A multi-locus chloroplast phylogeny for the Cucurbitaceae and its implications for character evolution and classification

Cucurbitaceae contain c. 800 species in 130 genera and are among the economically most important families of plants. We inferred their phylogeny based on chloroplast DNA sequences from two genes, one intron, and two spacers (rbcL, matK, trnL, trnL-trnF, rpl20-rps12) obtained for 171 species in 123 genera. Molecular data weakly support the traditional subfamilies Cucurbitoideae (111 genera) and Nhandiroboideae (19 genera, 60 species), and recover most of the eleven tribes, but almost none of the subtribes. Indofevillea khasiana is sister to all other Cucurbitoideae, and the genera of Joliffieae plus a few Trichosantheae form a grade near the base of Cucurbitoideae. A newly discovered large clade consists of the ancestrally Asian genera Nothoalsomitra, Luffa, Gymnopetalum, Hodgsonia, Trichosanthes, and the New World tribe Sicyeae. Genera that are poly- or paraphyletic include Ampelosicyos, Cucumis, Ibervillea, Neoachmandra, Psiguria, Trichosanthes, and Xerosicyos. Flower characters, especially number of free styles, fusion of filaments and/or anthers, tendril type, and pollen size, exine, and aperture number correlate well with the chloroplast phylogeny, while petal and fruit characters as well as karyotype exhibit much evolutionary flexibility.     

A petal‐specific InMYB1 promoter from Japanese morning glory: a useful tool for molecular breeding of floricultural crops

Production of novel transgenic floricultural crops with altered petal properties requires transgenes that confer a useful trait and petal‐specific promoters. Several promoters have been shown to control transgenes in petals. However, all suffer from inherent drawbacks such as low petal specificity and restricted activity during the flowering stage. In addition, the promoters were not examined for their ability to confer petal‐specific expression in a wide range of plant species. Here, we report the promoter of InMYB1 from Japanese morning glory as a novel petal‐specific promoter for molecular breeding of floricultural crops. First, we produced stable InMYB1_1kb::GUS transgenic Arabidopsis and Eustoma plants and characterized spatial and temporal expression patterns under the control of the InMYB1 promoter by histochemical β‐glucuronidase (GUS) staining. GUS staining patterns were observed only in petals. This result showed that the InMYB1 promoter functions as a petal‐specific promoter. Second, we transiently introduced the InMYB1_1 kb::GUS construct into Eustoma, chrysanthemum, carnation, Japanese gentian, stock, rose, dendrobium and lily petals by particle bombardment. GUS staining spots were observed in Eustoma, chrysanthemum, carnation, Japanese gentian and stock. These results showed that the InMYB1 promoter functions in most dicots. Third, to show the InMYB1 promoter utility in molecular breeding, a MIXTA‐like gene function was suppressed or enhanced under the control of InMYB1 promoter in Arabidopsis. The transgenic plant showed a conspicuous morphological change only in the form of wrinkled petals. Based on these results, the InMYB1 promoter can be used as a petal‐specific promoter in molecular breeding of floricultural crops.     

Evolution of petal epidermal micromorphology in Leguminosae and its use as a marker of petal identity

Background and Aims

The legume flower is highly variable in symmetry and differentiation of petal types. Most papilionoid flowers are zygomorphic with three types of petals: one dorsal, two lateral and two ventral petals. Mimosoids have radial flowers with reduced petals while caesalpinioids display a range from strongly zygomorphic to nearly radial symmetry. The aims are to characterize the petal micromorphology relative to flower morphology and evolution within the family and assess its use as a marker of petal identity (whether dorsal, lateral or ventral) as determined by the expression of developmental genes.

Methods

Petals were analysed using the scanning electron microscope and light microscope. A total of 175 species were studied representing 26 tribes and 89 genera in all three subfamilies of the Leguminosae.

Key Results

The papilionoids have the highest degree of variation of epidermal types along the dorsiventral axis within the flower. In Loteae and genistoids, in particular, it is common for each petal type to have a different major epidermal micromorphology. Papillose conical cells are mainly found on dorsal and lateral petals. Tabular rugose cells are mainly found on lateral petals and tabular flat cells are found only in ventral petals. Caesalpinioids lack strong micromorphological variation along this axis and usually have only a single major epidermal type within a flower, although the type maybe either tabular rugose cells, papillose conical cells or papillose knobby rugose cells, depending on the species.

