Arabidopsis PRC1 core component AtRING1 regulates stem cell-determining carpel development mainly through repression of class I <Emphasis Type="Italic">KNOX</Emphasis> genes

Background

Polycomb repressive complex 2 (PRC2)-catalyzed H3K27me3 marks are tightly associated with the WUS-AG negative feedback loop to terminate floral stem cell fate to promote carpel development, but the roles of Polycomb repressive complex 1 (PRC1) in this event remain largely uncharacterized.

Results

Here we show conspicuous variability in the morphology and number of carpels among individual flowers in the absence of the PRC1 core components AtRING1a and AtRING1b, which contrasts with the wild-type floral meristem consumed by uniform carpel production in Arabidopsis thaliana. Promoter-driven GUS reporter analysis showed that AtRING1a and AtRING1b display a largely similar expression pattern, except in the case of the exclusively maternal-preferred expression of AtRING1b, but not AtRING1a, in the endosperm. Indeterminate carpel development in the atring1a;atring1b double mutant is due to replum/ovule-to-carpel conversion in association with ectopic expression of class I KNOX (KNOX-I) genes. Moreover, AtRING1a and AtRING1b also play a critical role in ovule development, mainly through promoting the degeneration of non-functional megaspores and proper integument formation. Genetic interaction analysis indicates that the AtRING1a/b-regulated KNOX-I pathway acts largely in a complementary manner with the WUS-AG pathway in controlling floral stem cell maintenance and proper carpel development.

Conclusions

Our study uncovers a novel mechanistic pathway through which AtRING1a and AtRING1b repress KNOX-I expression to terminate floral stem cell activities and establish carpel cell fate identities.

     

Floral primordia-targeted ACS (1-aminocyclopropane-1-carboxylate synthase) expression in transgenic Cucumis melo implicates fine tuning of ethylene production mediating unisexual flower development

Main conclusion

Floral primordia-targeted expression of the ethylene biosynthetic gene, ACS , in melon suggests that differential timing and ethylene response thresholds combine to promote carpels, inhibit stamens, and prevent asexual bud formation. Typical angiosperm flowers produce both male and female reproductive organs. However, numerous species have evolved unisexuality. Melons (Cucumis melo L.) can produce varying combinations of male, female or bisexual flowers. Regardless of final sex, floral development begins with sequential initiation of all four floral whorls; unisexuality results from carpel or stamen primordia arrest regulated by the G and A loci, respectively. Ethylene, which promotes femaleness, is a key factor regulating sex expression. We sought to further understand the location, timing, level, and relationship to sex gene expression required for ethylene to promote carpel development or inhibit stamen development. Andromonoecious melons (GGaa) were transformed with the ethylene biosynthetic enzyme gene, ACS (1-aminocyclopropane-1-carboxylate synthase), targeted for expression in stamen and petal, or carpel and nectary, primordia using Arabidopsis APETALA3 (AP3) or CRABSCLAW (CRC) promoters, respectively. CRC::ACS plants did not exhibit altered sex phenotype. AP3::ACS melons showed increased femaleness manifested by gain of a bisexual-only phase not seen in wild type, decreased male buds and flowers, and loss of the initial male-only phase. In extreme cases, plants became phenotypically hermaphrodite, rather than andromonoecious. A reduced portion of buds progressed beyond initial whorl formation. Both the ACS transgene and exogenous ethylene reduced the expression of the native carpel-suppressing gene, G, while elevating expression of the stamen-suppressing gene, A. These results show ethylene-mediated regulation of key sex expression genes and suggest a mechanism by which temporally regulated ethylene production and differential ethylene response thresholds can promote carpels, inhibit stamens, and prevent the formation of asexual buds.     

