Generic recircumscriptions of Oncidiinae (Orchidaceae: Cymbidieae) based on maximum likelihood analysis of combined DNA datasets

Phylogenetic relationships within the orchid subtribe Oncidiinae sensu Chase were inferred using maximum likelihood analyses of single and multilocus DNA sequence data sets. Analyses included both nuclear ribosomal internal transcribed spacer DNA and plastid regions (matK exon, trnH‐psbA intergenic spacer and two portions of ycf1 exon) for 736 individuals representing approximately 590 species plus seven outgroup taxa. Based on the well resolved and highly supported results, we recognize 61 genera in Oncidiinae. Mimicry of oil‐secreting Malpighiaceae and other floral syndromes evolved in parallel across the subtribe, and many clades exhibit extensive variation in pollination‐related traits. Because previous classifications heavily emphasized these floral features, many genera recognized were not monophyletic. Our classification based on monophyly will facilitate focused monographs and clarifies the evolution of morphological and biochemical traits of interest within this highly diverse subtribe. © 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 168 , 117–146.     

Floral nectary morphology and evolution in Pedicularis (Orobanchaceae)

Intricate associations between floral morphology and pollinator foraging behaviour are common. In this context, the presence and form of floral nectaries can play a crucial role in driving floral evolution and diversity in flowering plants. However, the reconstruction of the ancestral state of nectary form is often hampered by a lack of anatomical studies and well‐resolved phylogenetic trees. Here, we studied 39 differentially pollinated Pedicularis spp., a genus with pronounced interspecific variation in colour, shape and size of the corolla. Anatomical and scanning electron microscopy observations revealed two nectary forms [bulged (N = 27) or elongated (N = 5)] or the absence of nectaries (N = 7). In a phylogenetic context, our data suggest that: (1) the bulged nectary should be the ancestral state; (2) nectaries were independently lost in some beaked species; and (3) elongated nectaries evolved independently in some clades of beakless species. Phylogenetic path analysis showed that nectary presence is indirectly correlated with beak length/pollinator behaviour through an intermediate factor, nectar production. No significant correlation was found between nectary type and nectar production, beak length or pollinator behaviour. Some beaked species had nectary structures, although they did not produce nectar. The nectary in beaked species may be a vestigial structure retained during a recent rapid radiation of Pedicularis, especially in the Himalaya–Hengduan Mountains of south‐western China. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 178 , 592–607.     

How do plants balance multiple mutualists? Correlations among traits for attracting protective bodyguards and pollinators in cotton (Gossypium)

Many species, both plants and animals, are simultaneously engaged in interactions with multiple mutualists. However, the extent to which separate traits that attract different mutualist guilds display negative or positive relationships remains largely unstudied. We asked whether correlations exist among extrafloral nectary traits to attract arthropod bodyguards and floral traits to attract pollinator mutualists. For 37 species in the cotton genus (Gossypium), we evaluated correlations among six extrafloral nectary traits and four floral traits in a common greenhouse environment, with and without correction for phylogenetic non-independence. Across Gossypium species, greater investment in extrafloral nectary traits was positively correlated with greater investment in floral traits. Positive correlations remained after accounting for the evolutionary history of the clade. Our results demonstrate that traits to maintain multiple mutualist guilds can be positively correlated across related species and build a more general understanding of the constraints on trait evolution in plants.     

