Póster presentado en el III Simposio Anual de Botánica Española celebrado en el Institut Botànic de Barcelona, 25-26 de noviembre de 202

Ackerfield, J.R.

Blanco-Gavaldà, C

Brochmann, C

Calleja, J.A.

Fujikawa, Kazumi

Galbany Casals, Mercedes

Garcia-Jacas, Núria

Gizaw, A

López-Alvarado, Javier

Moreyra, Lucía D.

Roquet Ruíz, C

Susanna de la Serna, Alfonso

Vilatersana, Roser

English

Blanco-Gavaldà, C.

Digital.CSIC

Carduus keniensis- Mount Kilimanjaro, Tanzania.The Afromontane region comprises humid and temperate isolated refuges in thehighest African mountains. A question of major interest is understanding the originsand evolution of their diverse and singular flora (Brochmann et al., 2021 andreferences therein). The complex Carduus-Cirsium (Cardueae-Carduinae,Compositae) has some species endemic to this region, with a distribution restrictedmainly to Tropical East Africa. These species show a combination of morphologicalcharacters that resemble Carduus and Cirsium; however, they do not fit properly withthe definition of any of the two genera. Likewise, molecular evidence has suggesteda high divergence between the Afromontane species and the species of thecomplex Carduus-Cirsium (Häffner and Hellwig, 1999; Barres et al., 2013; Ackerfield etal., 2020). Thus, a thorough study including a complete representation of the speciesof the subtribe and a morphological evaluation are needed to understand properlythe evolution of isolated and highly adapted mountain species.IntroductionAfrican mountain thistles: generic delimitation problems analyzed with NGSMoreyra,  L. D.1; Ackerfield, J. R.2; Blanco-Gavaldà , C.3; Brochmann,  C.4; Calleja,  J. A.5; Fujikawa, K.6; Galbany-Casals, M.3; Garcia-Jacas, N.¹;Gizaw, A.4; López-Alvarado,  J.3; Roquet, C.3; Susanna, A.¹; Vilatersana, R.¹1Botanic Institute of Barcelona (IBB), CSIC-Ajuntament de Barcelona; 2Department of Biology, Colorado State University, U.S.A.; 3Autonomous University of Barcelona, Systematics and Evolution of Vascular Plants (UAB) – Associated Unit to CSIC by IBB; 4University of Oslo, Norway; 5Autonomous University of Madrid; 6Kochi Prefectural Makino Botanical Garden, Japan.As in Herrando-Moraira et al. (2019), subtribe Carduinae was recovered as monophyletic with highresolution by the Hyb-Seq data analyses. Within the Carduus-Cirsium complex, Picnomon, Galactites, Silybumand Tyrimnus are natural genera. On the other hand, Carduus and Cirsium are both monophyletic only if theAfrican species are excluded. This result was partly expected from previous evidence both morphological andmolecular provided by Häffner and Hellwig (1999), Barres et al. (2013) and Ackerfield et al. (2020). Species ofCarduus subg. Afrocarduus and Cirsium from Tropical East Africa are nested at the base of the complexCarduus-Cirsium with high support (LPP=1), except for two species of Cirsium that were recovered out of thecomplex Carduus-Cirsium and appear closely related to Galactites and Lamyropsis.Morphological analysis has shown that the species of Carduus from Africa have more characters shared withCirsium than with Carduus. This evidence, along with our phylogenetic analyses, supports subg. Afrocarduusas a monophyletic natural group independent from Carduus and Cirsium. Hence, after analyzing themorphological evidence and our molecular results, we propose a new genus for the Carduus species of Africa,using the name provided by Kazmi (1963) for the subgenus: Afrocarduus.As to the African Cirsium, a close look at the morphology of these species has shown clear unique characters,such as phyllaries with well-developed appendages that are not present in Cirsium. This observation, alongwith the phylogenetic results, made us think of these species as a genus independent of Afrocarduus andCirsium. Here we propose a second genus, Afrocirsium, for the three African species of Cirsium that arenested in the base of Afrocarduus and constitute a monophyletic group.Results and DiscussionCarduus nyassanusA total of 198 samples representing all the genera of subtribe Carduinae wereselected for the phylogenetic analyses according to Herrando-Moraira et al. (2019).Eleven raw sequences were obtained from this study and the rest of the samples wereobtained from herbarium specimens or fresh material collected and preserved insilica-gel. DNA extraction, library, capture and sequencing were carried out in ourfacilities at the Botanic Institute of Barcelona. In some cases, DNA extractions weresent to Daicel Arbor Biosciences (Ann Arbor, Michigan, USA).Phylogenetic analyses were carried out using HybPhyloMaker pipeline (Fér &Schmickl, 2018) and Paralog Wizard (Ufimov et al., 2022). For coalescence analysis,gene trees were calculated using RAxML v.8.2.12 and the analysis was done usingASTRAL-III v.5.7.8, both implemented in the HybPhyloMaker pipeline.Material and MethodsCarduus leptacanthusReferences• Ackerfield, J. et al. 2020. A prickly puzzle: Generic delimitations in the Carduus-Cirsium group (Compositae: Cardueae: Carduinae). Taxon 69(4): 715‒738.• Barres, L. et al. 2013. Reconstructing the evolution and biogeographic history of tribe Cardueae (Compositae). American Journal of Botany 100(5): 867–882.• Brochmann, C. et al. 2021. History and evolution of the afroalpine flora: in the footsteps of Olov Hedberg. Alpine Botany, 1-23.• Fér, T. & Schmickl, R. E. 2018. HybPhyloMaker: target enrichment data analysis from raw reads to species trees. Evolutionary Bioinformatics 14:1176934317742613.• Häffner, E. & Hellwig, F. 1999. Phylogeny of the tribe Cardueae (Compositae) with emphasis on the subtribe Carduinae: An analysis based on ITS sequencedata. Willdenowia 29: 27‒39.• Herrando-Moraira, S. et al. 2019. Nuclear and plastid DNA phylogeny of tribe Cardueae (Compositae) with Hyb-Seq data: A new subtribal classification anda temporal diversification framework. Molecular Phylogenetics and Evolution 137: 313‒332.• Ufimov, R. et al. 2022. Utilizing paralogues for phylogenetic reconstruction has the potential to increase species tree support and reduce gene treediscordance in target enrichment data. Molecular Ecology Resources 22(8): 3018‒3034.a Achene pericarp with 10-15 longitudinal grooves………………………………..…..……Carduusa' Achene pericarp with 4 longitudinal lines………………………………..………………………………….bb Barbellate pappus…………………………………………………………………………………..…Afrocarduusb’ Plumose pappus…………………………………………………………………..…………………………………….…cc Phyllaries without appendages…………………………………………..……………………….……Cirsiumc’ Phyllaries with well-developed appendages………………….………………..……….Afrocirsium* Key built only with the main genera of the complex; monotypic genera are excluded.Key to Complex Carduus-CirsiumFinancial support from the Spanish Ministry of Science, Innovation and Universities (PID2019-105583GB-C21/ AEI /10.13039/501100011033) and the Catalan government (2017-SGR1116)Photos: https://www.malawiflora.com/; https://www.zambiaflora.com/; https://www.flagtails.com/; https://www.inaturalist.com/; Mercè Galbany-CasalsCirsium buchwaldiiCoalescence tree of subtribe Carduinae- 985 nuclear genes - LPP values are shown for main branches

African mountain thistles: generic delimitation problems analyzed with NGS

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African mountain thistles: generic delimitation problems analyzed with NGS

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