Petal Loss Research Articles

Functional domains of the PETAL LOSS protein, a trihelix transcription factor that represses regional growth in Arabidopsis thaliana.

PETAL LOSS (PTL) is a trihelix transcription factor that represses growth, especially between sepal primordia. As one of 30 trihelix proteins in Arabidopsis, it falls in the GT2 clade with duplicated trihelix DNA-binding domains and a long α-helical central domain. PTL orthologs occur in all angiosperm genomes examined except grasses, and sequence comparisons reveal that there are two further short conserved domains at each end. GT2 itself carries two nuclear localization sequences, but PTL has an additional nuclear localization sequence (NLS). We show that PTL can act as a transcriptional activator in yeast and in planta, with the latter tested by two different functional assays. Specific deletions revealed that the activation region is C-terminal. Site-directed mutagenesis of the DNA-binding domains has shown that a conserved tryptophan and two downstream acidic amino acids in the second trihelix, predicted to promote folding, are each required for PTL function. Also, three basic residues in the third helix, near the DNA interaction sites, support its function. PTL was found to dimerize in yeast. This was confirmed and extended by jointly expressing differentially tagged forms of PTL in a transient expression system in Nicotiana benthamiana leaves. Cytoplasmic PTL (with mutant NLS sequences) was carried into the nucleus upon binding with nuclear-localized PTL, providing each partner carried intact central domains. As this 90-amino acid domain is conserved in most trihelix family members, it seems likely that they all function in dimeric form.

Flower morphology and pollen germination in the genus Capsella (Brassicaceae)

Evolutive changes in mating systems are often accompanied by changes in flower morphology, such as the reduction in size or even loss of petals, changes in production of volatiles, pollen/ovule ratio, the position between anthers and stigma and the germination time of pollen after pollination. These changes have been merged under the term “selfing syndrome” and often result in new taxonomic species. The evolutionary shift frequently happens parallel within many families and genera, for example within the Brassicaceae family. Within the genus Capsella, which is closely related to the molecular model species pair Arabidopsis lyrata (SI)/A. thaliana (SC), we studied self-incompatible and self-compatible species. SC species C. rubella and C. bursa-pastoris produce in comparison with the SI species C. grandiflora (i) smaller petals as the result of decreased cell division and only less of decreasing cell volume, (ii) less production of pollen in one flower, (iii) show a lesser incision between the two valves of the fruits, in combination with a shorter style, and (iv) have a much quicker fertilization of SC pollen after pollination. Crossing success between the diploid species, between different provenances of the tetraploid C. bursa-pastoris, and between the two diploid species and particular individuals of the self-incompatible C. grandiflora has been proven.

Disruption of the petal identity gene APETALA3-3 is highly correlated with loss of petals within the buttercup family (Ranunculaceae)

Absence of petals, or being apetalous, is usually one of the most important features that characterizes a group of flowering plants at high taxonomic ranks (i.e., family and above). The apetalous condition, however, appears to be the result of parallel or convergent evolution with unknown genetic causes. Here we show that within the buttercup family (Ranunculaceae), apetalous genera in at least seven different lineages were all derived from petalous ancestors, indicative of parallel petal losses. We also show that independent petal losses within this family were strongly associated with decreased or eliminated expression of a single floral organ identity gene, APETALA3-3 (AP3-3), apparently owing to species-specific molecular lesions. In an apetalous mutant of Nigella, insertion of a transposable element into the second intron has led to silencing of the gene and transformation of petals into sepals. In several naturally occurring apetalous genera, such as Thalictrum, Beesia, and Enemion, the gene has either been lost altogether or disrupted by deletions in coding or regulatory regions. In Clematis, a large genus in which petalous species evolved secondarily from apetalous ones, the gene exhibits hallmarks of a pseudogene. These results suggest that, as a petal identity gene, AP3-3 has been silenced or down-regulated by different mechanisms in different evolutionary lineages. This also suggests that petal identity did not evolve many times independently across the Ranunculaceae but was lost in numerous instances. The genetic mechanisms underlying the independent petal losses, however, may be complex, with disruption of AP3-3 being either cause or effect.

