Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. nWs eta fia(eeo ta.22) h V opie ltasad1 omn volcanoes, dormant 11 and plateaus comprises CVL The refuge forest 2020). ancient an was al. chain volcanic Gulf et This the (Demenou from 2015). extends Africa al. that Central et feature West (Adams topographical Cameroon SW-NE in central km onshore volcanic 1800 to an substantial Kenya Guinea is and of was (CVL) Tanzania Line of there Volcanic Mountains Cameroon Arc epochs, The Eastern and the Pliocene-Pleistocene as the 2011). such Africa of Tolley Africa, Forest during Central East & West speciation Moreover, and 2007). (Measey in allopatric African patterns Central al. to West diversity 2012). et in current origin activity (Kebede to gave al. contributed Africa that rivers et East and driver in (Nicolas refugia a m forest was 6000 Likewise, and Pleistocene animals. as 3500 the such et between distribution, during (Duminil altitudes Africa species fragmentation lowland with Guinea, Afro-alpine mountains Lower of shaped and on expansion oscillations Guinea occurs climate and Upper These of fragmentation savannas 2015). and caused al. forests, oscillations mountain environmental forests, rain and tropical climate Pleistocene’s The sky South Afromontane establishing the Introduction of after age Cameroon the Bioko mainland concerning and in progression Cameroon inverse island the an sky was of each older history divergence from the biogeography the Bioko lowland The colonized islands. to of with columnaris islands. led populations depression sky L. oscillations the central younger separate climatic Bioko’s the keeps clades. Pleistocene Furthermore, that three The barrier into Cameroon. geographic populations. in populations young current mainland islands Bioko’s a the South sky is and from Bioko old conditions other South Pleistocene northern dry Bioko and and middle-late the South cold North the and during between in during Bioko isolated forest population likely occurred North was the history to Bioko split colonization South island Ma) of the sky along population Presumably, (1.54 first The event colonized refugia. divergent Bioko. columnaris plastome- forest North earliest L. three and in The that sky into mainland indicate the subdivided the islands. islands on origin, sky sky of those monophyletic the young from age a of Line’s have the age Volcanic columnaris data and reflect Cameroon varied L. results plastome size the of history phylogenomic The plastome plastid populations colonization complete systems. The The The the mountain clades. bp. Inference. five Does geographic 165,368 Bayesian from to (3) and columnaris bp likelihood L. Maximum 164,609 islands? using of populations? from aim sky analyses individuals Cameroon phylogenetic we 20 older and with Specifically, from Island the explored Bioko plastomes was Cameroon. on novel among and found assembled relationship Bioko We phylogenetic populations of the islands? older islands is estimate sky the What and the Are genomes (1) on chloroplast (2) questions: using colonization following columnaris historical the its answer of reconstruct to populations to of time relationships divergence phylogenetic the the infer to aimed We Abstract 2021 30, January 2 1 Perez Perez Miguel of islands sky Africa across Central West Lobelioideae) columnaris (: Lobelia f. of history Hook. colonization and origin Pleistocene ihagan rpclBtnclGarden Campus Botanical Mountain Tropical Flagstaff Xishuangbanna - University Arizona Northern 1 n e-i Yu Wen-Bin and 2 1 oei giberroa, Lobelia which Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. eetsuyo h eceityo ocncrcsfo ik ae h antresrt-ocne at strato-volcanoes three main the dates Bioko from end rocks shallow the volcanic of at of level km geochemistry sea 32 of the by rise <1.3 of 2004). the coast study by al. Cameroon recent ago et the years A (Schabetsberger 000 from period 10, separated glacial Africa is last mainland the and from of separated shelf was continental Bioko the (60m). water on lies Island islands? Bioko sky the the of age is populations the area What reflect older Study (1) history the colonization questions: Are the Methods following Does (2) & the (3) populations? Materials sky islands? answer Cameroon the sky to older on and aim the colonization Island on we historical Bioko found Specifically, its among reconstruct Cameroon. relationship to and phylogenetic of time Bioko populations divergence of of the relationships islands phylogenetic estimate the and infer genomes to and chloroplast is endangered objective study’s Africa, the present western of The like islands outlying bioregions, framework the to both a endangered mainland provides eastern the in the like mountains relatives from distribution, hypothesis African close widespread The geographically East among 2007). a and rat, al. origin Oku West et in Mount biogeographical the (Popp endemic present Africa the between currently West time discuss to animals over to dispersed and connections recently genus plants and recurrent Afromontane Mountains of of Rift the lineages However, Western example, Cameroon. the the and For and Bioko documented Cameroon. in have angiosperms and of studies Bioko histories phylogeographic colonization into and few origin transformed a the clearings about to known forest is 1000m grasslands, Little between submontane populations discontinuous including in with habitats overgrown grows ecological grasslands lobelia different giant in This 3000m Cameroon. and 2015). Guinea. Thulin Nigeria, of & Guinea), Gulf (Cheek 2015 and Species Africa Threatened tral of List Red disper- IUCN and subsequently the and in 2017). mountains sky Li species Africa the & vulnerable East inhabit (Knox in Africa that West diversification animals to rapid and sed experienced plants have of may and species endemic Giant 2016), of al. conservation 2009). the Konvicka et (Myers for & hotspot (Missoup critical (Tropek biodiversity possess is islands Guinea mammals Africa and the Central small 2000) of West part al. 1994), and is et Guinea (Figueiredo region Gvoˇzd´ık of 2014). African & angiosperms Gulf al. This (Zimkus et the laboratories 2013). of amphibians (Cox of natural diversity diversity archipelagos considered of island extraordinary accumulation are sky the islands the an and to Sky of islands lead islands. isolation sky that volcanic sky and the processes of or separation Moreover, and age sky geographical patterns decreasing marked the evolutionary late of The in studying to trend 2016). islands for middle a al. to with the et analogous north, from (Missoup the are the of ranging area mountains in in ages structure are southern unequal peaks geological mountains the with oldest volcanic The in origins The three activity volcanic Oku). 