Kun-Li Xiang

Andrey S Erst

Jian Yang

Huan-Wen Peng

Rosa Del C Ortiz

Florian Jabbour

Tatyana V Erst

Wei Wang

Rosa Del C Ortiz

Biogeographic diversification of Eranthis (Ranunculaceae) reflects the geological history of the three great Asian plateaus

Keywords: Asia, biogeography, Cenozoic, phylogeny, Ranunculaceae, tectonic evolution

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Introduction

The collisions of Indian Subcontinent with Eurasia in the east and Arabian plate with Eurasia in the west are two important tectonic events in Asia [START_REF] Yin | Cenozoic tectonic evolution of Asia: A preliminary synthesis[END_REF]. The India-Eurasia collision started at about 61 Ma [START_REF] Yuan | Rapid drift of the Tethyan Himalaya terrane before two-stage India-Asia collision[END_REF] and subsequently caused the deformation and uplift of the Qinghai-Tibetan Plateau (QTP) and the eastward lateral extrusion of Indochina [START_REF] Yin | Cenozoic tectonic evolution of Asia: A preliminary synthesis[END_REF][START_REF] Spicer | Why the 'Uplift of the Tibetan Plateau' is a myth[END_REF][START_REF] Tapponnier | The Ailao Shan/Red River metamorphic belt: tertiary left-lateral shear between Indochina and South China[END_REF].

In addition, the Mongolian Plateau (MP) is usually considered to be the consequence of the far-field effect of the continued India-Eurasia convergence [START_REF] Vassallo | Uplift age and rates of the Gurvan Bogd system (Gobi-Altay) by apatite fission track analysis[END_REF]. The Arabia-Eurasia collision started at c. [START_REF] Ebersbach | In and out of the Qinghai-Tibet Plateau: Divergence time estimation and historical biogeography of the large arctic-alpine genus Saxifraga L[END_REF] Ma and resulted in the deformation and uplift of the Iranian Plateau (IP) region and the westward lateral extrusion of the Anatolia block [START_REF] Mouthereau | Building the Zagros collisional orogen: Timing, strain distribution and the dynamics of Arabia/Eurasia plate convergence[END_REF]. The India-Eurasia and Arabia-Eurasia convergences have usually been regarded as separate collision processes, but they may have interacted by controlling the far-field Cenozoic deformation in Asia [START_REF] Yin | Cenozoic tectonic evolution of Asia: A preliminary synthesis[END_REF]. This interaction may have generated a north-east oriented zone of Central Asia right-slip faults (300-400-km wide and >1500-km long), which encompassed the Zagros mountains, Tian Shan and Altai Shan orogens, and western Mongolia (electronic supplementary material, figure S1). The uplifts and outward growths of the QTP, IP and MP regions and associated mountain belts have had a profound impact on regional and even global climate and environment [START_REF] Farnsworth | Past East Asian monsoon evolution controlled by paleogeography, not CO 2[END_REF][START_REF] Spicer | Cenozoic topography, monsoons and biodiversity conservation within the Tibetan Region: An evolving story[END_REF][START_REF] Deng | Major turnover of biotas across the Oligocene/Miocene boundary on the Tibetan Plateau[END_REF][START_REF] Li | Orographic evolution of northern Tibet shaped vegetation and plant diversity in eastern Asia[END_REF]. Yet, the evolution of these three great Asian plateaus remains considerably controversial and is still poorly understood [START_REF] Yin | Cenozoic tectonic evolution of Asia: A preliminary synthesis[END_REF][START_REF] Su | No high Tibetan Plateau until the Neogene[END_REF][START_REF] Renner | Available data point to a 4-km-high Tibetan Plateau by 40 Ma, but 100 molecular-clock papers have linked supposed recent uplift to young node ages[END_REF]. The tectonic events affected biotic evolution; in turn, evolutionary diversifications of organisms might could reflect the geological and related climate changes. However, with few exceptions [START_REF] Jabbour | A phylogeny of Delphinieae (Ranunculaceae) shows that Aconitum is nested within Delphinium and that Late Miocene transitions to long life cycles in the Himalayas and southwest China coincide with bursts in diversification[END_REF][START_REF] Manafzadeh | A tale of migrations from east to west: The Irano-Turanian floristic region as a source of Mediterranean xerophytes[END_REF], evolutionary studies of Asian organisms have focused on a single plateau, such as the QTP [START_REF] Favre | The role of the uplift of the Qinghai-Tibetan Plateau for the evolution of Tibetan biotas[END_REF][START_REF] Zhao | Extinction vs. rapid radiation: The juxtaposed evolutionary histories of coelotine spiders support the Eocene-Oligocene orogenesis of the Tibetan Plateau[END_REF]. This precludes broad comparisons of the geological evolution of Asian plateaus associated with the India-Eurasia and Arabia-Eurasia collisions as a whole.

