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To assess the clinical efficacy of a neurotonic component containing fixed quantities of two plant extracts (Crataegus oxyacantha and Eschscholtzia californica) and magnesium versus placebo in mild-to-moderate anxiety disorders with associated functional disturbances, under usual general practice prescription conditions.A total of 264 patients (81% female; mean age: 44.6 years) presenting with generalised anxiety (DSM-III-R) of mild-to-moderate intensity (total Hamilton anxiety scale score between 16 and 28) were included in a double-blind, randomised, placebo-controlled trial. Patients were randomly assigned to two groups: 130 received the study drug (Sympathyl), and 134 a placebo (two tablets twice daily for 3 months). Efficacy and safety data were recorded before first administration and 7, 14, 30, 60 and 90 days after start of treatment.Efficacy was assessed by (a) change in Hamilton anxiety scale total and somatic scores; (b) change in patient self-assessment; (c) number and percentage of responsive subjects (reduction of at least 50% in Hamilton or self-assessment score); and (d) the physician's clinical global impression. Tolerance was assessed by undesirable events spontaneously reported by the patients over the study period.Total and somatic Hamilton scale scores and subjective patient-rated anxiety fell during treatment, indicating clinical improvement. The decrease was greater in the study drug than in the placebo group. End of treatment clinical improvement, as measured by the mean difference between final and pre-treatment scores, was, for the study drug and placebo groups respectively: -10.6 and -8.9 on the total anxiety score (p = 0.005); -6.5 and -5.7 on the somatic score (p = 0.054); and -38.5 and -29.2 for subjectively assessed anxiety (p = 0.005). The risk/benefit ratio as judged by the investigating physicians was also significantly greater in the study drug than in the placebo group. In all, 15 patients (11.5%) in the study drug group and 13 patients (9.7%) in the placebo group experienced 22 and 15 adverse events, respectively. Undesirable events were mainly mild or moderate digestive or psychopathological disorders.The preparation containing fixed quantities of Crataegus oxyacantha, Eschscholtzia californica, and magnesium proved safe and more effective than placebo in treating mild-to-moderate anxiety disorders. Sympathyl is produced and marketed by Laboratoire Innotech International, Arcueil, France.

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

Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG geförderten) Allianz- bzw. Nationallizenz frei zugänglich. This publication is with permission of the rights owner freely accessible due to an Alliance licence and a national licence (funded by the DFG, German Research Foundation) respectively. A facile test system based on the accumulation of benzo[c]phenanthridine alkaloids in Eschscholzia californica cell suspension culture (an indicator of defense gene activation) has been used to analyze a series of synthetic compounds for elicitor-like activity. Of the 200 jasmonic acid and coronatine analogs tested with this system, representative results obtained with 49 of them are presented here. The following can be summarized concerning structure-activity relationships: there is a large degree of plasticity allowed at the C-3 of jasmonic acid in the activation of defense genes. The carbonyl moiety is not strictly required, but exocyclic double bond character appears necessary. The pentenyl side chain at C-2 cannot tolerate bulky groups at the terminal carbon and still be biologically active. Substitutions to the C-1′ position are tolerated if they can potentially undergo β-oxidation. Either an alkanoic acid or methyl ester is required at C-1, or a side chain that can be shortened by β-oxidation or by peptidase hydrolysis. Coronatine and various derivatives thereof are not as effective as jasmonic acid, and derivatives in inducing benzo[c]phenanthridine alkaloid accumulation. Jasmonic acid rather than the octadecanoic precursors is therefore considered to be a likely signal transducer of defense gene activation in planta.

California poppy ( Eschscholzia californica ) is grown in UK gardens as an annual for its colourful display. In October 2012, diseased samples of California poppies (cv. Summer Sun) collected in a garden in Shropshire (West Midlands) were…

The genera Eschscholtzia and Argemone(Papaveraceae) represent a rich source of pavinane alkaloids, the identification of which in alkaloid extracts is generally problematic without standards. The alkaloid extracts of three Argemone and four Eschscholtzia species were analyzed using GC-MS. The alkaloids were identified based on comparison of their mass spectra with commercial libraries, with reported data in the literature and with spectra of reference compounds. A total of 23 alkaloids of six structural types (pavinane, protopine, benzylisoquinoline, benzophenanthridine, aporphine and protoberberine) were identified. The fragmentation pathway of pavinane alkaloids was used for their identification. O-Methylneocaryachine has been reported for the first time from a natural sources and the alkaloid pattern of Eschscholzia pulchella has been analyzed and described for the first time.

