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Diversification of CYCLOIDEA-like TCP genes in the basal eudicot families Fumariaceae and Papaveraceae s.str.

CYCLOIDEA-like genes belong to the TCP family of transcriptional regulators and have been shown to control different aspects of shoot development in various angiosperm lineages, including flower monosymmetry in asterids and axillary meristem growth in monocots. Genes related to the CYC gene from ANTIRRHINUM show independent duplications in both asterids and rosids. However, it remains unclear to what extent this affected the evolution of flower symmetry and shoot branching in these and other eudicot lineages. Here, we show that CYC-like genes have also undergone duplications in two related Ranunculales families, Fumariaceae and Papaveraceae s.str. These families exhibit morphological diversity in flower symmetry and inflorescence architecture that is potentially related to functions of CYC-like genes. We present sequences of 14 CYC-related genes covering 9 genera. Phylogenetic analyses indicate the presence of three clades of CYC-like genes. Shared motifs in the region between the TCP and R domains of CYC-like genes between Fumariaceae, Papaveraceae s.str., and AQUILEGIA (Ranunculaceae) indicate that the observed duplications originated from a single CYC gene present in all Ranunculales. RT-PCR expression data suggest that gene duplication and diversification in Fumariaceae and Papaveraceae s.str. was accompanied by divergence in expression patterns.

Amino Acid Sequence↗

Diversity and evolution of CYCLOIDEA-like TCP genes in relation to flower development in Papaveraceae.

Monosymmetry evolved several times independently during flower evolution. In snapdragon (Antirrhinum majus), a key gene for monosymmetry is CYCLOIDEA (CYC), which belongs to the class II TCP gene family encoding transcriptional activators. We address the questions of the evolutionary history of this gene family and of possible recruitment of genes homologous to CYC in floral development and symmetry in the Papaveraceae. Two to three members of the class II TCP family were found in each species analyzed, two of which were CYC-like genes, on the basis of the presence of both the TCP and R conserved domains. The duplication that gave rise to these two paralogous lineages (named PAPACYL1 and PAPACYL2) probably predates the divergence of the two main clades within the Papaveraceae. Phylogenetic relationships among angiosperm class II TCP genes indicated that (1) PAPACYL genes were closest to Arabidopsis (Arabidopsis thaliana) AtTCP18, and a duplication at the base of the core eudicot would have given rise to two supplementary CYC-like lineages; and (2) at least three class II TCP genes were present in the ancestor of monocots and eudicots. Semiquantitative reverse transcription-polymerase chain reaction and in situ hybridization approaches in three species with different floral symmetry indicated that both PAPACYL paralogs were expressed during floral development. A pattern common to all three species was observed at organ junctions in inflorescences and flowers. Expression in the outer petals was specifically observed in the two species with nonactinomorphic flowers. Hypotheses concerning the ancestral pattern of expression and function of CYC-like genes and their possible role in floral development of Papaveraceae species leading to bisymmetric buds are discussed.

DNA-Binding Proteins↗

Expression patterns of STM-like KNOX and Histone H4 genes in shoot development of the dissected-leaved basal eudicot plants Chelidonium majus and Eschscholzia californica (Papaveraceae).

Knotted-like homeobox (KNOX) genes encode important regulators of shoot development in flowering plants. In Arabidopsis, class I KNOX genes are part of a regulatory system that contributes to indeterminacy of shoot development, delimitation of leaf primordia and internode development. In other species, class I KNOX genes have also been recruited in the control of marginal blastozone fractionation during dissected leaf development. Here we report the isolation of class I KNOX genes from two species of the basal eudicot family Papaveraceae, Chelidonium majus and Eschscholzia californica. Sequence comparisons and expression patterns indicate that these genes are orthologs of SHOOTMERISTEMLESS (STM), a class I KNOX gene from Arabidopsis. Both genes are expressed in the center of vegetative and floral shoot apical meristems (SAM), but downregulated at leaf or floral organ initiating sites. While Eschscholzia californica STM (EcSTM) is again upregulated during acropetal pinna formation, in situ hybridization could not detect Chelidonium majus STM (CmSTM) transcripts at any stage of basipetal leaf development, indicating divergent evolution of STM gene function in leaves within Papaveraceae. Immunolocalization of KNOX proteins indicate that other gene family members may control leaf dissection in both species. The contrasting direction of pinna initiation in the two species was also investigated using Histone H4 expression. Leaves at early stages of development did not reveal notable differences in cell division activity of the elongating leaf axis, suggesting that differential meristematic growth may not play a role in determining the observed dissection patterns.