Ranunculaceae, commonly known as the buttercup family, is an important herbaceous element of mountain ecosystems in the Northern Hemisphere [START_REF] Ziman | A geographical analysis of the family Ranunculaceae[END_REF]. The family originated in the Lower Cretaceous and experienced a burst of diversification during the Upper Cretaceous (c. 83-74 Ma) [START_REF] Wang | The rise of angiosperm-dominated herbaceous floras: Insights from Ranunculaceae[END_REF]. Owing to their seeds with poor dispersal abilities, the ranunculaceous species with follicles are often reported to be geographically highly isolated [START_REF] Wang | The rise of angiosperm-dominated herbaceous floras: Insights from Ranunculaceae[END_REF] and have been used to infer East Asian floristic evolution [START_REF] Xiang | A molecular phylogeny of Dichocarpum (Ranunculaceae): Implications for eastern Asian biogeography[END_REF][START_REF] Xiang | Biogeography of Coptis Salisb. (Ranunculales, Ranunculaceae, Coptidoideae), an Eastern Asian and North American genus[END_REF]. In this study, we focus on the biogeographic diversification of the genus Eranthis, an early-spring ephemeral, known as winter aconite in Europe. Recent molecular clock estimates suggest a stem age of 56-59 Ma for the genus [START_REF] Wang | The rise of angiosperm-dominated herbaceous floras: Insights from Ranunculaceae[END_REF]. Eranthis mainly inhabits humid environments under deciduous forests of temperate Eurasia (figure 1a). The fruits of Eranthis are dehiscent follicles (figure 1b). Seeds may be simply gravity-dispersed owing to lacking obvious adaptation to wind or animal dispersal, and hence are not expected to disperse over long distance or through physical barriers. The species of the genus are found throughout the Eurasian continent, but their distribution is discontinuous and each species is restricted to a particular biogeographic region, i.e., South Europe, South Turkey, the west and east edges of the QTP, MP, or Northeast Asia (figure 1c; electronic supplementary material, table S1).

Furthermore, species of the genus occur both in lowlands and at high elevations in different geographic regions (electronic supplementary material, figure S2). The present-day distribution of Eranthis may be linked to the geological evolution of Asia, especially the formation of the three great Asian plateaus and the extrusion of the Anatolia block.

Given the inferred late Paleocene age and extensive Eurasian distribution of the genus, a poor dispersal ability, as well as the restriction of each species to particular biogeographic regions, Eranthis is a key biological model to infer the geological history of Asia. Here, we present a molecular phylogeny for Eranthis that includes all currently recognized species.

By integrating phylogenetic, molecular dating, and biogeographic methods, we investigate the historical biogeography of the genus, and thereby aim to illuminate the likely evolutionary histories of the three great Asian plateaus.

Material and methods

(a) Samples and sequences

We sampled 34 individuals of Eranthis (Cimicifugeae, Ranunculaceae), representing all 12 species (including one undescribed new species) [START_REF] Erst | An integrative taxonomic approach reveals a new species of Eranthis (Ranunculaceae) in North Asia[END_REF]. We included nine species covering all three other genera of Cimicifugeae, as well as three species of Asteropyreae, Caltheae and Adonideae as the outgroups [START_REF] Wang | The rise of angiosperm-dominated herbaceous floras: Insights from Ranunculaceae[END_REF]. Materials used in this study were mainly collected from herbarium specimens except for a few from the field. Voucher information and GenBank accession numbers are listed in electronic supplementary material, table S3.

Five DNA regions, including plastid (trnL-F, trnH-psbA, matK, rbcL) and nuclear (ITS) loci were sequenced. Laboratory procedures, sequence handling and phylogenetic inference followed Xiang et al. [START_REF] Xiang | A molecular phylogeny of Dichocarpum (Ranunculaceae): Implications for eastern Asian biogeography[END_REF]. Sequences were aligned using Geneious 10.1.3 [START_REF] Kearse | Geneious basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data[END_REF], and adjusted manually. The final alignments contained 46 taxa with 1001 bp for trnL-F (excluding three ambiguous regions with 114 nucleotides), 251 bp for trnH-psbA (excluding one ambiguous region with 73 nucleotides), 1210 bp for matK, 1263 bp for rbcL, and 668 bp for ITS. Phylogenetic analyses were conducted using maximum likelihood and Bayesian inference methods in RAxML 8.0 [START_REF] Stamatakis | RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies[END_REF] and MrBayes 3.2.1 [START_REF] Ronquist | MrBayes 3.2: Efficient Bayesian phylogenetic inference and model choice across a large model space[END_REF], respectively. The plastid and nuclear datasets were each first analyzed independently. As there were no significant topological incongruences (≥70% bootstrap percentage and ≥0.95 posterior probability) between plastid and nuclear datasets (electronic supplementary material, figure S3 vs. figure S4), all subsequent analyses were performed for the combined five-locus dataset only.

