Floral Symmetry Research Articles

Genome-wide identification and expression pattern analysis of the TCP transcription factor family in Ginkgo biloba

ABSTRACT Plant-specific TCP transcription factors play an essential role in plant growth and development. They can regulate leaf curvature, flower symmetry and the synthesis of secondary metabolites. The flavonoids in Ginkgo biloba leaf are one of the main medicinally bioactivate compounds, which have pharmacological and beneficial health effects for humans. In this study, a total of 13 TCP genes were identified in G. biloba, and 5 of them belonged to PCF subclades (GbTCP03, GbTCP07, GbTCP05, GbTCP13, GbTCP02) while others belonged to CIN (GbTCP01, GbTCP04, GbTCP06, GbTCP08, GbTCP09, GbTCP10, GbTCP11, GbTCP12) subclades according to phylogenetic analysis. Numerous cis-acting elements related to various biotic and abiotic signals were predicted on the promoters by cis-element analysis, suggesting that the expression of GbTCPs might be co-regulated by multiple signals. Transcript abundance analysis exhibited that most of GbTCPs responded to multiple phytohormones. Among them, the relative expression levels of GbTCP06, GbTCP11, and GbTCP13 were found to be significantly influenced by exogenous ABA, SA and MeJA application. In addition, a total of 126 miRNAs were predicted to target 9 TCPs (including GbTCP01, GbTCP02, GbTCP04, GbTCP05, GbTCP06, GbTCP08, GbTCP11, GbTCP12, GbTCP13). The correlation analysis between the expression level of GbTCPs and the flavonoid contents showed that GbTCP03, GbTCP04, GbTCP07 might involve in flavonoid biosynthesis in G. biloba. In short, this study mainly provided a theoretical foundation for better understanding the potential function of TCPs in G. biloba.

Chromosome-level de novo genome assembly of Telopea speciosissima (New South Wales waratah) using long-reads, linked-reads and Hi-C.

Telopea speciosissima, the New South Wales waratah, is an Australian endemic woody shrub in the family Proteaceae. Waratahs have great potential as a model clade to better understand processes of speciation, introgression and adaptation, and are significant from a horticultural perspective. Here, we report the first chromosome-level genome for T. speciosissima. Combining Oxford Nanopore long-reads, 10x Genomics Chromium linked-reads and Hi-C data, the assembly spans 823Mb (scaffold N50 of 69.0Mb) with 97.8% of Embryophyta BUSCOs "Complete". We present a new method in Diploidocus (https://github.com/slimsuite/diploidocus) for classifying, curating and QC-filtering scaffolds, which combines read depths, k-mer frequencies and BUSCO predictions. We also present a new tool, DepthSizer (https://github.com/slimsuite/depthsizer), for genome size estimation from the read depth of single-copy orthologues and estimate the genome size to be approximately 900Mb. The largest 11scaffolds contained 94.1% of the assembly, conforming to the expected number of chromosomes (2n = 22). Genome annotation predicted 40,158 protein-coding genes, 351 rRNAs and 728 tRNAs. We investigated CYCLOIDEA (CYC) genes, which have a role in determination of floral symmetry, and confirm the presence of two copies in the genome. Read depth analysis of 180 "Duplicated" BUSCO genes using a new tool, DepthKopy (https://github.com/slimsuite/depthkopy), suggests almost all are real duplications, increasing confidence in the annotation and highlighting a possible need to revise the BUSCO set for this lineage. The chromosome-level T. speciosissima reference genome (Tspe_v1) provides an important new genomic resource of Proteaceae to support the conservation of flora in Australia and further afield.

Gene Duplication and Differential Expression of Flower Symmetry Genes in Rhododendron (Ericaceae).

Bilaterally symmetric flowers have evolved over a hundred times in angiosperms, yet orthologs of the transcription factors CYCLOIDEA (CYC), RADIALIS (RAD), and DIVARICATA (DIV) are repeatedly implicated in floral symmetry changes. We examined these candidate genes to elucidate the genetic underpinnings of floral symmetry changes in florally diverse Rhododendron, reconstructing gene trees and comparing gene expression across floral organs in representative species with radial and bilateral flower symmetries. Radially symmetric R. taxifolium Merr. and bilaterally symmetric R. beyerinckianum Koord. had four and five CYC orthologs, respectively, from shared tandem duplications. CYC orthologs were expressed in the longer dorsal petals and stamens and highly expressed in R. beyerinckianum pistils, whereas they were either ubiquitously expressed, lost from the genome, or weakly expressed in R. taxifolium. Both species had two RAD and DIV orthologs uniformly expressed across all floral organs. Differences in gene structure and expression of Rhododendron RAD compared to other asterids suggest that these genes may not be regulated by CYC orthologs. Our evidence supports CYC orthologs as the primary regulators of differential organ growth in Rhododendron flowers, while also suggesting certain deviations from the typical asterid gene regulatory network for flower symmetry.