Floral morphogenesis and flowering in aseptic cultures ofBrowallia demissa L

Floral buds ofBrowallia demissa, at three stages of development, were cultured on Nitsch and Nitsch basal medium. The supplements used include IAA; several cytokinins— benzyladenine, kinetin and 6-benzyl-9 tetrahydropyran-adenine (SD 8339); gibberellic acid (GA₃); 2, 3, 5-triiodobenzoic acid (TIBA); arginine and cysteine. All three stages of floral buds failed to complete development. In some treatments stages II and III produced callus and/or roots from the morphological basal end. Cytokinins promoted bud formation whereas both IAA and GA₃ depressed bud formation The shoots differentiatedin vitro were capable of setting flowers, fruits and seeds in all the treatments. The seeds were viable. Comparative studies of development of flowersin vivo andin vitro were made. In some treatments the flowers exhibited abnormal corolla, and roecium and gynoecium. Factors affecting normal bud initiation, organization and development are discussed.     

Isolation and characterization of the C-class MADS-box gene from the distylous pseudo-cereal <Emphasis Type="Italic">Fagopyrum esculentum</Emphasis>

In the model species Arabidopsis thaliana, the floral homeotic C-class gene AGAMOUS (AG) specifies reproductive organ (stamen and carpels) identity and floral meristem determinacy. Gene function analyses in other core eudicots species reveal functional conservation, subfunctionalization and function switch of the C-lineage in this clade. To identify the possible roles of AG-like genes in regulating floral development in distylous species with dimorphic flowers (pin and thrum) and the C function evolution, we isolated and identified an AG ortholog from Fagopyrum esculentum (buckwheat, Family Polygonaceae), an early diverging species of core eudicots preceding the rosids-asterids split. Protein sequence alignment and phylogenetic analysis grouped FaesAG into the euAG lineage. Expression analysis suggested that FaesAG expressed exclusively in developing stamens and gynoecium of pin and thrum flowers. Moreover, FaesAG expression reached a high level in both pin and thrum flowers at the time when the stamens were undergoing rapidly increased in size and microspore mother cells were in meiosis. FaesAG was able to substitute for the endogenous AG gene in specifying stamen and carpel identity and in an Arabidopsis ag-1 mutant. Ectopic expression of FaesAG led to very early flowering, and produced a misshapen inflorescence and abnormal flowers in which sepals had converted into carpels and petals were converted to stamens. Our results confirmed establishment of the complete C-function of the AG orthologous gene preceding the rosids-asterids split, despite the distinct floral traits present in early- and late-diverging lineages of core eudicot angiosperms.     

Relationships of the purple-fringed orchids Platanthera Psycodes and P. grandiflora

The vegetative and floral differences between two frequently confused species are discussed in terms of function.Platanthera psycodes is a widespread species with flowers adapted to visits by short-tongued lepidoptera, especially skippers of the genusPolites; the pollinaria are attached to the base of the proboscis.Platanthera grandiflora is a geographically more limited species with flowers adapted to visits by longer-tongued lepidoptera; the pollinaria are probably attached to compound eyes. The species overlap geographically and phenologically. Chromosome numbers are 2n = 42 in each species. Cultivated plants of each are partially self-incompatible but produce apparently normal embryos when intercrossed. Mechanical, seasonal, ethological, and distributional differences isolate the species.Habenaria andPlatanthera are considered distinct but closely related genera.Platanthera psycodes is based upon a type at L,P. grandiflora on a type at AMES.Platanthera grandiflora andP. psycodes have affinities with three other eastern North American species characterized by tripartite, fringed, or erose labella. Species pairs within this group, based on column structure, includegrandifloraperamoena andpsycodeslacera. The column ofP. leucophaea is morphologically intermediate in form.     

Flavonoids and phylogenetic reconstruction

Flavonoids have been used successfully for interpreting evolutionary relationships in many groups of angiosperms. These interpretations often have been presented in narrative fashion without specific indications of the kinds of relationships expressed. In this paper a method of phylogeny reconstruction with flavonoid data showing cladistic, patristic, and phenetic relationships is presented. Such a phylogram contains maximal information about flavonoid evolution. As an example, relationships in the North American species ofCoreopsis (Compositae), containing 46 species in 11 sections, are analyzed by this approach. A phylogeny of sections of the genus from previous morphological, chromosomal and hybridization data is compared with that from data on anthochlors (chalcones and aurones). Strong correspondence of these evolutionary interpretations gives support to the hypothesized evolutionary trends within the group.     

