Posted on Authorea 6 Apr 2020 | CC BY 4.0 | https://doi.org/10.22541/au.158620585.55411740 | This a preprint and has not been peer reviewed. Data may be preliminary. etdb the by cepted genus the Kosovo subgenera two In sylvestris the to 2010). belonging subgenus (Govaerts subspecies), The two 22 and of to species number (six 1933) taxa (Hayek The 15 accepted. from are varying species et (Eker 76 researchers for only various listed to the been according have consideration, species literature 40-150 into of from that ranges Taking body Tulipa the and large In defined 2012). a 2014). exactly Groot, of al., not existence de is that, the and still of despite Zonneveld species Because difficult, 2009; 2012). be Zonneveld, of Groot, to 2014; classification de considered and al., even is Zonneveld et sometimes genus 2013; (Eker (polythetic), al., this characterize plastic et of to be (Christenhusz taxonomy used to species the traditionally appear a were characters of which charac- floral populations characters and monocots, within floral vegetative bulbous and the vegetative are Furthermore, of They variability species. 2012). high Zonneveld, a and by (Veldkamp terized importance taxonomic and vational genus the of ITS species. the Species independent than as trnL-trnF complex the scardica of T. resolution the Introduction serbica, lower of taxa T. the of scardica, Despite separation (T. the complex taxa. support scardica Tulipa, independent not T. and sequence as do the Eriostemones on luanica) loci of subgenera T. Based both species the PAUP. and dataset, distinguish to kosovarica in to belonging T. evidence conducted clades, albanica, genetic main were sufficient T. Kosovo two datasets not into in trnL-trnF) is grouped population from There + species wild trees Tulipa respectively. from (ITS from Likelihood of obtained Maximum collected combined sequences trnL-trnF), and taxa and our Parsimony 26 eight analyses, Maximum trnL-trnF) and from Neighbor-Joining, ITS and generated GenBank. (29 (ITS newly from sequences independent were obtained 55 were sequences total sequences one In 14 Forty and used. originated which analysed. were been species characters, have Tulipa were sequences sometimes of floral taxa DNA which relationships and taxa, 14 trnL-trnF phylogenetic eight vegetative and by the of represented ITS investigate variability is Kosovo, To Tulipa high from genus misclassified. the a or Kosovo identified, with In erroneously characterized difficult. synonymized, genus are this (Liliaceae) of taxonomy L. the Tulipa makes genus the of Species Abstract 2020 28, April 3 2 1 Mustafa Behxhet sequences and DNA nuclear Hajdari and Avni plastid on (Liliaceae) based Kosovo, species in Tulipa present wild of analysis phylogenetic Molecular hriNtoa Park National Vienna Sharri Medicine Veterinary of University Prishtina of University u ny12tx aebe cetd hl nthe in while accepted, been have taxa 102 only but , subsp. ol hcls fSlce ln Families Plant Selected of Checklist World Eriostemones 1 ldrPulaj Bledar , Tulipa australis ol hcls fSlce ln Families Plant Selected of Checklist World Tulipa Tulipa a enrvsdsvrltms(uka ta. 03.Tettlnme of number total The 2013). al., et (Turktas times several revised been has 1 Ln)Pm acpe us. and subsp.) (accepted Pamp (Link) .(iica)hv ra cnmc otclua,eteia,eooia,conser- ecological, esthetical, horticultural, economic, great have (Liliaceae) L. n 2 aahv enacpe.Acrigt Christenhusz to According accepted. been have taxa 120 and srpeetdby represented is 1 oin Schmiderer Corinna , .sylvestris T. Tulipa 1 pce aiet h aknPnnuai uhless, much is Peninsula Balkan the to native species Gvet,21) nKsv h subgenus the Kosovo In 2019). (Govaerts, 2 Gvet,21) 1 ae aebe itdfor listed been have names 516 2019), (Govaerts, hc srpeetdb w subspecies, two by represented is which , hvtMala Xhavit , ln List Plant uiasylvestris Tulipa Eriostemones “h ln it”21)49names 499 2013) List,” Plant (“The 3 Tulipa ieeLua-Rizani Lluga Kimete , subsp and srpeetdb eight by represented is sylvestris . Tulipa, tal et respectively. (2013) . Tulipa nyac- only Tulipa Tulipa 1 , is Posted on Authorea 6 Apr 2020 | CC BY 4.0 | https://doi.org/10.22541/au.158620585.55411740 | This a preprint and has not been peer reviewed. Data may be preliminary. ie a o h .SrN eeadnn o h 6 DAgn,respectively. gene, rDNA 26S the informative for C parsimony none + potentially and G of gene average number rDNA The The rDNA. 5.8S indels. 26S the without in for characters 76.9% 1 partial alignment and was The 26 60.8%, 57.0% sites was respectively. only was it rDNA sites, included ITS2 5.8S informative gene respectively. The for in parsimony ITS2, rDNA content while and potentially 26S 59.5%, 2). 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Additionally, and species, used. 2010) these been al., of has relationships distribution been geographical and and have Shuka their taxonomy sometimes 2010; the species al., study these et To similarities, (Shuka misclassified. their features or of morphological 2018). identified Because al., similar erroneously et synonymized, Krivoˇsej, many 1997). Tati´c (Millaku and share 2012; too serpentine Deva kosovarica and Kit, of on Tan 2013) T. village village, Kosovar al., albanica, Surroj the et T. in to (Christenhusz serbica, found Kolshi recently from T. Kosovo was (Kuk¨esi scardica, district: but Southern Albania Tulipa 2010), Prizren, from 2015). al., the of species et Elezaj, in district (Shuka new the and as slopes) a in (Millaku well as located Albania as described is with 2012), time which al. Pashtriku border Mt. et the Llap¸cev¨e on (Shuka near 2018). and substrate villages al., limestone Mrasori et on between Dev¨e (Millaku buted and Kozniku, Prush Krivoˇsej, 1997). Qaf¨e (Tati´c Mt. – Guri¸c, and Llapushnik, river) of localities Ibar foot near near the Rogozna hill, at Knjaˇzevac: Mt. Laz (community (Beli Serbia Kosovo of kosovarica Northern South and the is Kamenica) in the it as soil by Donja Kosovo, synonymized serpentine species is In a on It as distributed Krivoˇsej 1970). Macedonia. accepted is 2016), Micevski, of