Katharina Schulte

Phylogeny, adaptive radiation, and historical biogeography in Bromeliaceae: Insights from an eight-locus plastid phylogeny

PREMISE Bromeliaceae form a large, ecologically diverse family of angiosperms native to the New World. We use a bromeliad phylogeny based on eight plastid regions to analyze relationships within the family, test a new, eight-subfamily classification, infer the chronology of bromeliad evolution and invasion of different regions, and provide the basis for future analyses of trait evolution and rates of diversification. METHODS We employed maximum-parsimony, maximum-likelihood, and Bayesian approaches to analyze 9341 aligned bases for four outgroups and 90 bromeliad species representing 46 of 58 described genera. We calibrate the resulting phylogeny against time using penalized likelihood applied to a monocot-wide tree based on plastid ndhF sequences and use it to analyze patterns of geographic spread using parsimony, Bayesian inference, and the program S-DIVA. RESULTS Bromeliad subfamilies are related to each other as follows: (Brocchinioideae, (Lindmanioideae, (Tillandsioideae, (Hechtioideae, (Navioideae, (Pitcairnioideae, (Puyoideae, Bromelioideae))))))). Bromeliads arose in the Guayana Shield ca. 100 million years ago (Ma), spread centrifugally in the New World beginning ca. 16-13 Ma, and dispersed to West Africa ca. 9.3 Ma. Modern lineages began to diverge from each other roughly 19 Ma. CONCLUSIONS Nearly two-thirds of extant bromeliads belong to two large radiations: the core tillandsioids, originating in the Andes ca. 14.2 Ma, and the Brazilian Shield bromelioids, originating in the Serro do Mar and adjacent regions ca. 9.1 Ma.

DISENTANGLING THE EFFECTS OF KEY INNOVATIONS ON THE DIVERSIFICATION OF BROMELIOIDEAE (BROMELIACEAE)

The evolution of key innovations, novel traits that promote diversification, is often seen as major driver for the unequal distribution of species richness within the tree of life. In this study, we aim to determine the factors underlying the extraordinary radiation of the subfamily Bromelioideae, one of the most diverse clades among the neotropical plant family Bromeliaceae. Based on an extended molecular phylogenetic data set, we examine the effect of two putative key innovations, that is, the Crassulacean acid metabolism (CAM) and the water‐impounding tank, on speciation and extinction rates. To this aim, we develop a novel Bayesian implementation of the phylogenetic comparative method, binary state speciation and extinction, which enables hypotheses testing by Bayes factors and accommodates the uncertainty on model selection by Bayesian model averaging. Both CAM and tank habit were found to correlate with increased net diversification, thus fulfilling the criteria for key innovations. Our analyses further revealed that CAM photosynthesis is correlated with a twofold increase in speciation rate, whereas the evolution of the tank had primarily an effect on extinction rates that were found five times lower in tank‐forming lineages compared to tank‐less clades. These differences are discussed in the light of biogeography, ecology, and past climate change.

Detection of recent hybridization between sympatric Chilean Puya species (Bromeliaceae) using AFLP markers and reconstruction of complex relationships

The Chilean Puya species constitute a monophyletic group, co-occurring in different species combinations within the country and displaying a remarkable morphological variability. Here, we studied the importance of recent hybridization and introgression in the group and reconstructed the complex inter- and intraspecific relationships. Amplified fragment length polymorphism (AFLP) analysis, including 109 accessions of all Chilean Puya species and four putative hybrids, yielded 984 characters. Three main genetic groups were revealed, with the chilensis group (P. chilensis, P. gilmartiniae, P. boliviensis) diverging first, and the alpestris (P. alpestris, P. berteroniana) and coerulea group (P. venusta, P. coerulea) forming sister groups. STRUCTURE analyses confirmed a hybrid origin of morphologically intermediate individuals, and detected several additional hybrids. Hybrids were found between the chilensis and alpestris group, and between the alpestris and coerulea group. Exclusion of hybrids improved phylogenetic reconstructions. The study demonstrates that the detection of hybrids within Bromeliaceae can be difficult based on morphological characters alone and that efficient reproductive barriers may only slowly establish, leading to hybridization between closely related sympatric species. The importance of hybridization for the rapid diversification of Puya is discussed.