EVIDENCE FOR A CLOSE RELATIONSHIP BETWEEN IOSTEPHANE AND VIGUIERA (ASTERACEAE: HELIANTHEAE)

The phylogenetic relationship of Iostephane is assessed using data from morphology, flavonoid chemistry, and chloroplast DNA and nuclear ribosomal DNA restriction fragment analysis. Morphological evidence supports placement of Iostephane in subtribe Helianthinae, but fails to clarify the placement of the genus within this assemblage. Further evidence for the placement of Iostephane in subtribe Helianthinae is provided by the presence in all species of the genus of floral flavonoids of the chalcone/aurone type, which provides a distinctive trait for the subtribe within the tribe Heliantheae. Analysis of chloroplast DNA from two species of Iostephane, I. heterophylla and I. madrensis, in comparison to Viguiera and related genera indicates that the restriction site patterns with 16 enzymes for the Iostephane species are virtually identical to one another as well as to those of Viguiera sect. Maculatae. Data from restriction fragment patterns of nuclear rDNA are concordant with the results from chloroplast DNA in suggesting a direct relationship between the two groups. The close phylogenetic relationship between Iostephane and Viguiera sect. Maculatae suggested by the DNA restriction fragment data was not suggested by any other set of data.     

Floral nectaries in Sapindaceae <Emphasis Type="Italic">s.s.</Emphasis>: morphological and structural diversity,and their systematic implications

We investigated the morphology and structure of the floral nectary in 11 Neotropical genera belonging to the subfamilies Dodonaeoideae and Paullinioideae (Sapindaceae) from southern South America representing three tribes (Dodonaeaeae, Paullinieae, and Melicocceae), in relation to other floral traits in species with contrasting morphological flower characteristics. Nectary organization was analyzed under light, stereoscopic, and scanning electron microscopes; Diplokeleba floribunda N.E. Br. was also observed using transmission electron microscopy. Our comparative data may contribute to the understanding of floral nectary evolution and systematic value in this family. The nectaries were studied in both staminate and pistillate flowers. All the floral nectaries are typical of Sapindaceae: extrastaminal, receptacular, structured, and persistent. The anatomical analysis revealed a differentiated secretory parenchyma and an inner non-secretory parenchyma; the nectary is supplied by phloem traces and, less frequently, by phloem and xylem traces. Nectar is secreted through nectarostomata of anomocytic type. The anatomical analysis showed the absence of nectary in the three morphs of Dodonaea viscosa flowers. Nectary ultrastructure is described in D. floribunda. In this species, the change in nectary color is related to progressive accumulation of anthocyanins during the functional phase. We found relatively small variation in the nectary structural characteristics compared with large variation in nectary morphology. The latter aspect agreed with the main infrafamilial groupings revealed by recent phylogenetic studies, so it is of current valuable systematic importance for Sapindaceae. In representatives of Paullinieae, the reduction of the floral nectary to 4–2 posterior lobes should be interpreted as a derived character state.     

Hidden biodiversity: total evidence phylogenetics and evolution of morphological traits in a highly diverse lineage of endogean ground beetles,Typhlocharis Dieck, 1869 (Carabidae,Trechinae, Anillini)

Typhlocharis is the most diverse eyeless endogean ground beetle genus known to date, with 62 species all endemic to the West Mediterranean region. The lineage is characterized by a conservative and singular body plan within Carabidae that contrasts with a high morphological diversity in many traits. We provide an exhaustive phylogeny of the lineage through the study of 92 morphological characters from all 62 described species and 45 potential new species from 70 additional populations, and the combination of morphological and available molecular data, in the first total evidence phylogenetic approach for a highly diverse endogean lineage. We tracked the evolution of morphological traits over the obtained phylogenies. Results suggest eight morphologically distinct clades, which do not correspond to the species groups proposed formerly. Ancestral state reconstructions and phylogenetic signal analyses of morphological traits revealed that some of the previously key characters to the classification of Typhlocharis, such as the umbilicate series or the apical denticles of elytra, are highly homoplasic, whereas other characters show stronger phylogenetic signal, including structures in the antennae, gula, pronotum and last abdominal ventrite. This evidence supports the split of Typhlocharis into three genera: Lusotyphlus gen. nov. ; Typhlocharis Dieck, 1869 and Microcharidius Coiffait, 1969 (revalidated), forming the subtribe Typhlocharina Jeanne, 1973.     

The evolution of floral nectaries in Disa (Orchidaceae: Disinae): recapitulation or diversifying innovation?

