Advances in Research on the Regulation of Floral Development by CYC-like Genes.

CYCLOIDEA (CYC)-like genes belong to the TCP transcription factor family and play important roles associated with flower development. The CYC-like genes in the CYC1, CYC2, and CYC3 clades resulted from gene duplication events. The CYC2 clade includes the largest number of members that are crucial regulators of floral symmetry. To date, studies on CYC-like genes have mainly focused on plants with actinomorphic and zygomorphic flowers, including Fabaceae, Asteraceae, Scrophulariaceae, and Gesneriaceae species and the effects of CYC-like gene duplication events and diverse spatiotemporal expression patterns on flower development. The CYC-like genes generally affect petal morphological characteristics and stamen development, as well as stem and leaf growth, flower differentiation and development, and branching in most angiosperms. As the relevant research scope has expanded, studies have increasingly focused on the molecular mechanisms regulating CYC-like genes with different functions related to flower development and the phylogenetic relationships among these genes. We summarize the status of research on the CYC-like genes in angiosperms, such as the limited research conducted on CYC1 and CYC3 clade members, the necessity to functionally characterize the CYC-like genes in more plant groups, the need for investigation of the regulatory elements upstream of CYC-like genes, and exploration of the phylogenetic relationships and expression of CYC-like genes with new techniques and methods. This review provides theoretical guidance and ideas for future research on CYC-like genes.

Effects of floral symmetry and orientation on the consistency of pollinator entry angle.

Since the publication of Sprengel's (1793) observations, it has been considered that flowers with zygomorphic (or bilaterally symmetrical) corollas evolved to restrict the movement of pollinators into the flower by limiting the pollinator's direction of approach. However, little empirical support has been accumulated so far. Our aim was to build on previous research that showed zygomorphy reduces variance in pollinator entry angle, aiming to observe whether floral symmetry or orientation had an impact on pollinator entry angle in a laboratory experiment using bumble bees, Bombus ignitus. Using nine different combinations of artificial flowers created from three symmetry types (radial, bilateral and disymmetrical) and three orientation types (upward, horizontal, and downward), we tested the effects of these two floral aspects on the consistency of bee entry angle. Our results show that horizontal orientation significantly reduced the variance in entry angle, while symmetry had little effect. We also found either little or no significant interactions between angle and symmetry in their effect on entry angle. Thus, our results suggest that horizontal orientation forces the bees to orient themselves relative to gravity rather than the corolla and stabilizes their flower entry. This stabilizing effect may have been mistaken for the effect of zygomorphic corolla as it is presented horizontally in most species. Consequently, we suggest that the evolution of horizontal orientation preceded that of zygomorphy as indicated by some authors, and that the reason behind the evolution of zygomorphy should be revisited.

Pollination-precision hypothesis: support from native honey bees and nectar bats.

The evolution of floral traits is often considered to reflect selection for increased pollination efficiency. Known as the pollination-precision hypothesis, increased pollination efficiency is achieved by enhancing pollen deposition on precise areas of the pollinator. Most research to date addressing this hypothesis has examined plant species that are a priori predicted to place pollen precisely, but we still lack comparisons with species predicted to have low pollination efficiency. We studied 39 plant species with diverse floral morphologies and measured the precision of pollen placement on two pollinator groups: honey bees (genus Apis) and nectar bats (family Pteropodidae). Pollen was collected from four locations of each pollinator's body (bees: dorsal thorax, ventral thorax, dorsal abdomen, ventral abdomen; bats: crown, face, chest, wing) to calculate pollen placement precision using Pielou's evenness index. We also quantified variation in floral design by scoring floral symmetry, corolla fusion, floral orientation and stamen number. We confirm the importance of four floral character states (bilateral symmetry, fused corollas, horizontal orientation and reduced stamen number) in promoting precise pollen placement on diverse pollinators. Our findings provide phylogenetically corrected, empirical support that the evolution of the four floral characters reflect selection for enhanced precision of pollen placed on pollinators.

Floral orientation affects outcross-pollen deposition in buzz-pollinated flowers with bilateral symmetry.