Conclusions

Strong micromorphological variation between different petals in the flower is exclusive to the subfamily Papilionoideae. Both major and minor epidermal types can be used as micromorphological markers of petal identity, at least in papilionoids, and they are important characters of flower evolution in the whole family. The molecular developmental pathway between specific epidermal micromorphology and the expression of petal identity genes has yet to be established.Key words: Epidermis, Fabaceae, Papilionoideae, Caesalpinioideae, Mimosoideae, petal surface, scanning electron microscopy, papillose conical cells, tabular rugose cells, tabular flat cells, organ identity     

Homeosis in floral development of Sanguinaria canadensis and S. canadensis ‘Multiplex’ (Papaveraceae)

Sanguinaria canadensis is a member of the Papaveraceae that normally has eight petals rather than four as is usual in the family. Using epi-illumination microscopy to study floral development, we show that the four additional petal primordia are initiated in positions that correspond to the first four stamen positions in species of the Papaveraceae with four petals. Also, these additional petal primordia share early developmental features with stamen primordia: at inception they are circular in outline, and the relationship between organ length and width while very young is similar. The developmental pathway of the additional petals combines both stamen and petal features: initially stamenlike in appearance, they develop into typical petals. The additional petals of S. canadensis can therefore be interpreted as homeotic because petal features are expressed in stamen positions. Organogenesis in the ‘Multiplex’ cultivar is similar to that of its wild progenitor, but during development all primordia in the androecial region become petals. This cultivar, as well as variants within natural populations, show that replacement of stamens with petals occurs within the species.     

Evolution of funnel-revolver flowers and ornithophily in nasa (loasaceae)

Floral morphology, distribution, and flower visitors for 60 taxa of Nasa are investigated and compared to molecular trees inferred both from a combined marker analysis (ITS1 and trnL (UAA)) and from a single marker (ITS1). Flowers conform to two different floral types: Firstly, "tilt-revolver flowers", with spreading to reflexed, white to yellow petals and small, brightly coloured floral scales contrasting with the petals and firmly enclosing the nectar (Saccatae and Carunculatae); secondly, "funnel-revolver flowers", with half-erect to erect, orange to red petals and floral scales not contrasting with the petals, or enclosed in the corolla, and nectar freely accessible by funnel-shaped floral scales ( Alatae, Grandiflorae, and N. venezuelensis species group). Phylogenetic analysis shows that "tilt-revolver flowers" represent the plesiomorphic condition by outgroup comparison. The two groups with tilt-revolver flowers in Nasa are not monophyletic ( Saccatae are paraphyletic, Carunculatae are polyphyletic). Most Saccatae fall into two monophyletic assemblages, the N. poissoniana species group and the N. triphylla species group. The remainder of Saccatae group either with Grandiflorae ( N. insignis species group) or with Alatae ( N. laxa species group). The clades retrieved in the molecular analysis contradict the traditional classification, but are congruent with vegetative morphology, details of the flower morphology, and biogeography. "Funnel-revolver flowers" represent the derived condition, but molecular data suggest a convergent development (at least twice independently), since the corresponding species do not constitute a monophyletic group. "Tilt-revolver flowers" are visited and pollinated by bees (especially Colletidae), whereas "funnel-revolver flowers" are mostly visited by hummingbirds. The transition from melittophily to ornithophily may have been the license for the colonization of, and the diversification in, both cloud forest and high Andean habitats.     

Original Paper Floral Organogenesis in Verbenaceae sensulato

The early floral ontogeny of three subfamilies, viz. Verbenoideae, Viticoideae and Caryopteridoideae of Verbenaceae (s.l.), was compared. Two differently initiated patterns were found. In the present species of Verbenoideae, there is a unidirectional sequence of organogenesis, from abaxial to adaxial side of the floral apex. While the abaxial paired sepal, petal and stamen arise sequentially, the adaxial paired sepal, petal and stamen do not appear or appear in a much earlier stage. The centripetal whorled sequence of organogenesis appears in Viticoideae and Caryopteridoideae, where sepal primordia arise simultaneously or successively (from adaxial to abaxial). After completion of sepal initiation a plastochron is indicated, during which time a change to the induction of petal takes place, and five petals appear simultaneously, followed by initiation of four stamens. Events of floral organogenesis support the phylogeny inferred from morphological data and rbcL sequence analysis, i.e. the subfamily Verbenoideae does not form a monophyletic group with the subfamilies Viticoideae and Caryopteridoideae.     