Background and Aims

The Orchidaceae have a history of recurring convergent evolution in floral function as nectar production has evolved repeatedly from an ancestral nectarless state. However, orchids exhibit considerable diversity in nectary type, position and morphology, indicating that this convergence arose from alternative adaptive solutions. Using the genus Disa, this study asks whether repeated evolution of floral nectaries involved recapitulation of the same nectary type or diversifying innovation. Epidermis morphology of closely related nectar-producing and nectarless species is also compared in order to identify histological changes that accompanied the gain or loss of nectar production.

Methods

The micromorphology of nectaries and positionally equivalent tissues in nectarless species was examined with light and scanning electron microscopy. This information was subjected to phylogenetic analyses to reconstruct nectary evolution and compare characteristics of nectar-producing and nectarless species.

Key Results

Two nectary types evolved in Disa. Nectar exudation by modified stomata in floral spurs evolved twice, whereas exudation by a secretory epidermis evolved six times in different perianth segments. The spur epidermis of nectarless species exhibited considerable micromorphological variation, including strongly textured surfaces and non-secreting stomata in some species. Epidermis morphology of nectar-producing species did not differ consistently from that of rewardless species at the magnifications used in this study, suggesting that transitions from rewardlessness to nectar production are not necessarily accompanied by visible morphological changes but only require sub-cellular modification.

Conclusions

Independent nectary evolution in Disa involved both repeated recapitulation of secretory epidermis, which is present in the sister genus Brownleea, and innovation of stomatal nectaries. These contrasting nectary types and positional diversity within types imply weak genetic, developmental or physiological constraints in ancestral, nectarless Disa. Such functional convergence generated by morphologically diverse solutions probably also underlies the extensive diversity of nectary types and positions in the Orchidaceae.     

Phylogeny of the Homalotina (Coleoptera: Staphylinidae: Aleocharinae) based on morphology

The aleocharine subtribe Homalotina Heer represents one of the most diverse lineages of Staphylinidae. Despite its wide distribution and diversity, the phylogenetic relationships of the subtribe remain poorly understood. Here, we present the first cladistic analysis of the Homalotina based on morphological data. The subtribe is hypothesized to be a monophyletic group consisting of seven genera (Anomognathus Solier, Cephaloxynum Bernhauer, Holisomimus Cameron, Homalota Mannerheim, Neomalota Cameron, Stenomastax Cameron, and Thecturota Casey). The dataset for phylogenetic analysis comprised 83 characters representing 245 character states derived from adult morphology. These data were analysed using equal weighting and implied weighting schemes (k = 1–6) and results support the monophyly of the subtribe based on two synapomorphic characters (complete postoccipital sutures on head, posterolateral margin of metacoxae with macrosetae) and three homoplastic characters (medial setae on prementum not extended to apex of ligula, medial setae on labium contiguous, posterolateral angle of elytron slightly sinuate). Generic relationships differ in each analysis within the Homalotina (EW, IW with k = 1, 2–4, 5–6) although there are some identical topologies among the IW trees. Clades A, B, C, D, H, J and G were resolved as monophyletic in all weighting regimes. The monophyly of the genera is relatively well supported except for the genera Homalota and Stenomastax. Homalota species were recovered in four independent clades (clade C, D, I, K) and the Stenomastax species were recovered in two independent lineages. Candidates for the possible new genera are discussed. We herein transfer Homalota flavomaculata Bernhauer to the genus Stenomastax, resulting in the new combination [Stenomastax flavomaculata (Bernhauer)]. Our preliminary character correlation tests using phylogenetic pairwise comparisons did not support the hypothesis of association between flattened body form, and subcortical habitat and anterior shift of antennal insertion in Homalotini.     