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.

PETAL LOSS is a boundary gene that inhibits growth between developing sepals in Arabidopsis thaliana

Flower primordia are partitioned by boundaries during their early development. Such boundaries occur between whorls of organs, and also between organs within whorls. PETAL LOSS (PTL) is a trihelix transcription factor gene that is expressed in boundaries between sepal primordia in the outer whorl. Over-expression of PTL results in growth suppression suggesting that PTL normally inhibits growth between newly arising sepals. We have tested this by examining the consequences of loss of PTL function using confocal imaging. The size of the inter-sepal zone in stage 4 buds expands radially by 35-40% in ptl-1 mutants as a consequence of additional cell proliferation. There is no change in the size of PTL-expressing cells. PTL expression does not overlap with the sites of petal initiation identified using the DR5 auxin response reporter. The latter are closer to the centre of the flower. Thus the consequence of loss of PTL function on petal initiation is indirect, perhaps through interference with a mobile petal-initiation signal or movement of the PTL protein. CUP-SHAPED COTYLEDON (CUC) genes are also involved in defining inter-sepal boundaries. However, genetic studies combining ptl with loss of cuc1 function, and gain of CUC function in extra early petals-1 (miR164c) mutants, have revealed that CUC and PTL act differently. CUC suppresses growth of sepal tissues from the boundary region whereas PTL acts to keep the size of the boundary in check.

A plastid DNA phylogeny ofDasymaschalon(Annonaceae) and allied genera: Evidence for generic non–monophyly and the parallel evolutionary loss of inner petals

AbstractDasymaschalonand the closely related generaDesmos, FriesodielsiaandMonanthotaxistogether comprise ca. 170 species of trees, shrubs and woody climbers distributed in tropical Africa and tropical Asia. These genera form the desmoid clade, which, because of the presence of diverse flower and fruit syndromes including different types of pollination chambers and moniliform monocarps, offers an opportunity to investigate potentially ecologically significant shifts in flower and fruit characters. Despite its morphological diversity, however, generic delimitation within the desmoid clade is problematic and the intergeneric relationships of the constituent genera are only poorly understood. Bayesian, maximum likelihood and maximum parsimony analyses of plastid DNA sequence data (matK, psbA–trnH, ndhF, rbcL, trnL–F; ca. 5.4 kb; 52 taxa) were used to clarify phylogenetic relationships within the desmoid clade. The evolution and taxonomic utility of selected fruit and flower characters was investigated with likelihood and parsimony ancestral character reconstructions. The results indicate problems in the current delimitations ofDasymaschalonandFriesodielsia. Friesodielsiaas currently circumscribed is polyphyletic, with AfricanFriesodielsiaspecies allied to the African genusMonanthotaxis, and only distantly related to Asian representatives. The majority ofDasymaschalonspecies form a strongly supported clade, but three species are more closely related to Asian species ofFriesodielsia. Ancestral character reconstructions indicate that seed number and monocarp shape are of limited value in generic circumscriptions, and that the three–petalled corolla characteristic ofDasymaschalonevolved independently twice within the desmoid clade. Disruptions to homeotic gene expression or strong selective pressure for a partial enclosure of the mature stamens and carpels by the corolla are hypothesised to underlie the parallel evolution of pollination chambers formed by outer petal homologues subsequent to inner petal loss.