Floral morphogenesis of the Maddenia and Pygeum groups of Prunus (Rosaceae), with an emphasis on the perianth

Although the vast majority of Prunus L. (Rosaceae) species have clearly differentiated sepals and petals, two former genera Maddenia and Pygeum have been described as having an undifferentiated perianth. However, floral morphological and morphogenetic data are scarce, and a renewed investigation is essential to understand the evolution of the perianth differentiation. Here, floral morphogenesis in Prunus hypoleuca (Koehne) J.Wen (=Maddenia hypoleuca Koehne) and Prunus topengii (Merr.) J. Wen & L. Zhao (=Pygeum topengii Merr.) were examined with scanning electron microscopy. The floral development demonstrates that the ten perianth parts can be distinguished as five sepals in an external whorl and five petals in an internal whorl. The sepal primordia are broad, crescent-shaped, and truncate. The petal primordia are rounded and initially resemble the androecium. However, at maturity petals and sepals look much the same in the two species, differing from other Prunus species. The ovule is anatropous and unitegmic, but there is a basal appendage near the ovule of P. hypoleuca which is absent in P. topengii. The direction of development of floral nectaries in the hypanthium is basipetal in P. hypoleuca but acropetal in P. topengii. Perianth segments are differentiated in the two groups and the similarity of the perianth parts is secondarily acquired. Our results support the separation of the Maddenia and Pygeum groups as well as their inclusion in a broader monophyletic Prunus based on molecular phylogenetic studies. We herein provide a new nomenclatural change: Prunus topengii (Merr.) J. Wen & L. Zhao, comb. nov.     

Insect Attracting Structures on Erodium Petals (Geraniaceae)

Abstract: Two types of structures previously unrecorded in Erodium petals are investigated. Spherical hairs filled with liquid resembling nectar droplets are exclusive to an ibero-mauritanic group of species included in Erodium subsect. Romana. Broad flat hairs which reflect light, shining as do nectar droplets, are restricted to most of the species included in Erodium sect. Malacoidea. Long, simple hairs in petals and sepals are involved in collection of nectar droplets. Some of them are arranged at the margin of the petal claw, just over the nectaries. Others are on the internal surface of sepals or on the upper surface of petals, serving apparently the same function. Their shape is aciculate or flattened. The nectar collected among the hairs forms shining spherical droplets, perceptible to insects. The glistening flat hairs and spheres shine in a similar way, probably mimicking nectar and attracting insects. Species with these special nectar-like structures produce nectar in quantities that can be observed by the naked eye, suggesting that these structures increase the attraction efficiency of flowers. Some taxonomic and biogeographic consequences are also discussed.     

Evolution of petaloid sepals independent of shifts in B-class MADS box gene expression

Attractive petals are an integral component of animal-pollinated flowers and in many flowering plant species are restricted to the second floral whorl. Interestingly, multiple times during angiosperm evolution, petaloid characteristics have expanded to adjacent floral whorls or to extra-floral organs. Here, we investigate developmental characteristics of petaloid sepals in Rhodochiton atrosanguineum, a close relative of the model species Antirrhinum majus (snapdragon). We undertook this in two ways, first using scanning electron microscopy we investigate the micromorphology of petals and sepals, followed by expression studies of genes usually responsible for the formation of petaloid structures. From our data, we conclude that R. atrosanguineum petaloid sepals lack micromorphological characteristics of petals and that petaloid sepals did not evolve through regulatory evolution of B-class MADS box genes, which have been shown to specify second whorl petal identity in a number of model flowering plant species including snapdragon. These data, in conjunction with other studies, suggests multiple convergent pathways for the evolution of showy sepals.     

A sensitive flower: mechanical stimulation induces rapid flower closure in Drosera spp. (Droseraceae)

Some plants rapidly close their leaves in response to mechanical stimulation, but no case is known in which mechanical stimulation causes rapid petal closure. In this study, we found that Drosera tokaiensis closes petals within 2–10 min after experimental stimulation of the calyx, closed flowers or scapes with a pair of tweezers. Although petal closure was induced more rapidly by touching a position closer to a flower, it was not induced by stimulating stamens and pistils. The habit of petal closure varies among species of Drosera: after experimental stimulations of the calyx or scapes, D. tokaiensis and D. spatulata often closed petals but D. rotundifolia and D. toyoakensis did not close them. The petal closure may function as defense against a specialist florivore.     

