Jonathan Kissling

Taxonomy and Classification

This chapter provides a description of all plant families and genera that include putative fully mycoheterotrophic species, excluding initial and partial mycoheterotrophs. The overview covers a total of 17 families, 101 genera, and ca. 880 species. For each family and genus (except for Orchidaceae) a short morphological description is provided followed by notes on taxonomy, distribution, evolution, and ecology. For most genera a line drawing of a representative species is provided. Included families are: Aneuraceae, Burmanniaceae, Corsiaceae, Ericaceae, Gentianaceae, Gleicheniaceae, Iridaceae, Lycopodiaceae, Ophioglossaceae, Orchidaceae, Petrosaviaceae, Podocarpaceae, Polygalaceae, Psilotaceae, Schizaeaceae, Thismiaceae, and Triuridaceae.

Phylogenetic relationships of the mycoheterotrophic genus Voyria and the implications for the biogeographic history of Gentianaceae

PREMISE OF THE STUDY The angiosperm family Gentianaceae comprises over 1700 species in 91 genera. Gentianaceae are distributed worldwide, but most species occur in temperate zones. Phylogenetic studies demonstrate that the family consists of six monophyletic tribes. However, the phylogenetic position of the mycoheterotrophic genus Voyria, with a remarkable trans-Atlantic distribution, remained unknown. METHODS We used nuclear ITS and 18S rDNA and mitochondrial apt1 and matR data to infer the phylogenetic position of Voyria in Gentianaceae. In addition, with Bayesian relaxed molecular clock analyses we obtained age estimates for the diversification of Voyria and Gentianaceae in general and used these results to reconstruct the ancestral areas associated with the early diversification events in Gentianaceae. KEY RESULTS Our results demonstrate that Voyria is an early diverging lineage within Gentianaceae with no close relationships to other mycoheterotrophic Gentianaceae lineages. Voyria originated in the neotropics during the Early Eocene but only reached its current transoceanic distribution around the end of the Oligocene. The neotropics were an important area for the early diversification events in Gentianaceae, most of which occurred during the Eocene. CONCLUSIONS Voyria is an old, phylogenetically isolated lineage within Gentianaceae, and the current distribution of the genus is indicative of the ancestral area in which the early diversification events of Gentianaceae occurred. In parallel with many other pantropical families, our results suggest that migration of tropical taxa through Laurasia during the Early Eocene has played an important role in shaping the current global distribution of Gentianaceae.

Ancestral and monophyletic presence of diplostigmaty in Sebaea (Gentianaceae) and its potential role as a morphological mixed mating strategy.

* Diplostigmaty, the presence of a primary (apical) stigma and secondary (mid-stylar) stigmas along the style, is only known from the genus Sebaea (Gentianaceae). Early work indicated that the secondary stigmas provide a mechanism of autogamy, suggesting that it might ensure reproductive assurance. * Here, we test the monophyly of this unique morphological trait. Using Bayesian methods, we infer a nuclear DNA phylogeny for 96 accessions, including c. 50% of the species from the genus Sebaea. With this phylogeny, we infer the distribution of ancestral states on critical nodes using parsimony and likelihood methods. * The inferred nrDNA phylogeny shows that the genus Sebaea is divided in two statistically well-supported clades, A and B, consistent with recent estimates. The most recent ancestor (MRCA) of clade A, except the most basal species (Sebaea pusilla), is resolved as diplostigmatic. No reversal to a single stigma is observed within this clade. * We suggest that diplostigmaty is evolutionarily stable through time. We also discuss why this reproductive system is not found elsewhere than in Gentianaceae and the potential advantage of diplostigmaty as a stable mixed mating strategy.

The polyphyletic genus Sebaea (Gentianaceae): a step forward in understanding the morphological and karyological evolution of the Exaceae.