Floral closure induced by pollination in gynodioecious Cyananthus delavayi (Campanulaceae): effects of pollen load and type, floral morph and fitness consequences

Background and aims

Pollination-induced floral changes, which have been widely documented in flowering plants, have been assumed to enhance the plant''s reproductive success. However, our understanding of the causes and consequences of these changes is still limited. Using an alpine gynodioecious species, Cyananthus delavayi, we investigated the factors affecting floral closure and estimated the fitness consequences of floral closure.

Methods

The timings of floral closure and fertilization were determined. The effects of pollen load, pollen type (cross- or self-pollen) and floral morph (female or perfect flower) on the occurrence of floral closure were examined. Ovule fertilization and seed production were examined to investigate the causes and consequences of floral closure. Flowers were manipulated to prevent closing to detect potential benefits for female fitness.

Key Results

Floral closure, which could be induced by a very low pollen load, occurred within 4–7 h after pollination, immediately following fertilization. The proportion of closed flowers was influenced by pollen load and floral morph, but not by pollen type. Floral closure was more likely to occur in flowers with a higher proportion of fertilized ovules, but there was no significant difference in seed production between closed and open flowers. Those flowers in which closure was induced by natural pollination had low fruit set and seed production. Additionally, seed production was not influenced by closing-prevented manipulation when sufficient pollen deposition was received.

Conclusions

The occurrence of floral closure may be determined by the proportion of fertilized ovules, but this response can be too sensitive to ensure sufficient pollen deposition and can, to some extent, lead to a cost in female fitness. These results implied that the control of floral receptivity by the recipient flowers does not lead to an optimal fitness gain in C. delavayi.     

Dependency on floral resources determines the animals' responses to floral scents

Animal-pollinated angiosperms either depend on cross-pollination or may also reproduce after self-pollination—the former are thus obligately, the latter facultatively dependent on the service of animal-pollinators. Analogously, flower visitors either solely feed on floral resources or complement their diet with these, and are hence dependent or not on the flowers they visit. We assume that obligate flower visitors evolved abilities that enable them to effectively forage on flowers including mechanisms to bypass or tolerate floral defences such as morphological barriers and repellent/deterrent secondary metabolites. Facultative flower visitors, in contrast, are supposed to lack these adaptations and are often prevented to consume floral resources by defence mechanisms. In cases where obligate flower visitors are mutualists and facultative ones are antagonists, this dichotomy provides a solution for the plants'' dilemma to attract pollinators and simultaneously repel exploiters. In a meta-analysis, we recently supported this hypothesis: obligate flower visitors are attracted to floral scents, while facultative ones are repelled. Here, we add empirical evidence to these results: bumblebees and ants, obligate and facultative flower visitors, respectively, responded as predicted by the results of the meta-analysis to synthetic floral scent compounds.Key words: antagonists, exploitation, floral defences, mutualism, nectar, pollinationThe mutualism between flowers and their pollinators is often exploited by cheaters that consume floral rewards but do not contribute to or even reduce the reproductive success of plants.¹ The classification into mutualistic and antagonistic flower visitors represents a phytocentric point of view and only considers the interaction''s net effect for the plant. However, the outcome of each plant-flower visitor interaction may be highly conditional and variable over time² and thus constitutes a continuum between beneficial and detrimental, and it may not be unequivocally assigned to be either positive or negative. Furthermore, many flower visitor species that function as effective pollinators of some plant species represent severe antagonists to other plant species.³ Thus, except for highly specialised systems, it is difficult to predict whether an interaction is mutualistic, commensalistic or antagonistic. We proposed a different classification of flower visitors based on the animals'' interest in flower visits.⁴ Animals visit flowers primarily in search for food; pollination is just a secondary effect.⁵ For some taxa nectar and pollen are the sole nutrient supply, others only supplement their more generalistic diets with floral resources. These different dependencies on floral resources can often be unequivocally assigned to each animal species. Bees, for example, strongly depend on pollen and nectar and are thus obligate flower visitors. In contrast, ants are omnivores and thus facultative flower visitors that consume large amounts of floral nectar of some plant species but obtain most of the nutrients required by the colony from non-floral resources.⁶Optimal foraging theory predicts that animals evolve physiological and behavioral features that allow them to exploit their resources as effectively as possible.^7,8 Therefore, a classification considering the animals'' dependencies on floral resources (obligate versus facultative) may be better suited to explain adaptations to flower visits than their effect on plants'' reproduction (mutualistic versus antagonistic). One very important adaptation to the consumption of floral resources is the ability to tolerate or overcome floral defences that are employed by the flowers to reduce the visitation frequency of detrimental flower visitors.⁹ Floral scents are innate attractants or reinforce floral visits due to associative learning but do also serve as effective repellents against antagonists.¹⁰ In a meta-analysis, we recently demonstrated that the dependency on floral resources determines the responses to floral scents.⁴ In the bioassay presented here, using bumblebees (Bombus terrestris) and ants (Lasius niger), we empirically tested the predictions deduced from the metaanalysis. We expected that bumblebees—as obligate flower visitors—are attracted to floral scent compounds, while ants—as facultative flower visitors—are repelled.     