Chilean Bromeliaceae: diversity, distribution and evaluation of conservation status

Chile is home to 23 species of Bromeliaceae, including 2 subspecies and 4 varieties. Twenty species are endemic to the country. We examined 883 herbarium specimens from 27 herbaria for our treatment of the Bromeliaceae for the “Flora de Chile”. These data and field observations resulted in a comprehensive database that we used to generate distribution maps for each species. We applied ecological niche modelling to reveal distribution areas and centers of Bromeliaceae diversity. We further analysed the collecting dates of the herbarium specimens to assess possible changes in species abundance. In this study we assess the conservation status of the bromeliad species in Chile. IUCN categories were assigned to the 27 bromeliad taxa as follows: Critically endangered: 4, Endangered: 6, Vulnerable: 11, Near threatened: 2, Least concern: 4. No species has become “Extinct” up to now. We also put forth a hypothesis about their biogeographic history.

Phylogenetic analysis of Fosterella L.B. Sm. (Pitcairnioideae, Bromeliaceae) based on four chloroplast DNA regions.

The about 31 species of Fosterella L.B. Sm. (Bromeliaceae) are terrestrial herbs with a centre of diversity in the central South American Andes. To resolve infra- and intergeneric relationships among Fosterella and their putative allies, we conducted a phylogenetic analysis based on sequence data from four chloroplast DNA regions (matK gene, rps16 intron, atpB-rbcL and psbB-psbH intergenic spacers). Sequences were generated for 96 accessions corresponding to 60 species from 18 genera. Among these, 57 accessions represented 22 of the 31 recognized Fosterella species and one undescribed morphospecies. Maximum parsimony and Bayesian inference methods yielded well-resolved phylogenies. The monophyly of Fosterella was strongly supported, as was its sister relationship with a clade comprising Deuterocohnia, Dyckia and Encholirium. Six distinct evolutionary lineages were distinguished within Fosterella. Character mapping indicated that parallel evolution of identical character states is common in the genus. Relationships between species and lineages are discussed in the context of morphological, ecological and biogeographical data as well as the results of a previous amplified fragment length polymorphism (AFLP) study.

Molecular phylogeny of the Brazilian endemic genus Orthophytum (Bromelioideae, Bromeliaceae) and its implications on morphological character evolution

The saxicolous genus Orthophytum (∼60 species, Bromeliaceae) is endemic to eastern Brazil and diversified in xeric habitats of the Atlantic Rainforest, Caatinga and campos rupestres. Within the genus, two main groups are discerned based on the presence or absence of a pedunculate inflorescence, which are further subdivided into several morphological subgroups. However, these systematic hypotheses have not yet been tested in a molecular phylogenetic framework. Here we present the first phylogenetic analysis of Orthophytum using nuclear and plastid markers (phytochrome C, and trnH-psbA and trnL-trnF spacers). Forty species representing the two main groups and all subgroups of Orthophytum, and the related genera Cryptanthus (8 spp.) and Lapanthus (2 spp.) were analyzed. The phylogenetic reconstruction revealed a well-supported clade termed Eu-Orthophytum, containing species with pedunculate inflorescences only. The Orthophytum species with sessile inflorescence formed two clades: (1) the amoenum group and (2) the vagans group plus O. foliosum, the only pedunculate Orthophytum species found outside Eu-Orthophytum. The vagans clade is in sister group position to Eu-Orthophytum. Within the latter, the subgroup mello-barretoi was sister to the most diversified clade, termed Core Orthophytum. Morphological character state reconstructions of floral characters used in previous taxonomic treatments as key diagnostic characters (penduncle presence, corolla form, and petal appendage form) showed different levels of homoplasy.

Rapid microsatellite marker development for African mahogany (Khaya senegalensis, Meliaceae) using next-generation sequencing and assessment of its intra-specific genetic diversity.

Khaya senegalensis (African mahogany or dry‐zone mahogany) is a high‐value hardwood timber species with great potential for forest plantations in northern Australia. The species is distributed across the sub‐Saharan belt from Senegal to Sudan and Uganda. Because of heavy exploitation and constraints on natural regeneration and sustainable planting, it is now classified as a vulnerable species. Here, we describe the development of microsatellite markers for K. senegalensis using next‐generation sequencing to assess its intra‐specific diversity across its natural range, which is a key for successful breeding programs and effective conservation management of the species. Next‐generation sequencing yielded 93 943 sequences with an average read length of 234 bp. The assembled sequences contained 1030 simple sequence repeats, with primers designed for 522 microsatellite loci. Twenty‐one microsatellite loci were tested with 11 showing reliable amplification and polymorphism in K. senegalensis. The 11 novel microsatellites, together with one previously published, were used to assess 73 accessions belonging to the Australian K. senegalensis domestication program, sampled from across the natural range of the species. STRUCTURE analysis shows two major clusters, one comprising mainly accessions from west Africa (Senegal to Benin) and the second based in the far eastern limits of the range in Sudan and Uganda. Higher levels of genetic diversity were found in material from western Africa. This suggests that new seed collections from this region may yield more diverse genotypes than those originating from Sudan and Uganda in eastern Africa.