Within the Gentianaceae-Exaceae, the most species-rich genus Sebaea has received very little attention in terms of phylogenetic or karyological investigations. As a result, the exact number of species remains vague and the relationships with the other members of the Exaceae poorly understood. In this paper, we provide the first comprehensive phylogeny of the Exaceae including most Sebaea species known so far based on four cpDNA sequence regions. In addition, morphological and karyological characters were mapped on the inferred phylogenetic trees to detect possible non-molecular synapomorphies. Our results reveal the paraphyly of Sebaea and highlight new generic relationships within the Exaceae. Sebaea pusilla (lineage S1--Lagenias) forms a highly supported and early diverging clade with Sebaeas.str. (clade S2 -Sebaea). A third clade of the former Sebaea s.l. (clade S3--Exochaenium) contains exclusively tropical African species, and is sister with a large clade containing all the remaining genera of Exaceae. Within the latter, the proposed sister relationships between the recently described Klackenbergia and Ornichia are highly supported. Optimization of several morphological characters onto the inferred phylogenetic trees reveals several synapomorphies for most highly supported clades. In particular, lineage S1 (Lagenias) is supported by medifixed anthers that are inserted at the base of the corolla tube and cubical seeds with polygonal testa cells; clade S2 (Sebaea) is supported by both the presence of secondary stigmas along the style and ridged seeds with rectangular testa cells arranged in row; clade S3 (Exochaenium) is supported by its particular gynoecium (stylar polymorphism and clavate, papillose stigma). Finally, karyological reconstructions suggest a basal number of x=7 for the Exaceae and several episodes of dysploidy leading to x=8 and 9.

Variation and evolution of herkogamy in Exochaenium (Gentianaceae): implications for the evolution of distyly

BACKGROUNDS AND AIMS The spatial separation of stigmas and anthers (herkogamy) in flowering plants functions to reduce self-pollination and avoid interference between pollen dispersal and receipt. Little is known about the evolutionary relationships among the three main forms of herkogamy - approach, reverse and reciprocal herkogamy (distyly) - or about transitions to and from a non-herkogamous condition. This problem was examined in Exochaenium (Gentianaceae), a genus of African herbs that exhibits considerable variation in floral morphology, including the three forms of herkogamy. METHODS Using maximum parsimony and maximum likelihood methods, the evolutionary history of herkogamic and non-herkogamic conditions was reconstructed from a molecular phylogeny of 15 species of Exochaenium and four outgroup taxa, based on three chloroplast regions, the nuclear ribosomal internal transcribed spacer (ITS1 and 2) and the 5·8S gene. Ancestral character states were determined and the reconstructions were used to evaluate competing models for the origin of reciprocal herkogamy. KEY RESULTS Reciprocal herkogamy originated once in Exochaenium from an ancestor with approach herkogamy. Reverse herkogamy and the non-herkogamic condition homostyly were derived from heterostyly. Distylous species possessed pendent, slightly zygomorphic flowers, and the single transition to reverse herkogamy was associated with the hawkmoth pollination syndrome. Reductions in flower size characterized three of four independent transitions from reciprocal herkogamy to homostyly. CONCLUSIONS The results support Lloyd and Webbs model in which distyly originated from an ancestor with approach herkogamy. They also demonstrate the lability of sex organ deployment and implicate pollinators, or their absence, as playing an important role in driving transitions among herkogamic and non-herkogamic conditions.

Back to Gondwanaland: can ancient vicariance explain (some) Indian Ocean disjunct plant distributions?

Oceans, or other wide expanses of inhospitable environment, interrupt present day distributions of many plant groups. Using molecular dating techniques, generally incorporating fossil evidence, we can estimate when such distributions originated. Numerous dating analyses have recently precipitated a paradigm shift in the general explanations for the phenomenon, away from older geological causes, such as continental drift, in favour of more recent, long-distance dispersal (LDD). For example, the ‘Gondwanan vicariance’ scenario has been dismissed in various studies of Indian Ocean disjunct distributions. We used the gentian tribe Exaceae to reassess this scenario using molecular dating with minimum (fossil), maximum (geological), secondary (from wider analyses) and hypothesis-driven age constraints. Our results indicate that ancient vicariance cannot be ruled out as an explanation for the early origins of Exaceae across Africa, Madagascar and the Indian subcontinent unless a strong assumption is made about the maximum age of Gentianales. However, both the Gondwanan scenario and the available evidence suggest that there were also several, more recent, intercontinental dispersals during the diversification of the group.