2005). and Only Mt. al. tectonic et and of 2005). (Jesus Tertiary combination Cameroon Basil´e, a al. Mt. is (Pico et CVL 3000m (Chauvel the above volcano elevations active have currently CVL a Cameroon, Mount and anadCmro opie eea footn k sad.I h otws on M. aeonis Cameroon (Mt.) Mount southwest in the In arranged islands. is sky Bioko Afromontane several of the at comprises vegetation and appearing Cameroon The elements Mainland (2009m) 2000). Afromontane 2004). Biao-Moka al. with al. maximum Pico et species et a (Fa the tropical elevations (Schabetsberger with and Guineo-Congolian higher South depression by North the central dominated the in rings a in elevational m) Furthermore, Basil´e (3011m) (2260 age. Pico Caldera in separates Gran difference 500m without of young elevation very are North) and L ± . .7M;KA Ymoo ta.21) hsdtn niae httesyilnso ik (South Bioko of islands sky the that indicates dating This 2016). al. et (Yamgouot K/Ar Ma; 0.07 barnsii xl Mbely17a r h w in oei pce nw rmtetoia etCen- West tropical the from known species lobelia giant two the are 1974a) (Mabberley Exell rnsafricana Prunus aotmsokuensis Lamottemys trdu aquilinum Pteridium L . columnaris oei columnaris Lobelia Dwo oel1999). Powell & (Dawson nteCL(isu ta.21) oee,sm aahave taxa some However, 2016). al. et (Missoup CVL the in sedmct ik’ onan n ihad (Equatorial highlands and mountains Bioko’s to endemic is L)Kuhn (L.) 2 okf Cmauaee oeiiee slse sa as listed is Lobelioideae) (Campanulaceae: f. Hook , n ln tem n uapn meadows. subalpine and streams along and Lynchis a eea iprasfo Ethiopia from dispersals several had .columnaris L. L . columnaris using Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. eoe r rmtosyilnsi aeon(iue1.Mroe,tepyoeei reicue two included tree plastid phylogenetic nine the and Moreover, Guinea), 1). (Equatorial (Figure Bioko Cameroon in in islands islands sky sky two from three of are from genomes genomes populations plastid individual represent 20 plastomes of 2017). Li total S15797 and & A no. Genomics (Knox TreeBASE platforms. from accessions for Campanulaceae MiSeq GenBank of Center Illumina and taxa University and 22 2014) of NextSeq (Knox Indiana sequences Illumina plastome the of using 22 downloaded at performed we individuals Additionally, was sequence done generation 20 construction, were Sequence Library of processing Bioinformatics. 1987). bioinformatics samples Doyle and & leaf (Doyle generation, silica-dried protocol (CTAB) from bromide cetyltrimethylammonium extracted was DNA Jard´ın Bot´anico Guinea-Real of data of collection Flora and and Locality (YA) 1. duplicates Table Cameroon Voucher of (PH). Herbarium Herbarium Innovation, Philadelphia National and (MA). the the of in Madrid Research samples deposited in Scientific are the deposited of specimens on were Voucher Ministry data 1. the collection Table Guinea in and from de given Nacional Locality and Universidad Cameroon. from Guinea, of permits 1975). Equatorial Republic with thousands Mabberley 2015 of with 1998; of Republic capsule March Palmer and a Ecuatorial, & February is (Knox in the fruit wind conducted as The by was such Fieldwork 2009). dispersed birds al. 1975) et attracts (Mabberley appro- (Givnish that seeds live bee-pollinated winged and considered of sugared season are produce flowers dry flowers The the The sects. in 2012). ( present al. sunbird are et Orange-tufted flowers (Bartos protandrous days ten The ximately panicle. multi-terminal or terminal 2015). columnaris al. by Lobelia sequencing et disturbed and (Ineich highly extraction been Cameroon DNA has Mt. plants, and on Study forest Gvoˇzd´ık except montane & activities, the (Zimkus 2013). human within Pleistocene and bands to fire, elevational grazing, Pliocene in like arranged the volcanoes is during southern vegetation origins Mainland The (2260m), with 2016). highlands youngest al. Bamenda-Banso et the and (Missoup are Oku Miocene) the Cameroon Mt. to with of Mt. as (Oligocene peak orography such Cenozoic high the Central, the represent another North during Mountains is uplifted the Bambutos Oku from and Mount Mountains Plateau northeast, region. Bamenda the North Likewise, In 3011m. Africa. of Central elevation West an of (4085m) volcano highest the aeon31 t k 2407 1798 1723 1977 2197 Bamenda Bamenda Mt. Mt. Oku Mt. 2381 2522 Oku Mt. Oku Mt. Oku Oku Mt. Mt. (2) 3118 1539 1157 3117 Cameroon Biao-Moka 1582 Biao-Moka 3116 Pico Pico Cameroon Biao-Moka 3114 1001 Pico 2836 2322 Cameroon 3113 (2) Cameroon Caldera 3112 Gran 2947 Cameroon Basil´e Basil´e 3111 Pico Pico Cameroon (2) 3110 Cameroon Basil´e (m) Pico Elevation 3109 Bioko South 3108 Bioko South (2) 3107 Bioko South (2) Island Bioko 3106 Sky South Bioko 3105 ind) North (# Population (2) Bioko 3104 North Bioko North Region sasmlaoshr rsrbu o1m ihcre upecrla raie na on organized corollas purple curved with m, 1 to up or herb semelparous a is inrsbouvieri Cinnyris oei columnaris Lobelia Jncke l 02 igr ta.21)adHmnpeain- Hymenoptera and 2011) al. et Riegert 2012; al. et (Janecek ) .columnaris L. 3 eenwysqecdadasmld Eleven assembled. and sequenced newly were .columnaris L. .columnaris L. sdi hssuyare study this in used sn modified a using Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. Mdletvrin211 Driae l 02 uno ace 03.Mro hi ot Carlo Monte each Chain and Markov runs, simultaneous two 2003). with Gascuel generations, & 10,000,000 Bull Guindon for & MrBayes 2012; using (Hillis in with (BIC) criterion 2003), run al. supported from information Huelsenbeck were well Bayesian et calculated & analyses the as (Darriba were (Ronquist by (MCMC) partition considered 3.2.6 2.1.10 clade gene version version is and each MrBayes for 70 jModeltest node using selected the performed models proportion [?] were substitution for analyses the DNA (BS) values BI and support Support The sites used Bootstrap data. 1993). analyses among replicates. the These heterogeneity from 2008). bootstrap rate al. estimated 1000 gamma-distributed et estimation sites (Stamatakis with bootstrap invariable 8.2.10 and model of version analyses substitution RAxML tree -cons with ML GTR 0.001 phylogenetic conducted The the reconstruct -st were analyses. to 0.4 from support used “-gt excluded clade were by were of Only methods 2009) positions (BI) nucleotide al. 2016). Inference