond to pathogens by the induction of phytoalexin biosynthesis, but the underlying changes of gene expression are often obscured by their close linkage to the complex rearrangements during pathogen defense, especially the hypersensitive cell death. In root-derived cell cultures of Eschscholzia californica, the overproduction of cytotoxic benzophenanthridine alkaloids can be triggered by a minimum of pathogen pressure that does not evoke hypersensitive reactions. Such conditions activate a signal chain that is initiated by a short contact to low concentrations of yeast glycoprotein elicitor and includes a transient acidification of the cytoplasm. In contrast, high elicitor concentrations signal via an increase of jasmonate and trigger hypersensitive cell death, preceded by a drastic decay of translatable mRNAs. The main changes in protein and mRNA patterns caused by either signal path were compared by 2D proteomic separation, MS/MS sequencing and mRNA-in vitro translation. The four proteins showing the highest overexpression were identical between cells that received low or high-elicitor treatment and overlapped with the three proteins most up-regulated by artificial pH shifts. They comprised one biosynthetic enzyme (norcoclaurine:SAM 4' O-methyl-transferase) plus a unique combination of stress-protective proteins: a heat shock protein (hsp 70); a peptidyl-prolyl-cis/trans isomerase (cyclophilin); and a glyceraldehyde-3-phosphate dehydrogenase. It appears that overproduction of the benzophenanthridine phytoalexins requires the up-regulation of a rate-limiting biosynthetic enzyme plus the coordinated expression of a specific set of protective enzymes and thus is managed like an oxidative stress. [ABSTRACT FROM PUBLISHER]

Higher plants produce a diverse array of secondary metabolites. These chemicals are synthesized from simple precursors through multistep reactions. To understand how plant cells developed such a complicated metabolism, we examined the plasticity of benzyl isoquinoline alkaloid biosynthesis in transgenic Eschscholzia californica cells with the ectopic expression of Coptis japonica scoulerine-9-O-methyltransferase (CjSMT). CjSMT catalyzes the O-methylation of scoulerine to produce tetrahydrocolumbamine (THC) in berberine biosynthesis and is not involved in benzophenanthridine alkaloid biosynthesis in E. californica. While a preliminary characterization confirmed that columbamine (oxidized product of THC) was produced in transgenic E. californica cells, many newly found peaks were not identified. Here, we report the identification of novel products, including allocryptopine and 10-hydroxychelerythrine. This result indicates that CjSMT reaction products were further converted by endogenous enzymes to produce double O-methylated compounds instead of a methylenedioxy ring at the 7,8-position of the original benzophenanthridine alkaloids. Further metabolite profiling revealed the enhanced diversification of the alkaloid profile in transgenic cells. Metabolic plasticity and the enzymes involved in metabolic diversity are discussed. [ABSTRACT FROM PUBLISHER]

Background: The floral homeotic C function gene AGAMOUS (AG) confers stamen and carpel identity and is involved in the regulation of floral meristem termination in Arabidopsis. Arabidopsis ag mutants show complete homeotic conversions of stamens into petals and carpels into sepals as well as indeterminacy of the floral meristem. Gene function analysis in model core eudicots and the monocots rice and maize suggest a conserved function for AG homologs in angiosperms. At the same time gene phylogenies reveal a complex history of gene duplications and repeated subfunctionalization of paralogs. Results: EScaAG1 and EScaAG2, duplicate AG homologs in the basal eudicot Eschscholzia californica show a high degree of similarity in sequence and expression, although EScaAG2 expression is lower than EScaAG1 expression. Functional studies employing virus-induced gene silencing (VIGS) demonstrate that knock down of EScaAG1 and 2 function leads to homeotic conversion of stamens into petaloid structures and defects in floral meristem termination. However, carpels are transformed into petaloid organs rather than sepaloid structures. We also show that a reduction of EScaAG1 and EScaAG2 expression leads to significantly increased expression of a subset of floral homeotic B genes. Conclusions: This work presents expression and functional analysis of the two basal eudicot AG homologs. The reduction of EScaAG1 and 2 functions results in the change of stamen to petal identity and a transformation of the central whorl organ identity from carpel into petal identity. Petal identity requires the presence of the floral homeotic B function and our results show that the expression of a subset of B function genes extends into the central whorl when the C function is reduced. We propose a model for the evolution of B function regulation by C function suggesting that the mode of B function gene regulation found in Eschscholzia is ancestral and the C-independent regulation as found in Arabidopsis is evolutionarily derived. [ABSTRACT FROM AUTHOR]