Taxonomy of Exochaenium and Lagenias: Two Resurrected Genera of Tribe Exaceae (Gentianaceae)

Abstract Morphological and molecular evidence has recently revealed the paraphyly of Sebaea (Gentianaceae: Exaceae), and support the establishment of three genera corresponding to highly supported clades. Consequently, the reinstatement of both Exochaenium and Lagenias as genera segregated from Sebaea is proposed here. Exochaenium consists of 22 species distributed in tropical continental Africa that are characterized by papillate stigmas and cubical seeds with star-shaped testa cells. Lagenias is a monotypic genus endemic to the Western Cape Province of South Africa, which differs from Exochaenium and Sebaea by its medifixed anthers, the insertion of the filaments at the base of the corolla tube, and cubical seeds with polygonal testa cells. The synapomorphies of Sebaea s. s. are the presence of secondary stigmas along the style and seeds with rectangular testa cells arranged in parallel rows. Differences with the other genera of tribe Exaceae (Exacum, Gentianothamnus, Klackenbergia, Ornichia, and Tachiadenus) are discussed. In addition, diagnostic characters as well as keys to the genera of the tribe and species of Exochaenium are provided. To account for the new circumscription of Exochaenium, 12 new combinations are also published here ( Exochaenium alatum, E. caudatum, E. clavatum, E. dimidiatum, E. fernandesianum, E. hockii, E. lineariforme, E. macropterum, E. oliganthum, E. perparvum, E. rotundifolium, and E. wildemanianum ).

Novel microsatellite loci for Sebaea aurea (Gentianaceae) and cross-amplification in related species.

Premise of the study: Microsatellite loci were developed in Sebaea aurea (Gentianaceae) to investigate the functional role of diplostigmaty (i.e., the presence of additional stigmas along the style). Methods and Results: One hundred seventy-four and 180 microsatellite loci were isolated through 454 shotgun sequencing of genomic and microsatellite-enriched DNA libraries, respectively. Sixteen polymorphic microsatellite loci were characterized, and 12 of them were selected to genotype individuals from two populations. Microsatellite amplification was conducted in two multiplex groups, each containing six microsatellite loci. Cross-species amplification was tested in seven other species of Sebaea. The 12 novel microsatellite loci amplified only in the two most closely related species to S. aurea (i.e., S. ambigua and S. minutiflora) and were also polymorphic in these two species. Conclusions: These results demonstrate the usefulness of this set of newly developed microsatellite loci to investigate the mating system and population genetic structure in S. aurea and related species.

Diplostigmaty in plants: a novel mechanism that provides reproductive assurance

Differentiation of female sexual organs in flowering plants is rare and contrasts with the wide range of male reproductive strategies. An unusual example involves diplostigmaty, the possession of spatially and temporally distinct stigmas in Sebaea (Gentianaceae). Here, the single pistil within a flower has an apical stigma, as occurs in most flowering plants, but also a secondary stigma that occurs midway down the style, which is physically discrete and receptive several days after the apical stigma. We examined the function of diplostigmaty in Sebaea aurea, an insect-pollinated species of the Western Cape of South Africa. Floral manipulations and measurements of fertility and mating patterns provided evidence that basal stigmas function to enable autonomous delayed self-pollination, without limiting opportunities for outcrossing and thus avoiding the costs of seed discounting. We suggest that delayed selfing serves as a mechanism of reproductive assurance in populations with low plant density. The possession of dimorphic stigma function provides a novel example of a flexible mixed-mating strategy in plants that is responsive to changing demographic conditions.