Zonneveld, border and 2013; the Families (Mayer List,” to Macedonia Plant close and (“The Krivenik, L. Kosovo village Southern the in near present distributed is Bornm. T from derived origin species: several by represented Tulipa Gvet,21) u o cetdb lr uee Ttne l 1980). al. et (Tutin Eurepea Flora by accepted not but 2019), (Govaerts, pce agdfo 4 o67b.Teaineto h nru nldd4 miuu positions, ambiguous 45 included ingroup the of alignment The bp. 657 to 644 from ranged species i a,Sua&Kanq sdsrbtdi ooo ntesretn rao iuh region Mirusha of area serpentine the in Kosovo, in distributed is Krasniqi & Shuka Tan, Kit Omn,21;Suae l,2010), al., et Shuka 2018; (Osmani, .luanica T. trnL-trnF trnL-trnF gnni,T rea .suaveolens T. armena, T. agenensis, . MlauadEea,21) swl sncergnm ie(N Cvle)for 2C-values) (DNA size genome nuclear as well as 2015), Elezaj, and (Millaku uiagesneriana Tulipa ein(eesne l,20)wr sdfrpyoeei analyses. phylogenetic for used were 2008) al., et (Peterson region n ula T ein otebs forkolde pt o ostudies no now to up knowledge, our of best the To region. ITS nuclear and Tulipa .scardica T. . p l:36(73 Mlaue l,21)hsahybrid a has 2018) al., et (Millaku (1753) 306 Pl.: Sp. L., 2 pce eereported. were species and Zneed 2009), (Zonneveld, uiaalbanica Tulipa .luanica T. n tes(oars 2019). (Govaerts, others and eogt the to belong ol hcls fSlce Plant Selected of Checklist World uialuanica Tulipa i a hk a h first the was Shuka & Tan Kit .kosovarica T. Tulipa Tulipa uiaserbica Tulipa .albanica, T. Thus, . .scardica T. pce fKosovo, of species uiagesneriana Tulipa ilk sdistri- is Millaku uiascardica Tulipa and Tulipa .luanica T. Suaet (Shuka complex ai & Tatic Tulipa DNA Posted on Authorea 6 Apr 2020 | CC BY 4.0 | https://doi.org/10.22541/au.158620585.55411740 | This a preprint and has not been peer reviewed. Data may be preliminary. yN 10 S,M 10 S n L(5) hl h eodcaeicue ebr ftesubgenus the of of samples australis members sequenced includes newly 1C clade the and second clade, the (MP), julia while 1B T. (95%), ML (NJ), suaveolens, and 1A BS) (100% Figure MP Tulipa in BS), (100% shown NJ are by subgenus analysed sequences the the of ITS that specimens show includes 29 trees on generated The based (ML). analyses phylogenetic The region ITS support. generated stronger those + a showed to ITS but combined compared sequences, the resolutions from better trees provided generated sequences ITS from from obtained trees and phylogenetic ITS (separated datasets different trnL–trnF the from generated trees Phylogenetic and ITS for characteristics analysis tree Phylogenetic was parsimony-based RI and 0.840, set was and Data CI the characters 2. 0.890. 44.5%, ambiguous.Table informative was was were potentially content index G+C 62 composite positions average the sites, The 91 and variable ingroup 2). 0.940, (Table 241 the indels sites, informative within conserved potentially positions, 179 1229 ambiguous included 93 alignment showed The outgroup the contentincluding + C ITS + combined G The the region. and this 0.897 for counted index were composite 0.928, for for RI bp 30.8% 0.769, 717 was The included informative were alignment CI potentially respectively) positions 2). bp, 47 (Table 51 58 characters, 31.2% was in-group informative was length the potentially tree within ten MP showed positions, indels, sequences ambiguous Analysed 86 contained ambiguous. outgroup) (including alignment The ihnthe Within . of sequences published already of with species sequenced newly hs on nteISsqecsaayi,bti eea ihlwrrslto.Tetesotie from obtained trees subgenus the The of members resolution. of consists lower clade with first The general too. in clades but major two Eriostemones analysis, into sequences divided are ITS sequences analysed the in found those from obtained data Plastid 64%). than trnL-trnF (less MP and 54%) 1. than Figure (less NJ in in supported not supported was weakly samples was sequenced newly the of of grouping specific Species except trees, (78%). all ML and (86%) MP by serbica T. scardica, T. luanica, T. of separation The suaveolens tschimganica .albanica T. trnL-trnF trnL-trnF ( .abnc,T ooaia .laia .sadc,T ebc,T lpyl,T shmaia T. tschimganica, T. ulophylla, T. serbica, T. scardica, T. luanica, T. kosovarica, T. albanica, T. eaae from separated trnL, and region aaes n sn ieetmtos(J PadM) otyrvae iia ooois The topologies. similar revealed mostly ML), and MP (NJ, methods different using and datasets) hlgntcte osrce rmISdtsto the of dataset ITS from constructed tree Phylogenetic Tulipa (section ( .gesneriana T. n n apeof sample one and .sylvestris T. eune of sequences eune.ISand ITS sequences. 45 for 44.5% .albanica T. .suaveolens T. ld,i l re h otdsic pce rmtenwygnrt eune was sequences generate newly the from species distinct most the trees all in clade, and trnL-trnF Spiranthera .sylvestris T. trnF .gesneriana T. hc a oeaeyspotdol nN 8%.Acaeicuigalsamples all including clade A (82%). NJ in only supported moderately was which trnL-trnF nldn ohsubspecies) both including .abnc,T ooaia .laia .scardica T. luanica, T. kosovarica, T. albanica, T. Tulipa trnL-trnF hc eetecoetrltdt el eune pce alsection (all species sequenced newly to related closest the were which , n 18 o h G,rsetvl.Tnptnilyifraiecharacters informative potentially Ten respectively. IGS, the for 11.8% and and eune agdfo 37t 40p nlnt.Tecmlt alignment complete The length. in pb 1430 to 1377 from ranged sequences ngnrlwssrnl upre yN 9% n oeaeysupported moderately and (97%) NJ by supported strongly was general in ) .kosovarica T. Eriostemones pce ntedtstrne rm70t 7 pi egh h complete The length. in bp 775 to 720 from ranged dataset the in species ,floe by followed ), .uohla .tcignc,T uvoes .julia, T. suaveolens, T. tschimganica, T. ulophylla, T. trnL-trnF .gesneriana T. subsp eune hw nFgr A(J,2 M)ad2 M)resemble (ML) 2C and (MP) 2B (NJ), 2A Figure in shown sequences .sylvestris T. ( trnL 0%spotdb l he aclto methods calculation three all by supported 100% ) trnL–trnF sylvestris . trnL, 3-9 bp, 631-692 eune eecnret(L etrvae .2,while 0.82), = p revealed test (ILD congruent