(b) Divergence time estimation

Divergence time was estimated in BEAST 2.4.0 [START_REF] Bouckaert | BEAST 2: a software platform for Bayesian evolutionary analysis[END_REF]. We set an 83 Ma (95% HPD: 77-88 Ma) constraint for the root, which is the estimated age for the split of Cimicifugeae and Adonideae [START_REF] Wang | The rise of angiosperm-dominated herbaceous floras: Insights from Ranunculaceae[END_REF], with a normal distribution. Paleoactaea nagelii, fruits from the late Paleocene Almont and Beicegel Creek floras of North Dakota, USA, is close to extant Actaea based on fruit size and shape and details of seed morphology and anatomy [START_REF] Pigg | Paleoactaea gen. nov (Ranunculaceae) fruits from the Paleogene of North Dakota and the London clay[END_REF][START_REF] Stevens | onwards Angiosperm Phylogeny Website[END_REF].

In accordance with previous studies [START_REF] Wang | The rise of angiosperm-dominated herbaceous floras: Insights from Ranunculaceae[END_REF][START_REF] Chen | Molecular phylogeny and historical biogeography of Caltha (Ranunculaceae) based on analyses of multiple nuclear and plastid sequences[END_REF][START_REF] Kadereit | Dual colonization of European high-altitude areas from Asia by Callianthemum (Ranunculaceae)[END_REF], we gave a lower bound of 56 Ma for the stem group age of Actaea. Dating analyses were conducted under the GTR + I + Γ model for each DNA partition separately. Two independent runs of 100 million generations were carried out, sampling every 5000 generations and discarding the first 25% as burn-in. The maximum clade credibility (MCC) tree with median node ages was produced using TreeAnnotator 2.4.0 [START_REF] Bouckaert | BEAST 2: a software platform for Bayesian evolutionary analysis[END_REF].

We used Bayes factors [START_REF] Kass | Bayes factors[END_REF] calculated by marginal likelihoods derived from path sampling [START_REF] Baele | Accurate model selection of relaxed molecular clocks in Bayesian phylogenetics[END_REF] to compare four clock models (strict, exponential, lognormal, and random) and then two distinct tree speciation priors (Yule and birth-death). Under the optimal clock model (electronic supplementary material, table S4) and tree prior (electronic supplementary material, table S5), we carried out four dating analyses by using four distinct prior distributions for the calibrating fossil (exponential, lognormal, normal, and uniform).

These four analyses yielded highly similar time estimates for Eranthis (electronic supplementary material, table S6). Bayes factors suggested normal prior distribution as optimal for the calibrating fossil (electronic supplementary material, table S7); results from this analysis were thus reported and used for subsequent analyses. For comparative purposes, we also estimated divergence times using a pruned dataset consisting of 24 taxa (one accession per species). The run strategy was the same as the one for the full-data analysis. Both the full-data and species-level analyses highly agree in time estimates for Eranthis (electronic supplementary material, table S6).

To test the robustness of the age estimates, we also used the molecular dataset of Eranthis in combination with DNA substitution rate. Four plastid loci were linked in a single clock and a general plastid rate calculated by Zurawski et al. [START_REF] Zurawski | The nature of nucleotide sequence divergence between barley and maize chloroplast DNA[END_REF] was used. A normal distribution with mean value and standard deviation of 0.001 and 0.0001 were specified in ucld.mean. For the nuclear ITS, we selected the substitution rate of herbs used by Wojciechowski et al. [START_REF] Wojciechowski | Evidence on the monophyly of Astragalus (Fabaceae) and its major subgroups based on nuclear ribosomal DNA ITS and chloroplast DNA trnL intron data[END_REF]: a normal distribution with mean value and standard deviation of 0.0035 and 0.0002. The run settings were the same as the one using the full dataset and the fossil calibration under the optimal prior distribution. The dating results in combination with DNA substitution rate are highly congruent with those that uses only the calibrating fossil (electronic supplementary material, table S8).