including (Capella-Guti´errez Bayesian No et one trimAl and al. trees. prepared, (ML) using were et Likelihood matrixes (Yamada gaps Two Maximum removed 7.1 40”. analyses. another version phylogenetic and the MAFFT in gaps, using used all was aligned region was repeat matrix inverted one genome 2012). chloroplast al. exported whole CpGAVAS et The sequence using (Kearse final annotated 9.0 version the automatically Geneious was and using analyses genome viewed Phylogenetic adjusted chloroplast was then The graph 2012), “slim” the 2015). Brujin al. using De et filtered al. were (Liu filtered et files The (Wick using “fastg” final Bandage 3.9 the toolkit. using version contigs, SPAdes chloroplast GetOrganelle pure the of retain To using script assembled 2012). were al. et reads and (Bankevich filtered filtered The were reads 2020). paired-end raw genomes The chloroplast the of annotation of and (22 Assembling plastomes 44 included dataset total of The plastomes of distribution Geographic 1. Figure . 2017) Li & (Knox GenBank from obtained MF061187) 2085, (Muasya Cameroon Mt. of from sequences plastome more .columnaris L. rmSuhBoo(ee ee 13 F618 n another and MF061188) 3103, Perez (Perez Bioko South from enovo de .columnaris L. oei columnaris. Lobelia 4 sebe sn h eOgnletokt(i tal. et (Jin toolkit GetOrganelle the using assembled n 2fo te apnlca taxa). Campanulaceae other from 22 and k-mer 1 3 5 5ad85 and 65 45, 33, 21, Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. iooa N rN)gns h Rrgoshd1 D,svntN,adfu RAgenes. rRNA four 4 and and tRNA, 117 tRNA, seven contained 30 CDS, plastome 14 (CDS), The had 37,028 regions %. from regions 39.2 protein-coding IRs IR was and predicted The plastome bp, three whole 8025 genes. to the and (rRNA) bp of recognized RNA 7,950 content ribosomal 80 from GC SSC including The bp, genes, 164,609 2). 82,904 from to (Figure unique varied bp bp of 82,421 size 37636 from plastome to LSC complete bp with The bp, regions. 165,368 (IR) to repeated bp inverted two and (SSC), copy single of plastome The 1.4.2 version FigTree plastomes using of visualized Characteristics was tree MCC the The that Results by indicating TreeAnnotator heights”. 200, using using “Node than evaluated generated the more was was first were for tree analysis (http://tree.bio.ed.ac.uk/software/figtree/). The parameters (MCC) BEAST heights” credibility all the “Mean clade generations. of of setting Maximum (ESS) 20,000 output sizes confident. every Log were sample sampled estimations Effective Burnin”. and “Pre 1.6. generations as version Tracer removed 200,000,000 were ran generations MCMC 5,000 dataset, history phylogeographical each the reconstructing For For 2014). Knox 2016; of al. The et (1) (Chen 0.0) of 2016). calibrations offset: for al. 2.0, nodes (sigma et crown Mya (Matschiner Normal” two package Log selected CladeAge Clock we “Relaxed the Meanwhile, columnaris model”, using speciation. “Site data for for published Model” test” from “Yule The and model (http://www.phylo.org). model”, “BEAST 2.4 Gateway chose “Clock version Science we for CIPRES BEAST BEAUti, the in at in methods performed parameters (MCMC) was setting Carlo which 2014), Monte Chain al. Markov well et (Bouckaert using as conducted analyses, were BI analyses and history Dating phylogeographical ML and Both (http://www.phylo.org). a times supported Gateway 2007). generate divergent Science well Thornton to of CIPRES as & used Estimation Kolaczkowski the considered were 2003; at were trees performed al. was 0.95 remaining Potential were et the datasets jModeltest, [?] Erixon and and the as (PP) 2003; burn-in, 0.005, for values as al. than probability 1992) discarded et Posterior lower were Rubin (Alfaro because trees was & tree. and sufficient the datasets consensus (Gelman of tree was majority-rule the Diagnostic 25% datasets random Convergence first for three a The of frequencies the with 1.00. (PSRF) for split started Factor generations of were Reduction of deviation analyses Scale number BI standard The average The the generations. chains. 1000 heated every incrementally sampled four comprising run .columnaris L. .columnaris L. ld a . y sga10 ffe:00,ad(2) and 0.0), offset: 1.0, (sigma Mya 8.0 was clade .columnaris L. lsoepyoeyo the of phylogeny plastome a , n v ugop a siae nBAT ih“es-lsi akg”o BEAUti. of package” “beast-classic with BEAST2 in estimated was outgroups five and scrua n udiatt yhvn ag igecp LC,asmall a (LSC), copy single large a having by quadripartite and circular is .tuiin .columnaris L. – thuliniana L. 5 Lobelia aioa- laxiflora .columnaris L. ld nldn 2samples 22 including clade oei hlnaa–L. – thuliniana Lobelia ld a 21 was clade Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. ihNrhBoo(ioBsle itrt hs lds h ld fsuhr ik Pc ioMk and Biao-Moka (Pico Bioko 3). (Figure southern subclades of previous Oku) clade the The Mt. to and sister clades. Bamenda, and A these distinct Cameroon, tree. to is phylogenetic Basil´e) (Mt. sister Caldera) the clades (Pico Gran of in Cameroon Bioko clades populations mainland North supported the the highly with relates identical among three closely generated relationship distinguished ancestor (BI) we monophyletic common However, inference a Bayesian Cameroon. recovered and and We Bioko (ML) likelihood topologies. maximum tree using results phylogenetic Our of plastome relationships Phylogenetic sequenced newly of Map 2. Figure oei columnaris Lobelia 6 . .columnaris L. from Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. fmiln aeonaemr lsl eae.Ec i rp hw h rprino lsoesequences islands plastome sky of three proportion the 4). the in (Figure shows system populations graph contrast, pie island Each In sky populations geographic study. related. the each closely this between more from in relationship are populations distant Cameroon other the mainland the shows of and genomes Bioko plastid South the from of distribution geographic The history phylogeographical the of Reconstruction of tree Phylogenomic 3. Figure oei columnaris Lobelia . 