PREMISE: Floral orientation is central to plant-pollinator interactions and is commonly associated with floral symmetry. Bilaterally symmetrical flowers are often oriented horizontally for optimal pollinator positioning and pollen transfer efficiency, while the orientation of radially symmetrical flowers is variable. Buzz-pollinated species (pollinated by vibration-producing bees) include bilateral, horizontally oriented flowers, and radial, pendant flowers. The effect of floral orientation on pollen transfer has never been tested in buzz-pollinated species. METHODS: Here, we examined the effect of floral orientation on bumblebee-mediated pollen deposition in three buzz-pollinated Solanum species with different floral symmetry and natural orientations: S. lycopersicum and S. seaforthianum (radial, pendant), and S. rostratum (bilateral, horizontal). We tested whether orientation affects total stigmatic pollen deposition (both self and outcross pollen) when all flowers have the same orientation (either pendant or horizontal). In a second experiment, we evaluated whether different orientations of donor and recipient flowers affects the receipt of outcross pollen by S. rostratum. RESULTS: For the three Solanum species studied, there was no effect of floral orientation on total pollen deposition (both self and outcross) when flowers shared the same orientation. In contrast, in our experiment with S. rostratum, we found that pendant flowers received fewer outcross-pollen grains when paired with pendant donors. CONCLUSIONS: We suggest that floral orientation influences the quality of pollen transferred, with more outcross pollen transferred to horizontally oriented recipients in the bilaterally symmetrical S. rostratum. Whether other bilaterally symmetrical, buzz-pollinated flowers also benefit from increased cross-pollination when presented horizontally remains to be established.

Evolution and development of inflorescences and floral symmetry in Solanaceae.

PREMISE: The inflorescences of Solanaceae are unique and complex, which has led to long-standing disputes over floral symmetry mainly due to different interpretations of the cyme-like inflorescence structure. The main disagreements have been over how the phyllomes associated with the flower were arranged relative to the inflorescence axis especially during early flower initiation. METHODS: Here we investigated the evolution of inflorescences in Solanaceae by analyzing inflorescence structure in the context of phylogeny using ancestral state reconstruction (ASR) to determine the evolutionary transitions between loosely arranged and tightly clustered inflorescences and between monochasial-like and dichasial-like cymes. We also reconstructed two- and three-dimensional models for 12 solanaceous species that represent both inflorescence and phylogenetic diversity in the family. RESULTS: Our results indicate that the most recent common ancestor of Solanaceae had a loosely arranged and monochasial-like cyme, while tightly clustered inflorescences and dichasial-like cymes were derived. Compared to the known process of scorpioid cyme evolution, Solanaceae achieved their scorpioid cyme-like inflorescences through a previously undescribed way. Along the pedicel, the two flower-preceding prophylls are not in the typical transverse position of dicotyledonous plants; they frequently have axillary buds, and the main inflorescence axis continues in a sympodial fashion. As a result, the plane of symmetry of the flower is 36° from the median, and the inflorescence axis and the two flower-preceding prophylls are symmetrically located along that plane. CONCLUSIONS: A better understanding of the morphological evolution of solanaceous inflorescence structure helped clarify the floral symmetry of Solanaceae.

Transcriptomic Analysis Suggests Auxin Regulation in Dorsal-Ventral Petal Asymmetry of Wild Progenitor Sinningia speciosa.

The establishment of dorsal-ventral (DV) petal asymmetry is accompanied by differential growth of DV petal size, shape, and color differences, which enhance ornamental values. Genes involved in flower symmetry in Sinningia speciosa have been identified as CYCLOIDEA (SsCYC), but which gene regulatory network (GRN) is associated with SsCYC to establish DV petal asymmetry is still unknown. To uncover the GRN of DV petal asymmetry, we identified 630 DV differentially expressed genes (DV-DEGs) from the RNA-Seq of dorsal and ventral petals in the wild progenitor, S. speciosa 'ES'. Validated by qRT-PCR, genes in the auxin signaling transduction pathway, SsCYC, and a major regulator of anthocyanin biosynthesis were upregulated in dorsal petals. These genes correlated with a higher endogenous auxin level in dorsal petals, with longer tube length growth through cell expansion and a purple dorsal color. Over-expression of SsCYC in Nicotiana reduced petal size by regulating cell growth, suggesting that SsCYC also controls cell expansion. This suggests that auxin and SsCYC both regulate DV petal asymmetry. Transiently over-expressed SsCYC, however, could not activate most major auxin signaling genes, suggesting that SsCYC may not trigger auxin regulation. Whether auxin can activate SsCYC or whether they act independently to regulate DV petal asymmetry remains to be explored in the future.

Evolutionary trends and diversity of major floral nectary types across Solanaceae.