Fenologia da aceroleira cv. Olivier em Junqueirópolis-SP

A aceroleira é uma planta de clima tropical que produz frutos muito ricos em vitamina C. Apesar de ser amplamente difundida, são poucos os trabalhos realizados com essa cultura no País. O presente trabalho objetivou avaliar a fenologia e o crescimento dos frutos da aceroleira cv. Olivier em duas épocas. Para tanto, foi implantado um experimento em pomar comercial no município de Junqueirópolis-SP. Foram avaliados dois ciclos reprodutivos: setembro de 2009 e janeiro de 2010, sendo que, em cada época, foram marcados 400 botões florais distribuídos em 10 plantas. Para avaliação fenológica, o ciclo foi dividido em estádios, sendo avaliados a duração destes, do ciclo reprodutivo, a taxa de frutificação efetiva e o crescimento dos frutos. A duração dos estádios e do ciclo reprodutivo, a taxa de frutificação efetiva e o crescimento dos frutos foram influenciados pelo fator época, sendo que, no mês de janeiro, a duração dos estádios fenológicos foi menor, resultando no desenvolvimento mais rápido dos frutos, que também apresentaram maior diâmetro. Porém, no que se refere à taxa de frutificação, a época de setembro apresentou maior fixação dos frutos. Também foi verificado que o maior índice de abortamento se deu no estádio de perda de pétala em ambas as avaliações. Apesar da diferença na velocidade de crescimento e tamanho dos frutos, ambas as épocas apresentaram crescimento mais acentuado nos primeiros oito dias, mostrando-se linear no restante do ciclo.

Morphological, histological and ultrastructural changes in the olive pistil during flowering

Sexual reproduction is essential for the propagation of higher plants. From an agronomical point of view, this is a particularly key process because fertilization guarantees fruit formation in most cultivated fruit species. In the olive, however, in spite of its agricultural importance, little attention has been paid to the study of sexual reproduction. In order to investigate the cellular and molecular mechanisms that regulate pollen-pistil interactions in the olive during the progamic phase, it is essential to first have a good knowledge of the reproductive structures involved in such interactions. This study characterizes the anatomical and ultrastructural changes in the olive pistil, beginning from the young pistil developing within the bud until the time of petal loss and visible stigma senescence. We have correlated changes in the pistil with a series of defined floral developmental stages and determined that olive pistil structures cannot be considered completely mature and ready to be pollinated and fertilized until the onset of anthesis. Our results clearly show histological and ultrastructural variation during the diverse flowering events. We discuss whether the changes observed might influence or result from pollen-pistil interactions during the progamic phase.

Evolution and loss of long-fringed petals

Background: The Cucurbitaceae genus Trichosanthes comprises 90–100 species that occur from India to Japan and southeast to Australia and Fiji. Most species have large white or pale yellow petals with conspicuously fringed margins, the fringes sometimes several cm long. Pollination is usually by hawkmoths. Previous molecular data for a small number of species suggested that a monophyletic Trichosanthes might include the Asian genera Gymnopetalum (four species, lacking long petal fringes) and Hodgsonia (two species with petals fringed). Here we test these groups’ relationships using a species sampling of c. 60% and 4759 nucleotides of nuclear and plastid DNA. To infer the time and direction of the geographic expansion of the Trichosanthes clade we employ molecular clock dating and statistical biogeographic reconstruction, and we also address the gain or loss of petal fringes. Results: Trichosanthes is monophyletic as long as it includes Gymnopetalum, which itself is polyphyletic. The closest relative of Trichosanthes appears to be the sponge gourds, Luffa, while Hodgsonia is more distantly related. Of six morphology-based sections in Trichosanthes with more than one species, three are supported by the molecular results; two new sections appear warranted. Molecular dating and biogeographic analyses suggest an Oligocene origin of Trichosanthes in Eurasia or East Asia, followed by diversification and spread throughout the Malesian biogeographic region and into the Australian continent. Conclusions: Long-fringed corollas evolved independently in Hodgsonia and Trichosanthes, followed by two losses in the latter coincident with shifts to other pollinators but not with long-distance ispersal events. Together with the Caribbean Linnaeosicyos, the Madagascan Ampelosicyos and the tropical African Telfairia, these cucurbit lineages represent an ideal system for more detailed studies of the evolution and function of petal fringes in plant-pollinator mutualisms.

Evaluation of fertilization and flower abscission following over-expression of an apple NAK-type protein kinase in tomato

With 6 figures Abstract The Ser/Thr protein kinase subfamily of the novel Arabidopsis protein kinase (NAK) family is unique to plants and is not found in animals or prokaryotes, suggesting that NAKs play specific signalling roles in the control of plant growth and development. However, little is known about the function of NAK-type protein kinases. Here, we used a transgenic tomato plant system to examine the biological functions of AFSK, a NAK-type protein kinase isolated from apple fruits. Gain-of-function transgenic tomato plants containing the AFSK gene were developed by Agrobacterium-mediated transformation and plant regeneration. The plants had normal flower phenotype with respect to their overall shapes and sizes. However, the plants displayed decreased self-fertilization and early flower drop at the floral abscission zone following petal loss. These results suggest that AFSK affects pollen development and promotes floral abscission.