7 Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. fiaadNrhBoofo ot ik.Fnly h hr vn setmtdt e09 adisplaying Ma 0.99 be to estimated is event populations. Central West third Cameroon in mainland the populations Finally, and of Bioko Bioko. separation North South the of marked split from and the Bioko Ma North 1.54 Ma, and approximately 5.58 Africa occurred dated of is second event ancestor The first the The 5). between tree. divergence genomic-dating the the on marking noticeable are events divergent important Three times divergence of Events of plastomes of columnaris. Lobelia relationship phylogenomic and Geographic 4. Figure .columnaris L. 8 n in oeisfo atArc (Figure Africa East from lobelias giant and Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. nmiln aeonatretbihn ot ik’ one k sad.Mroe,teSuhBioko South the Moreover, islands. Their sky Cameroon). progression. younger clear Bioko’s Mt. a Bioko. South and have North establishing (Bioko studied predated after mountains we colonization young that Cameroon that to to indicated 2009). mainland islands east interpretation Cameroon) sky in from al. (West-Central phylogeographic six age et old chronological plastome the simple (Givnish from Nevertheless, Our col- a lobeliads run their have 2008). Hawaiian ages and not the estimated islands al. does as the archipelago et such of sky (Suh Africa age lineages, west West-Central between plants’ the correlation various contrast, the of In for radiation documented and well onization is archipelago Cameroon Hawaiian in The clade third The age island’s ancestor. Sky Bioko. common North the recent Remarkably, in more populations islands. colonization a with sky Historical ancestor share six common not the recent do on more Bioko distributed a clades in shares three located with clades second pattern populations two phylogeographic A two island a the Bioko’s of found Bioko. result, history We this South evolutionary to in the According that results event complex. suggesting our Cameroon. colonization more group However, and monophyletic early is Bioko a an ancestor. not North shows are common of populations clearly same populations outcome the the Our separated shared divergence Bioko hypothesis. on phylogenetic the robust populations a refute the show that to these marker hypothesized because right We possibly the scale. of probabilities, be fine posterior not populations this may or among at values plastome resolution the relationships bootstrap and high genealogical recently, have diverged The not populations do 2017). subclade Bioko’s Li South & (Knox Ma columnaris 5.5 approximately Africa of estimation of ancestor and The relationships phylogenomic for intervals confident Discussion HPD BEAST. 95% in with obtained tree times divergence MCC 5. Figure ntepeetdywr eovdwt h nlsso lsoe.Hwvr ouain within populations However, plastomes. of analyses the with resolved were day present the in hlgntcrltosi fpopulations of relationship Phylogenetic .columnaris L. pedfo atArc i h og’ ai oetbihi West-Central in establish to Basin Congo’s the via Africa East from spread 9 .columnaris L. ooie h le k islands sky older the colonized L. Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. rsneo oetbigsta once k sad n ihad Kbd ta.20)ad saresult, 2017). Gvoˇzd´ık a & al. as (Zimkus 2013). mountains et and, adjacent (Mairal 2007) in elevations isolated al. be lowland et to through and the (Kebede disperse forest like highlands to patterns 2011) Afromontane and species vegetation al. allowed the islands dynamic past et of This sky (Kadu the expansions connected processes changed periodic that historical Africa fluctuations facilitated reconstruct bridges Central climatic to West forest that possible of in known is presence Pleistocene is analysis the it during However, With dramatically. vegetation known. Afromontane well altitude, the not this of are At meadows. scenarios >3000m. subalpine paleoclimatic to and The up for elevation of Through 5000m habitat middle composed likely to from is 2500m. most 3000 range habitat the to from its the are 1000 elevations extends elevations Afroalpine population from middle Bioko) inhospitable that ranges Basil´e’s (North found the elevation We colonized at 1969). taxa forests (Hedberg some Afromontane adaptations, in tolerant evolved frost lobelias giant Eastern provide contraction will and Nigeria expansion from forest samples Afromontane with history. analyses colonization Future of populations. interpretation mainland better and a island the between links of populations Nigerian of lack The respon- between are correlation links forces Missing the evolutionary Bioko. which explore in find to differentiation and population analyses populations for genetic of islands sible differentiation population sky genetic with and old developed distance Cameroon’s geographic be mainland should from Bioko. explanation likely of in This was events islands sky Further- history colonization young colonization independent groups. southern two plastome the subsequent the other suggest or scenario, to the 3) bottleneck this to (Figure genetic Under relative and distinct splits island. very event early are founder two plastomes a the Bioko via South more, in occurred that split so possibly early drift Bioko The genetic South Africa. Central and of West mainland history of colonization the islands the sky for Afromontane explanation the alternative in an caused be could populations processes of Microevolutionary islands. extinction sky local the the on for habitat differentiation possibly Nigeria, Afromontane Population from Cameroon, of data in transformation population human populations have intense more not plastomes the find do by the we not Furthermore, because did trees populations. we ML/BI/Beast of per and the history individuals on biogeographic 1-2 the only position of sequenced basal view a skewed in a are provide populations Bioko South Perhaps, Line Volcanic Cameroon the suggest of results islands islands. Our sky sky six 2). Afromontane (Table of the Ma) age of 1.3 estimated age (ca. and the Elevation islands concerning sky 2. progression Bioko’s Table estimated inverse than with an older islands with are sky Ma history Cameroon’s 31.0 biogeography because and a hypothesis 22.0, age 3.0, island of sky ages our contradicts result this Overall, t k 013-2She l,2008 al., et Suh 2008 2013 al., Gvozdik, et & Suh 2016 Zimkus al., et Yamgouot 31-22 2016 al., et Yamgouot 2016 al., 22-21 et Yamgouot 3 1.3 3011 1.3 1.3 2621 Oku Mt. 4085 3011 Bamenda Mt. Reference Cameroon 2260 Mt. (Myr) Age Basil´e Pico (m) Elevation Caldera 2009 Gran Biao-Moka Pico Island Sky .columnaris L. ntepyoeoi eosrcinlkl scuigmissing causing is likely reconstruction phylogenomic the in 10 .columnaris L. rteei apigerrbcuewe because error sampling is there or , .columnaris L. .columnaris L. .columnaris L. ouain between populations .columnaris L. nyPico Only . nthe on Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. uprstehgetnmeso ln n nmlseiso h sad(e oe 1994). Jones (see island the on islands