were sequences ( (T6), .sylvestris T. 4b for bp 64 .jla .ulophylla T. julia, T. rmnwysqecdseis( species sequenced newly from Tulipa , 3 aae eemr iia otesotie rmITS from obtained trees to similar more were dataset .sylvestris T. , rvddfo h eBn.I h eodcae the clade, second the In GenBank. the from provided .luanica T. hc smdrtl upre yN 8% n ML and (86%) NJ by supported moderately is which trnF aadvddit w ancae.Tefis clade first The clades. main two into divided taxa trnF nldn ohsbpce)srnl supported strongly subspecies) both including 76 padIS2 pfrec sequence, each for bp 25 IGS and bp 57-64 n 5b o h G.G+Ccnetwas content C + G IGS. the for bp 25 and Tulipa subsp T3 and (T13) trnL-trnF sylvestris . taxa. bt section (both trnL–trnF .serbica T. and .abnc,T kosovarica, T. albanica, T. analyses. and .serbica T. n obndIS+ ITS combined and Tulipanum .sylvestris T. T9 respectively, (T19) and . .gesneriana T. r grouped are ntefirst the In and ) Tulipa subsp T. T. ). . Posted on Authorea 6 Apr 2020 | CC BY 4.0 | https://doi.org/10.22541/au.158620585.55411740 | This a preprint and has not been peer reviewed. Data may be preliminary. hlgntcaaye ae on based and analyses 2013; Phylogenetic ITS al., combined as et well (Christenhusz scardica as ITS, findings subgenus on of previous based species section with trees analyzed phylogenetic the agreed suaveolens, obtained grouped T. which All datasets, tschimganica, methods, 2013). T. al., applied et ulophylla, all Turktas T. by serbica, generated T. phyly scardica, T. julia luanica, T. T. kosovarica, T. albanica, support. stronger with general subgenus of + in sequences ITS but analysed combined sequences, The the ITS from from from generated obtained trees those trees generated with The with Turk- larities compared 2013). 2006; al., resolution al., et et better Turktas Fay and provided 2013; 2012) sequences al., al., ITS trnF et et from (Christenhusz Yanagisawa obtained ITS 2013; trees of al., use et successfully tas for and of findings ITS of relationships previous and feasibility intra-generic confirming the ITS evaluate revealed to using data datasets analyses Our these phylogenetic Kosovo. of molecular combination out the carried as we study, this In grouping. Discussion specific species no but 3. 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ML by ulophylla, and T. supported julia, not T. but scardica, (86%) of NJ subgenus of consists by specimens subclade the Furthermore, first (89%). the ML and (89%) MP suaveolens (87%), T. serbica, T. ulophylla scardica, subgenus the T. luanica, of T. members of consists the clade In (<50%). MP subsp by supported not but (74%) eune,wihaei codnewt rvosrprs(eesne l,20;Sn ta. 2015; al., et Sang 2008; al., et (Peterson reports previous with accordance in are which sequences, sylvestris . Tulipanum and and n section and ) (section rotmns(.sylvestris (T. Eriostemones hlgntcte osrce rmIS+ ITS from constructed tree Phylogenetic from constructed tree Phylogenetic .gesneriana T. and .serbica T. hwdsqec ieecst h el bandsqecso h pce.Tesecond The species. the of sequences obtained newly the to differences sequence showed .serbica T. 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Tulipa, n section and ) Eriostemones Tulipa Tulipa a h lss eae otenwysqecdseis sall as species, sequenced newly the to related closest the was and trnL-trnF trnL-trnF Eriostemones Eriostemones eogn osection to belonging .tschimganica T. .kosovarica T. and Tulipa trnL-trnF ld,tespecies the clade, ssrnl upre 9-0% naltesto The too. trees all in (97-100%) supported strongly is ) ) 4 ITS trnL-trnF ihnteN 8% n P(5)tes h other The trees. (85%) MP and (87%) NJ the within . .gesneriana T. h subgenus The aae fthe of dataset and Tulipa and nldn section including , eune Ptro ta. 08.Phylogenetic 2008). al., et (Peterson sequences and trnL-trnF .serbica, T. clade eune o h hlgn of phylogeny the for sequences Tulipa ssrnl upre naltes(99-100%) trees all in supported strongly is ) aao the of data ( ape 6adT,wal upre by supported weakly T7, and T6 samples .abnc,T ooaia .laia T. luanica, T. kosovarica, T. albanica, T. Tulipa a oeaeyspotdb J(84%) NJ by supported moderately was , .abnc,T ooaia .luanica, T. kosovarica, T. albanica, T. h ulse euneof sequence published the Tulipa ,section ), trnL-trnF hwdsrn upr o h mono- the for support strong showed .tschimganica T. Tulipa Tulipa aae r hw n3 N) 3B (NJ), 3A in shown are dataset pce r iie notomain two into divided are species hc a oeaeysupported moderately was which Tulipa Tulipa Spiranther smdrtl upre yNJ by supported moderately is trnL-trnF Tulipa taxa. ( Tulipanum .abnc,T kosovarica, T. albanica, T. .albanica T. aae hwdmr simi- more showed dataset iie notosubclades, two into divides taxa. pce rwn idin wild growing species (section ( alignments .tschimganica T. .julia T. ( .abnc,T. albanica, T. eaae from separated .julia T. Tulipa .kosovarika T. tal. et .sylvestris T. Spiranthera uia(T. Tulipa trnL-trnF , species, and (2013). swell as and trnL- T. T. ), Posted on Authorea 6 Apr 2020 | CC BY 4.0 | https://doi.org/10.22541/au.158620585.55411740 | This a preprint and has not been peer reviewed. Data may be preliminary. oeseiti w oorfrs elwt odnylo rcriesalttrigde eds maroon, reddish deep turning al., carmine-scarlet et or (Millaku golden-yellow too Kosovo to in yellow found forms, recently colour was two it in but exist time, flowers first the for Albania 2018). Northeast in species new between the difference genetic of no species is there analyses sequence our ( substrate albanica (2015) T. 