Diversity and evolution of pollinator rewards and protection by <Emphasis Type="Italic">Macaranga</Emphasis> (Euphorbiaceae) bracteoles

Flowering plants have modified their floral organs in remarkably diverse ways to optimize their interaction with pollinators. Although floral organs represent a major source of floral diversity, many plants also use extrafloral organs, such as bracts and bracteoles, in interacting with pollinators; however, the evolutionary dynamics of non-floral organs involved in pollination are poorly studied. The genus Macaranga is characterized by protective mutualisms with ants that potentially interfere with pollinators on flowers. Macaranga flowers lack perianths and, notably, bracteoles serve the dual function of rewarding pollinators and protecting them from guarding ants; in one group of species, bracteoles provide a nectar reward to generalist pollinators, while in another group, bracteole “chambers” protect thrips or hemipteran pollinators that use these structures as feeding and breeding sites. We examined the diversity and evolutionary dynamics of inflorescence morphology in Macaranga, focusing on bracteoles. We recognized three inflorescence types based on examination of herbarium materials: Discoid-gland, which possess disc-shaped glands on the bracteole surfaces (including all the generalist-pollinated species); Enclosing, in which bracteoles cover flowers (including all the thrips- and hemipteran-pollinated species); and Inconspicuous, in which bracteoles are small, narrow or absent. Ancestral state reconstruction indicated that inflorescence morphologies have changed multiple times in the genus. These findings suggest that morphological changes in non-floral characters (bracteoles) of Macaranga species have occurred as frequently as in the floral structures of many flowering plants. The multiple evolutions of the Enclosing bracteoles, which protect pollinators, might have been facilitated by pollination interference from mutualistic ants.     

Floral structure and development in Nartheciaceae (Dioscoreales), with special reference to ovary position and septal nectaries

We present a comparative study of the floral structure and development of Nartheciaceae, a small dioscorealean family consisting of five genera (Aletris, Lophiola, Metanarthecium, Narthecium, and Nietneria). A noticeable diversity existed in nine floral characters. Analyses of their respective character states in the light of a phylogenetic context revealed that the flowers of Nartheciaceae, whose plesiomorphies occur in Aletris and Metanarthecium, have evolved toward in all or part of Lophiola, Narthecium, and Nietneria: (1) loss of a perianth tube; (2) stamen insertion at the perianth base; (3) congenital carpel fusion; (4) loss of the septal nectaries; (5) unilocular style; (6) unfused lateral carpellary margins in the style; (7) flower with the median outer tepal on the abaxial side; (8) flower with moniliform hairs; and (9) flower with weak monosymmetry. We further found that, as the flowers developed, the ovary shifted its position from inferior to superior. As a whole, their structure changes suggest that the Nartheciaceae flowers have evolved in close association with pollination and seed dispersal. By considering inferior ovaries and the presence of septal nectaries as plesiomorphies of Nartheciaceae, we discussed evolution of the ovary position and septal nectaries in all the monocots.     