species and sky density animal three population and the human of low of two of numbers in and because highest procedure lobelia giant conservation the undisturbed this active is supports for an Bioko Bioko Cameroon, has South in Bioko Mt. Moreover, scenario P´ocs on better. (M¨uller 2007). The & is populations plans. forest protection the conservation Afromontane for have the Park, Only National fragments. a forest is montane which on pressure anthropogenic excessive height, of greater populations with colour. The spectacular length. flower is leaf variation and and the leaves, phenotypic 2600m, colour, inflorescence, to number, observed the 2000 inflorescence we of between Elevations size Indeed, grow (1000m). height, gradients. elevation populations plant lower elevational that in observed and example, study latitudinal, for Our columnaris of traits, habitats, studies. morphology some ecological future the in in of on variation developed effect mosaic be possible a to a in has have observation may This habitat 1974b). ecological of type among The flow gene and dispersal conservation facilitating and and high Ecology 2000). islands, Cameroon, al. In sky et islands. connecting sky (Smith corridor across forest populations latitudinally natural contemporaneous and a altitudinally as seeds act tiny ridges dispersing in role essential of an recolonization and the colonization, between expansion, isolation et The reproductive (Schabetsberger maintains Bioko probably South that and barrier of North geographic between populations another masl) dispersal exemplifies (0-500 recent 2004) forest of Bioko lowland al. evidence separates a with have Cameroon. channel not when depression mainland do central shallow ago) We and The years A 2004). Bioko 10,000 1994). al. North (ca. et (Jones between (Schabetsberger period mainland flow Additional km glacial African gene 32 landscape. last the and by the the coast from of Cameroon Bioko changed the end isolated radically from the level at Pleistocene sea region a in the the rise be during in might occurred occurred changes Bioko that landscape contraction South forest in The Biao-Moka Pico However, Africa. barriers species. Geographical Central plant Afromontane West to of refuge forest islands of Pleistocene populations sky the the that environmental between the possible Once is refugia. It 2007). in al. periods, as drought et such (Assefa interglacial and 2017) Africa the temperature al. East-Central of during et (Pineiro oscillations changed are trees Pleistocene conditions refugia different Forest the in flora. survived detected distinctive diversity (L.) its genetic for high refuge genus the forest the of Pleistocene because a considered supported view is also holistic line a Cameroon entire in The considered be to have and lobelia. use refugia giant forest land Pleistocene this Afromontane and of volcanism of distribution active current isolation study, the and the of in of retraction explored disjunction of not and current product Although niches. warm Pleistocene the empty during to a Therefore, expanded dispersal was 2015). post-Pleistocene populations probably continuous al. Later, islands et refugia. sky forest (Gao fluctuations in environmental into alternations Also, populations environmental fragmentation. of humid Afromontane constriction and the oscillations caused climatic repeated by caused scenario, this Under Greenwayodendron ssalri egt noecne n eflnt tahge lvto apo.30m and 3000m) (approx. elevation higher a at length leaf and inflorescence, height, in smaller is .columnaris L. .columnaris L. .columnaris L. nBooIln.Tecnrldpeso ossso oln oetvegetation. forest lowland of consists depression central The Island. Bioko in Mgir ta.21) n te oeigpat.Frexample, For plants. flowering other and 2018), al. et (Migliore nmiln aeonaea ihrs flcletrainbcueof because extirpation local of risk high at are Cameroon mainland in rbbyhdcagsi ouainsz uigtePesoee mainly Pleistocene, the during size population in changes had probably .columnaris L. .columnaris L. .alpina A. 11 eednmclyioae n xaddwti and within expanded and isolated dynamically were ooie rrclnzdnwsyilnsin islands sky new recolonized or colonized a osbeb iddsesl idplayed Wind dispersal. wind by possible was .columnaris L. .columnaris L. rbsalpina Arabis (Mabberley L. Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. ael-uierz . il-atıe,J . aado,T 20) rml olfrauto- for tool a trimAl: analyses. phylogenetic (2009). large-scale T. Gabald´on, in & trimming M., alignment J. Silla-Mart´ınez, mated A. Drummond, analysis. . S., . evolutionary A.,. Capella-Guti´errez, bayesian M. for Suchard, patform D., software Xie, a C-H., 10 2: Wu, BEAST T., Vaughan, (2014). other K¨uhnert, D., J. six J., with Heled, R. comparison R., Tropek, A Bouckaert, & sakeriana: J., Kantorova, Impatiens sequencing. plant M., Pesata, sunbird-pollinated single-cell species. J., the to co-flowering Altman, of E., applications properties Patacova, Nectar its E., Padysakova, (2012). and S., Janecek, algorithm Pevzner, M., assembly . Bartos, . . genome 19 M., coloniza- Biology, V. new Lesin, Computational S., A Pleistocene of A. alpina. Kulikov, SPAdes: Journal M., (2007). Dvorkin, (2012). Arabis A., C. A. A. Gurevich, arctic-alpine P. D., Brochmann, Antipov, the S., & Nurk, A., by Bankevich, S., islands’ Nemomissa, the ’sky P., comparing afro-alpine doi:10.1038/sj.hdy.6800974 Taberlet, study simulation of D., A phylogenetic Ehrich, tion assessing bootstrap? A., in or bootstrapping Bayes Assefa, and (2003). sampling F. Carlo Monte Lutzoni, chain confidence. & Markov Bayesian S., of Zoller, insta- performance Lithospheric E., tomography. (2015). velocity M. R. phase Tibi, wave Alfaro, & Rayleigh Earth,120 J., from P. Solid Evidence Research: Shore, line: Geophysical G., volcanic G. of Cameroon Euler, the Journal of A., source A. the Nyblade, and A, bility D. Wiens, N., A. Adams, References GeneBank in deposited be with accessibility patches Data forest Afromontane the threatening isolate further are grazing, Moreover, to Cameroon. Bioko-Cameroon). deforestation barrier North geographic versus and Bioko a (South as burning, analysis functions our likely in discovered depression groups of central population forest islands. Bioko’s history was sky times, biogeographic recent Bioko our the more South of In Likely, that ages clades. show of three we progression led into Here, the oscillations populations follow periods. climatic Bioko not interglacial Pleistocene and the The Cameroon uplift. during the volcanoes’ refugia