Efficient production of novel floral traits in torenia by collective transformation with chimeric repressors of Arabidopsis transcription factors

Molecular breeding with genetic modification enables the production of novel floral traits in floricultural plants that could not be obtained by traditional breeding. To facilitate novel flower production, we collectively introduced 2 sets of 42 and 50 chimeric repressors of Arabidopsis transcription factors into Agrobacterium and then used these to co-transform torenia (Torenia fournieri). We generated 750 transgenic torenias, and identification of the transgenes revealed that more than 80% of the transgenic torenias had a single transgene. A total of 264 plants showed phenotypic modification, and 91.2% displayed modified flower colors and/or shapes, such as altered color patterns, curled petal margins, and wavy petals. These results indicated that the collective transformation system can be applied to molecular breeding of flowers. Detailed analysis of the phenotypes revealed that PETAL LOSS could control blotch sizes and that modification of cell shape could change the texture of petals. We found that the chimeric repressors of functionally unknown transcription factors also induced novel floral traits, and therefore, the transgenic torenias provide an understanding of the functions of transcription factors that could not be revealed by previous studies in Arabidopsis.

Do pollinators respond in a dose-dependent manner to flower herbivory?: An experimental assessment in Loasa tricolor (Loasaceae)

Pollinators are able to discriminate among different floral phenotypes. Thus, flowers more attractive to pollinators usually receive more visits to their reproductive structures. However, this effect of attraction does not necessarily occur in a binary way, all-or-nothing. Loasa tricolor KerGawl. is an annual herb covered with stinging hairs. Flowers present a corolla with five yellow petals and red nectarine scales. In this study, we assess whether the variation in the floral phenotype of L. tricolor affects discrimination by pollinators, following a dose-dependent type of response. This study was carried out in a protected area of Chile where L. tricolor shows high levels of florivory, probably affecting its attraction to pollinators. We experimentally modified the corolla phenotype, removing three or all petals. We measured visitation rate, richness and diversity of pollinator species in treated and control flowers. Results show that three-petal-removal did not reduce the visitation rate compared to control, while complete-petal-removal significantly reduced the visitation rate. Most pollinator species were recorded in the three-petal-removal treatment (nine species), while complete-petal-removal treatment obtained the lowest richness value (four species). These results suggest that species diversity, richness and visitation rates are affected by flower phenotype variation, but probably for complete petal loss only. We suggest that different levels of florivory on L. tricolor do not affect pollinator discrimination in a dose-dependent way.

Systematics of the Aquatic Angiosperm Genus Myriophyllum (Haloragaceae)

Abstract The angiosperm genus Myriophyllum (Haloragaceae) is among the most species-rich genera of aquatic core-eudicots. Myriophyllum has a cosmopolitan distribution with its center of diversity in Australia (> 37 endemics). The widespread invasive species of the genus (M. aquaticum, M. heterophyllum, and M. spicatum) have drawn attention from international natural resource managers. Myriophyllum species are notoriously difficult to identify using vegetative morphology alone, which commonly is all that is available for these highly clonal plants. The relationships among taxa have been difficult to determine with suspected parallelisms in sex expression, sepal and petal loss, and reduced stamen number. A molecular phylogenetic approach was taken to examine relationships among taxa and to employ molecular markers for the reliable identification of Myriophyllum species. This study included ≈ 80% of the known Myriophyllum species. Both nrDNA ITS and cpDNA matK and trnK data were used to examine phylogenetic ...

Morphological evolution in the variable resin-producing Detarieae (Fabaceae): do morphological characters retain a phylogenetic signal?