Plant cell and tissue cultures are a scalable and controllable alternative to whole plants for obtaining natural products of medical relevance. Cultures can be optimized for high yields of desired metabolites using rapid profiling assays such as HPLC. We describe an approach to establishing a rapid assay for profiling cell culture expression systems using a novel microscale LC-UV-MS-NMR platform, designed to acquire both MS and NMR each at their optimal sensitivity, by using nanosplitter MS from 4 mm analytical HPLC columns, and offline microdroplet NMR. The approach is demonstrated in the analysis of elicited Eschscholzia californica cell cultures induced with purified yeast extract to produce benzophenanthridine alkaloids. Preliminary HPLC-UV provides an overview of the changes in the production of alkaloids with time after elicitation. At the time point corresponding to the production of the most alkaloids, the integrated LC-MS-microcoil NMR platform is used for structural identification of extracted alkaloids. Eight benzophenanthridine alkaloids were identified at the sub-microgram level. This paper demonstrates the utility of the nanosplitter LC-MS/microdroplet NMR platform when establishing cell culture expression systems.

Eschsholzia californica Cham. is a traditional medicinal plant of the Indians used by the rural population of California for its analgesic and sedative properties. Our study on the aqueous extract shows that this plant reduced the behavioural parameters measured in a familiar environment test in mice (novelty preference, locomotion and rearings in two compartments test) at doses above 100 mg/kg and in non-familiar environment tests (staircase test) at doses above 200 mg/kg. This finding validates its traditional sedative properties confirmed by the sleeping induction at doses above 100 mg/kg. Furthermore, when administered at a dose a of 25 mg/kg, E. californica appeared to also have an anxiolytic action since it produced an increase of the number of steps climbed by mice in the staircase test (anticonflict effect) and that of the time spent by animals in the lit box when they were confronted with the light/dark choice situation. Before evaluation of the behavioural effects, it was verified that our aqueous extract did not induce any toxic effect when administered i.p. and p.o.

In twelve populations ofEschscholzia californica from central Chile, where it was introduced in the latter part of the 19th century, there is a regular gradient in frequency of the allelesJ andj, which control orange vs. yellow flowers, ranging from a maximum value of 0.531 to a minimum of 0.160 forJ. Populations having higher values ofJ occupy more mesic sites. In addition, the great majority of the populations have significant excess of heterozygotes over those expected on the basis of theHardy-Weinberg equilibrium, and so exhibit balanced polymorphism for theJ-j pair of alleles, probably associated with heterosis ofJj genotypes. Evidence is presented which suggests that this locus serves as a marker for adaptative gene complexes rather than being itself associated with adaptation. In Chile, the association ofJJ genotypes with cooler, moister habitats is the opposite of the condition existing in California, and suggests that extensive reorganization of the gene pool, through changed linkage relationships, took place asE. californica populations were being established in Chile.

ResumeLa quantite de cyanoglucosides et leur localisation chez Eschscholzia mexicana varient en fonction du developpement individuel. Au debut de la saison, les feuilles des rosettes sont tres fortement cyanogeniques. En phase reproductrice, les feuilles plus âgees deviennent graduellement acyanogeniques; en revanche, les structures reproductrices et surtout l'ovaire contiennent beaucoup de cyanoglueosides. Au moment de la dispersion des graines, le pericarpe est fortement cyanogenique, mais les graines sont acyanogeniques.Le role de la cyanogenese est envisage dans ses rapports avec la defense des plantes contre les herbivores et dans ses relations avec l'assimilation et l'economie de l'azote.

Summary Somewhat comparable to the dormant embryo in seed, the in vitro-produced embryoids of Eschscholzia californica could develop into plantlets when given a chilling treatment.

Laticifers of Papaver bracteatum Lindl., population Arya II, seedlings were examined by electron microscopy. Laticifers were first differentiated in procambium of the radicle associated with phloem about 72 hr after seeds were sown. Proliferation of membrane-bound vesicles of apparent endoplasmic reticulum origin distinguished laticifers from adjacent cells. Vesicles developed electron-dense caps from the internal condensation of small particles. Laticifer initials possessed the usual complement of organelles that became obscured in mature cells by the large, closely packed vesicles. Plastids contained an electron-dense, membrane-bound inclusion, but never developed lamellae or starch grains. Articulation and anastomoses between laticifer elements resulted from gradual removal of wall materials by both cells on opposite sides of the common walls at a perforation site. Differentiation of the laticifer initials and the micromorphology of the protoplast of P. bracteatum is similar to that reported for P. somniferum. THE ARTICULATED character of laticifers in the Papaveraceae was recognized in the classical literature. Members of this family show a wide range in development of this cell which may be of importance for determining the pattern of alkaloid accumulation frequently associated with this cell and for interpreting the phylogenetic significance of laticifers in the family (Esau, 1965). The laticifer system of Papaver bracteatum Lindl. was recognized early as being of the articulated, anastomosing type (Trecul, 1865), whereas this system in Chelidonium is nonanastomosing in character (Schacht, 1851). Sanguinaria is reported to develop perforations between laticifer members in the stem, but not in the leaf (Holm, 1908). Glaucium and Eschscholzia possess articulated laticifers that lack perforations between laticifer members (Leger, 1894