of youngest divergence line’s of Cameroon the the ancestor to during the Ma) calibration, 1.5 phylogenomic (ca. recon- our colonization to partial According but reconstruction phylogenetic informative of an struction provided analysis analysis plastome the sequences. with robust The genomic history nuclear more partial include a a should presents allow analysis study and robust Conclusions this more Finally, to effort Park. A challenging sampling National marker. was the non-recombinant the it in a expand because work of under-collected will to were permits populations Cameroon the more Mt. get samples. collect from individual Populations and the study. all find phylogeographic with of to studies samples Nigeria no future sizes reason in sample conduct main available to (1-2) the The was individuals hope studies. instability We future few Social for individuals. with baseline ten conducted a is was considered population be study should every preliminary such for This as and factors. population, several each by for limited are results Our limits Study 1057 doi:10.1371/journal.pcbi.1003537 e1003537. , oeua ilg n vlto,20 Evolution, and Biology Molecular oei columnaris Lobelia ot fia ora fBotany of Journal African South eaewrigtedt trg rmtepatmsgnrtdi hssuy aawill Data study. this in generated plastomes the from storage data the working are We vrl,tregop r nWs-eta Africa. West-Central in are groups three Overall, . 5–7.doi:10.1089/cmb.2012.0021 455–477. , 5–6.doi:10.1093/molbev/msg028 255–266. , 7812.doi:10.1002/2014jb011580 1708–1727. , .columnaris L. 12 , 78 37.doi:10.1016/j.sajb.2011.05.015 63–74. , .columnaris L. eecletdi iei.Ftr fieldwork Future Nigeria. in collected were iifrais 25 Bioinformatics, eeiy(dn) 99 (Edinb), Heredity rie nWs eta Africa Central West in arrived LSCmuainlBiology, Computational PLOS .columnaris L. .columnaris L. 1972–1973. , nmainland in 133–142. , does Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. uno,S,&Gsul .(03.Asml,fs,adacrt loih oetmt ag hlgne by phylogenies large estimate to algorithm accurate and fast, simple, A (2003). likelihood. Campanu- O. maximum (: Gascuel, Sytsma, . lobeliads & . . Hawaiian S., M., the J. Guindon, Henss, of L., diversification A. Hipp, and J., radiation T. Theim, adaptive B., laceae). Origin, T. Paterson, (2009). R., A. J. Mast, K. C., sequences. K. multiple Millam, J., using T. simulation Givnish, iterative from expansion Inference (1992). demographic B. 7 glaciations, D. Science, Pleistocene Rubin, & (2015). A., Rosa M. Gelman, alpine Z. the of Zhu, study & phylogeographic (Rosaceae). A F., complex heterogeneity: X. sericea morphological Gao, promoted isolation Y., subsequent of Zhang, and islands. measure D., poste- Guinea a Y. of as Gao, Gulf Bayesian the Island in Bioko angiosperms of on of 3 enemism markets Conservation, and Reliability Bushmeat Diversity (1994). (2000). E. (2003). R. Figueiredo, Castelo, B. & phylogenetics. E., Oxelman, J. pressure. hunting in Garcia-Yuste, E., & frequencies J. Fa, T., bootstrap Britton, and B., doi:10.1080/10635150390235485 probabilities Svennblad, (2015). rior hypothesis. African P., J. in refuge O. changes Hardy, Erixon, forest demographic & the and J-L., supports fragmentation Doucet, species population F., doi:10.1111/jbi.12510 tree past Walmacq, of C., forest tissue. leaf dating Doumenge, rain fresh P., molecular Dauby, of Mardulyn, Pleistocene quantities J., small S., Late for Mona, J. procedure J., isolation DNA Wieringa, Duminil, rapid A I., (1987). that 19 L. D. reveals J. Bulletin, trees Doyle, Phytochemical Ojeda, & legume J., K., forest J. line. Doyle, rain volcanic F. African Cameroon two Monthe, the in from M., phylogeography doi:10.1016/j.ympev.2020.106854 originate Plastome Heuertz, populations (2020). Gap J., J. Dahomey endan- O. Migliore, an Hardy, africana, . . B., Prunus G.,. tree B. Afromontane the Demenou, in variation Genetic and (1999). species. heuristics W. medicinal new Powell, gered models, & more K., 2: I. jModelTest Dawson, (2012). D. Posada, R., Doallo, computing. promotes L., parallel divergence Zosteropidae). G. Niche Taboada, (Aves: (2014). D., white-eyes J. Darriba, J. island Day, & sky A., African B. doi:10.1111/mec.12840 Amakobe, East 4103–4118. giant 2015. C., of a J. species of Habel, diversification P., divergence threatened rapid R. ancient Prys-Jones, and of C., Lobelioideae S. Asian list Cox, East red Bhutan. (2016). Himalayan S. IUCN in S. Lobelia Renner, The & rosette columnaris. Q-F., Wang, L-Y., Lobelia Chen, (2015). M. Thulin, Came- doi:10.2305/IUCN.UK.2015-4.RLTS.T39505A2928168.en & Mount I. in M., flows Ngounouno, lava . . basaltic Cheek, M.,. of Gerard, compositions P., chemical Ildefonse, the S., affect Durand, rainfall F., roon? tropical Chabaux, of M., decades Bulourde, Do (2005). N., A. Dia, C., Chauvel, doi:10.1093/bioinformatics/btp348 ora fVlaooyadGohra eerh 141 Research, Geothermal and Volcanology of Journal rceig fteRylSceyB ilgclSine,276 Sciences, Biological B: Society Royal the of Proceedings 5–7.doi:10.1214/ss/1177011136 457–472. , 785–793. , osrainBooy 14 Biology, Conservation aueMtos 9 Methods, Nature ytmtcBooy 52 Biology, Systematic oeua clg 8 Ecology Molecular 11–15. , cetfi eot,5 Reports, Scientific ao,65 Taxon, 7.di 10.1038/nmeth.2109 doi: 772. , 6211.di 10.1111/j.1523-1739.2000.99067.x doi: 1602–1613. , 9–0.Didoi:10.1080/10635150390235520 Doi 696–704. , 5–5.di 10.1046/j.1365-294X.1999.00518.x doi: 151–156. , 9–0.doi:10.12705/652.6 293–304. , 19.doi:10.1038/srep11698 11698. , 13 9–2.doi:10.1016/j.jvolgeores.2004.10.008 195–223. , ora fBoegah,42 Biogeography, of Journal 0–1.doi:10.1098/rspb.2008.1204 407–416. , ytmtcBooy 52 Biology, Systematic o hlgntEo,150 Evol, Phylogenet Mol oeua clg,23 Ecology, Molecular idvriyand Biodiversity 1443–1454. , 665–673. , Statistical 106854. , , Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. abre,D .