Structural diversity of elaiophores in Argentine species of Malpighiaceae: morphology, anatomy, and interaction with pollinators.

Most Neotropical Malpighiaceae species are characterized by having zygomorphic flowers and oil glands in the sepals called elaiophores; these floral characteristics are associated with a particular pollination syndrome through oil-collecting bees. This work proposes a study about the structural characteristics of elaiophores in 18 species of Malpighiaceae present in Argentina. The main objectives are to describe the morphology and anatomy of the elaiophores, to detect variation in the number of glands, to compare similarities or differences in elaiophores of species belonging to different lineages, and to know about the potential pollinators and their association with the structural traits of the elaiophores. The morphology and the anatomy were studied using traditional methods of scanning electron and bright-field microscopes. Field trips were carried out to capture oil-collecting bee species on flowers, in different natural populations. Different measurements were taken in the flowers, elaiophores, and oil-collecting bees and were statistically analyzed. Although elaiophores showed a common pattern, some particularities in number, morphology, and anatomy were detected; few of these seem to be restricted to some groups of species phylogenetically related. As regards pollinators, a positive tendency was observed between the size of the flowers, elaiophores, and oil-collecting bees. However, the thickness of the cuticle presented a negative association with the size of the elaiophore and consequently with the floral diameter, which could be presumably related to the foraging behavior and/or the structure of oil-collecting apparatus of the bee species.

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

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

Increased CG, CHG and CHH methylation at the cycloidea gene in the “Peloria” mutant of Linaria vulgaris

Heritable DNA methylation variation is frequently observed in natural populations of plants, but is thought mostly to be functionally inconsequential. An exception to this is the “Peloria” mutant of Linaria vulgaris, which was originally described by Carl von Linné in 1744. A study in 1999 found that the Peloria phenotype is caused by an epiallele of the L. vulgaris cycloidea homolog Lcyc that showed increased levels of DNA methylation compared to wild-type. The DNA methylation results in silencing of Lcyc, which causes radial flower symmetry in the peloric mutant, whereas wild-type plants have flowers with bilateral symmetry. However, a detailed view of DNA methylation at Lcyc at the single-nucleotide level has not been available.In this study, we investigated DNA methylation at Lcyc and, as a control, at the LvHIRZ gene in wild-type and peloric plants of L. vulgaris using DNA bisulfite treatment coupled to next-generation sequencing. We found strong increases in CHG and CHH methylation at Lcyc, but not LvHIRZ, in Peloria. CG methylation was also increased, but wild-type Lcyc also showed moderate levels of CG methylation. Our results suggest that DNA methylation in all three sequence contexts has been maintained, and potentially transgenerationally inherited, in the peloric L. vulgaris population over decades or even centuries.

Confirmation of distyly in Perovskia and floral dimorphism of P. abrotanoides (Salviinae: Lamiaceae)

Heterostyly is a specialised reproductive system with a reciprocal arrangement between the stamen and style, and often associated with tubular actinomorphic flowers. Accordingly, heterostyly is extremely rare in Lamiaceae, a representative zygomorphic flower group. However, some evidence indicates the occurrence of heterostyly in Perovskia. To confirm the existence of heterostyly, 288 herbarium specimens of Perovskia were observed, and a detailed floral morphological study was conducted on P. abrotanoides as a representative to diagnose the evolution of heterostyly. All examined specimens could be divided into two groups: a long-styled morph (L-morph) and a short-styled morph (S-morph), while P. abrotanoides, P. angustifolia, P. atriplicifolia, P. botschantzevii, and P. scrophulariifolia were recognized as distylous taxa. In P. abrotanoides, L-morph flowers possessed a larger calyx, a longer corolla, stigma, and anther, while significantly larger pollen grains and wider corolla were detected in S-morphs. Micromorphological structure and distribution patterns of trichomes were identical to both floral morphs, but the stomatal size on leaves and density of specific trichomes were somewhat different between morphs. Collectively, all data indicate a stable distylous condition of P. abrotanoides as the first case showing general heterostylous syndrome within Lamiaceae.