Previous molecular phylogenetic studies disagree with the informal generic-level taxonomic groups based on morphology. In this study morphological characters in the caesalpinioid clade Detarieae are evaluated within a phylogenetic framework as a means of better understanding phylogenetic relationships and morphological evolution. Morphological characters were observed and scored for representative species of Detarieae focusing on the resin-producing genera. Phylogenetic analyses were carried out with morphological characters alone and then combined with DNA sequences. Despite a high level of homoplasy, morphological data support several clades corresponding to those recovered in molecular phylogenetic analyses. The more strongly supported clades are each defined by at least one morphological synapomorphy. Several characters (e.g. apetaly) previously used to define informal generic groups evolved several times independently, leading to the differences observed with the molecular phylogenetic analyses. Although floral evolution is complex in Detarieae some patterns are recovered. New informal taxonomic groupings are proposed based on the present findings. Floral evolution in the diverse Detarieae clade is characterized by a repeated tendency toward zygomorphy through the reduction of lateral petals and toward complete loss of petals.

Origin choice and petal loss in the flower garden of spiral wave tip trajectories.

Rotating spiral waves have been observed in numerous biological and physical systems. These spiral waves can be stationary, meander, or even degenerate into multiple unstable rotating waves. The spatiotemporal behavior of spiral waves has been extensively quantified by tracking spiral wave tip trajectories. However, the precise methodology of identifying the spiral wave tip and its influence on the specific patterns of behavior remains a largely unexplored topic of research. Here we use a two-state variable FitzHugh-Nagumo model to simulate stationary and meandering spiral waves and examine the spatiotemporal representation of the system's state variables in both the real (i.e., physical) and state spaces. We show that mapping between these two spaces provides a method to demarcate the spiral wave tip as the center of rotation of the solution to the underlying nonlinear partial differential equations. This approach leads to the simplest tip trajectories by eliminating portions resulting from the rotational component of the spiral wave.

Homology and Evolution of Petals in the Core Eudicots

Seventeen morphological characters are described and plotted on a phylogeny of the eudicots. The distribution of perianth characters demonstrates that the currently held view that petals originated from stamens in the core eudicots is not consistent with the predominance of bract-derived petals (bracteopetals). Petals in the core eudicots have the same morphological affinities and structure as sepals or bracts, with a few notable exceptions. Depending on the most likely topology, Berberidopsidales may represent the plesiomorphic perianth condition in the core eudicots, giving rise to pentamerous flowers with sepals and petals of a similar tepaline origin. Within the core eudicots several divergent trends have led to a wide variety of perianth morphologies involving to different degrees the presence of a calyx and corolla. Different factors contribute to the development of a biseriate perianth, including the upward shift of bracts and their absorption in the flower and a progressive differentiation of tepals through shifts in petaloidy. A number of correlations in perianth evolution are highlighted, such as petaloid sepals linked with loss of petals and presence of a hypanthium, a reversal to tepal-like petals with secondary polymery in the androecium and gynoecium, or the loss of sepals linked with the development of stamen-petal tubes. The relationship between petal morphology and the molecular basis for petaloidy is discussed.

Arabidopsis GenesAS1,AS2, andJAGNegatively Regulate Boundary-Specifying Genes to Promote Sepal and Petal Development

Boundary formation is crucial for organ development in multicellular eukaryotes. In higher plants, boundaries that separate the organ primordia from their surroundings have relatively low rates of cell proliferation. This cellular feature is regulated by the actions of certain boundary-specifying genes, whose ectopic expression in organs can cause inhibition of organ growth. Here, we show that the Arabidopsis thaliana ASYMMETRIC LEAVES1 and 2 (AS1 and AS2) and JAGGED (JAG) genes function in the sepal and petal primordia to repress boundary-specifying genes for normal development of the organs. Loss-of-function as1 jag and as2 jag double mutants produced extremely tiny sepals and petals. Analysis of a cell-cycle marker HISTONE4 revealed that cell division in sepal primordia of the double mutant was inhibited. Moreover, these abnormal sepals and petals exhibited ectopic overexpression of the boundary-specifying genes PETAL LOSS (PTL) and CUP-SHAPED COTYLEDON1 [corrected] and 2 (CUC1 and CUC2). Loss of PTL or CUC1 and CUC2 functions in the as1 jag background could partially rescue the tiny sepal and petal phenotypes, supporting the model that the tiny sepal/petal phenotypes are caused, at least in part, by ectopic expression of boundary-specifying genes. Together, our data reveal a previously unrecognized fundamental regulation by which AS1, AS2, and JAG act to define sepal and petal from their boundaries.