MAIN CONCLUSION: In a co-evolutionary crosstalk amid plants and their pollinators, nectaries serve as a labile link between the relatively fixed structural domains of divergent flower forms and associated pollination syndromes. Floral nectary plays a crucial role in sexual plant reproduction by enabling interaction between plants and their pollinators. It is known to associate with different floral whorls, and exhibits variations in structure and location in different clades across angiosperms. To infer evolutionary patterns, it is important to map key features associated with the trait at various taxonomic ranks. In the present study, we analysed variability and distribution of floral nectaries in Solanaceae for the first time. Floral nectaries of 23 taxa representing different clades in the family were studied using bright-field and scanning electron microscopy. The study reveals that although floral nectaries share anatomical similarity, they differ in morphology, composition within cells, and locations within a flower across the clades. The analysis suggests that (i) there is a shift from symmetric, lobed type nectary in the early branching sub-families to asymmetric, annular type in the late branching ones, (ii) floral organization has shifted from asymmetry (zygomorphy) to symmetry (actinomorphy) in corolla, and (iii) the lobed nectary correlates with zygomorphic floral forms that are pollinated by birds and long-tongued vectors, while the annular nectary is predominant among species with bee-pollinated actinomorphic flowers.

Evolution of pollination syndromes and corolla symmetry in Balsaminaceae reconstructed using phylogenetic comparative analyses.

BACKGROUND AND AIMS: Floral diversity as a result of plant-pollinator interactions can evolve by two distinct processes: shifts between pollination systems or divergent use of the same pollinator. Although both are pollinator driven, the mode, relative importance and interdependence of these different processes are rarely studied simultaneously. Here we apply a phylogenetic approach using the Balsaminaceae (including the species-rich genus Impatiens) to simultaneously quantify shifts in pollination syndromes (as inferred from the shape and colour of the perianth), as well as divergent use of the same pollinator (inferred from corolla symmetry). METHODS: For 282 species we coded pollination syndromes based on associations between floral traits and known pollination systems, and assessed corolla symmetry. The evolution of these traits was reconstructed using parsimony- and model-based approaches, using phylogenetic trees derived from phylogenetic analyses of nuclear ribosomal and plastid DNA sequence data. KEY RESULTS: A total of 71 % of studied species have a bee pollination syndrome, 22 % a bimodal syndrome (Lepidoptera and bees), 3 % a bird pollination syndrome and 5 % a syndrome of autogamy, while 19 % of species have an asymmetrical corolla. Although floral symmetry and pollination syndromes are both evolutionarily labile, the latter shifts more frequently. Shifts in floral symmetry occurred mainly in the direction towards asymmetry, but there was considerable uncertainty in the pattern of shift direction for pollination syndrome. Shifts towards asymmetrical flowers were associated with a bee pollination syndrome. CONCLUSION: Floral evolution in Impatiens has occurred through both pollination syndrome shifts and divergent use of the same pollinator. Although the former appears more frequent, the latter is likely to be underestimated. Shifts in floral symmetry and pollination syndromes depend on each other but also partly on the region in which these shifts take place, suggesting that the occurrence of pollinator-driven evolution may be determined by the availability of pollinator species at large geographical scales.

Molecular framework underlying floral bilateral symmetry and nectar spur development in Tropaeolum, an atypical member of the Brassicales.

PREMISE: Floral spurs are key innovations associated with elaborate pollination mechanisms that have evolved independently several times across angiosperms. Spur formation can shift the floral symmetry from radial to bilateral, as it is the case in Tropaeolum, the only member of the Brassicales with floral nectar spurs. The genetic mechanisms underlying both spur and bilateral symmetry in the family have not yet been investigated. METHODS: We studied flower development and morphoanatomy of Tropaeolum longifolium. We also generated a reference transcriptome and isolated all candidate genes involved in adaxial-abaxial differential growth during spur formation. Finally, we evaluated the evolution of the targeted genes across Brassicales and examined their expression in dissected floral parts. RESULTS: Five sepals initiate spirally, followed by five petals alternate to the sepals, five antesepalous stamens, three antepetalous stamens, and three carpels. Intercalary growth at the common base of sepals and petals forms a floral tube. The spur is an outgrowth from the adaxial region of the tube, lined up with the medial sepal. We identified Tropaeolum specific duplications in the TCP3/4L and STM gene lineages, which are critical for spur formation in other taxa. In addition, we found that TM6 (MADS-box), RL2 (RAD-like7), and KN2/6L2 and OSH6L (KNOX1 genes), have been lost in core Brassicales but retained in Tropaeolum. CONCLUSIONS: Three genes are pivotal during the extreme adaxial-abaxial asymmetry of the floral tube, namely, TlTCP4L2 restricted to the adaxial side where the spur is formed, and TlTCP12 and TlSTM1 to the abaxial side, lacking a spur.