Broad geographic covariation between floral traits and the mating system in Camissoniopsis cheiranthifolia (Onagraceae): multiple stable mixed mating systems across the species' range?

Background and Aims

Plants vary widely in the extent to which seeds are produced via self-fertilization vs. outcrossing, and evolutionary change in the mating system is thought to be accompanied by genetic differentiation in a syndrome of floral traits. We quantified the pattern of variation and covariation in floral traits and the proportion of seeds outcrossed (t) to better understand the evolutionary processes involved in mating system differentiation among and within populations of the short-lived Pacific coastal dune endemic Camissoniopsis cheiranthifolia across its geographic range in western North America.

Methods

We quantified corolla width and herkogamy, two traits expected to influence the mating system, for 48 populations sampled in the field and for a sub-sample of 29 populations grown from seed in a glasshouse. We also measured several other floral traits for 9–19 populations, estimated t for 16 populations using seven allozyme polymorphisms, and measured the strength of self-incompatibility for nine populations.

Key Results

Floral morphology and self-incompatibility varied widely but non-randomly, such that populations could be assigned to three phenotypically and geographically divergent groups. Populations spanned the full range of outcrossing (t = 0·001–0·992), which covaried with corolla width, herkogamy and floral life span. Outcrossing also correlated with floral morphology within two populations that exhibited exceptional floral variation.

Conclusions

Populations of C. cheiranthifolia seem to have differentiated into three modal mating systems: (1) predominant outcrossing associated with self-incompatibility and large flowers; (2) moderate selfing associated with large but self-compatible flowers; and (3) higher but not complete selfing associated with small, autogamous, self-compatible flowers. The transition to complete selfing has not occurred even though the species appears to possess the required genetic capacity. We hypothesize that outcrossing populations in this species have evolved to different stable states of mixed mating.     

A double-flowered variety of lesser periwinkle (Vinca minor fl. pl.) that has persisted in the wild for more than 160 years

Background and Aims

Homeotic transitions are usually dismissed by population geneticists as credible modes of evolution due to their assumed negative impact on fitness. However, several lines of evidence suggest that such changes in organ identity have played an important role during the origin and subsequent evolution of the angiosperm flower. Better understanding of the performance of wild populations of floral homeotic varieties should help to clarify the evolutionary potential of homeotic mutants. Wild populations of plants with changes in floral symmetry, or with reproductive organs replacing perianth organs or sepals replacing petals have already been documented. However, although double-flowered varieties are quite popular as ornamental and garden plants, they are rarely found in the wild and, if they are, usually occur only as rare mutant individuals, probably because of their low fitness relative to the wild-type. We therefore investigated a double-flowered variety of lesser periwinkle, Vinca minor flore pleno (fl. pl.), that is reported to have existed in the wild for at least 160 years. To assess the merits of this plant as a new model system for investigations on the evolutionary potential of double-flowered varieties we explored the morphological details and distribution of the mutant phenotype.

Methods

The floral morphology of the double-flowered variety and of a nearby population of wild-type plants was investigated by means of visual inspection and light microscopy of flowers, the latter involving dissected or sectioned floral organs.