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

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

Nocturnal Bees as Crop Pollinators

Bees are typically diurnal but around 1% of described species have nocturnal activity. Nocturnal bees are still poorly studied due to bias towards studying diurnal insects. However, knowledge concerning their biology and role as crop pollinators has increased. We review the literature on nocturnal bees’ traits and their host plants, and assess the crop pollination effectiveness of this neglected group. Nocturnal bees have visual adaptations to cope with low light intensities, and floral scents are a key sensory cue used to find their host flowers. Nocturnal bees generally show high flower constancy, the ability to vibrate flowers, and high transfer rates of pollen grains to stigmas. The flowers visited by nocturnal bees range from small radial and zygomorphic flowers to large brush blossoms; moreover, they visit plants with different flowering strategies. Nocturnal bees are effective pollinators of regional fruit crops in Brazil, such as cambuci (Campomanesia phaea), guaraná (Paullinia cupana), cajá (Spondias mombin), and in North America of cultivated pumpkins (Cucurbita species). However, they most likely are pollinators of several other crops. Strategies to host high numbers of nocturnal bees around cropping areas should be taken, such as preserving adjacent native forests, restricting soil management, providing food resources beyond crop flowers, and avoiding light pollution.

Asymmetric expression patterns of B- and C-class MADS-box genes correspond to the asymmetrically specified androecial identities of Canna indica.

Canna indica is a common ornamental plant with asymmetric flowers having colourful petaloid staminodes. The only fertile stamen comprises a one-theca anther and a petaloid appendage and represents the lowest stamen number in the order Zingiberales. The molecular mechanism for the asymmetric androecial petaloidy remains poorly understood. Here, we studied the identity specification in Canna stamen. We observed four types of abnormal flower in terms of androecium identity transformation and analysed the corresponding floral symmetry changes. We further tested the expression patterns of B- and C-class MADS-box genes using in situ hybridization in normal Canna stamen. Homeotic conversions in the androecium were accompanied by floral symmetry changes, and the asymmetric stamen is key in contributing to the floral asymmetry. Both B- and C-class genes exhibited higher expression levels in the anther primordium than in other androecial parts. This asymmetric expression pattern precisely corresponded to the asymmetric identities of the Canna androecium. We identified C. indica as a model species for studying androecial organ identity and floral symmetry synthetically in Zingiberales. We hypothesized that homeotic genes specify floral organ identity in a putative dose-dependent manner. The results add to the current understanding of organ identity-related floral symmetry.

Mechanism of Leaf Vein Coloration and Inheritance of Leaf Vein Color, Flower Form, and Floral Symmetry in Gloxinia

Double-flowered gloxinia (Sinningia speciosa) cultivars with foliar variegation might have a greater market appeal as flowering foliage plants. Crosses were made among 16 gloxinia cultivars and their progenies were analyzed to determine the inheritance of leaf vein color, flower form, and floral symmetry. All plants from self-pollinating white-veined cultivars or crosses between white-veined and green-veined cultivars produced white veins. Progeny derived from self-pollinating plants of white-veined cultivars × green-veined cultivars segregated into a ratio of 3 white-veined:1 green-veined. All plants from self-pollinating or cross-pollinating single-flowered cultivars produced single flowers. Progeny of self-pollination or crosses between double-flowered cultivars segregated into a ratio of 3 double flowers:1 single flower. Contingency chi-square tests revealed that leaf vein color and flower form were inherited independently. New gloxinia progenies with homozygous white veins and double flowers were successfully developed from the F2 segregating population. Plants from self-pollinating or cross-pollinating cultivars with actinomorphic flowers produced actinomorphic flowers. A single dominant gene expressed in the homozygous or heterozygous state resulted in the zygomorphic flowers. Independent inheritance was observed between vein color and floral symmetry. Air spaces between the epidermis and the mesophyll cells were observed in the white, but not in the green, leaf vein portions. Net photosynthesis did not differ significantly between the white vein and adjacent green portion of the same leaf.

An Optimized Transformation System and Functional Test of CYC-Like TCP Gene CpCYC in Chirita pumila (Gesneriaceae).

The development of an ideal model plant located at a key phylogenetic node is critically important to advance functional and regulatory studies of key regulatory genes in the evolutionary developmental (evo-devo) biology field. In this study, we selected Chirita pumila in the family Gesneriaceae, a basal group in Lamiales, as a model plant to optimize its genetic transformation system established previously by us through investigating a series of factors and further conduct functional test of the CYC-like floral symmetry gene CpCYC. By transforming a RNAi:CpCYC vector, we successfully achieved the desired phenotypes of upright actinomorphic flowers, which suggest that CpCYC actually determines the establishment of floral zygomorphy and the horizontal orientation of flowers in C. pumila. We also confirmed the activities of CpCYC promoter in dorsal petals, dorsal/lateral staminodes, as well as the pedicel by transferring a CpCYC promoter:GUS vector into C. pumila. Furthermore, we testified the availability of a transient gene expression system using C. pumila mesophyll protoplasts. The improved transformation system together with the inherent biological features would make C. pumila an attractive new model in functional and regulatory studies for a broad range of evo-devo issues.