Kauriphanes n. gen., a new genus of braconid parasitoid wasp (Hymenoptera: Braconidae: Doryctinae) from New Zealand

A new genus belonging to the braconid wasp subfamily Doryctinae, Kauriphanes n. gen. (type species K. khalaimi n. sp.), is described from New Zealand. This genus is placed within the doryctine subtribe Caenophanina. The extent of this subtribe is discussed and the phylogenetic relationships of three of its genera were investigated using one mitochondrial and one nuclear DNA sequence markers. Similar to previous studies, the Bayesian analyses performed significantly support a clade with the included members of Caenophanina as a sister group of a clade with the examined species of Spathiini sensu stricto. The placement of the Caenophanini within Doryctini, however, is left pendant to further exhaustive phylogenetic studies. A key to genera and subgenera belonging to Caenophanina is given.     

Recruitment of CRABS CLAW to promote nectary development within the eudicot clade

Nectaries are secretory organs that are widely present in flowering plants that function to attract floral pollinators. Owing to diversity in nectary positions and structures, they are thought to have originated multiple times during angiosperm evolution, with their potential contribution to the diversification of flowering plants and pollinating animals being considerable. We investigated the genetic basis of diverse nectary forms in eudicot angiosperm species using CRABS CLAW (CRC), a gene required for nectaries in Arabidopsis. CRC expression is conserved in morphologically different nectaries from several core eudicot species and is required for nectary development in both rosids and asterids, two major phylogenetic lineages of eudicots. However, in a basal eudicot species, no evidence of CRC expression in nectaries was found. Considering the phylogenetic distribution of nectary positions and CRC expression analyses in eudicots, we propose that diverse nectaries in core eudicots share conserved CRC gene regulation, and that derived nectary positions in eudicots have altered regulation of CRC. As the ancestral function of CRC lies in the regulation of carpel development, it may have been co-opted as a regulator of nectary development within the eudicots, concomitant with the association of nectaries with reproductive organs in derived lineages.     

苦苣苔科镜像花的多样性及演化

泛热带分布的苦苣苔科(Gesneriaceae)在我国南方具有极高的物种丰富度与特有率,花部特征变化丰富,是研究物种形成与适应演化的代表类群。镜像花(mirror-image flowers)是极为特化的传粉系统,在苦苣苔科中出现了较多的不同类型,可能与苦苣苔科物种多样性形成与维持有关。该研究总结与分析了苦苣苔科镜像花的类型多样性以及系统分布与适应演化等,讨论了镜像花对苦苣苔科物种形成与维持的积极意义。结果表明:镜像花仅分布在亚洲和非洲的苦苣苔亚科(Didymocarpoideae)的7个属,在历史上就至少发生了5次独立起源。长冠苣苔属(Rhabdothamnopsis)、南洋苣苔属(Henckelia)及长蒴苣苔属(Didymocarpus)镜像花的花柱与可育雄蕊分别向左、右两侧偏转,形成互补镜像花;蛛毛苣苔属(Paraboea)、喜鹊苣苔属(Ornithoboea)、非洲堇属(Saintpaulia)镜像花缺乏与花柱对应侧偏的可育雄蕊(非互补镜像花);而海角苣苔属(Streptocarpus)直立堇兰亚属(subg.Streptocarpella Engler)则同时出现了互补、非互补镜像花。不同于其他被子植物(离瓣花、缺乏花冠筒),苦苣苔科中的镜像花大多伴随着明显的花冠筒、内藏的雄蕊、合生的花药,以非互补镜像花为主;传粉者以小型的无垫蜂(Amegilla spp.)和熊蜂(Bombus spp.)为主。这些特殊的花部综合征与特化的传粉机制,提高了传粉精确性,可能促进了传粉隔离与物种适应辐射。今后的一个研究重点应通过分子系统发育方法,进一步揭示苦苣苔亚科互补与非互补镜像花的进化顺序及其在物种分化与长距离扩散过程中的可能作用。     

Molecular phylogenetics of subtribe Aeridinae (Orchidaceae): insights from plastid matK and nuclear ribosomal ITS sequences