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ITS this and (Tati´c supported ITS anthers weakly acute from and of generated yellowish) separation results the (not support phylogenetic violet dull Our filaments, Krivoˇsej, staminal 1997). blackish) and (not pale segments, periapt osdrdt ecoeyrltdt ahother each to related closely be Ernest, to as (Mayer species considered distribution described new its and the 1962) of (Pavlovic areas as Rogozna different as Mt such in at characters anthers recorded morphological and several filaments in of 1970). variation length Micevski, great colour, the show flower in species form, this species leaf of new accepted Individuals a but 1970). as 1980), Micevski, described al. species et first (Tutin the Eurepea of Flora that Because by confirm the accepted sequences by not too. species 2016), exist a Zonneveld, as similarities 2013; morphological List,” Plant the “The similarities molecular similarities except their species, of those specimens Between sequenced newly between resolution Micevski, albanica species-specific Ernest, show Mayer not 2013; the did analyses al., phylogenetic et Our Christenhusz However, the of 1998; the populations Raamsdonk, methods. within within 2009). and polymorphism Zonneveld, molecular detected (Booy The 1970; by was too pool. resolved variation previously gene clearly the intra-specific reported in be cases polymorphism cannot of the presence which of 2013), ( most section species al., in to of complex et belongs a (Christenhusz to species Balkans belong these species these of that all confirmed species as the of species and grouping sequenced Furthermore, newly 2016). the Zonneveld, to closest was the sequence generated by newly followed the from species the Within .luanica T. .scardica T. .scardica T. uiaalbanica Tulipa uialuanica Tulipa hc a oeaeyspotd(iue1)o oeaeyt togyspotd(iue3A). (Figure supported strongly to moderately or 1A) (Figure supported moderately was which , .luanica T. aae i o hwtedvrec of divergence the show not did dataset Tulipa .julia T. noesbld (section subclade one in .scardica T. ope ( complex .scardica T. uiaserbica, Tulipa yiswieo hts einhbs hti oeie akdb brlaepthsof patches obtrullate by masked sometimes is that base perianth whitish or white its by ol hcls fSlce ln Families Plant Selected of Checklist World ld,gnrtdb h T n obndITS combined and ITS the by generated clade, .serbica T. and rw xlsvl nlmsoe hl te pce rwol nsretn) ae on Based serpentine). on only grow species other while limestone, on exclusively grows stems eetseisdsrbda ebro the of member as described species recent most the is hw ra aito nsvrlmrhlgclcaatr,freapeiscampanulate its example for characters, morphological several in variation great shows .kosovarica T. .gesneriana T. .serbica T. .ulophylla T. .albanica T. .abnc,T cria .sria .kosovarica T. serbica, T. scardica, T. albanica, T. , ope Crsehs ta. 2013). al., et (Christenhusz complex T oeie aebe yoyie as synonymized been have sometimes . kosovarica Tt´ n rvˇe,19) ohseis( species Both Krivoˇsej, 1997). (Tati´c and loblnigt hscmlx(hitnuze l,21) a h rttime first the was 2013), al., et (Christenhusz complex this to belonging also sidpnetseis hl eut bandb the by obtained results while species, independent as olce o h rttm ln rsr ie Mrsargo)a the at region) (Mirusha river Mrasori along time first the for collected a roeul sda yoy for synonym a as used erroneously was .sadc,T scardica, T. iesfo hsseisb t obnto fylo einhbases perianth yellow of combination its by species this from differs .kosovarica T. Tulipa (section and uiaserbica Tulipa . T ,cereiecdacoerltosi ewe hs aaand taxa those between relationship close a evidenced clear ), Tulipanum . .tschimganica, T. .kosovarica T. serbica .scardica T. Suae l 02.Ltr hsseiswsrcre in recorded was species this Later, 2012). al. et (Shuka . 5 .luanica T. serbica .serbica T. .sadc,T ebc,T laia .kosovarica T. albanica, T. serbica, T. scardica, T. u lodffr nsvrlcaatr,icuigthe including characters, several in differs also but , .scardica T. .luanica T. .scardica T. ,while ), iesfrom differs .scardica T. and Gvet,21) u nig ae nITS on based findings Our 2019). (Govaerts, ope Brnler 93,(ae Ernest, (Mayer 1923), (Bornnullejr, complex eune rmohrtx fthe of taxa other from sequences uialuanica Tulipa hrsmn opooia hrceswith characters morphological many shares .gesneriana T. .albanica T. trnL-trnF + hc eog osection to belongs which ol o ecnre sindependent as confirmed be not could .suaveolens T. and u a ae eie n described and revised later was but , ope)dsrbtdi h southern the in distributed complex) .scardica T. .scardica T. Suae l,21) n21 the 2012 In 2010). al., et (Shuka .scardica T. .scardica T. .scardica T. .kosovarica T. .sadc.Tlp scardica Tulipa scardica. T. Suae l,21) tdiffers It 2010). al., et (Shuka and trnL-trnF ilk a eodda a as recorded was Millaku aaes h otdistinct most the datasets, Crsehs ta. 2013; al., et (Christenhusz .luanica T. ope niaigthe indicating complex and nisplr unspotted paler, its in ope Mlauand (Millaku complex trnL-trnF . and .albanica, T. Tulipa .gesneriana T. trnL-trnF sa independent an as eune i not did sequences Tulipa uiakosovarica Tulipa .serbica T. Spiranthera ta.2012; al. et ,except ), pce was species .scardica T. n T + ITS and sequences another are ) were was T. , Posted on Authorea 6 Apr 2020 | CC BY 4.0 | https://doi.org/10.22541/au.158620585.55411740 | This a preprint and has not been peer reviewed. Data may be preliminary. fncergnm ie(sdDA2-aus ilb eesr o etrudrtnigo h natural the of understanding better for necessary be will determination the the 2C-values) as of DNA well taxa as (used the markers, within size additional variability using genome sampling, nuclear extensive more of with analysis Further pool. the the to of the related of taxa resolution the lower of the Despite taxa. independent of the sequences of subgenera our species the analyses, to sequence presented belonging the on Based within artefacts species technical of by relationships about caused Conclusions information in probably provide out or to carried Kosovo not 2005), in were present analyses al., complex. cytogenetic species et and the content unconvincing (Beaulieu of DNA an used habitat all However, phenotype the (2010) be 2005). in plant Greilhuber, al. to differences (2018) with and 2004), seems et correlated Osmani (Obermayer Shuka be what al., (2018). could while populations, species et Osmani populations, within natural by (Jakob sizes wild from genome material of from collected differences plant time, seeds Such the explanation. from blooming of germinated in origin leaves plants the adult from by collected explained leaves were used (2018) Osmani and for pg 2013). 