赤芍和白芍新鲜花瓣正己烷提取成分GC-MS分析

目的:赤芍和白芍新鲜花瓣正己烷提取成分比较分析。方法:正己烷浸出法提取新鲜花瓣挥发性物质,GC-MS分析、鉴定其化学成分。结果:从赤芍新鲜花瓣挥发性物质中鉴定出33种化学成分;白芍新鲜花瓣挥发性物质中鉴定出35种化学成分。二者中的主要成分为棕榈酸,二十三烷,二十五烷,二十七烷,二十九烷等。赤芍花瓣含有更多小分子芳香类成分,如苯乙醇、法尼醇类等;白芍缺少这些成分,其芳香类成分有β-沉香醇、反式-橙花椒醇等。结论:赤芍(野生芍药)与白芍(栽培芍药)花瓣芳香气成分有差异。     

Floral ontogeny in Sophoreae (Leguminosae: Papilionoideae): II. Sophora sensu lato (Sophora group)

Floral ontogeny is described in eight species of Sophora sensu lato, representing the Sophora group, as part of a comparative ontogenetic analysis of Polhill's eight groups of tribe Sophoreae, subfamily Papilionoideae. This tribe includes taxa having relatively unspecialized floral structure. Flowers have a five-lobed calyx, a corolla of five free petals, ten mostly unfused, identical stamens, and a carpel. Order of initiation is predominantly acropetal (except for the carpel): sepals, petals, outer stamens plus carpel, inner stamens. Order of initiation within each whorl is unidirectional from the abaxial side. Overlapping initiation among whorls occurs only in S. chrysophylla. Keel petals are slightly fused in six species, and wing petals are fused in 5. tomentosa. Two bird-pollinated species (S. chrysophylla, S. microphylla) lack the papilionaceous corolla of other species, and their petals are unusually long and lack wing sculpturing found in the others. Other floral differences among species mostly involve flower color, differing absolute or relative sizes among organs, and degree of reflexing of vexillum. All but S. davidii have a hypanthium, which develops very late, starting when the bud is about 5 mm long. The distinctions among species (petal size, degree of reflexed position of vexillum, petal sculpturing, color, anther shape, filament hairs, hypanthium presence, calyx lobing) tend to be expressed late in ontogeny.     

Petal Development in Lotus japonicus

Previous studies have demonstrated that petal shape and size in legume flowers are determined by two separate mechanisms, dorsoventral (DV) and organ internal (IN) asymmetric mechanisms, respectively. However, little is known about the molecular mechanisms controlling petal development in legumes. To address this question, we investigated petal development along the floral DV axis in Lotus japonicus with respect to cell and developmental biology by comparing wild‐type legumes to mutants. Based on morphological markers, the entire course of petal development, from initiation to maturity, was grouped to define 3 phases or 13 stages. In terms of epidermal micromorphology from adaxial surface, mature petals were divided into several distinct domains, and characteristic epidermal cells of each petal differentiated at stage 9, while epidermal cells of all domains were observed until stage 12. TCP and MIXTA‐like genes were found to be differentially expressed in various domains of petals at stages 9 and 12. Our results suggest that DV and IN mechanisms interplay at different stages of petal development, and their interaction at the cellular and molecular level guides the elaboration of domains within petals to achieve their ideal shape, and further suggest that TCP genes determine petal identity along the DV axis by regulating MIXTA‐like gene expression.     

Patterns of flower morphology and structural changes during interconversion between chasmogamous and cleistogamous flowers in Viola philippica北大核心CSCD