We conducted phylogenetic analyses using two DNA sequence data sets derived from matK, the maturase-coding gene located in an intron of the plastid gene trnK, and the internal transcribed spacer region of 18S–26S nuclear ribosomal DNA to examine relationships in subtribe Aeridinae (Orchidaceae). Specifically, we investigated (1) phylogenetic relationships among genera in the subtribe, (2) the congruence between previous classifications of the subtribe and the phylogenetic relationships inferred from the molecular data, and (3) evolutionary trends of taxonomically important characters of the subtribe, such as pollinia, a spurred lip, and a column foot. In all, 75 species representing 62 genera in subtribe Aeridinae were examined. Our analyses provided the following insights: (1) monophyly of subtribe Aeridinae was tentatively supported in which 14 subclades reflecting phylogenetic relationships can be recognized, (2) results are inconsistent with previous classifications of the subtribe, and (3) repeated evolution of previously emphasized characters such as pollinia number and apertures, length of spur, and column foot was confirmed. It was found that the inconsistencies are mainly caused by homoplasy of these characters. At the genus level, Phalaenopsis, Cleisostoma, and Sarcochilus are shown to be non-monophyletic.     

Pollen morphology of the Swertiinae (Gentianaceae): phylogenetic implications

Recent molecular surveys of the Swertiinae (Gentianaceae–Gentianeae) revealed unexpected phylogenetic relationships, including polyphyly of the genera Gentianella , Jaeschkea , Lomatogonium and Swertia . To find new non-molecular characters supporting the phylogeny, we examined the exine variation of 73 species of all major lineages of subtribe Swertiinae using environmental scanning electron miscroscopy supplementing older, mainly light microscopical, studies. In contrast to previous studies, we were able to pick out taxa from phylogenetic key positions with particular focus on Swertia . Many distantly related taxa such as parts of Frasera , Gentianopsis , Halenia , Gentianella , Megacodon and several lineages of Swertia share a striate–reticulate or reticulate exine pattern. This is interpreted as the plesiomorphic character state of Swertiinae. There is also considerable variation of derived patterns; for example, different types of microechinate or almost smooth pollen was repeatedly observed in distantly related groups. Another extreme was the ring-shaped reticulation found in a North American species of Gentianopsis . Unfortunately, major relationships as revealed by molecular analyses were rarely supported because of the abundance of the plesiomorphic type and homoplasy even on low taxonomic levels; for example, within Lomatogonium . Exine variation was particularly useful in characterizing independent lineages of Swertia . For example, according to pollen characters and in agreement with other data, the Asian Swertia cuneata is a sister group of a strongly diversified African lineage and Swertia yunnanensis , which is rather aberrant in flower morphology, seems close to parts of Lomatogonium .  © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society , 2008, 157 , 323–341.     

Hybrid genera in Liatrinae (Asteraceae: Eupatorieae)

Liatrinae is a small subtribe of Eupatorieae that occurs in North America with a center of generic-level diversity in the southeastern United States. Molecular phylogenetic data were sought to assess whether two monotypic genera, Garberia and Hartwrightia, are accurately placed in the subtribe, and to resolve questions of the generic-level classification of Carphephorus. Phylogenetic analyses of nuclear ITS/ETS and plastid DNA data indicated that Garberia is the basalmost diverging lineage, and that Hartwrightia is phylogenetically embedded in the subtribe. There was significant incongruence between the ITS/ETS and plastid DNA datasets in the placement of Hartwrightia and another monotypic genus, Litrisa, suggesting that both are of original hybrid origin. The results also showed that Carphephorus s.l. is not monophyletic, and even after removal of the two species of Trilisa, it is still paraphyletic to Liatris. The apparent hybrid origin of Hartwrightia, which is morphologically transgressive relative to its inferred parental lineages, suggests that reticulation between phylogenetically distinct lineages may be a recurrent problem for phylogenetic estimation in Asteraceae.     