(94) rnhn nPcyalSnco:TeDra hoyadteeouino angio- of evolution the and Theory Durian The herbs. Senecios: and chloroplast Pachycaul trees in sequenced Branching spermous completely server (1974a). web J. of integrated D. an submission CpGAVAS, Mabberley, (2012). GenBank X. pos- and Guan, Bayesian X., analysis, Lin, sequences. on J., visualization, genome Zhang, annotation, uncertainty H., Chen, the length Y., for Zhu, branch L., Shi, of C., Liu, hypotheses. Effects (2007). phylogenetic W. for doi:10.1093/molbev/msm141 J. probabilities Lobelias Thornton, giant of terior radiation & and origin B., the Kolaczkowski, on evidence DNA Chloroplast (1998). Africa. lobelias. D. eastern giant J. in Palmer, the & of B., origin E. Knox, Asian unique East is The (2017). lato doi:10.3732/ajb.1700025 C. sensu 924–938. Li, Campanulaceae B., the E. in Knox, genomes plastid of history 11097–11102. dynamic The angiosperms. (2014). among B. E. mountains. African Knox, East Tropical and Ethiopian in conserva- and giberroa) Phylogeography 16 (Lobelia (2007). Ecology, lobelia C. and giant Brochmann, a organization & of S., genetics the Nemomissa, tion P., for Taberlet, D., A. platform Ehrich, Drummond, M., software . Kebede, . desktop S.,. Buxton, extendable data. S., and sequence Sturrock, integrated M., of an Cheung, analysis S., Basic: Stones-Havas, Geneious A., V. Wilson, (2012). R., Williams, Moir, migration A., M., former Muchugi, Kearse, a highlands. O., reveals Eyog-Matig, africana African Prunus M., West Afromontane M. and the 294X.2010.04931.x G. of East Muluvi, Phylogeography between H., (2011). corridor T. Konrad, Geburek, S., . overview. . Schueler, . An C., L., Guinea: A. of C. Kadu, Gulf the in GetOrganelle: Biodiversity (2020). (1994). 772–784. D-Z. genomes. T. Li, organelle P. & of T-S., Jones, assembly Yi, novo de W., accurate C. for dePamphilis, toolkit Y., doi:10.1186/s13059-020-02154-5 versatile from Song, and J-B., geckos fast Yang, estimated a Hemidactylus W-B., introductions Yu, of anthropogenic J-J., Jin, relationships and colonizations Phylogenetic sequences. natural (2005). DNA of J. nuclear doi:10.1016/j.ympev.2004.11.006 patterns and D. islands: mitochondrial Harris, Guinea from & of 2012. Gulf A., J. the Dolezal, Brehm, . . Africa. E.,. West J., Padysakova, in interactions J., Jesus, visitor Society plant-flower Reif, Linnean of D., the aspect Horak, important of an M., Journal visitors: Biological Bartos, floral Mount avian O., by of Sedlacek, selection summits Food J., the Riegert, of confidence reptiles S., The assessing Janecek, Cameroon.pdf. (2015). for Highlands, L. method Bamenda Chirio, a the & as and N., Lhermitte-Vallarino, bootstrapping Oku M., of Lebreton, test I., empirical Ineich, An (1993). J. analysis. J. phylogenetic flora. Bull, in mountain & M., high D. tropical Hillis, a in London, speciation of and Society Evolution Linnean (1969). O. Hedberg, 2314.di10.1111/J.1365-294x.2007.03232.X doi 1233–1243. , ytmtcBotany, Systematic M eois 13 Genomics, BMC rceig fteNtoa cdm fSine fteUie ttso America,111 of States United the of Sciences of Academy National the of Proceedings ytmtcBooy 42 Biology, Systematic iifrais 28 Bioinformatics, e htlgs,73 Phytologist, New ,1518 o:10.1111/j.1095-8312.1969.tb01816.x doi: 135–148. 1, 3 109–149. 23, 715. , 0:5–6.doi:10.1111/j.1095-8312.2012.01943.x 107:355–367. 6714.doi:10.1093/bioinformatics/bts199 1647–1649. , 967–975. , 8–9.doi:10.1093/sysbio/42.2.182 182–192. , mhba etl osrain 9 Conservation, Reptile & Amphibian oeua hlgnttc n vlto,34 Evolution, and Phylogenetetics Molecular 14 oeua ilg n vlto,24 Evolution, and Biology Molecular oeua clg,20 Ecology, Molecular mrcnJunlo oay 104 Botany, of Journal American idvriyadCnevto,3 Conservation, and Biodiversity 6–7.doi:10.1111/j.1365- 165–178. , eoeBiology, Genome ilgclJunlo the of Journal Biological 15–38. , 2108–2118. , ril 21. Article 480–485. , Molecular , , , Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. mt,T . odr . imn . ’ef,K,Lrsn . hn .(00.Cmaaieavian Comparative (2000). Y. Chan, conservation. & for (Equatorial B., implications Bioko Larison, of mountains: K., Guinea island O’Keefe, Equatorial Limmnologi- the 9 D., Ecology, and on Girman, Cameroon (2004). Loreto) K., of E. Lago Holder, phylogeography and Rott, B., Biao T. & (Lago Smith, D., lakes C. crater Jersabek, tropical two I., Guinea). of Drozdowski, aspects G., cal Drozdowski, models. mixed R., under inference Schabetsberger, phylogenetic Bayesian mountains. 3: MrBayes Cameroon (2003). the P. Bioinformatics,19 J. from Huelsenbeck, & study niche F., case Food Ronquist, A (2011). Peˇsata, M. species: & sunbird 152 J., Hoˇr´ak, syntopic Ornithology, Reif, Sedlaˇcek, O., two D., M., in Antczak, differentiation D., Fainova, in J., diversification Riegert, and Colonization (Caryophyllaceae). (2007). L. C. Lychnis Brochmann, shade- Eurasian doi:10.1111/j.1365-2699.2008.01902.x & by of J., islands’ Suda, expansions ‘sky S., African population the Nemomissa, A., Pleistocene Gizaw, Ages. M., Ice Popp, (2017). the during J. 284 rainforest Sciences, O. Biological African Hardy, the B: of Society E., fragmentation Kaymak, indicate trees G., tolerant Dauby, Pleistocene R., mouse. African West-Central -furred Pineiro, soft (2012). Missonne’s C. of Denys, phylogeography & the C., by conservation47 Cruaud, revealed for evolution M., hotspots forest Colyn, Biodiversity lowland A-D., (2000). Missoup, J. V., Kent, Nicolas, & B., A. G. Island Fonseca, Bioko A, priorities. of R. bryophytes Mittermeier, epiphyllous N., of Myers, species. knowledge non-epiphyllous the the on to of remarks doi:10.1179/174328207X186803 contribution position additional A 81–94. Phylogenetic including (2007). Guinea), (2016). T. (Equatorial Pocs, C. & Denys, morphological F., & and Muller, molecular K., on E. based Chung, Muridae), S., okuensis(Rodentia: data. Eiseb, rat,Lamottemys Oku V., Mount Nicolas, endemic East D., Greenway- from A. an insights (2018). New Missoup, J. in trees: O. forest Hardy forests rain & Pi˜neiro, history. R., J., African phylogenomics. B. in Pleistocene Lissambou, forest odendron(Annonaceae) breaks P., phylogeographical L. of track of T. origin Couvreur, fragmentation to C., Pre-Pleistocene Mariac, E., Sequential model Kaymak, J., (2011). a Migliore, A. as Chameleons K. Tolley, doi:10.1371/journal.pone.0026606 hotspot: of & biodiversity dispersal trans-Atlantic J., Africa R. supports Bouckaert, G. sampling & fossil Measey, M., of Steel, probabilities W., (2017). on Eastern Salzburger, based fishes. Starostov´a, M. the Z., cichlid dating I., Alarcon, in node M. variation & Bayesian J. genetic (2016). J., Barth, of Musilov´a, J. Z., M., patterns Aldasoro, Matschiner, P., shaped Vargas, change L., climate hotspot. Pokorny, biodiversity Pleistocene Afromontane A., and Herrero, barriers I., Geographic Sanmartin, M., Mairal, Drift. Continental and Ornithophily Biogeography, 74 Pachycauly, Helena. Phytologist, Lobelias: St. Giant New and The Africa (1975). J. of D. Lobelias Mabberley, Pachycaul The (1974b). J. D. Mabberley, 100–112. , olgclJunlo h ina oit,177 Society, Linnean the of Journal Zoological yrbooi,524 Hydrobiologia, Nature,403 5510.di 10.1046/j.1365-294x.2000.01032.x doi: 1505–1506. , bioRxiv 1–2.doi:10.1007/s10336-011-0650-0 819–825. , 5217.doi:10.1093/bioinformatics/btg180 1572–1574. , 365–374. , doi:10.1101/038455 . 