Petal Initiation Research Articles

The ontogeny of flowers and inflorescences in the andromonoecious annual Iranian endemic Dicyclophora persica (Apiaceae)

Species of Apiaceae, despite uniform floral structure, show great variation in floral morphs and sexual distribution. In the present study, the ontogeny of the diclinous flowers within the cage‐shaped inflorescences of the andromonoecious Dicyclophora persica Boiss. was studied using scanning electron microscopy (SEM), to determine if they follow the same ontogenetic processes. The development of an interesting, unusual club‐like organ in the center of the umbel was also studied. Our findings show that the floral organs in diclinous flowers follow the same pattern. Petals initiate in a clockwise direction, while stamens form in a helical pattern. The perfect central sessile flower in the umbellet, despite sepal suppression, was synchronized with inner staminate flowers in the initiation of petals, stamens, and carpels. In staminate flowers, two prominent sepals initiate after the inception of the stamens. Ovules only get aborted in the late stages. The club‐like organ initiates from a naked receptacle that has no resemblances to umbellets or flowers. In perfect flowers, the delayed initiation and prolonged spatial constraints are caused by bracts and massive staminate flowers which suppress the sepals and make them sessile. Considering the ovule suppression, staminate flowers may be adapted to prevent the perfect flowers from being exposed to extreme environmental conditions.

Perianth phyllotaxis and symmetry in Lamiales

ABSTRACT Genetic studies of floral development have essentially focused on events within the floral meristem. By contrast, phyllotaxis models have shown that sepal arrangement is determined by the structure of the inflorescence. In this study, we assess the interplay between the genetic program underlying floral symmetry and phyllotaxic determination, which together shape the flower. We used a phyllotaxic model for the initiation of sepals and petals in six species of Lamiales and studied the impact of the zygomorphic genetic program on their pattern at anthesis. The initiation pattern of floral organs is determined by hormone signaling and contact pressure acting during organogenesis, while the genetic program underlying zygomorphy forces the bilateral symmetry of the initiation of organ primordia. The integration of phyllotaxic and genetic studies allows to deepen our understanding of the mechanisms producing actinomorphic or zygomorphic flowers. In any case, these mechanisms respect Curie’s symmetry principle which states that “when certain causes (here, expression of CYC-like genes) produce certain effects, the elements of symmetry of the causes must be found in the effects produced (here, perianth symmetry)”.

The Iranian enigmatic endemic Salvia majdae (Lamiaceae): its taxonomic status based on floral ontogeny

Since the first description of Salvia majdae (syn. Zhumeria majdae) in 1967, its taxonomic status has remained an enigma within Salvia senso lato due to its unusual characters, in particular the floral structure. Based on recent molecular phylogenetic data, two defensible opinions on its status have been proposed. Here, we studied the floral development of this species using scanning electron microscopy to resolve this ambiguity. The aspects of floral symmetry were also studied. Like species of Salvia and representative genera of Lamiaceae, the initiation sequence of sepals and stamens in S. majdae is unidirectional from adaxial to abaxial and reverse, respectively. The flowers show temporal overlaps in initiation of petals and stamens. However, in contrast, the bidirectional initiation of the petals in S. majdae is reversed, starting with the abaxial middle petal, followed by the two adaxial ones, and then finally ending with the two lateral ones. The filaments of the sterile stamens are elongated and exposed as staminodes thorough development. We conclude that subequal growth of the calyx lips, weak cochlear aestivation and bidirectional initiation of the petals and well‐developed sterile stamen primordia are responsible for the weak floral asymmetry in S. majdae. Based on these data, S. majdae shows significant differences from Salvia.

A MITE insertion abolishes the AP3-3 self-maintenance regulatory loop in apetalous flowers of Nigella damascena.

MADS-box transcription factors are important regulators of floral organ identity through their binding to specific motifs, termed CArG, in the promoter of their target genes. Petal initiation and development depend on class A and B genes, but MADS-box genes of the APETALA3 (AP3) clade are key regulators of this process. In the early diverging eudicot Nigella damascena, an apetalous [T] morph is characterized by the lack of expression of the NdAP3-3 gene, with its expression being petal-specific in the wild-type [P] morph. All [T] morph plants are homozygous for an NdAP3-3 allele with a Miniature Inverted-repeat Transposable Element (MITE) insertion in the second intron of the gene. Here, we investigated to which extent the MITE insertion impairs regulation of the NdAP3-3 gene. We found that expression of NdAP3-3 is initiated in the [T] morph, but the MITE insertion prevents its positive self-maintenance by affecting the correct splicing of the mRNA. We also found specific CArG features in the promoter of the NdAP3-3 genes with petal-specific expression. However, they are not sufficient to drive expression only in petals of transgenic Arabidopsis, highlighting the existence of Nigella-specific cis/trans-acting factors in regulating AP3 paralogs.