Nectary structure and nectar characteristics in someBignoniaceae

The nectary structure and chemical nectar composition of 15 species belonging to 12 genera ofBignoniaceae are analyzed. All taxa bear a conspicuous nuptial nectary surrounding the ovary base. The secretory tissue is mostly supplied by phloem branches. The stomata are located in the middle and upper part of the nectary epidermis with an homogeneous distribution. The nuptial nectary is proportionally large in relation to the ovary (15–30%), disregarding the nectary volume. Most species have extranuptial nectaries in both inner and outer surfaces of the calyx. Both kinds of nectaries lack a vascular tissue that straightly supplies them. Nuptial nectar concentration (wt/wt) ranges from 19 to 68%. Sugars and amino acids are found in all species. Half of the species have hexose predominant nectars, the remaining sucrose predominant. Phenols are detected in only three species, whereas reducing acids exclusively inTecoma stans. Alkaloids and lipids were never detected. Extranuptial nectar chemical composition is analyzed in two species:Dolichandra cynanchoides andPodranea ricasoliana. Bees constitute the main flower visitors of the species studied whereas hummingbirds were seen visiting three species. A correlation analysis is performed with the data obtained. There are a few significant correlations which indicate a parallel increase of three parameters: the longer the flower length, the more voluminous the nectary and the higher stomata number, independently of the floral biotype. Phenograms are obtained using 24 floral characters including nectary and nectar data. The clusters obtained do not reflect taxonomic relationships but are useful in the understanding of animal-plant interactions when the flower biotype is considered.This paper is based on a chapter of a doctoral thesis presented at the University of Córdoba (Argentina).     

Analysis of Microsatellite Loci of the Chloroplast Genome in the Genus Capsicum (Pepper)

Six plastome microsatellites were examined in 43 accessions of the genus Capsicum. In total, 33 allelic variants were detected. A specific haplotype of chloroplast DNA was identified for eachCapsicum species. Species-specific allelic variants were found for most wild Capsicum species. The highest intraspecific variation was observed for the C. baccatum plastome. Low cpDNA polymorphism was characteristic of C. annuum:the cpSSRs were either monomorphic or dimorphic. The vast majority of C. annuum accessions each had alleles of one type. Another allele type was rare and occurred only in wild accessions. The results testified again to genetic conservation of C. anuum and especially its cultivated forms. The phylogenetic relationships established for the Capsicum species on the basis of plastome analysis were similar to those inferred from the morphological traits, isozyme patterns, and molecular analysis of the nuclear genome.     

Chromosome studies in Orchidaceae: karyotype divergence in Neotropical genera in subtribe Maxillariinae

Here, we study karyotype divergence in the closely related genera Brasiliorchis, Christensonella and Trigonidium belonging to subtribe Maxillariinae of subfamily Epidendroideae (Orchidaceae). We compare karyotypes in 15 species by (1) measuring 1C genome sizes, (2) mapping the distribution of 4′,6‐diamidino‐2‐phenylindole and chromomycin A3 chromosome bands and (3) localizing 5S and 45S nuclear ribosomal DNA (rDNA) sequences using fluorescent in situ hybridization. Recently, phylogenetic studies have been conducted to resolve species and genera relationships in subtribe Maxillariinae. We used these phylogenetic trees to map the cytogenetic characters in an evolutionary framework. This has enabled a better understanding of the patterns of genomic divergence in the group. Genome sizes range from 1C = 1.85 to 4.1 pg. The largest, B. schunkeana, shows evidence of genome upsizing, probably through the acquisition of tandem repeats that now form large 4′,6‐diamidino‐2‐phenylindole‐positive blocks of heterochromatin. Our cytogenetic data are consistent with a base chromosome number of 2n = 40, although Christensonella is characterized by a dysploid reduction in chromosome number to 2n = 36. The number of 5S and 45S rDNA sites is variable between species, consistent with high rates of karyotype divergence. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 170 , 29–39.