CmCYC2-like transcription factors may interact with each other or bind to the promoter to regulate floral symmetry development in Chrysanthemum morifolium.

The complex capitulum of Chrysanthemum morifolium is often comprised of bilaterally symmetrical ray florets and radially symmetrical disc florets. The TCP transcription factor clade CYCLOIDEA2 (CYC2) appears to play a vital role in determining floral symmetry and in regulating floral organ development in Asteraceae. Our previous study identified six CmCYC2 genes from chrysanthemum and showed that CmCYC2c participated in the regulation of ray floret identity. However, the functions of other CmCYC2 genes and the underlying molecular mechanism of CmCYC2-mediated floral development regulation in chrysanthemums have not been elucidated. In this study, we analysed the function of CmCYC2 genes by ectopic expression of CmCYC2 in Arabidopsis. Then, we examined the protein-protein interaction using yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays. Finally, we analysed the protein-DNA interaction using yeast one-hybrid (Y1H) and dual-luciferase reporter assays. We found that ectopic expression of CmCYC2 genes in the Arabidopsis tcp1 mutant changed its floral symmetry and flowering time. Y2H and BiFC assays confirmed three pairs of interactions between CmCYC2 proteins, that is, CmCYC2b-CmCYC2d, CmCYC2b-CmCYC2e and CmCYC2c-CmCYC2d, suggesting that heterodimeric complexes may form between CmCYC2 proteins to increase their functional specificity. The results of Y1H and dual-luciferase reporter assays indicate that CmCYC2c can bind to the promoter of ClCYC2f. Our findings provided clues that CmCYC2-like transcription factors may interact with each other or bind to the promoter to regulate floral symmetry development in C. morifolium. KEY MESSAGE: CmCYC2-like transcription factors may interact with each other or bind to the promoter to regulate floral symmetry development in Chrysanthemum morifolium.

Evolution of Class II TCP genes in perianth bearing Piperales and their contribution to the bilateral calyx in Aristolochia.

Controlled spatiotemporal cell division and expansion are responsible for floral bilateral symmetry. Genetic studies have pointed to class II TCP genes as major regulators of cell division and floral patterning in model core eudicots. Here we study their evolution in perianth-bearing Piperales and their expression in Aristolochia, a rare occurrence of bilateral perianth outside eudicots and monocots. The evolution of class II TCP genes reveals single-copy CYCLOIDEA-like genes and three paralogs of CINCINNATA (CIN) in early diverging angiosperms. All class II TCP genes have independently duplicated in Aristolochia subgenus Siphisia. Also CIN2 genes duplicated before the diversification of Saruma and Asarum. Sequence analysis shows that CIN1 and CIN3 share motifs with Cyclin proteins and CIN2 genes have lost the miRNA319a binding site. Expression analyses of all paralogs of class II TCP genes in Aristolochia fimbriata point to a role of CYC and CIN genes in maintaining differential perianth expansion during mid- and late flower developmental stages by promoting cell division in the distal and ventral portion of the limb. It is likely that class II TCP genes also contribute to cell division in the leaf, the gynoecium and the ovules in A. fimbriata.

A lever action hypothesis for pendulous hummingbird flowers: experimental evidence from a columbine.

BACKGROUND AND AIMS: Pendulous flowers (due to a flexible pedicel) are a common, convergent trait of hummingbird-pollinated flowers. However, the role of flexible pedicels remains uncertain despite several functional hypotheses. Here we present and test the 'lever action hypothesis': flexible pedicels allow pendulous flowers to move upwards from all sides, pushing the stigma and anthers against the underside of the feeding hummingbird regardless of which nectary is being visited. METHODS: To test whether this lever action increased pollination success, we wired emasculated flowers of serpentine columbine, Aquilegia eximia, to prevent levering and compared pollination success of immobilized flowers with emasculated unwired and wire controls. KEY RESULTS: Seed set was significantly lower in wire-immobilized flowers than unwired control and wire control flowers. Video analysis of visits to wire-immobilized and unwired flowers demonstrated that birds contacted the stigmas and anthers of immobilized flowers less often than those of flowers with flexible pedicels. CONCLUSIONS: We conclude that flexible pedicels permit the levering of reproductive structures onto a hovering bird. Hummingbirds, as uniquely large, hovering pollinators, differ from flies or bees which are too small to cause levering of flowers while hovering. Thus, flexible pedicels may be an adaptation to hummingbird pollination, in particular due to hummingbird size. We further speculate that this mechanism is effective only in radially symmetric flowers; in contrast, zygomorphic hummingbird-pollinated flowers are usually more or less horizontally oriented rather than having pendulous flowers and flexible pedicels.