Non-cell-autonomous regulation of petal initiation in Arabidopsis thaliana

In many flowering plants, petals initiate in alternate positions from first whorl sepals, suggesting possible signaling between sepal boundaries and petal initiation sites. PETAL LOSS (PTL) and RABBIT EARS (RBE) regulate petal initiation in Arabidopsis thaliana and their transcripts are expressed in sepal boundary and petal initiation sites, respectively, suggesting that PTL acts in a non-cell-autonomous manner. Here, we determined that cells expressing PTL and RBE fusion proteins did not overlap but were adjacent, confirming the non-cell-autonomous function of PTL. Genetic ablation of intersepal cells by expressing the diphtheria toxin-A chain gene driven by the PTL promoter resulted in flowers lacking petals, suggesting these cells are required for petal initiation. Transcriptome analysis combined with a PTL induction system revealed 42 genes that were upregulated under PTL activation, including UNUSUAL FLORAL ORGANS (UFO), which likely plays an important role in petal initiation. These findings suggest a molecular mechanism in which PTL indirectly regulates petal initiation and UFO mediates positional signaling between the sepal boundary and petal initiation sites.

Lepidium-like, a Naturally Occurring Mutant of Capsella bursa-pastoris, and Its Implications on the Evolution of Petal Loss in Cruciferae.

Naturally occurring mutants whose phenotype recapitulates the changes that distinguish closely related species are of special interest from the evolutionary point of view. They can give a key about the genetic control of the changes that led to speciation. In this study, we described lepidium-like (lel), a naturally occurring variety of an allotetraploid species Capsella bursa-pastoris that is characterized by the typical loss of all four petals. In some cases, one or two basal flowers in the raceme had one or two small petals. The number and structure of other floral organs are not affected. Our study of flower development in the mutant showed that once initiated, petals either cease further development and cannot be traced in anthetic flowers or sometimes develop to various degrees. lel plants showed an earlier beginning of floral organ initiation and delayed petal initiation compared to the wild-type plants. lel phenotype has a wide geographical distribution, being found at the northern extremity of the species range as well as in the central part. The genetic analysis of inheritance demonstrated that lel phenotype is controlled by two independent loci. While the flower in the family Cruciferae generally has a very stable structure (i.e., four sepals, four petals, six stamens, and two carpels), several deviations from this ground plan are known, in particular in the genus Lepidium, C. bursa-pastoris is an emerging model for the study of polyploidy (which is also very widespread in Cruciferae); the identification and characterization of the apetalous mutant lays a foundation for further research of morphological evolution in polyploids.

The role of heterochrony in Schizanthus flower evolution- a quantitative analysis

Changes in the timing of development within a lineage (heterochrony) have an enormous impact on floral diversification. However, comparative quantitative studies in a broad range of ontogenetic stages are still needed to identify the role of different heterochronic processes shaping diversification patterns. In this study, changes in petal development were compared across species in the genus Schizanthus showing floral diversification mediated by pollinators. In this genus, transitions from bee- to hummingbird- and moth- pollination are associated with lower lip diversification. Variation in size and shape of petals was quantified at key ontogenetic stages by using a traditional and a geometric morphometric approach. Buds from petal initiation until shortly before anthesis were considered. Lower lip reduction can arise by different combinations of ontogenetic processes such as a delay in organ initiation, a decrease in growth rate, and/or by terminal deletion of the final shape. These processes may interact having an additive effect along ontogeny. Compensatory effects can also occur, as an increase in growth rates can offset the effect of a delay in organ initiation. Heterochronic changes underlying independent reductions of the lower lip varied between lower lip structures: the wing and the keel. While wing reduction was achieved by similar ontogenetic process in both clades, keel reduction was achieved mostly by terminal deletion of the final shape in the moth-pollinated clade and by a decrease in growth rates in the hummingbird-pollinated clade. Therefore, the lower lip structures (keel and wings) originating from the same primordia are governed by different processes indicating that they are discrete developmental units. In conclusion, comparative quantitative studies along a broad range of ontogenetic stages are key to understand how different combinations of ontogenetic processes shape morphological variation and promote adaptive evolution.