Male sterility significantly elevates shape variation and fluctuating asymmetry of zygomorphic corolla in gynodioecious Glechoma hederacea (Lamiaceae)

Female flowers of gynodioecious plants usually have smaller corollas than bisexual flowers. This difference is explained as a developmental consequence of stamen abortion and as a result of stronger selection for larger corolla size in hermaphroditic flowers that solely ensure male function within populations. This study evaluated whether the size difference of zygomorphic corollas in a widely distributed gynodioecious herb Glechoma hederacea is accompanied by variation in shape and bilateral fluctuating asymmetry of sexually differentiated flowers. Geometric morphometric analyses of bilateral symmetry in the shapes of corolla lower lips showed that male-sterile flowers were significantly more plastic and asymmetric, implying that they may be subjected to weaker stabilizing selection for corolla shape in comparison to hermaphrodites. These results illustrated that sexual differentiation may be an important factor contributing to bilateral fluctuating asymmetry in the shape of zygomorphic flowers.

Evolution: Cleistogamy to the rescue of zygomorphic flowers

Cleistogamous flowers never fully bloom and are thought to have evolved as a means to promote self-fertilisation. A new study reveals that this curious feature arose more frequently in flowers with bilateral symmetry.

Bopopia, a new monotypic genus of Gesneriaceae (Gesnerioideae, Coronanthereae) from New Caledonia

A new genus of Gesneriaceae, Bopopia Munzinger & J.R.Morel gen. nov., is described from New Caledonia. The genus is based on B. parviflora Munzinger & J.R.Morel gen. et sp. nov., a new species collected during an expedition on Mt Katalupaik, in the North Province of New Caledonia’s main island. Originally considered as a species of Coronanthera, our phylogenetic analysis – including 19 species within Coronanthereae and two individuals of B. parviflora gen. et sp. nov., and using three molecular markers (nuclear rDNA ITS, and chloroplast regions trnL-trnF and trnE-trnT) – showed that the new species is not close to Coronanthera in subtribe Coronantherinae, but belongs to subtribe Negriinae where it is sister to Depanthus. From that genus Bopopia gen. nov. differs in floral symmetry (zygomorphic vs actinomorphic) and the number of stamens (4 vs 5). From the other genera of Negriinae the new genus differs in the white corolla and its indeterminate thyrse with 3 to 5 levels of branching. The morphological circumscription of the subtribe Negriinae is amended to include Bopopia gen. nov. Two keys are provided, one to the subtribes in the tribe Coronanthereae, and one to the genera in subtribe Negriinae. Following the IUCN Red List categories and criteria, the conservation status of B. parviflora gen. et sp. nov. is provisionally assessed as Endangered (EN).

Plant communities exhibit low resource partitioning for pollinator guilds under sub-tropical conditions of Pakistan.

Assessment of resource partitioning in pollinators at a particular place can be used to conserve plant communities by minimizing their inter-specific competition. Current study was conducted to investigate the occurrence of this phenomenon among plant communities under sub-tropical conditions for the first time in Pakistan. We considered the entire available flowering plant and floral visitor communities in the study area—Lal Suhanra forest of Bahawalpur, Pakistan- along with different variations among them based on morphology, color and symmetry (functional groups) i.e. four functional groups among insects and nine among plants. Weekly floral visitor censuses were conducted during spring season -from the first week of March to the fourth week of May 2018. Thirty individuals of each plant species -in bloom- were observed for floral visitors in each census. Plant species with different floral shapes, colors and symmetry did not show any significant resource partitioning. The Non-metric multidimensional scaling analysis followed by one-way ANOSIM test showed non- significant differences among all the pair of floral shapes, colors (except white and yellow) and symmetry (R-value < 0.168). However, SIMPER test suggested that flies were the most common group that contributed more towards within group similarities of different floral shapes (19 to 21% similarity), colors (16 to 30%) and symmetry (19%) followed by long-tongue bees i.e. 14 to 21%, 9 to 19% and 18%, respectively. Our results suggest that plant communities under sub-tropical conditions of Pakistan exhibit a generalist pollination system with no significant resource partitioning in pollinator species. Therefore, plant communities may have high competition for pollinator species which exhibits fewer implications of species loss on overall pollination process. Our study provides the basis for understanding the partitioning of pollinator guilds under sub-tropical conditions. Future studies should focus on functional traits in more detail at the community and the population scales for their possible impact on resource partitioning.