Evolution of CYCLOIDEA-like genes in Fabales: Insights into duplication patterns and the control of floral symmetry.

Cycloidea-like (CYC-like) genes are the key regulatory factors in the development of flower symmetry. Duplication and/or reduction of CYC-like genes have occurred several times in various angiosperm groups and are hypothesized to be correlated with the evolution of flower symmetry, which in turn has contributed to the evolutionary success of these groups. However, less is known about the evolutionary scenario of CYC-like genes in the whole Fabales, which contains four families with either symmetric or actinomorphic flowers. Here we investigated the evolution of CYC-like genes in all the four families of Fabales and recovered one to nine CYC-like genes (CYC1, CYC2, and CYC3) depending on which lineages, but the CYC3 genes were most likely lost in the ancestor of Leguminosae. Phylogenetic analysis suggested that the CYC-like genes could have undergone multiple duplications and losses in different plant lineages and formed distinct paralogous/orthologous clades. The ancestor of the Papilionoideae and Caesalpinioideae may possess two paralogs of CYC1 genes but one of them was subsequently lost in Papilionoideae and was retained only in several species of Caesalpinioideae. CYC2 genes were more frequently duplicated in Papilionoideae than in other legumes. We propose that the diversification patterns of both CYC1 and CYC2 genes are not related to the floral symmetry in non-papilionoid Fabales groups, however, gene duplication and functional divergence of CYC2 are essential for the floral zygomorphy of Papilionoideae. This is the first systematic analysis of the CYC-like genes in Fabales and could form the basis for further study of molecular mechanisms controlling floral symmetry in non-model plants of Fabales.

Evolution of RADIALIS and DIVARICATA gene lineages in flowering plants with an expanded sampling in non-core eudicots.

PREMISE OF THE STUDY: Bilateral symmetry in core eudicot flowers is established by the differential expression of CYCLOIDEA (CYC), DICHOTOMA (DICH), and RADIALIS (RAD), which are restricted to the dorsal portion of the flower, and DIVARICATA (DIV), restricted to the ventral and lateral petals. Little is known regarding the evolution of these gene lineages in non-core eudicots, and there are no reports on gene expression that can be used to assess whether the network predates the diversification of core eudicots. METHODS: Homologs of the RAD and DIV lineages were isolated from available genomes and transcriptomes, including those of three selected non-core eudicot species, the magnoliid Aristolochia fimbriata and the monocots Cattleya trianae and Hypoxis decumbens. Phylogenetic analyses for each gene lineage were performed. RT-PCR was used to evaluate the expression and putative contribution to floral symmetry in dissected floral organs of the selected species. KEY RESULTS: RAD-like genes have undergone at least two duplication events before eudicot diversification, three before monocots and at least four in Orchidaceae. DIV-like genes also duplicated twice before eudicot diversification and underwent independent duplications specific to Orchidaceae. RAD-like and DIV-like genes have differential dorsiventral expression only in C. trianae, which contrasts with the homogeneous expression in the perianth of A. fimbriata. CONCLUSIONS: Our results point to a common genetic regulatory network for floral symmetry in monocots and core eudicots, while alternative genetic mechanisms are likely driving the bilateral perianth symmetry in the early-diverging angiosperm Aristolochia.

Dosage imbalance of B- and C-class genes causes petaloid-stamen relating to F1 hybrid variation.