Elucidating phylogenetic relationships in the Aechmea alliance: AFLP analysis of Portea and the Gravisia complex (Bromeliaceae, Bromelioideae)

Abstract Genetic concepts within the Bromelioideae are highly problematic, in particular within the tank-forming “core bromelioid” clade. Previous molecular studies showed that the largest genus, Aechmea, and allied genera are polyphyletic and require revision. Here we focus on one group within the Aechmea alliance, the Portea/Gravisia group. To assess whether species of this group form a distinct lineage within the core bromelioid clade, and to clarify generic limits and interspecific relationships within the group, we generated and analyzed amplified fragment length polymorphisms (AFLP). In total, 69 species were sampled, including 26 species previously assigned to the Portea/Gravisia group. Neighbor joining, maximum likelihood and Bayesian analyses of the AFLP data consistently retrieved the Portea/Gravisia group as a monophyletic clade with several subclades, comprising species from the genera Aechmea, Canistrum and Portea. The phylogenetic distribution of polyporate pollen of the resulting trees indicates this character state arose once within the core bromelioids and thus can be regarded as a synapomorphy for the Portea/Gravisia clade. Further, our study shows that within the Portea/Gravisia group, subclades are characterized by petal color and geographic distribution. Thus, the present study is a further advance in the challenging task of elucidating phylogenetic relationships within the core bromelioids as the basis for a revised taxonomy of this ecologically important group.

Molecular evidence for hybridization and introgression in the neotropical coastal desert-endemic Palaua (Malveae, Malvaceae).

Recent years have seen an increasing awareness of the importance of hybridization and introgression in plant evolution. Both processes were also invoked to have had a strong impact in the evolution of Palaua, based on morphological intermediacy. Here, we used nuclear ITS and cellulose synthase DNA sequences to assess previous phylogenetic hypotheses and to uncover previously undescribed reticulate evolution in Palaua. Both nuclear regions recovered largely the same reticulate pattern, showing that interspecific crosses occurred between distantly related species of the Dissecta and Integrifolia clade. Hybridization between species occurred only in a narrow contact zone. Moreover, many sequences of Palaua dissecta were placed in the strongly divergent Integrifolia clade, a pattern that is best explained by introgression. These results support our view that the strong interannual fluctuations of humidity and habitat expansion provide opportunities for the isolation of taxa, establishment of hybrid species, and for introgression between incompletely reproductively isolated taxa following recurrent contact.

Taxonomic revision of the Chilean Puya species (Puyoideae, Bromeliaceae), with special notes on the Puya alpestris-Puya berteroniana species complex

The Chilean Puya species (Puyoideae, Bromeliaceae) constitute an early diverging group within the genus and thus are key taxa for the understanding of the early evolution and biogeography of Puya. However, a modern taxonomic treatment including information from molecular phylogenetic studies is still lacking. Here, a taxonomic revision of the Chilean species of Puya is presented based on morphological, molecular, and biogeographic data. A re-evaluation of the widely applied concept of P. berteroniana led us to the conclusion that the type of P. berteroniana is of hybrid origin and is maintained as Puya × berteroniana. Our studies revealed that the name P. berteroniana has been widely misapplied to what in fact is the northern metapopulation of P. alpestris, which is here described as a new subspecies, Puya alpestris subsp. zoellneri, a member of subgenus Puya. A lectotype is designated for Puya copiapina and a neotype for P. chilensis. Altogether, six Puya species, two subspecies, four varieties and one hybrid taxon are recognized for the Chilean flora in this revision: P. alpestris subsp. alpestris and subsp. zoellneri; P. boliviensis; P. chilensis; P. gilmartiniae; P. coerulea var. coerulea, var. intermedia, var. monteroana, and var. violacea; P. venusta; and P.×berteroniana. A key is provided for their identification.