45.71 2018), for al., (Osmani, et pg (Christenhusz 43.86 influenced or of be (2C) radiation easily species analyses with can size experiments some based colour mutation Genome within Experiments flower as secondly, colours. well and light as and margin provenances, very and blotch intensity natural or blotch that colour from yellow showed the obtained in in first accessions resulting and of aspects, accessions selection size two certain on in in in differ differences lacking may show are species anthocyanidins may within margins colour colour blotch flower yellow the the in (1995) Vries base and perigon Raamsdonk the of half gesneriana (Millaku like areas characters distribution from mixed colour sympatric trait with al., from flower in individuals trait et species intermediate inherited example, different (Shuka with (an For of maroon hybrids individuals 2018). natural reddish al. Later as of et deep colour. et explained population of turning Shuka and intermediates one classifications reported carmine-scarlet 2010; with the within were al., to even for individuals yellow/golden-yellow et suitable colour including Shuka flower form very 2010), the 2015; be be the example, to Elezaj, to For discriminate not and reported 2013). seems to Millaku al., it used 1970; et Hence, characters Micevski, (Christenhusz 2009). main Ernest, Zonneveld, the Mayer 2012; of the 2014; al. one of al., was et taxon colour (Eker a flower the to the of analysis, belongs species morphological Prishtina, the University In the of Herbarium the complex. of unidentified the material that herbarium showed analyses phylogenetic taxon. Our this of pool gene the in confirm Millaku necessarily 2012, not ITS, al. on does between et divergences based (Shuka supported results pollen moderate phylogenetic to violet-purple weak Our and 2015). filaments an yellow Elezaj, have blotches, and black individuals without some bases Furthermore perianth T 2010). serbica, al., T. maroon. et scardica, reddish (Shuka T. to flowers yellow golden-yellow of the colour intermediate of dominance a with .scardica T. .scardica T. .kosovarica T. ,wieters fteprgnwspn lk to (like pink was perigon the of rest the while ), .scardica T. .scardica T. Zneed 09.Teicnretrslsfor results incongruent The 2009). (Zonneveld, ope hwitaseicvrain niaigtepeec fplmrhs ntegene the in polymorphism of presence the indicating variation, intra-specific show complex .gesneriana T. eercre,o nemdaeidvdaswt elwprgnbs a ti in is it (as base perigon yellow with individuals intermediate or recorded, were ) .scardica T. complex ope ( complex . kosovarica Eriostemones .albanica T. Tulipa .scardica T. , u h oe oorapae ob eyvral ihnoespecies one within variable very be to appeared colour flower the but .sadc,T ebc,T laia .kosovarica T. albanica, T. serbica, T. scardica, T. pce eeldta Cof 2C that revealed species ope sidpnetseisto eei ieecsbtentaxa between differences Genetic too. species independent as complex n h te afo h eio aei ht oor(ninherited (an colour white in base perigon the of half other the and ) and sidpnetseis u niaetepeec fpolymorphism of presence the indicate but species, independent as .kosovarica T. or .luanica, T. complex. Tulipa uiaalbanica Tulipa .albanica T. 6 hr sntsffiin eei vdnet confirm to evidence genetic sufficient not is There . trnL-trnF u tdffr rmte yiscmiaino yellow of combination its by them from differs it but n 74 gfor pg 47.49 and uiaspecies Tulipa trnL-trnF .luanica T. n te aao the of taxa other and Tulipa .albanica T. .albanica T. hrsmn opooia iiaiiswith similarities morphological many shares aae,i osntspotteseparation the support not does it dataset, pce rue notomi clades, main two into grouped species n T + ITS and Mlaue l,21) codn to According 2018). al., et (Millaku ) .luanica T. sml 9 al )otie from obtained 1) Table T9, (sample a 41 g(hk ta. 2010) al., et (Shuka pg 54.15 was eotdb hk ta.(2010) al. et Shuka by reported trnL-trnF .scardica T. Omn 08 n 9pg 69 and 2018) (Osmani and aaesshowed datasets .albanica T. .luanica T. ope,what complex, Tulipa .scardica T. .scardica T. species as ) was T. Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. nttl5 eune bandfo 4tx eeaaye,4 fte eenwygnrtdsqecspro- sequences generated newly were them of 41 analysed, were taxa 14 eight from from obtained vided sequences 55 total In to sequences subjected analyses all for Phylogenetic were numbers submitted sequences accession were 1. GenBank Table sequences Edited (NCBI). generated in 2018). Information Newly given Biotechnology al. 1990). are for Fisher al., et Center et National (Kumar (Thermo (Altschul the software analysis to analyzer preliminary X DNA for Brisbane, MEGA searches South capillary (Technelysium, with BLAST 2.6.6 96 aligned vers. CHROMAS 3730xl and with edited Australia) Microsynth Biosystems manually by was Applied performed Sequence was Every using Sequencing Scientific). company. Austria) sequencing the (Vienna, of Austria requirements the to according 85 primers in min 15 and Tem ihrSinicBlis inu,Ltuna eepeie nterto14adsoe ntefreezer. the in stored and 1:4 ratio the in premixed were Lithuania) 13 Vilnius, Baltics, Scientific Fisher (Thermo sn lcrpoei n1 grs es(o etn on grs,SgaAdih afice,Germany), DNA. from Taufkirchen, genomic Aldrich, I for Sigma Exonuclease above agarose, described point as melting an conditions (low gels similar with agarose using 72 performed 1% at in were min electrophoresis qPCR amplifications 7 using MIC of PCR step a elongation in 95 Australia). final performed at were Coomera, step PCRs denaturation Upper The initial Germany). systems, (ITS4 Taufkirchen, (Biomloceular Aldrich, primers [5’-ATTTGAACTGGTGACACGAG-3’; (Sigma reverse cycler f 1991]) and or al., 1991]) White et 1990] al., [5’-GGAAGGAGAAGTCGTAACAAGG-3’; Taberlet et al., (ITS5 Taberlet [5’-CGAAATCGGTAGACGCTACG-3’; et forward c White [(5’-TCCTTCCGCTTATTGATATGC-3; nM or 14 1990] 300 to al., and 15 added at Estonia) a was Tartu, For ng) Biodyne, subspecies. 1–50 MgCl two mM and approx. 