Amino Acid Sequence↗

Isolation and chemistry of alkaloids from plants of the family Papaveraceae LXVII: Corydalis cava (L.) Sch. et K. (C. tuberosa DC).

From Corydalis cava (L.) Sch. et K. (C. tuberosa DC) (Papaveraceae: genus Corydalis Med.), a mixture of alkaloids (0.53%) was isolated. The main alkaloid was (+)-bulbocapnine, and the minor alkaloids were coptisine, (+)-domestine, adlumidiceine, (+)-predicentrine, protopine, (-)-capnoidine, (+)-stylopine, (+)-isoboldine, 8-oxocoptisine, 1,2-methylenedioxy-6a,7-dehydroaporphine-10,11-quinone, and corysamine. In addition, fumaric acid was obtained. Coptisine, adlumidiceine, predicentrine, 8-oxocoptisine, corysamine, 1,2-methylenedioxy-6a, 7-dehydroaporphine-10,11-quinone, and isoboldine were found in C. cava for the first time, but rhoeadine and papaverrubine alkaloids were not detected. Predicentrine and isoboldine were identified on the basis of the UV, IR, mass, and PMR spectra.

Acetylation↗

Pharmacological activities of Chelidonium majus L. (Papaveraceae).

Chelidonium majus L. (Papaveraceae) has a long history as being useful for the treatment of many diseases in European countries. This plant is of great interest for its use also in Chinese herbal medicine. The plant contains, as major secondary metabolites, isoquinoline alkaloids, such as sanguinarine, chelidonine, chelerythrine, berberine and coptisine. Other compounds structurally unrelated to the alkaloids have been isolated from the aerial parts: several flavonoids and phenolic acids. C. majus extracts and its purified compounds exhibit interesting antiviral, antitumour and antimicrobial properties both in vitro and in vivo.

Alkaloids↗

Potential preventive effects of Chelidonium majis L. (Papaveraceae) herb extract on glandular stomach tumor development in rats treated with N-methyl-N'-nitro-N nitrosoguanidine (MNNG) and hypertonic sodium chloride.

The modifying effects of Chelidonium majis L. (Papaveraceae) herb extract (CH), an analgesic traditionally prescribed for gastric and duodenal ulcer patients, on gastric tumor development were studied in rats given N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Sixty-four male 6-week-old Wistar rats were used. Group 1 rats were initially given MNNG (200 mg/kg b.w.) by gavage at days 0 and 14 as well as saturated sodium chloride solution (S-NaCl, 1 ml per rat) every 3 days during weeks 0-3 (six times), and then placed on basal diet containing 0.1 or 0.2% CH for 16 weeks from week 4. Rats of Group 2 and 3 were treated with MNNG together with S-NaCl or saline (0.9% NaCl, 1 ml per rat), respectively, timed as in Group 1 but without further treatment. All surviving animals were killed at week 20 and histopathologically investigated. In the glandular stomach, the number of preneoplastic pepsinogen 1 altered pyloric glands (PAPGs) in the MNNG + S-NaCl-->CH (0.1%) group (Group 1) was significantly smaller than in the MNNG + S-NaCl group (Group 2) (P < 0.02). The incidences of forestomach neoplastic lesions (papillomas and squamous cell carcinomas) also showed a tendency to decrease with the CH treatment. The results thus indicate that CH exerts inhibitory effects on glandular stomach carcinogenesis in the rat, so that it may have potential as a chemopreventive agent for stomach cancer in man.

Animals↗

Fungitoxic alkaloids from Hokkaido Papaveraceae.

Papaver orientale, Dicentra spectabilis and Chelidonium majus, three Papaveraceae collected from Hokkaido area, were studied for their fungitoxic alkaloids. Ten alkaloids were isolated, identified and tested for their fungitoxic activity. Two of these isolates were found to be new compounds.

Alkaloids↗

Transformations of (+-)-salsolinol into optically active O- and/or N-methylated derivatives by several Papaveraceae plants and their tissue-cultured cells.

(+-)-Salsolinol [3], a substance possibly inducing parkinsonism, was biotransformed into optically active O- and/or N-methylated salsolinols 5, 6, and 7 by several Papaveraceae plants and tissue-culture cells derived from these plants. The bioconversion of racemic salsolinol into optically active tetrahydroisoquinolines has never been shown to occur, either in animals or in plants.