Evolution of the process underlying floral zygomorphy development in pentapetalous angiosperms.

Observations of floral ontogeny indicated that floral organ initiation in pentapetalous flowers most commonly results in a median-abaxial (MAB) petal during early development, a median-adaxial (MAD) petal being less common. Such different patterns of floral organ initiation might be linked with different morphologies of floral zygomorphy that have evolved in Asteridae. Here, we provide the first study of zygomorphy in pentapetalous angiosperms placed in a phylogenetic framework, the goal being to find if the different patterns of floral organ initiation are connected with particular patterns of zygomorphy. We analyzed patterns of floral organ initiation and displays of zygomorphy, extracted from floral diagrams representing 405 taxa in 330 genera, covering 83% of orders (30 out of 36) and 37% of families (116 out of 313) in core eudicots in the context of a phylogeny using ancestral state reconstructions. The MAB petal initiation is the ancestral state of the pattern of floral organ initiation in pentapetalous angiosperms. Taxa with MAD petal initiation represent ∼30 independent origins from the ancestral MAB initiation. There are distinct developmental processes that give rise to zygomorphy in different lineages of pentapetalous angiosperms, closely related lineages being likely to share similar developmental processes. We have demonstrated that development indeed constrains the processes that give rise to floral zygomorphy, while phylogenetic distance allows relaxation of these constraints, which provides novel insights on the role that development plays in the evolution of floral zygomorphy.

Inflorescence and floral traits of the Colombian species of <em>Tristerix (Loranthaceae)</em> related to hummingbird pollination

Floral diversification in Loranthaceae reaches its highest peak in the Andes. The flowers of the exclusively Andean genus Tristerix exhibit tubular and vividly coloured flowers pollinated by hummingbirds. We studied inflorescence and flower morphoanatomy of the two Colombian species, T. longebracteatus and the highly endangered T. secundus. Both species have terminal racemes with up to 26 ebracteolate flowers, of which the proximal one opens and sets fruits first. The slightly irregular calyx initiation is followed by the simultaneous initiation of petals and the successive initiation of stamens. Anthesis is fenestrate, explosive, and triggered by the tension of the style against the abaxial petals, a mode so far not reported in Loranthaceae. Anthetic petals spread symmetrically in T. longebracteatus and asymmetrically in T. secundus. Nectar is produced by a supraovarial disk and by the petal mesophyll. Floral lifespan lasts up to 20 days. The hummingbirds Eriocnemis vestita and Pterophanes cyanopterus are the likely pollinators of T. secundus. Morphological traits are inconclusive to support one of the two competing sister group relationships that involve Tristerix, as the lack of cataphylls in renewal shoots links Ligaria and Tristerix, whereas the terminal inflorescences support its relationship with Desmaria and Tupeia.

Seasonal Regulation of Petal Number.

Four petals characterize the flowers of most species in the Brassicaceae family, and this phenotype is generally robust to genetic and environmental variation. A variable petal number distinguishes the flowers of Cardamine hirsuta from those of its close relative Arabidopsis (Arabidopsis thaliana), and allelic variation at many loci contribute to this trait. However, it is less clear whether C. hirsuta petal number varies in response to seasonal changes in environment. To address this question, we assessed whether petal number responds to a suite of environmental and endogenous cues that regulate flowering time in C. hirsuta We found that petal number showed seasonal variation in C. hirsuta, such that spring flowering plants developed more petals than those flowering in summer. Conditions associated with spring flowering, including cool ambient temperature, short photoperiod, and vernalization, all increased petal number in C. hirsuta Cool temperature caused the strongest increase in petal number and lengthened the time interval over which floral meristems matured. We performed live imaging of early flower development and showed that floral buds developed more slowly at 15°C versus 20°C. This extended phase of floral meristem formation, coupled with slower growth of sepals at 15°C, produced larger intersepal regions with more space available for petal initiation. In summary, the growth and maturation of floral buds is associated with variable petal number in C. hirsuta and responds to seasonal changes in ambient temperature.

PETAL LOSS and ROXY1 Interact to Limit Growth Within and between Sepals But to Promote Petal Initiation in Arabidopsis thaliana.