Stomata are present on the outer and inner fruit walls and seed coats of Eschscholzia californica, E. covillei, E. glyptosperma, E. lemmonii and E. minutiflora. The stomata on the inner fruit wall and seed coat remain constantly open, even under plasmolyzing conditions, whereas those of the outer fruit wall are able to open and close. This allows for gas exchange in these chlorophyllous structures. Fibrous bundle caps in the costal regions of the fruit act as windows allowing light transmission to the photosynthetic seeds within. Preliminary results show that the total photosynthesis by the fruits and seeds of Eschscholzia californica together appears to at least balance respiratory losses, and under favorable conditions might significantly contribute to seed and fruit development. STOMATA are known to occur on the seeds of species in a number of apparently unrelated families: Amaryllidaceae, Bixaceae, Bombacaceae, Cannaceae, Capparidaceae, Dipterocarpaceae, Euphorbiaceae, Fagaceae, Flacourtiaceae, Geraniaceae, Iridaceae, Juglandaceae, Krameriaceae, Liliaceae, Magnoliaceae, Malvaceae, Meliaceae, Myrsinaceae, Polygalaceae, Rosaceae, Sterculiaceae, and Violaceae (Macloskie, 1884; Netolitzky, 1926; Corner, 1976). In the Papaveraceae, three genera, Argemone (Sachar, 1955), Eschscholzia (first reported by Meunier, 1891), and Bocconia (our observations of B. frutescens) are known to have seed coat stomata. Considerations of the function of seed coat stomata have appeared in the literature over the years. Netolitzky (1926) suggested that seed coat stomata function in some species in water uptake by the seed during germination, rather than in gas exchange. However Flint and Moreland (1943) found considerable photosynthetic activity in the seeds of Hymenocallis occidentalis after they are shed from the fruit and inferred that seed coat stomata allow for gas exchange within this green seed. Our work has led us to believe that the seed coat stomata in Eschscholzia function in photosynthetic gas exchange by the seed while it is I Received for publication 3 October 1978; revision accepted 10 January 1979. This work was supported in part by NSF Grant DEB7702234 to the second author. We thank Robert Pearcy and David Randall for assistance with measurements of photosynthesis; Donald Kyhos and Thomas Rost for critical reading of the manuscript; Barbara Rugeley for translating the French paper; and Rudolf Schmid for helpful suggestions both during early stages of this study and in reviewing the manuscript. still contained within the fruit. In this paper we will (1) describe the structure of Eschscholzia fruits and seeds with particular attention to features that are important for photosynthesis and gas exchange, and (2) provide preliminary data showing the occurrence of photosynthesis in the fruits and seeds. MATERIALS AND METHODS-For SEM studies fresh immature seeds and fruits of Eschscholzia californica, E. glyptosperma, E. minutiflora, E. lemmonii and E. covillei were fixed in FAA, dehydrated in an ethanol series, dried using the critical point method, attached to aluminum stubs with silver conductive paint and coated with gold in a vacuum evaporator or sputter coater. Unimbibed mature seeds of these and the remaining eight species in the genus were likewise attached to stubs and coated with gold. Specimens were observed with a Cambridge Stereoscan S4 scanning electron microscope. Fresh material was hand-sectioned for anatomical study and observed either unstained or stained with 0.25% toluidine blue 0. Unstained guard cells were plasmolyzed with glycerine. Additional material was fixed in FAA, dehydrated through a tertiary butyl alcohol series, embedded in "Paraplast," sectioned at 10 ,um, mounted and stained with safranin, fast green and crystal violet. Using an infra-red gas analyzer, we measured photosynthesis and respiration as changes in CO, concentration in a reference gas mixture as it passed through a chamber enclosing several capsules attached to a single plant of E. californica. The temperature was 24-25 C, and the experiments were repeated at various light intensities. After completion of measurements the seeds