A gain-of-function mutation of transcriptional factor PTL results in curly leaves, dwarfism and male sterility by affecting auxin homeostasis

GT factors are plant-specific trihelix DNA-binding transcription factors, which are involved in light responses and other developmental processes in plant. We identified a gain-of-function mutant of a GT-2 factor gene, PETAL LOSS (PTL), which displayed pleiotropic phenotypes including dwarfism, curly leaves, retarded growth and male sterility. We found that constitutive and ectopic over-expression of PTL driven by the CaMV 35S promoter could not recapitulate the phenotypes of the 35S enhancer-driven mutant ptl-D, and was lethal in some of the transgenic plants at the cotyledon developmental stage, suggesting that accurate temporal and spatial expression of PTL is essential for its proper functional implementation during plant development. Further analysis showed that ptl-D was defective in auxin action and that the alteration of auxin distribution corresponded to the curly leaf phenotype. The fact that degeneration of septum cells and subsequent breakage along the stomium was not observed in ptl-D anthers suggests that defective anther dehiscence was the cause for male sterility. Identification and characterization of the gain-of-function mutant ptl-D will improve our understanding of the diverse functions of GT factors during plant development.

Generic level phylogeny of Thalictroideae (Ranunculaceae) — implications for the taxonomic status of Paropyrum and petal evolution

The subfamily Thalictroideae, which is clearly defined by its chromosome characters, contains nine or ten genera. Here we use sequences of chloroplast rbcL and matK genes as well as non‐coding trnL‐F and nuclear ITS regions to reconstruct the generic level phylogeny of this subfamily. Our data recognize three major groups within Thalictroideae: (1) Thalictrum, Leptopyrum, Paropyrum, and Paraquilegia; (2) Urophysa, Semiaquilegia, and Aquilegia; and (3) Dichocarpum, Enemion, and Isopyrum. Isopyrum anemonoides, the sole species of Paropyrum, and Paraquilegia form a clade sister to Leptopyrum; together they make up the first group. Isopyrum manshuricum and I. thalictroides form another clade within the third group. Isopyrum anemonoides is sister to the monophyletic Paraquilegia. Our data suggest that Isopyrum is polyphyletic and support the segregate genus Paropyrum. Reconstruction of petal evolution suggests a secondary loss of petals in Enemion and Thalictrum, in contrast with the view that apetalous taxa are the most primitive. Petals with longer stalks in Dichocarpum are shown to be derivative, refuting the viewpoint that this type is a primitive condition. The flat petals in Paraquilegia may be derived from the tubular‐type petals in Paropyrum and Leptopyrum, and spurred petals are thought to have originated from a single ancestor.

Molecular evolution of the petal and stamen identity genes, APETALA3 and PISTILLATA, after petal loss in the Piperales

Organ loss is an evolutionary phenomenon commonly observed in all kinds of multicellular organisms. Across the angiosperms, petals have been lost several times over the course of their diversification. We examined the evolution of petal and stamen identity genes in the Piperales, a basal lineage of angiosperms that includes the perianthless (with no petals or sepals) families Piperaceae and Saururaceae as well as the Aristolochiaceae, which exhibit a well-developed perianth. Here, we provide evidence for relaxation of selection on the putative petal and stamen identity genes, homologs of APETALA3 and PISTILLATA, following the loss of petals in the Piperales. Our results are particularly interesting as the B-class genes are not only responsible for the production of petals but are also central to stamen identity, the male reproductive organs that show no modification in these plants. Relaxed purifying selection after the loss of only one of these organs suggests that there has been dissociation of the functional roles of these genes in the Piperales.