Aims: Viola philippica is a species with a typical chasmogamous-cleistogamous (CH-CL) mixed breeding system. It provides a flower model system to investigate floral organs development under different photoperiods. Morphological changes of intermediate cleistogamous (inCL) flowers have been observed, the trends in variation of changes from CH flowers to CL flowers or from CL flowers to CH flowers have been analyzed, the localized effects of poorly developed stamens and petals in CL and inCL flowers have been identified. This research provided morphology and structural changes with implication for the evolutionary significance of the dimorphic flower formation for further study in dimorphic flower development. Methods: We used methods of anatomy and structural analysis to observe the morphological structures of flowers under different photoperiods. Important findings: Photoperiod played an important role in the development of CH and CL flowers in V. philippica. Under short-day light and intermediate-day light, both CH and inCL flowers developed simultaneously. Most of the floral buds were CH flowers under a photoperiod of short-day light, but most of the floral buds were inCL flowers under mid-day light. Complete CL flowers formed under long-day lights. However, there were a series of transitional types in the number and morphology of stamens and petals among inCL flowers, including five stamens with three petals related to CH flowers and two stamens with one petal related to CL flowers. The former type was dominant under short-day light conditions, and the latter type was dominant under mid-day light. Further more, there were localized effects in stamen and petal development for CL and inCL flowers. The development of ventral lower petal (corresponding to the lower petal with spur of CH flower) and the adjacent two stamens in inCL flowers were best, and the back petal was similar to that of CL flowers, an organ primordium structure. The adjacent stamens with the back petals tended to be poorly developed. In extreme cases, these stamens in inCL flowers had no pollen sac, only a membranous appendage or even a primordium structure. When the plants with CL or CH flowers were placed under short-day light or long-day light, the newly induced flowers all showed a series of inCL flower types, finally the CL flowers transformed into CH flowers, and the CH flowers transformed into CL flowers. This result indicates the gradual effects of different photoperiods on dimorphic flowers development of V. philippica. A long photoperiod could inhibit the development of partial stamens and petals, and a short photoperiod could prevent the suppression of long-day light and promote the development of stamens and petals.     

Molecular phylogenetics of the Macaronesian-endemic genus Bystropogon (Lamiaceae): palaeo-islands, ecological shifts and interisland colonizations

Abstract A molecular phylogenetic study of Bystropogon L'Her. (Lamiaceae) is presented. We performed a cladistic analysis of nucleotide sequences of the internal transcribed spacers (ITS), of the nuclear ribosomal DNA, and of the trnL gene and trnL-trnF intergenic spacer of the chloroplast DNA. Bystropogon odoratissimus is the only species endemic to the Canary Islands that occurs in the three palaeo-islands of Tenerife. This species is not part of an early diverging lineage of Bystropogon and we suggest that it has a recent origin. This phylogenetic pattern is followed by most of the species endemic to the palaeo-islands of Tenerife. The two sections currently recognized in Bystropogon form two monophyletic groups. Taxa belonging to the section Bystropogon clade show interisland colonization limited to the Canary Islands with ecological shifts among three ecological zones. Taxa from the section Canariense clade show interisland colonization both within the Canary Islands and between the Canary Islands and Madeira. Speciation events within this clade are mostly limited to the laurel forest. The genus has followed a colonization route from the Canaries towards Madeira. This route has also been followed by at least five other plant genera with species endemic to Macaronesia. Major incongruences were found between the current infrasectional classification and the molecular phylogeny, because the varieties of Bystropogon origanifolius and Bystropogon canariensis do not form two monophyletic groups. The widespread B. origanifolius appears as progenitor of the other species in section Bystropogon with a more restricted distribution.     

Continuous petal variation in Epimedium tianmenshanense (Berberidaceae), a taxon endemic to western Hunan,China

In the protologue, Epimedium tianmenshanense (Berberidaceae), a species endemic to western Hunan, China, was described as ‘flower small, 0.2–0.4 cm diam., inner sepals white, petals as long as inner sepals or a little shorter than the latter, spur very short, ca 5.0 mm’. However, both morphological characteristics and molecular evidence suggest that E. tianmenshanense is closely related to E. baojingense, a taxon with a long spur, thus suggesting that the size of the floral parts is not as reliable as previously believed. When investigating the variability of E. tianmenshanense in more detail, in the field as well as in cultivation, we found that the petals were are highly variable in morphology (both shape and size), being cucullate, subulate, short to long spurred, and with various transitions. The flowers size varied from small to large accordingly. The flowers with cucullate and subulate petals, which were a little shorter than the inner sepals or almost as long as the latter, were small (about 0.8 cm in diameter). The flowers with long spurs, which were much longer than the inner sepals, were also large (about 2.5–3.5 cm in diameter). Finally, the flowers with short spurs, which were a little longer than the inner sepals, were medium-sized (about 1.0–1.2 cm in diameter). In addition, the color of inner sepals was revised from ‘white, occasionally light mulberry-purple’ to yellowish green or yellowish white. Epimedium tianmenshanense is a perfect example of natural petal evolution, which could be used for further taxonomic and evolutionary studies. The reason for the variation and the taxonomic treatment of the species still need further study.