5–5.di 10.1038/35002501 doi: 853–858. , 79–90. , cetfi Reports,7 Scientific doi:10.1098/rspb.2017.1800 . ora fBoegah,46 Biogeography, of Journal 0–2.doi:10.1111/zoj.12361 209–226. , 15 54.doi:10.1038/srep45749 45749. , ora fBiogeography,35 of Journal 212–223.doi:10.1111/jbi.13476 , e Bulletin Kew ora fBylg,29 Bryology, of Journal LSOe 6 One, PLoS rceig fteRoyal the of Proceedings , fia Zoology, African 29 1016–1029. , ,535–584. ora of Journal Molecular e26606. , , Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. iks .M,&Goz´k .(03.SyIlnso h aeonVlai ie iesfiainhtso for spot hot diversification a (Phrynobatrachidae:Phrynobatrachus). Line: Volcanic frogs Cameroon the interaction.puddle of Gvoˇzd´ık, plume-lithosphere Islands & for Sky M., (2013). Evidence B. V. Zimkus, Line”): Hot (“Cameroon Island 7 Bioko Geochemistry Frontiers, (2016). from Geoscience D. rocks Demaiffe, volcanic & program the I., trees. alignment Ngounouno, of B., sequence guide Gbambi´e D´eruelle, I. B., MAFFT Mbowou, N., chained F. the Yamgouot, of of usefulness Application novo the de (2016). of of K. doi:10.1093/bioinformatics/btw412 visualization Katoh, datareexamination interactive & large Bandage: K., to (2015). Tomii, E. D., highlands, K. K. Holt, Yamada, Bamenda & the J., Zobel, of B., assemblies. butterflies M. genome Schultz, Endemic R., R. Wick, eternal? Forest in forest. erupted (2009). close-canopy Scoriae 2028.2009.01129.x M. from avoid inclusions Konvicka, glass Cameroon, & Olivine-hosted R., (2008). Tropek, S. web M. RAxML doi:10.1016/J.Jvolgeores.2007.07.004 Africa. Njome, the West 1–33. & volcano, for , Cameroon F., algorithm Mount J. bootstrap at Luhr, 1954-2000 rapid E., A C. Suh, (2008). J. Rougemont, & P., servers. Hoover, A., Stamatakis, ytmtcBiology,57 Systematic iifrais 31 Bioinformatics, 4–5.doi:10.1016/j.gsf.2015.06.003 743–757. , 5–7.doi:10.1080/10635150802429642 758–771. , 3035.doi:10.1093/bioinformatics/btv383 3350–3352. , fia ora fEooy 48 Ecology, of Journal African 16 olgc Scripta Zoologica ora fVlaooyadGohra Research,169 Geothermal and Volcanology of Journal , 42 iifrais 32 Bioinformatics, 2–3.doi:10.1111/j.1365- 428–437. , 9–1.doi:10.1111/zsc.12029 591–611. , 3246–3251. , Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. 1.00 100 1.00 100 1.00 100 1.00 100 1.00 100 1.00 100 1.00 95 1.00 100 1.00 1.00 100 100 1.00 1.00 1.00 100 100 100 1.00 1.00 1.00 1.00 0.96 100 100 100 58 85 1.00 1.00 100 99 1.00 1.00 100 100 19 17 1.00 1.00 1.00 1.00 1.00 1.00 100 100 100 55 52 78 68 1.00 100 0.82 1.00 1.00 1.00 1.00 1.00 1.00 100 100 100 100 100 100 100 64 70 66 Lobelia jasionoidesKY354220 Lobelia laxifloraKY354221 Lobelia siphiliticaKY354225 Lobelia inflataKY354219 Lithotoma petraeaKY354215 Lobelia polyphyllaKY354224 Siphocampylus krauseanusKY045477 Centropogon nigricans KY045478 Burmeistera auriculataKY045495 Burmeistera resupinata KY045482 Lobelia boninensisKY354217 Cyanea fissaKY354213 Brighamia insignisKT372780 Delissea rhytidosperma KY354214 Lobelia stricklandiaeKY354226 Lobelia longisepalaKY354222 Lobelia morogoroensis KY354223 Lobelia thulinianaKY354227 Lobelia gregoriana KY354218 Lobelia mildbraediiK426 Lobelia giberroa K118 Lobelia stulmanniiK120 Lobelia columnarisPz3109VI3 Lobelia columnarisPz3107IV 2 Lobelia columnarisPz3103 Lobelia columnarisPz3110 VII1 Lobelia columnarisPz3110 VII3 Lobelia columnarisPz3108V 1 Lobelia columnarisPz3107IV 3 Lobelia columnarisPz3104I2 Lobelia columnarisPz3104I3 Lobelia columnarisPz3106III2 Lobelia columnarisPz3105II1 Lobelia columnarisPz3106III1 Lobelia columnarisPz3118 XIV 3 Lobelia columnarisPz3114 XII1 Lobelia columnarisPz3113 XI1 Lobelia columnarisPz3116 VIII Lobelia columnarisM2085 Lobelia columnarisPz3118 XIV 2 Lobelia columnarisPz3112 X2 Lobelia columnarisPz3111 IX1 Lobelia columnarisPz3112 X3 Lobelia columnarisPz3117 XIII1 Posted on Authorea 30 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161201731.10223625/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. 2 2 2 2 2 Oligocene 5 5 5 5 5 Mt. Cameron Kilum Bamenda N. Bioko S. Bioko E. Africa 20.83 2 2 2 2 2 0 0 0 0 0 18.6 14.88 12.56 14.76 1 1 1 1 1 Miocene 5 5 5 5 5 11.37 9.37 0 10.86 10.83 18 1 1 1 1 1 0 0 0 0 0 8.5 7.79 7.63 5.81 5.58 5.5 4.6 3.09 5 5 5 5 5 3.64 Pliocene 2.79 2.54 1.54 1.79 Pleistocene 0.99 0.48 Lobelia columnarisPz3112.X.3 Lobelia columnarisPz3111.IX.1 Lobelia columnarisPz3118.XIV.2 Lobelia columnarisPz3113.XI.1 Lobelia columnarisPz3104.I.3 Lobelia columnarisPz3108.V.1 Lobelia columnarisPz3107.IV.2 Lobelia columnarisPz3109.VI.3 Lobelia mildbraediiK426 Lobelia thulinianaKY354227 Lobelia columnarisPz3117.XIII.1 Lobelia columnarisPz3112.X.2 Lobelia columnarisPz3118.XIV.3 Lobelia columnarisPz3116.VIII Lobelia columnarisM2085 Lobelia columnarisPz3114XII.1 Lobelia columnarisPz3106.III.2 Lobelia columnarisPz3106.III.1 Lobelia columnarisPz3105.II.1 Lobelia columnarisPz3104.I.2 Lobelia columnarisPz3110.VII.1 Lobelia columnarisPz3107.IV.3 Lobelia columnarisPz3110.VII.3 Lobelia columnarisPz3103 Lobelia stulmanniiK120 Lobelia giberroa_K118 Lobelia gregorianaKY354218 0.61 0.44 0.35 0.33 0.23 0.25 0.19 0.15 0.05 0.06 0.09 0.08 0.04 0.15 0.05 0.02 0.01 0.1 0 0 0 0 0 Lobelia columnarisPz3117_XIII_1 Lobelia columnarisPz3112_X_3 Lobelia columnarisPz3112_X_2 Lobelia columnarisPz3111_IX_1 Lobelia columnarisPz3118_XIV_2 Lobelia columnarisPz3116_VIII Lobelia columnarisM2085 Lobelia columnarisPz3113_XI_1 Lobelia columnarisPz3114_XII_1 Lobelia columnarisPz3118_XIV_3 Lobelia columnarisPz3106_III_1 Lobelia columnarisPz3105_II_1 Lobelia columnarisPz3106_III_2 Lobelia columnarisPz3104_I_3 Lobelia columnarisPz3104_I_2 Lobelia columnarisPz3110_VII_1 Lobelia columnarisPz3107_IV_3 Lobelia columnarisPz3110_VII_3 Lobelia columnarisPz3108_V_1 Lobelia columnarisPz3107_IV_2 Lobelia columnarisPz3103 Lobelia columnarisPz3109_VI_3 Lobelia stulmanniiK120 Lobelia giberroaK118 Lobelia mildbraediiK426 Lobelia gregorianaKY354218 Lobelia thulinianaKY354227 Lobelia morogoroensisKY354223 Lobelia longisepalaKY354222 Lobelia stricklandiaeKY354226 Delissea rhytidospermaKY354214 Brighamia insignisKT372780 Cyanea fissaKY354213 Lobelia boninensisKY354217 Burmeistera resupinata_KY045482 Burmeistera auriculataKY045495 Centropogon nigricansKY045478 Siphocampylus krauseanusKY045477 Lobelia polyphyllaKY354224 Lithotoma petraeaKY354215 Lobelia siphiliticaKY354225 Lobelia inflataKY354219 Lobelia laxifloraKY354221 Lobelia jasionoidesKY354220 Holo. Mya