The perplexing variation and striking adaptability in the California poppy, Eschscholzia californica Cham., have long attracted interest. The present experimental study was undertaken to elucidate the causes of this polymorphism and ecological success. Techniques of mass sampling, growth of plants in more or less uniform conditions, and experimentation with breeding system have been employed to study the nature of morphological and physiological variation and to relate this variation to the reproductive behavior and environmental relations of the species. Eschscholzia californica is an annual or perennial herb of the family Papaveraceae. It is native in western North America from the Columbia River to Baja California and from the Pacific Coast eastward into the Great Basin. Within this area it grows at altitudes from sea level to 6,500 feet on open, well-drained soils of dunes, alluvial fans, river terraces, and hillsides. It is both widespread and abundant within its native region; man has extended its range by introducing it to Chile, New Zealand, Tasmania, and Australia, where it has become naturalized.

A B S T R A C T Both species of Dendromecon are diplochorous, that is, the seeds are first scattered from the mother plant by violently dehiscing capsules and then transported further away by ants, which are strongly attracted to the e]aiosome at the micropylar end of the seed. The tensions that cause the capsule to explode develop between the two carpels and also between lateral and central fiber strands in the capsular valves. The elaiosome is unusual in that it contains large quntities of starch in addition to the commonly present fatty oils and that it is more drought resistant than most known elaiosomes. Seed dispersal is very efficient over short distances, but migration barriers a few hundred meters or more cannot be crossed except by chance dispersal. Dendromecon differs strikingly from most ant-dispersed plants in being woody and xerophytic; so far, D. rigida is the only chaparral plant known to be habitually dispersed by ants. The presence of an autochorous-myrmecochorous type of seed dispersal in D. rigida may partly explain the polymorphism of this species, since this type of seed dispersal tends to keep local populations isolated. The nearly complete correspondence, anatomically, morphologically, and functionally, between the fruit of Dendromecon and that of Eschscholzia strongly supports the idea of a close taxonomic relationship between these two genera. The seed appendage is believed to be a "recent" specialization in Dendroniecon, phylogenetically independent from seed appendages facilitating dispersal by ants in other genera of the Papaveraceae. DISPERSAL ECOLOGY, the study of dispersal processes and mechanisms, is an underdeveloped, mostly neglected sector of botany, despite the fact that ecologists, evolutionists, plant geographers, and evolutionary taxonomists all would profit from increased teaching and research activity in this classical field. A particularly interesting and rewarding aspect for study is myrmecochory, i.e., seed dispersal by ants. Only a very few plants in North America are known to be regularly dispersed by ants: Uvularia grandiflora Sm. (Robertson, 1897), Sanguinaria canadensis L. (Robertson, 1897; Kerner, 1898

A detailed morphological study has led to the conclusion that Entyloma Glaucii should be included in E. fuscum. The symptoms of the smut on Papaver Rhoeas and P. somniferum are described. Study of limited material suggests that the fungus on Eschscholzia and Dendromecon in California should be retained as E. Eschscholtziae. The production of conidia on these hosts is reported for the first time Entyloma fuscum from Papaver Rhoeas has been shown to attack P. atlanticum, P. dubium, P. glaucum, P. monanthum, P. orientale, P. setigerum, and P. somniferum.

The roots and aerial parts of Eschscholzia californica Cham. (Papaveraceae) were extracted with ethanol. Repeated column chromatography, preparative TLC, and crystallization led to the isolation of fourteen isoquinoline alkaloids, the structures of which were determined on the basis of spectroscopic techniques and by comparison with literature values. One of the compounds isolated, 1-(3-hydroxy-4-methoxybenzyl)-2-methyl-6,7-methylenedioxy-1,2,3,4-tetrahydroisoquinoline (14), has not previously been isolated from a natural source. All isolated compounds were tested for human blood acetylcholinesterase (HuAChE) and human plasma butyrylcholinesterase (HuBuChE) inhibition activity. None of the compounds isolated significantly inhibited both HuAChE and HuBuChE, but the two benzylisoquinoline alkaloids, reticuline 9 and 14, showed promising inhibitory activity against HuBuChE.

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.