The activity of genes controlling organ development may be associated with the redox state of subregions within the meristem. Glutaredoxins react to the level of oxidative potential and can reduce cysteine dithiols, in some cases to activate specific transcription factors. In Arabidopsis, loss of function of the glutaredoxin ROXY1 or the trihelix transcription factor PETAL LOSS (PTL) each results in reduced numbers of petals. Here, genetic studies have revealed that loss of petals in ptl mutant plants depends on ROXY1 function. The two genes also act together to restrain stamen-identifying C function from entering the outer whorls. On the other hand, they suppress growth between sepals and in sepal margins, with ROXY1 action partially redundant to that of PTL. Genetic interactions with aux1 mutations indicate that auxin activity is reduced in the petal whorl of roxy1 mutants as in ptl mutants. However, it is apparently increased in the sepal whorl of triple mutants associated with the ectopic outgrowth of sepal margins, and of finger-like extensions of inter-sepal zones that in 20% of cases are topped with bunches of ectopic sepals. These interactions may be indirect, although PTL and ROXY1 proteins can interact directly when co-expressed in a transient assay. Changes of conserved cysteines within PTL to similar amino acids that cannot be oxidized did not block its function. It may be in some cases that under reducing conditions ROXY1 binds PTL and activates it by reducing specific conserved cysteines, thus resulting in growth suppression.

Genome-wide transcriptomic analysis uncovers the molecular basis underlying early flowering and apetalous characteristic in Brassica napus L.

Floral transition and petal onset, as two main aspects of flower development, are crucial to rapeseed evolutionary success and yield formation. Currently, very little is known regarding the genetic architecture that regulates flowering time and petal morphogenesis in Brassica napus. In the present study, a genome-wide transcriptomic analysis was performed with an absolutely apetalous and early flowering line, APL01, and a normally petalled line, PL01, using high-throughput RNA sequencing. In total, 13,205 differential expressed genes were detected, of which 6111 genes were significantly down-regulated, while 7094 genes were significantly up-regulated in the young inflorescences of APL01 compared with PL01. The expression levels of a vast number of genes involved in protein biosynthesis were altered in response to the early flowering and apetalous character. Based on the putative rapeseed flowering genes, an early flowering network, mainly comprised of vernalization and photoperiod pathways, was built. Additionally, 36 putative upstream genes possibly governing the apetalous character of line APL01 were identified, and six genes potentially regulating petal origination were obtained by combining with three petal-related quantitative trait loci. These findings will facilitate understanding of the molecular mechanisms underlying floral transition and petal initiation in B. napus.

Floral organogenesis in Urophysa rockii, a rediscovered endangered and rare species of Ranunculaceae

Urophysa is an Asian endemic genus in the Ranunculaceae, but data on floral organogenesis, which would be a useful complement to molecular data in clarifying the relationship with closely related taxa (Aquilegia and Semiaquilegia) in Ranunculaceae, are completely lacking. We used scanning electron microscopy and light microscopy to study the floral development of Urophysa rockii Ulbrich, a recently rediscovered species in this genus. The sepals are initiated spirally, whereas other organs are nonsimultaneously whorled; the floral phyllotaxis is whorled. Primordia of the sepals are lunular and truncate, but those of the petals and stamens are hemispherical and rounded. After sepal initiation, there is a delay in development, but the initiation of petals and stamens is continuous. The developmental sequence of the microspores in the stamens is centrifugal, although the stamens are initiated centripetally. The early developmental stages of the staminodes are similar to those of the stamens, although much smaller, so they may be phylogenetically homologous organs. The carpel primordia are lunular in shape and plicate. The mature ovule is anatropous and bitegmic. Urophysa shows similar floral development features as Aquilegia and Semiaquilegia, although with some differences, which supports the relationship inferred by DNA sequence data.

Temporal Control of Plant Organ Growth by TCP Transcription Factors

The Arabidopsis petal is a simple laminar organ whose development is largely impervious to environmental effects, making it an excellent model for dissecting the regulation of cell-cycle progression and post-mitotic cell expansion that together sculpt organ form. Arabidopsis petals grow via basipetal waves of cell division, followed by a phase of cell expansion. RABBIT EARS (RBE) encodes a C2H2 zinc finger transcriptional repressor and is required for petal development. During the early phase of petal initiation, RBE regulates a microRNA164-dependent pathway that controls cell proliferation at the petal primordium boundaries. The effects of rbe mutations on petal lamina growth suggest that RBE is also required to regulate later developmental events during petal organogenesis. Here, we demonstrate that, early in petal development, RBE represses the transcription of a suite of CIN-TCP genes that in turn act to inhibit the number and duration of cell divisions; the temporal alleviation of that repression results in the transition from cell division to post-mitotic cell expansion and concomitant petal maturation.