The Arabidopsis transcription factor CRABS CLAW ( CRC) is a major determinant of carpel growth and fusion, and, in concert with other redundantly acting genes, of floral meristem termination. Its rice ortholog, however, has additional functions in specifying carpel organ identity. We were interested in understanding the history of gene function modulation of CRC-like genes during angiosperm evolution. Here, we report the identification and functional characterization of EcCRC, the Californica poppy ( Eschscholzia californica) CRC ortholog. The downregulation of EcCRC by virus-induced gene silencing (VIGS) produces additional organ whorls that develop exclusively into gynoecia, resulting in a reiteration of the fourth whorl. Additionally, defects in carpel polarity and ovule initiation are apparent, and the observed phenotype is restricted to the gynoecium. Our results further show that the history of CRC-like genes during angiosperm evolution is characterized by gains of function, independent of duplication processes in this gene subfamily. Moreover, our data indicate that the ancestral angiosperm CRC-like gene was involved in floral meristem termination and the promotion of abaxial cell fate in the gynoecium, and that in the lineage leading to Arabidopsis, additional genes have been recruited to adopt some of these functions, resulting in a high degree of redundancy. [ABSTRACT FROM AUTHOR]

Abstract   Eschscholzia californica produces various types of isoquinoline alkaloids. The structural diversity of these chemicals is often due to cytochrome P450 (P450) activities. Members of the CYP719A subfamily, which are found only in isoquinoline alkaloid-producing plant species, catalyze methylenedioxy bridge-forming reactions. In this study, we isolated four kinds of CYP719A genes from E. californica to characterize their functions. These four cDNAs encoded amino acid sequences that were highly homologous to Coptis japonica CYP719A1 and E. californica CYP719A2 and CYP719A3, which suggested that these gene products may be involved in isoquinoline alkaloid biosynthesis in E. californica, especially in methylenedioxy bridge-forming reactions. Expression analysis of these genes showed that two genes (CYP719A9 and CYP719A11) were preferentially expressed in plant leaf, where pavine-type alkaloids accumulate, whereas the other two showed higher expression in root than in other tissues. They were suggested to have distinct physiological functions in isoquinoline alkaloid biosynthesis. Enzyme assay analysis using recombinant proteins expressed in yeast showed that CYP719A5 had cheilanthifoline synthase activity, which was expected based on the similarity of its primary structure to that of Argemone mexicana cheilanthifoline synthase (deposited at DDBJ/GenBank™/EMBL). In addition, enzyme assay analysis of recombinant CYP719A9 suggested that it has methylenedioxy bridge-forming activity toward (R,S)-reticuline. CYP719A9 might be involved in the biosynthesis of pavine- and/or simple benzylisoquinoline-type alkaloids, which have a methylenedioxy bridge in an isoquinoline ring, in E. californica leaf. [ABSTRACT FROM AUTHOR]

In cultured cells of California poppy ( Eschscholzia californica), lysophosphatidylcholine (LPC) triggers a signal path that finally induces alkaloid biosynthesis. LPC is transiently generated by elicitor-activated phospholipase A2 of the plasma membrane. Externally added LPC is rapidly acylated by a membrane-bound enzyme that shows the highest specific activity in the purified plasma membrane. The fatty acid incorporated into the sn-2 position of LPC is preferentially linoleic (18:2), which is the most abundant acyl component in the PC species of Eschscholzia cells, but a minor component of the pool of free fatty acids. The fatty acid at the sn-1 position of LPC is less important for substrate specificity. The capacity of LPC acylation by intact cells or isolated plasma membranes by far exceeds the rate of LPC generation by activated phospholipase A2 and is not limited by the availability of acyl donors. Metabolites other than phosphatidylcholine (PC) were not significantly produced from labeled LPC within 20 min, indicating that lysophospholipases are not significantly contributing to the short-time metabolism of LPC. It is concluded that reacylation to PC is the dominating process in the detoxication of LPC and ensures the transient character of its steady state concentrations, even at maximum phospholipase A2 activities. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Production of the benzophenanthridine alkaloids in Eschscholtzia californica suspension cell cultures was optimized by adding 0.5 mg methyl jasmonate (MJ) and 0.02 mg salicylic acid (SA)/g FCW after 7 days cultivation. Sanguinarine reached 24 mg/g DCW by such treatment; 10 times higher than in control cell cultures. MJ and SA induced expression of berberine bridge enzyme and 3′-hydroxy-( S)- N-methylcoclaurine-4′- O-methyltransferase, respectively. MJ plus SA induced over-expression of both enzymes. [ABSTRACT FROM AUTHOR]

Insight into the speed and predictability of local adaptation can be gained by studying organisms, such as invasive species, that have recently expanded their geographical ranges. Common garden studies were designed to address these issues with the California poppy, Eschscholzia californica, collected from a wide range of environments in both its native (California) and invasive (Chile) ranges. We found similar patterns of plant trait variation along similar abiotic gradients in plants collected from both areas. Multivariate analysis demonstrated that coastal plants from both areas tended to be shorter, smaller plants with smaller seeds and flowers that germinate and flower later than plants collected from inland locations. In addition, size and fecundity traits in both native and invasive poppies were correlated with average rainfall totals; the plants that grew the largest and were the most fecund during the first year of growth originated from the driest areas. This parallel variation suggests that these traits are adaptive and that these patterns have evolved in Chile during the 110–150 years since introduction. [ABSTRACT FROM AUTHOR]