2.5 species to transcri- six internal (equivalent nuclear of The DNA PCR. samples chloroplast for the 23 used and then from transilluminator and (ITS) UV water region deionized with spacer with system bed diluted microDOC 1:50 wavelength. was using nm DNA documented 312 Extracted was using which UK) Erlangen, Warwickshire, V, (VWR, Rugby, PeqGreen LTD, 120 x Scientific at 0.4 (Cleaver containing min gels 40 was agarose quality for 1.0% DNA The with Germany) instructions. DNeasy electrophoresis manufacturer’s the gel the using agarose to specimens using according checked herbarium Germany) or Hilden, materials (Qiagen Kit gel-dried Mini silica Plant from sequencing extracted was and DNA genomic (PCR) Total reaction chain polymerase extraction, DNA the Table of in Prishtina, given gel University is silica the 1 information Table of in sample Herbarium dried Detailed the were USA. leaves Atlanta, at Herbarium, young deposited University 1. of were Emory part specimens the voucher and and Kosovo The herbarised extraction. were DNA specimens for Plant Prishtina. each. University localities the of different of and specimen Herbarium Koznik) the plant Gori¸c and by unidentified provided (locations material One All herbarised 2019. Albania. from and in obtained genus 2018 collected the 2017, was of May which subspecies) to two April and in species (six taxa Eight material Plant methods and Materials μ C rdcswr ie ih1.3 with mixed were products PCR L Tulipa ai hrceitc ftecleto ie,vuhrifrain n eBn ceso numbers accession GenBank and information, voucher sites, collection the of characteristics Basic . pcmn sdfrti study this for used specimens 2 130 , Tulipa ° .PrfidPRpout eedltdwt itle ae n die ihsequencing with admixed and water distilled with diluted were products PCR Purified C. .coli E. μ NPmx . a O IEo N oyeae(l egnsfo Solis from reagents (all Polymerase DNA FIREPol HOT Taq U 0.6 mix, dNTP M aa(i pce n w useis olce rmwl ouain nKsv and Kosovo in populations wild from collected subspecies) two and species (six taxa 0U/ 20 ° o 43 i,floe y4 ylsa 95/58/72 at cycles 40 by followed min, 14:30 for C μ .luanica T. EOI n hroestv laiePopaae1U/ 1 Phosphatase Alkaline Thermosensitive and I) (EXO l ° .Teapie C rget (2 fragments PCR amplified The C. μ X n atPmxueadicbtda 37 at incubated and mixture FastAP and I EXO L Tulip trnL-trnF lctosPsti n a¨ rs)wr olce rmtwo from collected were Qaf¨e Prush) and Pashtrik (locations pce eecletdi ooo except Kosovo, in collected were species 7 negncsae eeapie n hnsequenced then and amplified were spacer intergenic Tulipa μ atrmxcnann 1 containing mix master L μ C ecin 1 reaction, PCR L eecletdfo hi idpopulations wild their from collected were Tulipa μ fPRpout)wr checked were products) PCR of L p sml 9 al )was 1) Table T9, (sample sp. ° o 03/0s n a and s, 30/30/90 for C μ fdltdgenomic diluted of L × uiakosovarica Tulipa uiaalbanica Tulipa C ue B, buffer PCR ° o 5min 15 for C μ (FastAP) l ® , Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. yteRylBtncGres e.Pbihdo h nent tp/ws.cec.e.r/Rtivd13 Retrieved http://wcsp.science.kew.org/ Internet; the Facilitated on Arecaceae. Published of Checklist Kew. 2019. World A. Gardens, August Access. Henderson, Botanic & Royal D.R. [Internet]. the Hodel, Kew S., by Zona, Gardens, http://wcsp.science.kew.org/qsearch.do J., Dransfield, Botanic 2006;22(November from: R., Royal Govaerts, Available Families: 13]. [Internet]. Plant Aug Aliso evidence 2019 Selected summarized [cited of 2019 loci. Liliales: Checklist World mitochondrial of R. Phylogenetics one Govaerts Y. and plastid . . five Pillon. DS, of 2015):559–65. Devey analyses N, combined Ronsted from MW, Chase MF, Fay genus the p. of 1–112 Revision M. 2014. Koyuncu tulips MTI, the Babac Il, through I Tiptoe Eker of al. et classification PS, and Roberts phylogenetics K, 172(3):280–328. molecular 19:199—200. Borland Ecol. history, T, 1923; Hall Syst cultural Repert. JC, Feddes - Biochem David ; Mazedoniens. R, Govaerts Flora species MJM, levels. der Tulipa Christenhusz Tulpe taxonomical between neue different eine and Uber at within J. relationships Bornnullejr esterase genetic enzyme of the analysis in 1998;26(2):199-224 Variation the Van. for of LWD genome usefulness Journal Raamsdonk of evolution tool. G, Correlated al. search Booy et alignment CA, 2005;422–37. Knight local JB, mass. Dickie seed Basic MD, and Bennett DJ. size IJ, Leitch AT, Lipman Moles EW, JM, Beaulieu Myers 403–410. W, 215: Miller 1990: Biology. W, Molecular Gish scores SF, and Altschul support moderate as 89% and References 75% between scores the support. In support, strong 2002). weak (Swofford, as 4.0 PAUP defined in scores and were conducted ITS bootstrap between methods, trees, incongruence (ML) no phylogenetic Likelihood revealed Maximum test and ILD (MP) (ITS The Parsimony combined test). The (ILD 2). testing (Table compatibility trnL-trnF GenBank) for from used were obtained for and six while and GenBank, generated into from newly + obtained loaded (20 were were sequences eight 26 datasets and included The generated 2018). newly and were al. (ITS them et separate as (Kumar analysed software then one trnF X and only of MEGA 2002) by (Swofford, amplification represented using 4.0 were the PAUP aligned species of were some failure sequences T18), The the and T16 of T9, Because trnL-trnF sequences. species. T10; two and per or T8 specimens (ITS two specimens from some successfully sequenced and the while species, each using and rooted ITS were trees The GenBank. from GenBank: obtained were (HF952989) tschimganica for T. sequences ITS The 1). (HF952964), (Table GenBank tschimganica from T. obtained were sequences 14 trnL–trnF aaes o T nlss nttl2 eune eeaindt eemn eunesaitc,2 of 21 statistics, sequence determine to aligned were sequences 29 total in analyses, ITS For datasets. ) trnL-trnF eune p=08) hlgntcteswr osrce sn egbrJiig(J,Maximum (NJ), Neighbor-Joining using constructed were trees Phylogenetic 0.82). = (p sequences trnL-trnF .gesneriana T. aae nldd2 eune bandfo h aawihwr rsn nbt datasets both in present were which taxa the from obtained sequences 24 included dataset ) (HF952976), (HF953001), euneo oto aawr mlfidadte eune rmtreseiesfor specimens three from sequenced then and amplified were taxa of most of sequence F598adISK855)a outgroup. as KX865057) ITS and KF850988 .kosovarica T. M351,M352)adthe and MK335224) (MK335217, .sylvestris T. .sylvestris T. < lclt Gori¸c) and (locality 0 eenttkni osdrto,soe ewe 0 n 74% and 50% between scores consideration, in taken not were 50% subsp. subsp. sylvestris sylvestris 8 Tulipa (HF952999), (HF952974), .luanica T. trnL-trnF .