Aim Oceanic islands have played an important role in our understanding of the diversification of organisms, and phylogenetic estimates have been used in this context to investigate the origin of island diversity and its relationship to the continent. Using a typical orchid genus rich in island endemics and with widespread continental relatives, we aim to compare alternative hypotheses of diversification with a focus on island endemism. Location Tropical Africa and the Gulf of Guinea Islands of São Tomé & Príncipe, Central Africa. Taxon Tridactyle genus (Orchidaceae). Methods We used genome skimming to sequence the whole chloroplast genome and nuclear ribosomal genes from 157 individuals of 34 Tridactyle species and 15 individuals of 12 other orchid genera (outgroups) to infer a time-calibrated phylogenetic tree of the genus Tridactyle. We also used multiple statistical methods to infer the geographical ranges of the ancestral nodes from the estimated phylogeny. Alternative hypotheses for the origins of endemism on the islands of São Tomé and Príncipe were investigated based on the biogeographical reconstruction of the genus. Results The estimated phylogeny of Tridactyle and reconstruction of geographical ranges for the ancestral nodes suggested a general history of allopatric speciation for the genus, in particular via colonization of the islands of the Gulf of Guinea that induced a long period of geographical isolation. The most parsimonious hypothesis to explain island endemism in Tridactyle involved six independent colonizations of the islands from the continent. Main conclusions In contrast to other cases of oceanic island endemism that involved adaptive radiation on an island or within an archipelago, endemism in Tridactyle is better explained by multiple colonization events from the continent to São Tomé and Príncipe, with subsequent divergences due to geographical isolation but only one potential instance of further diversification on the islands.

Background and aims – Brachystegia is a species-rich tree genus found in tropical Africa and a typical element of Miombo woodlands, a widely distributed subtype of the Zambezian savanna. Plastid DNA was shown to be largely uninformative to assess species phylogenetic relationships due to widespread chloroplast capture among species. Here, we aim to assess the capacity of nuclear ribosomal DNA (rDNA) to clarify the phylogeny of Brachystegia species while accounting for intra-individual site polymorphisms (2ISPs), which are often present in rDNA and potentially phylogenetically informative. Material and methods – Genome skimming sequencing on 47 samples representing 27 of the 29 currently recognized Brachystegia species, allowed us to retrieve complete nuclear ribosomal cistrons encoding for 18S, 5.8S, and 25S rRNA genes (35S rDNA). We reconstructed the Brachystegia phylogeny using Maximum Likelihood methods based on the standard substitution model or integrating 2ISPs (GENOTYPE implementation in RAxML-NG). We additionally tested the effect of partitioning the data (one partition for rDNA genes and one for the ITS1+ITS2). We also conducted network inferences (Neighbor-Net splits graph), as a strict bifurcative approach might not properly model topological uncertainty at shallow phylogenetic depth. Key results – 2ISPs-aware and standard phylogenetic reconstructions are largely congruent. We identified several well-supported main clades clarifying the species relationships, including two clades of Miombo woodlands species. Miombo Group A includes species with ovoid to globose axillary dormant buds, while Miombo Group B species have flattened ones. Two morphologically close Brachystegia species (B. kennedyi and B. leonensis) found in Guineo-Congolian rain forests form also a robustly supported clade. 2ISPs coding allowed to identify an additional Guineo-Congolian clade (B. eurycoma and B. nigerica). Ribosomal DNA therefore proves more useful to explore the generic phylogeny than plastid DNA but the species relationships within and among the main clades remain poorly resolved, probably due to recent diversification and/or recurrent hybridization, so that the diversification of Brachystegia remains to be more properly characterised. Conclusion – Nuclear and plastid phylogenetic reconstructions of Brachystegia species are discordant. Even if not well-resolved, rDNA phylograms and networks are characterized by taxonomic sorting, while we observe a strictly geographic sorting in the plastid dataset. Most of the species’ relationships remain to be characterized using additional nuclear markers combined with in-depth morphological investigations.

Background and objectives—Podocarpus latifolius (synonym of P. milanjianus) is a key tree representative of Afromontane forests where it is highly threatened by climate and land-use changes. While large populations occur in East Africa, only a few isolated and usually small populations remain in western Central Africa (Cameroon to Angola). Studying the evolutionary history of such relictual populations can thus be relevant to understand their resilience under changing environments. Materials and Methods—we developed nine polymorphic nuclear microsatellites (nSSRs) to estimate genetic variability, (historical) gene flow, and demographic changes among natural populations from Central to East Africa. Results—despite the extended distribution range of P. latifolius, a strong isolation-by-distance pattern emerges at the intra-population scale, indicating low seed and pollen dispersal capacities. Central African populations display a lower genetic diversity (He = 0.34 to 0.61) and are more differentiated from each other (F_ST = 0.28) than are East African populations (He = 0.65 to 0.71; F_ST = 0.10), suggesting high genetic drift in the Central African populations. Spatial genetic structure reveals past connections between East and West Africa but also a gene flow barrier across the equator in western Central Africa. Demographic modelling anchors the history of current lineages in the Pleistocene and supports a strong demographic decline in most western populations during the last glacial period. By contrast, no signature of demographic change was detected in East African populations. Conclusions—in Cameroon, our results exclude a recent (re)colonization from one source population of all mountain ranges, but rather indicate long-term persistence of populations in each mountain with fluctuating sizes. A higher impact of genetic drift and further loss of diversity can be expected by survival through climatically unfavorable periods in such small refugial populations. Tracking the Quaternary legacy of podocarp populations is thus essential for their conservation since there is a temporal gap between environment crises and an ecological/genetic answer at the population level.

Characterizing the feeding ecology of threatened species is essential to establish appropriate conservation strategies. We focused our study on the proboscis monkey (Nasalis larvatus), an endangered primate species which is endemic to the island of Borneo. Our survey was conducted in the Lower Kinabatangan Wildlife Sanctuary (LKWS), a riverine protected area that is surrounded by oil palm plantations. We aimed to determine the diet of multiple proboscis monkey groups by using two methods. First, we conducted boat-based direct observations (scan and ad libitum sampling) and identified 67 plant species consumed by the monkeys at their sleeping sites in early mornings and late afternoons. Secondly, we used the DNA metabarcoding approach, based on next-generation sequencing (NGS, MiSeq Illumina) of faecal samples (n = 155), using the short chloroplast sequence, the trnL (UAA) P6 loop. In addition, we built a DNA reference database with the local plants available in the LKWS. When combining feeding data from both methods, we reported a diverse dietary ecology in proboscis monkeys, with at least 89 consumed plant taxa, belonging to 76 genera and 45 families. Moreover, we were able to add 22 new genera as part of the diet of this endangered colobine primate in the LKWS. The two methods provided congruent and complementary results, both having their advantages and limitations. This study contributed to enhance the knowledge on the feeding ecology of proboscis monkeys, highlighting the significance of several plant species that should further be considered in habitat restoration plans or corridor establishment.