Biotransformation↗

Invasive Argentine ants (Linepithema humile) do not replace native ants as seed dispersers of Dendromecon rigida (Papaveraceae) in California, USA.

We investigated the indirect effects of Argentine ant (Linepithema humile) invasions on patterns of seed dispersal and predation in the myrmecochorous tree poppy Dendromecon rigida in coastal San Diego County, California. Significantly more seeds were removed from ant-accessible seed stations at sites numerically dominated by a common harvester ant (Pogonomyrmex subnitidus), a native disperser of these seeds and a species sensitive to displacement by L. humile, than from those stations at sites where L. humile was in the majority. Predation of seeds was high, but variable, across sites, suggesting that reduced dispersal could result in increased seed predation in some habitats. Removal of elaiosomes did not affect the frequency with which predators removed seeds, but ants removed significantly more seeds with elaiosomes than without. In behavior trials, only P. subnitidus was able to carry seeds of Dendromecon rigida effectively. L. humile and a small native ant species, Dorymyrmex insanus, while displaying interest in the diaspores, were seldom able to carry whole seeds and, when they did, only carried them a few centimeters. Displacement of native harvester ants by L. humile appears to decrease the dispersal of Dendromecon rigida seeds and may be increasing loss of seeds due to predation.

Animals↗

EST database for early flower development in California poppy (Eschscholzia californica Cham., Papaveraceae) tags over 6,000 genes from a basal eudicot.

The Floral Genome Project (FGP) selected California poppy (Eschscholzia californica Cham. ssp. Californica) to help identify new florally-expressed genes related to floral diversity in basal eudicots. A large, non-normalized cDNA library was constructed from premeiotic and meiotic floral buds and sequenced to generate a database of 9,079 high quality Expressed Sequence Tags (ESTs). These sequences clustered into 5,713 unigenes, including 1,414 contigs and 4,299 singletons. Homologs of genes regulating many aspects of flower development were identified, including those for organ identity and development, cell and tissue differentiation, cell cycle control, and secondary metabolism. Over 5% of the transcriptome consisted of homologs to known floral gene families. Most are the first representatives of their respective gene families in basal eudicots and their conservation suggests they are important for floral development and/or function. App. 10% of the transcripts encoded transcription factors and other regulatory genes, including nine genes from the seven major lineages of the important MADS-box family of developmental regulators. Homologs of alkaloid pathway genes were also recovered, providing opportunities to explore adaptive evolution in secondary products. Furthermore, comparison of the poppy ESTs with the Arabidopsis genome provided support for putative Arabidopsis genes that previously lacked annotation. Finally, over 1,800 unique sequences had no observable homology in the public databases. The California poppy EST database and library will help bridge our understanding of flower initiation and development among higher eudicot and monocot model plants and provide new opportunities for comparative analysis of gene families across angiosperm species.

DNA, Complementary↗

[The confirmation of putative natural hybrid species Meconopsis x cookei G. Taylor (Papaveraceae) based on nuclear ribosomal DNA ITS region sequence].

The Nuclear Ribosomal DNA internal transcribed spacers (ITS) region sequences from a putative natural hybrid species Meconopsis x cookei and its possible parents M. punicea and M. quintuplinervia were obtained by using direct sequencing method. The sequence length of ITS region (including ITS1, 5.8S and ITS2) is 667 bp for M. punicea, 668 bp for M. x cookei, and 668 bp for M. quintuplinervia. The sequences were aligned by the software Clustal X, and the bases per locus were compared by using software with manual method. The aligned sequence length is 688 bp, of which ITS1 is 254 bp, 5.8S is 162 bp, and ITS2 is 252 bp. 16 variable loci were detected from the aligned sequence, approximately 2.40% to the whole sequence length, of which ITS1 has nine variable loci (56.25%), ITS2 has six variable loci (37.50%), and 5.8S has one variable locus (6.25%). The results show that M. x cookei have two kinds of ITS sequences contributed from two species M. puniceaa and M. quintuplinervia, i.e. the variation of ITS gene among M. x cookei, M. punicea and M. quintuplinervia is congruence with the Mendel's genetics law. Therefore, the molecular evidences indicate that M. x cookei is a hybrid origin from M. punicea and M. quintuplinervia.

Base Sequence↗