Eschscholzia californica Cham. (Papaveraceae) is traditionally used by the Indians as a medicinal plant for its anxiolytic, anticonflict, analgesic and sedative properties. The mechanisms of action for the sedative and anxiolytic activities have not been clearly established and so to further investigate the pharmacological profile of E. californica in some painful conditions, a 70% v/v ethanol extract, DERnative=5:1, was tested in rat models of neuropathy induced by chronic constriction injury of the sciatic nerve (CCI), with chemotherapeutic oxaliplatin, and osteoarthritis caused by intrarticular injection of monoiodoacetate. In the CCI model evaluated in the rat paw-pressure test, the examined extract (100 mg kg−1 p.o.) showed an antihyperalgesic effect. Eschscholzia extract, after single injection at a dose of 100–300 mg kg−1 p.o., produced also a statistically significant decrease of pain perception on hyperalgesia induced by oxaliplatin and osteoarthritis, while in the same condition gabapentin did not display any antihyperalgesic effect. Furthermore, in the range of antihyperalgesic doses, the extract was efficacious in the hot-plate (thermal stimulus) and carrageenan tests (inflammatory model) without producing any behavioral impairment, as evaluated by the Irwin test. The analgesic effect exhibited by Eschscholzia extract in the mouse hot-plate test was not antagonized by naloxone, indicating that opioid neurotransmission is not involved in the effect. The above reported results suggest that a 70% (v/v) ethanol dried extract (DERnative=5:1) of E. californica might represent a promising product for the therapy of acute and chronic pain.

Scanning electron microscope examination of seeds of all thirteen species of Eschscholzia, a genus of western North America, reveals distinguishing features which can be used to separate species, provides evidence of seed flotation as an adaptation to runoff dispersal in some species, and provides additional data toward the resolution of taxonomic and evolutionary problems in the genus. Seed surfaces generally consist of ridges or tubercles of radially elongate cells surrounding facets of tabular cells. Surface features of the mature seed are a function of the collapse of these cells during desiccation. The study of seed coat microsculpturing with the scanning electron microscope has provided taxonomic, evolutionary, and ecological insights in a number of groups. In many cases it has been possible to interpret seed dormancy (Gutterman & Heydecker, 1973), to separate tribes (Whiffin & Tomb, 1972), genera (Mulligan & Bailey, 1976), sections, and even species (Chuang & Heckard, 1972; Ehler, 1976; Hill, 1976; Seavey et al., 1977), and to postulate evolutionary relationships (Chuang & Heckard, 1972; Hill, 1976; Seavey et al., 1977; Whiffin & Tomb, 1972) on the basis of seed coat characteristics. These techniques can be applied profitably to Eschscholzia, a genus of 13 species of western North America. In contrast to seeds of the large genus Papaver, which are exceedingly uniform (Tomb, 1974; but see Gunn & Seldin, 1976), those of Eschscholzia are elaborately sculptured and quite diverse. Seed characters have been used to distinguish species (e.g., E. glyptosperma, E. lobbii) in the past (Greene, 1885, 1905; Jepson, 1922; Munz, 1959). Seeds of E. californica were first examined anatomically by Godfrin (1880), who looked only at mature seeds. Brandza (1891) included this species in his broad study of seed integuments. The first detailed study of the ontogeny of the seed coat was undertaken by Meunier (1891). Shaw (1904) studied the histology of seed coats of E. californica and certain other Papaveraceae. Sachar and Mohan Ram (1958) and Roder (1958) also examined the seed coat of E. californica, the former concurring with previous investigators, the latter differing somewhat, perhaps through inadequacy of observation. Gunn and Seldin (1976) included twelve species (eleven by our reckoning) of Eschscholzia in their light-microscopic study of papaveraceous seeds. They provide a key which separates nine of the species; it includes embryo and color characters, as well as external morphological characBotany, University of California, Davis, CA 95616.

Eschscholzia californica Cham., the California poppy, occurs naturally in western North America from the Columbia River south to Baja California and from the coast of California eastward into the Great Basin. Within this extensive region it grows in open sites, frequently associated with grasses. Both the poppy and the grasses require ample light for optimal development and begin growth contemporaneously after the advent of winter rains. In dry years, when grass develops poorly, poppies apparently do well; in wet years the reverse is true. The poppy grows on well-drained soils of dunes, alluvial fans, river terraces, steep banks, and rocky places. These soils may be derived from a wide range of parent materials, including serpentine. It has also been observed to colonize burned-over areas and endure on them for a number of years (Bowerman 1944). The poppy has behaved as a benign weed under conditions created by man. Small populations arise atop gravel piles, in gravelly ballast of railroads, or on road cuts and other areas where the established vegetation has been destroyed by excavation. Occasionally poppies may be seen in grain fields, where their growth is apparently favoured by mowing the overtopping grain. Although the plants themselves are cut back, new shoots emerge from ground level and flower from mid-summer to autumn. These observations suggested the following hypotheses: (1) Genetic adaptation to specific soil type is involved in the poppy's adaptation to a broad range of soil types. Such response has been reported for other herbaceous species (Kruckeberg 1951; Jowett 1958, 1959; Bradshaw & Snaydon 1959; Bradshaw 1960). (2) Growth of associated grass is antagonistic to Eschscholzia. (3) Eschscholzia cannot tolerate soils with high water content but can become established on porous soils high above the water table. These hypotheses were tested experimentally at the Botanical Garden of the University of California at Berkeley during 1959 and 1960.