Floral Evolution in the Detarieae (Leguminosae): Phylogenetic Evidence for Labile Floral Development in an Early-Diverging Legume Lineage

Premise of research. We examine floral evolution and phylogenetic relationships in the monophyletic Detarieae and related lineages of Caesalpinioideae. Tribe Detarieae (82 genera) includes nearly half of the genera in subfamily Caesalpinioideae and represents some of the most diverse legumes with respect to floral morphology.Methodology. A total of 67 floral ontogenetic and morphological characters were combined with DNA sequences from the plastid trnL-F and matK regions of 34 Detarieae species and representatives of Cassieae, Cercideae, Caesalpinieae, Papilionoideae, and Mimosoideae, for which we have near-complete ontogenetic series. The morphological and ontogenetic characters were optimized onto the resulting most parsimonious phylogenetic trees and Bayesian topology to study character evolution.Pivotal results. Our study supports previously proposed relationships within the tribe Detarieae and among caesalpinioid lineages and indicates that certain features (bracteole and hypanthium characters, sepal initiation, anther position in bud, overlap in timing of initiation of organ whorls) are phylogenetically informative for particular clades whereas others (reductions in petal and stamen number, sepal and petal initiation patterns) have evolved multiple times in parallel in the Detarieae and other Caesalpinioideae. These analyses suggest that modifications that occur early in ontogeny can be good phylogenetic characters for distinguishing both major taxonomic groups and more closely related taxa and that morphological differences that differentiate species within genera can be caused by changes that occur at all stages of ontogeny. Phylogenetic distribution of character states and ontogenetic evidence suggest that in the Caesalpinioideae, loss or suppression of organs within a whorl, both of which are very common, usually does not affect development in subsequent whorls.Conclusions. Our analyses reveal several switches from zygomorphy to actinomorphy (and vice versa), but in Caesalpinioideae (contrary to Papilionoideae), zygomorphy is likely not clearly associated with higher diversification rates. This study suggests that floral initiation patterns are much more variable in Caesalpinioideae than in the other two subfamilies. Although particular patterns may be canalized in certain lineages of the subfamily (Cercideae, Caesalpinieae), in other clades (Detarieae, Dialiinae) floral development is more labile, explaining the high diversity in floral morphology encountered overall in the paraphyletic Caesalpinioideae.

Comparative floral development in the tribe Mentheae (Nepetoideae: Lamiaceae) and its bearing on the evolution of floral patterns in asterids

AbstractA comparative developmental study of flowers was carried out using epi‐illumination light microscopy on four genera of Lamiaceae (Nepeta, Rosmarinus, Salvia, and Ziziphora), representing all three subtribes of Mentheae. All species examined share unidirectional (adaxial to abaxial) sepal initiation, except Rosmarinus, which has the reverse unidirectional sequence, starting abaxially. Initiated but suppressed bracteoles were detected only in Rosmarinus. In Rosmarinus, Salvia, and Ziziphora, initiation of petals and stamens proceeds unidirectionally from the abaxial side. Floral initiation of Nepeta has bidirectional inception of petals and unidirectional stamen initiation from the adaxial side. Temporal overlap in organ initiation between petal and stamen whorls occurs in all taxa, though this feature is more prominent in Rosmarinus. Significant structural and developmental features that distinguish the four genera include: (1) polysymmetric calyx tube, highly tomentose corolla and deeply four‐partitioned ovary in Nepeta; (2) monosymmetric two‐lipped calyx and shallowly four‐partitioned ovary in Ziziphora; and (3) suppression of adaxial stamens in Salvia and Rosmarinus. Adaxial stamens are absent from Rosmarinus, but reduced stamens remain as staminodia in Salvia. In a phylogenetic context, the late monosymmetry of Nepeta and very early monosymmetry of Rosmarinus could both be regarded as derived conditions compared with the early monosymmetry of Salvia and Ziziphora.