BACKGROUND: Great advances have been achieved in our understanding of flower development and evolution since the establishment of the ABC model. However, it remains a challenge to define the exact context of organ identity in the component interactions of the ABC model. RESULTS: Through hybridization, we detected a homeotic mutant in Petrocosmea (Gesneriaceae) uniquely displayed by the 'petaloid-stamen' in the third whorl with petal identity. Comparative Real-time PCR analyses demonstrate that both two B-class genes DEF2 and GLO are excessively expressed while the transcripts of the C-class gene PLE are reduced in the third floral whorl in the mutant compared to that in the wild-type F1 hybrids. Further allele-specific expression (ASE) analyses indicate that an allele-specific change in PgPLE might be responsible for up-regulation of both B-class genes and down-regulation of the C-class gene in the petaloid-stamen mutants. CONCLUSIONS: Our findings suggest that the petaloid-stamen is consequent upon an evident dosage imbalance between B- and C-class products that is probably triggered by a cis-regulatory change. In addition, the genetic pathway for the floral organ identity might be in parallel with that for the floral symmetry. The extreme variation in hybrids further suggests that interspecific hybridization may represent a major factor for evolutionary innovation and diversification in plants.

Evolution and genetic control of the floral ground plan.

Contents Summary 70 I. Introduction 70 II. What is the floral ground plan? 71 III. Diversity and evolution of the floral ground plan 72 IV. Genetic mechanisms 77 V. What's next? 82 Acknowledgements 83 References 83 SUMMARY: The floral ground plan is a map of where and when floral organ primordia arise. New results combining the defined phylogeny of flowering plants with extensive character mapping have predicted that the angiosperm ancestor had whorls rather than spirals of floral organs in large numbers, and was bisexual. More confidently, the monocot ancestor likely had three organs in each whorl, whereas the rosid and asterid ancestor (Pentapetalae) had five, with the perianth now divided into sepals and petals. Genetic mechanisms underlying the establishment of the floral ground plan are being deduced using model species, the rosid Arabidopsis, the asterid Antirrhinum, and in grasses such as rice. In this review, evolutionary and genetic conclusions are drawn together, especially considering how known genes may control individual processes in the development and evolution of ground plans. These components include organ phyllotaxis, boundary formation, organ identity, merism (the number or organs per whorl), variation in the form of primordia, organ fusion, intercalary growth, floral symmetry, determinacy and, finally, cases where the distinction between flowers and inflorescences is blurred. It seems likely that new pathways of ground plan evolution, and new signalling mechanisms, will soon be uncovered by integrating morphological and genetic approaches.

Geometric morphometrics reveals shifts in flower shape symmetry and size following gene knockdown of CYCLOIDEA and ANTHOCYANIDIN SYNTHASE.

BACKGROUND: While floral symmetry has traditionally been assessed qualitatively, recent advances in geometric morphometrics have opened up new avenues to specifically quantify flower shape and size using robust multivariate statistical methods. In this study, we examine, for the first time, the ability of geometric morphometrics to detect morphological differences in floral dorsoventral asymmetry following virus-induced gene silencing (VIGS). Using Fedia graciliflora Fisch. & Meyer (Valerianaceae) as a model, corolla shape of untreated flowers was compared using canonical variate analysis to knockdown phenotypes of CYCLOIDEA2A (FgCYC2A), ANTHOCYANIDIN SYNTHASE (FgANS), and empty vector controls. RESULTS: Untreated flowers and all VIGS treatments were morphologically distinct from each other, suggesting that VIGS may cause subtle shifts in floral shape. Knockdowns of FgCYC2A were the most dramatic, affecting the position of dorsal petals in relation to lateral petals, thereby resulting in more actinomorphic-like flowers. Additionally, FgANS knockdowns developed larger flowers with wider corolla tube openings. CONCLUSIONS: These results provide a method to quantify the role that specific genes play in the developmental pathway affecting the dorsoventral axis of symmetry in zygomorphic flowers. Additionally, they suggest that ANS may have an unintended effect on floral size and shape.

Genetics of flower development in Ranunculales - a new, basal eudicot model order for studying flower evolution.

Contents 361 I. 361 II. 362 III. 363 IV. 364 V. 364 Acknowledgements 365 References 365 SUMMARY: Ranunculales, the sister group to all other eudicots, encompasses species with a remarkable floral diversity, which are currently emerging as new model organisms to address questions relating to the genetic architecture of flower morphology and its evolution. These questions concern either traits only found in members of the Ranunculales or traits that have convergently evolved in other large clades of flowering plants. We present recent results obtained on floral organ identity and number, symmetry evolution and spur formation in Ranunculales species. We discuss benefits and future prospects of evo-devo studies in Ranunculales, which can provide the opportunity to decipher the genetic architecture of novel floral traits and also to appraise the degree of conservation of genetic mechanisms involved in homoplasious traits.