IN an earlier communication one of us1 described the results of intra-ovarian pollination in Papaver rhoeas. The present work deals with some observations of a similar nature on Eschscholzia californica, Argemone mexicana and A. ochroleuca.

Background Molecular genetic studies of floral development have concentrated on several core eudicots and grasses (monocots), which have canalized floral forms. Basal eudicots possess a wider range of floral morphologies than the core eudicots and grasses and can serve as an evolutionary link between core eudicots and monocots, and provide a reference for studies of other basal angiosperms. Recent advances in genomics have enabled researchers to profile gene activities during floral development, primarily in the eudicot Arabidopsis thaliana and the monocots rice and maize. However, our understanding of floral developmental processes among the basal eudicots remains limited. Results Using a recently generated expressed sequence tag (EST) set, we have designed an oligonucleotide microarray for the basal eudicot Eschscholzia californica (California poppy). We performed microarray experiments with an interwoven-loop design in order to characterize the E. californica floral transcriptome and to identify differentially expressed genes in flower buds with pre-meiotic and meiotic cells, four floral organs at pre-anthesis stages (sepals, petals, stamens and carpels), developing fruits, and leaves. Conclusions Our results provide a foundation for comparative gene expression studies between eudicots and basal angiosperms. We identified whorl-specific gene expression patterns in E. californica and examined the floral expression of several gene families. Interestingly, most E. californica homologs of Arabidopsis genes important for flower development, except for genes encoding MADS-box transcription factors, show different expression patterns between the two species. Our comparative transcriptomics study highlights the unique evolutionary position of E. californica compared with basal angiosperms and core eudicots.

Background We have developed a simulation approach to help determine the optimal mixture of sequencing methods for most complete and cost effective transcriptome sequencing. We compared simulation results for traditional capillary sequencing with "Next Generation" (NG) ultra high-throughput technologies. The simulation model was parameterized using mappings of 130,000 cDNA sequence reads to the Arabidopsis genome (NCBI Accession SRA008180.19). We also generated 454-GS20 sequences and de novo assemblies for the basal eudicot California poppy (Eschscholzia californica) and the magnoliid avocado (Persea americana) using a variety of methods for cDNA synthesis. Results The Arabidopsis reads tagged more than 15,000 genes, including new splice variants and extended UTR regions. Of the total 134,791 reads (13.8 MB), 119,518 (88.7%) mapped exactly to known exons, while 1,117 (0.8%) mapped to introns, 11,524 (8.6%) spanned annotated intron/exon boundaries, and 3,066 (2.3%) extended beyond the end of annotated UTRs. Sequence-based inference of relative gene expression levels correlated significantly with microarray data. As expected, NG sequencing of normalized libraries tagged more genes than non-normalized libraries, although non-normalized libraries yielded more full-length cDNA sequences. The Arabidopsis data were used to simulate additional rounds of NG and traditional EST sequencing, and various combinations of each. Our simulations suggest a combination of FLX and Solexa sequencing for optimal transcriptome coverage at modest cost. We have also developed ESTcalc http://fgp.huck.psu.edu/NG_Sims/ngsim.pl, an online webtool, which allows users to explore the results of this study by specifying individualized costs and sequencing characteristics. Conclusion NG sequencing technologies are a highly flexible set of platforms that can be scaled to suit different project goals. In terms of sequence coverage alone, the NG sequencing is a dramatic advance over capillary-based sequencing, but NG sequencing also presents significant challenges in assembly and sequence accuracy due to short read lengths, method-specific sequencing errors, and the absence of physical clones. These problems may be overcome by hybrid sequencing strategies using a mixture of sequencing methodologies, by new assemblers, and by sequencing more deeply. Sequencing and microarray outcomes from multiple experiments suggest that our simulator will be useful for guiding NG transcriptome sequencing projects in a wide range of organisms.