Spatial distribution of the RABBIT EARS protein and effects of its ectopic expression in Arabidopsis thaliana flowers

In many flowering plants, flowers consist of two peripheral organs, sepals and petals, occurring in outer two whorls, and two inner reproductive organs, stamens and carpels. These organs are arranged in a concentric pattern in a floral meristem, and the organ identity is established by the combined action of floral homeotic genes expressed along the whorls. Floral organ primordia arise at fixed positions in the floral meristem within each whorl. The RABBIT EARS (RBE) gene is transcribed in the petal precursor cells and primordia, and regulates petal initiation and early growth in Arabidopsis thaliana. We investigated the spatial and temporal expression pattern of a RBE protein fused to the green fluorescent protein (GFP). Expression of the GFP:RBE fusion gene under the RBE cis-regulatory genomic fragment rescues the rbe petal defects, indicating that the fusion protein is functional. The GFP signal is located to the cells where RBE is transcribed, suggesting that RBE function is cell-autonomous. Ectopic expression of GFP:RBE under the APETALA1 promoter causes the homeotic conversion of floral organs, resulting in sterile flowers. In these plants, the class B homeotic genes APETALA3 and PISTILLATA are down-regulated, suggesting that the restriction of the RBE expression to the petal precursor cells is crucial for flower development.

Comparative study of floral development in Onobrychis melanotricha, Hedysarum varium and Alhagi persarum (Leguminosae: Papilionoideae: Hedysareae)

Floral initiation and development of Hedysarum varium, Onobrychis melanotricha and Alhagi persarum was studied using epi-illumination light-microscopy techniques. The studied species belong to the tribe Hedysareae of the inverted repeat loss clade (IRLC clade), which is characterized by missing the large inverted repeat in the chloroplast genome. The main aim of our study was to determine developmental bases for similarities and differences among the three taxa and to verify the position of Alhagi relative to other genera of the IRLC clade. According to our observations, bracteoles are missing in Onobrychis melanotricha, but are present in the other two species. All three species share unidirectional sepal initiation starting with a median abaxial sepal and bidirectional petal initiation. Stamen initiation is unidirectional in all except in the outer stamen whorl of Hedysarum varium, where it is bidirectional. An important ontogenetic feature in O. melanotricha is the existence of five common primordia, which give rise to petal and stamen primordia. Although in H. varium and O. melanotricha common primordia are observed at some stages in floral organ initiations, in Alhagi all organs are initiated separately. Moreover, overlap in time of floral organs initiation occurs in H. varium and O. melanotricha, but not in A. persarum. The carpel initiates concurrently with the petal primordia in all. It might be presumed that Alhagi is primitive in relation to the other studied Hedysareae taxa, due to the presence of bracteoles, the absence of common primordia, and the lack of overlap in time of different organ initiations.

Silencing ofEcFLO, AFLORICAULA/LEAFYGene of the California Poppy (Eschscholzia californica), Affects Flower Specification in a Perigynous Flower Context

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

Auxin controls petal initiation in Arabidopsis

Floral organs are usually arranged in concentric whorls of sepals, petals, stamens and carpels. How founder cells of these organs are specified is unknown. In Arabidopsis, the PETAL LOSS (PTL) transcription factor functions in the sepal whorl, where it restricts the size of the inter-sepal zone. Genetic evidence suggests that PTL acts to support a petal initiation signal active in the adjacent whorl. Here we aimed to characterise the signal by identifying enhancers that disrupt initiation of the remaining petals in ptl mutants. One such enhancer encodes the auxin influx protein AUX1. We have established that auxin is a direct and mobile petal initiation signal by promoting its biosynthesis in the inter-sepal zone in ptl mutant plants and restoring nearby petal initiation. Consistent with this, loss of PTL function disrupts DR5 expression, an auxin-inducible indicator of petal-initiation sites. The signalling network was extended by demonstrating that: (1) loss of RABBIT EARS (RBE) function apparently disrupts the same auxin influx process as PTL; (2) the action of AUX1 is supported by AXR4, its upstream partner in auxin influx; (3) polar auxin transport, which is controlled by PINOID (PID) and PIN-FORMED1 (PIN1), functions downstream of PTL; and (4) the action of pmd-1d, a dominant modifier of the ptl mutant phenotype, is dependent on auxin transport. Thus, loss of PTL function disrupts auxin dynamics, allowing the role of auxin in promoting petal initiation to be revealed.