Key Results

The double-flowered variety was found in several patches covering dozens of square metres in a forest within the city limits of Jena (Germany). It appears to produce fewer flowers than the wild-type, and its flowers are purple rather than blue. Most sepals in the first floral whorl resemble those in the wild-type, although occasionally one sepal is broadened and twisted. The structure of second-whorl petals is very similar to that of the wild-type, but their number per flower is more variable. The double-flowered character is due to partial or complete transformation of stamens in the third whorl into petaloid organs. Occasionally, ‘flowers within flowers’ also develop on elongated pedicels in the double-flowered variety.

Conclusions

The flowers of V. minor fl. pl. show meristic as well as homeotic changes, and occasionally other developmental abnormalities such as mis-shaped sepals or loss of floral determinacy. V. minor fl. pl. thus adds to a growing list of natural floral homeotic varieties that have established persistent populations in the wild. Our case study documents that even mutant varieties that have reproductive organs partially transformed into perianth organs can persist in the wild for centuries. This finding makes it at least conceivable that even double-flowered varieties have the potential to establish new evolutionary lineages, and hence may contribute to macroevolutionary transitions and cladogenesis.     

Variation and taxonomy of Aesculus pavia L. (Hippocastanaceae) in Texas

The status of Aesculus pavia var. flavescens, a set of yellow-flowered populations endemic to the Edwards Plateau of central Texas, was evaluated on the basis of comparative morphology, geography, flowering phenology, and pollination ecology. Plants of typical A. pavia have red, tubular flowers with exserted stamens and are effectively pollinated by ruby-throated hummingbirds. Plants of var. flavescens have yellow, campanulate flowers with included stamens that can be pollinated effectively by large bees. These ethological reproductive isolating mechanisms are further augmented by a difference in flowering time between the two sets of populations and by geographical restriction of the yellow-flowered plants to localities on the Edwards Plateau. The yellow and red-flowered plants constitute two morphologically distinct groups, despite evidence of limited introgression in counties along the Balcones Escarpment. A yellow-flowered plant discovered in coastal east Texas may indicate the mechanism by which var. flavescens originated, but this plant is much more closely comparable to typical red-flowered var. pavia than to the yellow-flowered plants on the Edwards Plateau. An evolutionary history of section Pavia of Aesculus is presented, and it is concluded that the yellow-flowered plants of central Texas are best regarded as a taxonomic variety.     

Unusual alternations of floral organs in <Emphasis Type="Italic">Paeonia</Emphasis>: Structure and possible mechanism of formation

Morphological analysis of flowers was carried out in Paeonia L. cultivars. Some unusual alternations of floral organs were described: sepal-(petal-stamen) × n-carpel, where 2 ≤ n ≤ 4 (appearance of an additional zone of petal and stamen formation in the medial flower part). The identity of floral organs was not affected in the flowers with this unusual alternation. It was shown on the basis of mathematical simulation of the genes responsible for flower development that these alternations may be determined by increased pool of stem cells, which may lead to delayed termination of flower development.     

Revision of Dicorynia (Cassieae,Caesalpiniaceae)

A combined taxonomic revision and wood anatomical study ofDicorynia, a South American genus of legumes that is unusual in both its floral and wood anatomy, is presented. Two possible evolutionary pathways leading to the unique 8-to 10-thecate anthers are discussed, and it is concluded that apical proliferation of the thecal primordia is the more likely. Gross and microscopic features of this silica-accumulating wood are given in detail and contrasted with other legume woods especially those containing silica. Economic uses of the trees, especially the wood ofD. guianensis, are discussed particularly in regard to marine construction. It is suggested that the high resistance of this wood to marine borers lies not so much in its silica content as to the presence of some other chemical. Based on the similarity of floral and fruit structures, the number of species is reduced from 7 to 2:D. guianensis in the Guianas andD. paraensis in the Amazonian region. Vegetative variations in the Amazonian population, previously given species rank, are reduced to 5 new varieties ofD. paraensis.     