Repeated evolution of a reproductive polyphenism in plants is strongly associated with bilateral flower symmetry

Polyphenisms are a special type of phenotypic plasticity in which the products of development are not continuous but instead are separate and distinct phenotypes produced in the same genetic background. One of the most widespread polyphenisms in the flowering plants is cleistogamy, in which the same individual plant produces both open, cross-pollinated flowers as well as highly reduced and closed, self-pollinated (cleistogamous) flowers.1-5 Cleistogamy is not a rare evolutionary phenomenon. It has evolved independently at least 41 times.1 But what favors the evolution of cleistogamy is still largely unknown.1 Darwin6 proposed a hypothesis that has never been properly tested. He observed that cleistogamy is more common in taxa with bilaterally symmetric (zygomorphic) flowers than in those with radially symmetric (actinomorphic) flowers. Moreover, Darwin suggested that cleistogamous flowers help to ensure pollination, which he postulated is less certain in zygomorphic taxa that rely on more specialized groups of pollinators. Here, we combined the largest datasets on floral symmetry and cleistogamy and used phylogenetic approaches to show that cleistogamy is indeed disproportionately associated with zygomorphic flowers and that zygomorphic species are more likely to evolve cleistogamy than actinomorphic species. We also show that zygomorphic species are less capable of autonomous open-flower self-pollination (lower autofertility), suggesting that selection of cleistogamy via reproductive assurance in zygomorphic taxa could help account for Darwin's observation. Our results provide support for the hypothesis that polyphenisms are favored when organisms encounter contrasting environments.

Increasing Profit Margins by Substituting Species in Floral Arrangements

Flower species is one of the key determinants of the aesthetic and economic value of floral products. This research study sought to evaluate whether consumer perceptions of the aesthetic appeal and monetary valuations of floral arrangements change by substituting high-cost species with low-cost species of similar appearance. In addition, the researchers explored consumer preferences for flower symmetry, which provides information to assist floral designers in choosing and using species to increase profit margins and improve the economic efficiency of the floral industry. Two experiments were administered through an online survey. For the first experiment, no difference was shown in both willingness to pay and attractiveness ratings for flowers in the high-dollar value vs. low-dollar value comparison groups. For the second experiment, roses (Rosa hybrida) were rated the highest on attractiveness, followed by dahlia (Dahlia hybrida), ranunculus (Ranunculus asiaticus), and anthurium (Anthurium sp.). Radial flowers were considered most appealing, followed by asymmetrical flowers, and last, bilaterally symmetrical flowers. The results of this study lend insight into how the general floral consumer does not differentiate between flower species that are similar in design features such as color, size, or symmetry. This information can be used by floral business operators to sell their bouquets at a higher margin by strategically using lower-cost flower inputs.

Historical note on the taxonomy of the genus Delphinium L. (Ranunculaceae) with an amended description of its floral morphology

Since its first valid description by Linnaeus in 1753, the genus Delphinium L. has undergone numerous taxonomical changes that we synthesize here. The knowledge of the taxonomic history of the genus is essential to clarify its circumscription. For this purpose, we outline its pre-Linnaean taxonomic history, from Dioscorides' translated works dating back to the 16th century to Tournefort's classification in 1694. Regarding its post-Linnaean history, we discuss the three different lectotypifications proposed during in the 20th century. In addition, we amend the description of the typical flower of Delphinium by including the characteristics of the species D. ecalcaratum S.Y. Wang & K.F. Zhou and D. turcicum (H.Duman, Vural, Aytaç & Adigüzel) Espinosa with actinomorphic flowers.

Spatial orientation of gynoecium in legumes and beyond: Commentary to the paper of Wang et al. (2021)

The third largest angiosperm family, Leguminosae, exhibits a relatively wide range of variation in morphology of gynoecium. Some of gynoecial patterns found in this taxon are of special interest, as they resemble ones previously described in the earliest angiosperms. The different orientations of carpels in legumes appear easily switchable through changes in flower symmetry and floral meristem sizes. Regardless of orientation of a single carpel with respect to the inflorescence axis, the placenta-bearing suture invariably remains adaxial as related to the floral axis. This conclusion relaxes the existing controversies between the supposed megasporophyll- derived nature of the carpel and observed diversity of placentation in known Mesozoic angiosperms.