(iica)i uky o.17 Phytotaxa. 157, Vol. Turkey. in (Liliaceae) L. Tulipa trnL-trnF eune for sequences lclt a¨ rs)wr amplified were Qaf¨e Prush) (locality .suaveolens T. Llaee.BtJLn o.2013; Soc. Linn J Bot (Liliaceae). .suaveolens T. iimmartagon Lilium trnL-trnF uiaulophylla Tulipa n obnd(T + (ITS combined and ) .ulophylla T. (MK334468), (HF952998), ttsia analyses statistical otie from (obtained > (HF952978), (HF953003), 0 Sas BS 90% .julia T. .julia T. trnL– Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. ui G ewo H ugsN,VlnieD.FoaErpe.FoaErpe.18.3132p. of 381–382 nomenclature 1980. Europaea. infrageneric Flora The 298(1):87–92. Europaea. BJM. Flora Zonneveld DH. JF, Valentine Veldkamp Linn NA, J Burges Bot Turkey. VH, on Heywood emphasis TG, an Tutin with sequences DNA of 172(3):270–9. nuclear analysis 2013; and phylogenetic plastid Soc. Molecular noncoding http://www.theplantlist.org/. on E. Kaya based from: Ba¸stuˇg Sara¸c Available (Liliaceae) YI, Ertuˇgrul OK, F, 15]. B, Metin Aug 2019 M, [cited Turktas 2013; Online. Rev Choice List. Plant The regions non-coding three Z. of Krivosej 17:1105–1109. amplification B, 1991; for Tatic Biology primers Molecular Universal Plant J. 4. DNA. Bouvet Version chloroplast G., 2002. of Pautou L., Methods). Gielly Other P., Taberlet (*and Parsimony Massachusetts. Using Sunderland, Analysis Associates, Phylogenetic Sinauer PAUP*. L. D. Swofford, Albania. northeastern 9:1–9. from KE.2012; species Kit Tan new Lulezim, a Shuka (Liliaceae), albanica Tulipa S. 2010;10(1):17. Siljak-Yakovlev Phytotaxa. K, of Tan Biogeography L, and Shuka Evolution, Reticulate Phylogeny, DNA Chloroplast TF. Paeonia Stuessy DJ, Crawford T, in Sang taxonomy 46(2):446–65. and 2008; relationships Evol. Species 1995;195:13–44. Phylogenet DE. Mol ). T Vries data. Van, morphological LWD and Raamsdonk 3-20. molecular on 8: of based 1962; Systematics Gagea Beogradu, J. of Muz. Peterson cation Prir. IG, Levichev Glasn. A, Srbije. Peterson flore serpentinske elementi Hredity gjinise Karakteristicni te Z. colour? llojeve Pavlovic, tre fruit te with biokimike dhe vary citogjenetike villosum varica adaptime Dasypyrum Disa M. in Osmani size genome Does some 2005:91–5. J. of (Edinb), Greilhuber ecology R, and area Obermayer 2018;28(1):35–47. Sympatric Bot. N. J Berisha Thaiszia ISA, Peninsula. Elezaj F, Millaku I. 2015;52(5–6):315–20. Elezaj F, Millaku von Chorologie 591–8. und Taxonomie 1970; 35:1547-1549. Analysis Zur Genetics 2018. K. Evolutionary 55(4):291–307. Micevski Evolution. Molecular Ernest, 2017; 860-9 X: Mayer and MEGA Evol. (5), Biology K. Syst 21 Tamura Molecular J and platforms. C., 2004; future. Knyaz computing M., the Evol. across Li in Biol G., and Mol Stecher today S., Rates. Kumar Poaceae Applications Speciation ( barcodes: and Species DNA , Hordeum Plant Ecology of WJ. , Variation Form Kress Size Life Genome Phylogeny, Considerable to The Linked FR. Is specierum Blattner ) A, Repertorium Meister SS, (Monocotyledoneae). Jakob 3 Band 30:472 1933; Balcanicae, Vegetabilis, peninsulae Regni Novarum Florae Prodromus A. Hayek dhe Penaee.A o.19;8 (9),1120–36. 84 1997; Bot. J Am (Paeoniaceae). .luanica T. uiaserbica Tulipa uialuanica Tulipa ehbtttetr ayoe h israin nvriyo rstn;2018. Prishtina; of University dissertation. PhD natyrore. tyre e habitatet ne ) uiakosovarica Tulipa Llaee,anwseisfo eba ocna 1997;5(2):733–6. Bocconea. Serbia. from species new a (Liliaceae), iica ,aNwSeisfo otenKsv n o Fenn. Bot Ann . Kosovo Southern from Species New a ), Liliaceae ( Llaee,anwseiso ui rmKsv.Phytotaxa. Kosovo. from tulip of species new a (Liliaceae), Gagea 9 and uiascardica Tulipa Lloydia Tulipa Llaee n nrgnrcclassifi- infrageneric and (Liliaceae) Llaee.PatSs vl 2012; Evol. Syst Plant (Liliaceae). Tulipa Tulipa on.Fde Repertorium. Feddes Bornm. Tulipa pce ntews Balkan west the in species ug. subg ( .abnc,T koso- T. albanica, T. Tulipa Liliaceae ( Tulipa Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. varica koso- T. varica koso- T. albanica T. albanica T. banica al- T. species Potential the of ). Liliaceae of ( 1 species Table . “all” for L size Tulipa 281(1–2):217–45. genome of 2009; nuclear species Evol. of Syst ” value systematic all The “ BJM. for Zonneveld size 281(1):217-245 genome 2009; nuclear Evol. of Syst value Plant systematic The BJM. Zonneveld 953:103–10. 2012; Phylogenetic Hortic. K. 298(7):1293–6. Acta Okazaki 2012; DNA. JJ. Evol. chloroplast A, fungal Groot of Marasek-Ciolakowska de sequences of BJM, using D, sequencing Zonneveld tulips Sochacki direct garden T, and USA. and wild California, Nishikawa Amplification Diego, of T, San analysis 1990. Press, Kuhara Academic W. 315–322. R, J. applications, Yanagisawa Taylor and methods B., to guide S. A Lee protocols: phylogenetics. PCR D., for genes T. RNA Bruns ribosomal J., T. White Tulipa ai hrceitc ftecleto ie,vuhrifrain n eBn ceso numbers accession GenBank and information, voucher sites, collection the of characteristics Basic . T. T. flower) /yellow maroon (reddish T3 albanica T. flower) maroon (reddish T2 albanica T. flower) (yellow ica ban- T. Sequence T5 varica koso- T4 varica koso- ID ape sdfrti study this for used samples - - al- - - T1 - - - - - urjAbna42 Albania Surroj 42 Albania Surroj 42 Albania Surroj oicKsv 42 Kosovo 42 Gori¸c Kosovo Gori¸c Altitude Latitude Longitude Country locality Collection uiakolbintsevii Tulipa In .A ni,D .Glad .J nnk,adT .Wie[eds.], White J. T. and Sninsky, J. J. Gelfand, H. D. Innis, A. M. on,anwseisfo atr aahtn ln Syst Plant Kazakhstan. Eastern from species new a Zonn., 10 2.744’N 2.744’N 2.744’N 26.689’N 26.689’N ° ° ° ° ° 20.037’E 20 20.037’E 20 20.037’E 20 45.337’E 20 45.337’E 20 ° ° ° ° ° Tulipa 2 N321M469 00000156 MN446899 MN336201 622 00000157 MN446898 MN336200 00000158 622 MN446897 MN336199 622 5 N323M460 00000154 MN446901 MN336203 00000155 MN446900 659 MN336202 659 .