TL;DR: The circumscription of Coffeeae (Rubiaceae) and phylogenetic relationships within the tribe were evaluated using sequence data from four plastid regions and a morphological data set and support an enlarged circumscription for Coffeeae, containing 11 genera.
Abstract: The circumscription of Coffeeae (Rubiaceae) and phylogenetic relationships within the tribe were evaluated using sequence data from four plastid regions (trnL-F intron, trnL-F intergenic spacer [IGS], rpl16 intron, and accD-psa1 IGS) and a morphological data set. Eleven candidates for inclusion in Coffeeae were examined using plastid data, and a further three were investigated using morphology alone. Based on previous phylogenetic analysis of the subfamily Ixoroideae, nine genera representing five tribes were used as outgroups. Our results support an enlarged circumscription for Coffeeae, containing 11 genera, viz. Argocoffeopsis, Belonophora, Calycosiphonia, Coffea, Diplospora, Discospermum, Nostolachma, Psilanthus, Tricalysia, Sericanthe, and Xantonnea. The inclusion of Diplospora and Tricalysia within Coffeeae, based on published molecular data, and the inclusion of Argocoffeopsis, Belonophora, Calycosiphonia, Discospermum, and Sericanthe, based on morphological evidence, are well supported. Nostolachma is newly transferred from Gardenieae subtribe Diplosporinae to Coffeeae, and Xantonnea from Octotropideae to Coffeeae. The exclusion of Bertiera from Coffeeae and placement in tribe Bertiereae is supported on the basis of molecular and morphological data. The removal of Diplospora and all other genera from Gardenieae subtribe Diplosporinae to Coffeeae and Octotropideae renders Diplosporinae superfluous. It is proposed that Xantonneopsis be transferred to Octotropideae; Petitiocodon is tentatively placed in Gardenieae. The monophyly of seven genera is supported, but Coffea is identified as paraphyletic in relation to Psilanthus on the basis of molecular and combined molecular and morphological data.
TL;DR: The results suggest that the endosymbionts of Pavetta represent novel species, which can be classified as 'candidatus Burkholderia hispidae', 'CandidatusBurkholderia rigidae' and 'Candidia schumannianae'.
Abstract: Three genera in the Rubiaceae (Pavetta, Psychotria and Sericanthe) harbour bacterial endosymbionts within leaf nodules or galls. The present paper identifies the bacterial endophytes in three leaf-nodulating Pavetta species. In order to reveal their identity and assess their phylogenetic position, 16S rRNA, recA and gyrB genes were sequenced from an extensive sampling of Burkholderia strains. This multigene approach results in a robust phylogeny, which places the bacterial endosymbionts of Pavetta at two distinct positions within the genus Burkholderia (class Betaproteobacteria), suggesting that leaf-nodulating endosymbionts within Pavetta have different origins. The endophytes of nodulated Psychotria species were recognized as the closest relatives to the Pavetta endosymbionts. Our results suggest that the endosymbionts of Pavetta represent novel species, which can be classified as ‘Candidatus Burkholderia hispidae’, ‘Candidatus Burkholderia rigidae’ and ‘Candidatus Burkholderia schumannianae’.
TL;DR: The phylogenetic analyses revealed a pattern of strict host specificity and placed the two investigated endosymbionts at two distinct positions in the topology of the tree, suggesting at least two different evolutionary origins.
Abstract: Bacterial leaf symbiosis is an intimate association between bacteria and plants in which endosymbionts are housed within leaf nodules. This phenomenon has been reported in three genera of Rubiaceae (Pavetta, Psychotria, and Sericanthe), but the bacterial partner has only been identified in Psychotria and Pavetta. Here we report the identification of symbiotic bacteria in two leaf nodulating Sericanthe species. Using 16S rRNA data and common housekeeping genetic markers (recA and gyrB) we studied the phylogenetic relationships of bacterial endosymbionts in Rubiaceae. Endosymbionts of leaf nodulating Rubiaceae were found to be closely related and were placed as a monophyletic group within the genus Burkholderia (β-Proteobacteria). The phylogenetic analyses revealed a pattern of strict host specificity and placed the two investigated endosymbionts at two distinct positions in the topology of the tree, suggesting at least two different evolutionary origins. The degree of sequence divergence between the Sericanthe endosymbionts and their relatives was large enough to propose the Sericanthe endosymbionts as new species (‘Candidatus Burkholderia andongensis’ and ‘Candidatus Burkholderia petitii’). In a second part of this study, the pylogenetic relationships among nodulating and non-nodulating Sericanthe species were investigated using sequence data from six chloroplast regions (rps16, trnG, trnL-trnF, petD, petA-psbJ, and atpI-atpH). Overall, genetic variation among the plastid markers was insufficient to enable phylogenetic estimation. However, our results could not rule out the possibility that bacterial leaf symbiosis originated once in a common ancestor of the Sericanthe species.