Carpeloidy in flower evolution and diversification: a comparative study in Carica papaya and Arabidopsis thaliana

Background and Aims

Bisexual flowers of Carica papaya range from highly regular flowers to morphs with various fusions of stamens to the ovary. Arabidopsis thaliana sup1 mutants have carpels replaced by chimeric carpel–stamen structures. Comparative analysis of stamen to carpel conversions in the two different plant systems was used to understand the stage and origin of carpeloidy when derived from stamen tissues, and consequently to understand how carpeloidy contributes to innovations in flower evolution.

Methods

Floral development of bisexual flowers of Carica was studied by scanning electron microscopy and was compared with teratological sup mutants of A. thaliana.

Key Results

In Carica development of bisexual flowers was similar to wild (unisexual) forms up to locule initiation. Feminization ranges from fusion of stamen tissue to the gynoecium to complete carpeloidy of antepetalous stamens. In A. thaliana, partial stamen feminization occurs exclusively at the flower apex, with normal stamens forming at the periphery. Such transformations take place relatively late in development, indicating strong developmental plasticity of most stamen tissues. These results are compared with evo-devo theories on flower bisexuality, as derived from unisexual ancestors. The Arabidopsis data highlight possible early evolutionary events in the acquisition of bisexuality by a patchy transformation of stamen parts into female parts linked to a flower axis-position effect. The Carica results highlight tissue-fusion mechanisms in angiosperms leading to carpeloidy once bisexual flowers have evolved.

Conclusions

We show two different developmental routes leading to stamen to carpel conversions by late re-specification. The process may be a fundamental aspect of flower development that is hidden in most instances by developmental homeostasis.     

Floral development of <Emphasis Type="Italic">Gonocaryum</Emphasis> with emphasis on the gynoecium

We investigated the floral development of Gonocaryum, a genus of Cardiopteridaceae that was segregated from Icacinaceae s.l., using scanning electron microscopy to clarify its gynoecial structure and facilitate morphological comparisons of Cardiopteridaceae. The key floral developmental characters include sepal initiation that follows a quincuncial spiral sequence; petals that are valvate with inflexed tips and are postgenitally fused at the base; a petal and stamen initiation sequence that is almost simultaneous; a globular protuberance on top of the connective; a gynoecium that is tricarpellate and pseudomonomerous, with the stigma produced by one abaxial lateral carpel; and two ovules that are unitegmic and anatropous with an obturator on the funicle. The floral developmental characters of Gonocaryum are discussed relative to Cardiopteris, which has been well studied and whose gynoecial vasculature is reinterpreted here, and are briefly compared to other members of Aquifoliales and Icacinaceae s.l. The imbricate sepals, initiated in a quincuncial spiral sequence, and the tricarpellate, pseudomonomerous gynoecium are common characters of Cardiopteridaceae. Unisexual flowers are an autapomorphy of Gonocaryum in Cardiopteridaceae.     

Functional analysis of an <Emphasis Type="Italic">APETALA1</Emphasis>-like MADS box gene from <Emphasis Type="Italic">Eustoma grandiflorum</Emphasis> in regulating floral transition and formation

An Eustoma grandiflorum APETALA1 (EgAP1) gene showing high homology to the SQUA subfamily of MADS-box genes was isolated and characterized. EgAP1, containing a conserved euAP1 motif at the C-terminus, showed high sequence identity to Antirrhinum majus SQUAMOSA in the SQUA subfamily. EgAP1 mRNA was detected in the leaf and expressed significantly higher in young flower buds than in mature flower buds. In flowers, EgAP1 mRNA was strongly detected in sepal, weakly detected in petal and was absent in stamen and carpel. Transgenic Arabidopsis plants ectopically expressing EgAP1 flowered early and produced terminal flowers. In addition, the conversion of petals into stamen-like structures was also observed in 35S::EgAP1 flowers. 35S::EgAP1 was able to complement the ap1 flower defects by restoring the defect for sepal formation and significantly increasing second whorl petal production in Arabidopsis ap1 mutant plants. These results revealed that EgAP1 is the APETALA1 homolog in E. grandiflorum and that the function of EgAP1 is involved in floral induction and flower formation.