(iica) Plant (Liliaceae). L. number cession ac- ITS number sion acces- trnF trnL- no. acces. herb. ishtina Pr- of Uni. Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. serbica T. serbica T. scardica T. scardica T. scardica T. luanica T. luanica T. luanica T. luanica T. luanica T. species T. varica koso- T. varica koso- T. varica koso- T. species Potential T. T. T. scardica T. T15 scardica T. T14 anica T. T13 anica T. T12 anica T. T11 anica T. T10 anica T. T9 species T. T8 varica koso- T. T7 varica koso- T. T6 varica koso- T. Sequence T19 bica T18 bica T17 scardica T16 ID - - lu- lu- lu- lu- lu- - - - - ser- ser------rvnkKsv 42 00000150 Kosovo Krivenik 42 /// Kosovo MN336206 Qaf¨e 42 Prush 42 Kosovo Qaf¨e Prush /// Kosovo 42 Pashtrik /// Kosovo 42 Pashtrik Kosovo /// Pashtrik Kosovo 42 Krojmir Kosovo 42 Koznik Kosovo 42 Koznik Kosovo Koznik ebcKsv 42 Kosovo 42 Serboc Kosovo 42 Serboc Kosovo 42 Krivenik Kosovo Krivenik Altitude Latitude Longitude Country locality Collection 11 6.254’N 18.275’N 18.275’N 14.966’N 14.966’N 14.966’N 30.334’N 30.334’N 30.334’N 58.067’N 58.067’N 6.254’N 6.254’N ° ° ° ° ° ° ° ° ° ° ° ° ° 14.958’E 21 23.529’E 20 23.529’E 20 30.399’E 20 30.399’E 20 30.399’E 20 33.987’E 20 33.987’E 20 33.987’E 20 49.757’E 20 49.757’E 20 14.958’E 21 14.958’E 21 ° ° ° ° ° ° ° ° ° ° ° ° ° 7 N321M461 00000167 MN446910 MN336211 00000145 MN446909 MN336210 575 00000148 MN446908 MN336209 580 00000147 MN446907 MN336208 580 00000146 00000149 MN446906 MN446905 MN336207 1041 /// 1041 00000151 MN446904 1041 /// 00000152 MN446903 425 MN336205 00000153 MN446902 425 MN336204 425 9 N325M461 00000163 MN446912 MN336215 00000164 00000165 MN446911 MN336214 596 00000166 /// MN336213 596 /// MN336212 575 575 number cession ac- ITS number sion acces- trnF trnL- no. acces. herb. ishtina Pr- of Uni. Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. iana gesner- T. iana gesner- T. julia T. olens suave- T. sylvestris ssp. sylvestri T. ganica tschim- T. phylla ulo- T. sylvestris ssp sylvestris T. tralis aus- ssp sylvestris T. sylvestris T. serbica T. species Potential . . T30 T. T29 T. T28 T. T27 olens suave- T. T26 sylvestris subsp. sylvestris T. T25 ica gan- tschim- T. T24 phylla T. T23 sylvestris ssp. sylvestris T. T22 tralis aus- ssp. sylvestris T. T21 sylvestris T. T20 bica T. Sequence ana neri- ana neri- julia ID - - - ulo- - - - ser- ges- ges------/ / / / / K324///// /// /// MK335224 /// /// MK335217 /// HF952989 HF952964 /// /// /// HF952998 /// /// MK33446 /// /// /// /// /// HF952999 /// HF952974 /// /// /// /// /// /// HF953001 HF952976 /// /// /// /// HF953003 /// HF952978 /// /// /// /// /// /// /// /// /// /// /// /// /// /// /// /// 42 Kosovo Dev¨e 42 Kosovo 42 Dev¨e Kosovo 42 Gori¸c Kosovo Serboc oaiyCutyLniueLttd Altitude Latitude Longitude Country locality Collection 12 19.950’N 19.950’N 26.747’N 58.067’N ° ° ° ° 20.517’E 20 20.517’E 20 45.293’E 20 49.757’E 20 ° ° ° ° 0 N329M461 00000159 MN446916 MN336219 700 00000160 MN446915 MN336218 00000161 MN446914 MN336217 700 00000162 MN446913 MN336216 665 596 number cession ac- ITS number sion acces- trnF trnL- no. acces. herb. ishtina Pr- of Uni. Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. otns6.%3.%44.5% 273 0.940 31.2% 0.890 241 60 60.1% 0.928 (0.714) 0.880 (0.808) 1229 0.897 12 62 0.945 213 24 57 179 749 1471 1430 12 10 93 0.840 1377 184 26 91 47 480 806 0.769 775 14 52 86 720 29 132 51 contents C + G ITS 664 0.855 index composite index) index) (retention 657 (consistency RI only) characters 644 (informative CI 7 length 45 tree indels MP informative potentially of Number characters informative Potentially characters Variable characters outgroup Conserved positions: ambiguous of ingroup Number positions: ambiguous of Number (bp) length maximum Sequence (bp) length minimum Sequence (bp) length Alignment sequences No. taxa No. and ITS for Parameters characteristics tree parsimony-based and set Data 2. Table martagon Lilium species Potential martagon L01 L. Sequence ID - - - / / / / / X607K808 /// KF850988 KX865057 /// /// /// /// /// oaiyCutyLniueLttd Altitude Latitude Longitude Country locality Collection 13 trnL-trnF trnL-trnF analyses. obndIS+ ITS Combined number cession ac- ITS trnL-trnF number sion acces- trnF trnL- no. acces. herb. ishtina Pr- of Uni. Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. 14 Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. B. A. 15 Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. elcts .Niho-onn re .Mxmmprioyte.C aiu ieiodtree. likelihood Maximum C. tree. parsimony Maximum B. tree. Neighbor-joining A. replicates. 1. Figure C. ieetpyoeei re ae nISsqecsicuigBosrpvle ae n1000 on based values Bootstrap including sequences ITS on based trees phylogenetic Different 16 Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. 17 Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. .B. A. 18 Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. 00rpiae.A egbrjiigte.B aiu asmn re .Mxmmlklho tree. likelihood Maximum C. tree. parsimony Maximum B. tree. Neighbor-joining A. replicates. 1000 2. Figure C. ieetpyoeei re ae on based trees phylogenetic Different trnL-trnF 19 eune nldn otta ausbsdon based values Bootstrap including sequences Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. 20 Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. .B. A. 21 Posted on Authorea 6 Apr 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158620585.55411740 — This a preprint and has not been peer reviewed. Data may be preliminary. ta ausbsdo 00rpiae.A egbrjiigte.B aiu asmn re .Maximum C. tree. parsimony Maximum B. tree. Neighbor-joining A. tree. replicates. likelihood 1000 on based values strap 3 Figure C. ieetpyoeei re ae nacmie ITS+ combined a on based trees phylogenetic Different . 22 trnL-trnF eunesticuigBoot- including set sequence