(c) Ancestral range reconstruction

We estimated the ancestral ranges of Eranthis under the DEC and DIVALIKE models, implemented in the R package BioGeoBEARS [START_REF] Matzke | Probabilistic historical biogeography: new models for founder-event speciation, imperfect detection, and fossils allow improved accuracy and model-testing[END_REF]. The three outgroups were pruned.

Based on the present-day distribution of Eranthis and the floristic regions it covers, we defined five bioregions (figure 1a): Mediterranean region (including South Europe and the Anatolia peninsula), west QTP, east QTP, Northeast Asia (including eastern Siberia, northeastern China, Korea, and Japan), and MP. North America was also coded as a bioregion, in which a few species of Actaea occur. We performed the analyses in several different ways: without the maximum number of ancestral areas (maxarea) constrained and with the maxarea constrained to four, three, or two. Using the Akaike information criterion, DIVALIKE was selected to be the best-fitting model for each of these four analyses (electronic supplementary material, table S9). Under the DIVALIKE model, the results of the four analyses were largely similar based on cumulative ancestral range reconstructions [START_REF] Ebersbach | In and out of the Qinghai-Tibet Plateau: Divergence time estimation and historical biogeography of the large arctic-alpine genus Saxifraga L[END_REF]. Each species of Eranthis is distributed in one bioregion, the ancestral range reconstruction constraining maxarea to 2 was thus reported and discussed. To exclude the impact of outgroup areas, we also performed biogeographic analyses by pruning all outgroups. Both biogeographic estimations with or without outgroups entirely agree with the assigned ancestral areas for Eranthis (figure 2 vs. electronic supplementary material, figure S6).

(d) Ancestral elevation reconstruction

We obtained locality data of the 11 known Eranthis species from the following herbaria: B, E, IRK, K, LE, MO, NS, NSK, P, PE, TI, and TK (herbarium acronyms follow Index Herbariorum: http://sciweb.nybg.org/science2/Index-Herbariorum.asp), as well as from our field investigations. Localities without accurate coordinates were omitted. In total, we obtained 279 occurrence records with locality information (electronic supplementary material, table S2). For each record, we extracted the elevational variable from the calibration dataset for global land areas at 1-km resolution [START_REF] Ficka | WorldClim 2: New 1-km spatial resolution climate surfaces for global land areas[END_REF] using the 'raster' function in the 'raster' package of R [START_REF] Hijmans | raster: Geographic data analysis and modeling. R package version 3[END_REF]. The mean and boundaries of the 95% elevational range for each species was then calculated. We inferred the ancestral elevation of Eranthis using a squared-change parsimony method in Mesquite 3.61 [START_REF] Maddison | Mesquite: A modular system for evolutionary analysis[END_REF]. This method is relevant for analyzing continuous characters [START_REF] Paradis | APE: Analyses of phylogenetics and evolution in R language[END_REF]. The MCC tree was used and a single accession per species of Eranthis was kept. For comparative purposes, we also inferred the ancestral elevation using phylogenetic independent contrast (PIC) method, which is a Brownian-motion based estimator. The PIC analysis was conducted using the 'ace' function in the 'ape' package of R [START_REF] Wojciechowski | Evidence on the monophyly of Astragalus (Fabaceae) and its major subgroups based on nuclear ribosomal DNA ITS and chloroplast DNA trnL intron data[END_REF]. These two analytical methods generated highly congruent results (electronic supplementary material, figure S7). The parsimony reconstruction was reported and used for discussion (figure 3).

To test the robustness of the ancestral elevation inferences, we also used elevation information from the literature. The evolutionary reconstructions for boundaries of the elevational interval were carried out using the parsimony method in Mesquite. Both elevation reconstructions using the information extracted based on the occurrence records and from the literature yielded similar results (figure 3 vs. electronic supplementary material, figure S8).

Results

(a) Phylogeny

Maximum likelihood and Bayesian inference analyses resulted in identical trees (figure 1).

Both supported the monophyly of Eranthis and recovered Actaea as its sister group.

Eranthis includes two major monophyletic groups (I and II) and four well-supported clades (I-1, east QTP; I-2, North Asia; II-1, west QTP; II-2, Mediterranean), each defined largely by geography. Within clade I-2, the MP taxa clustered together and were nested within a clade of Northeast Asian taxa. The relationships at the species level within Eranthis are well resolved (figure 1).

(b) Divergence times

The divergence time estimates for Eranthis from the BEAST analysis under the optimal clock model (electronic supplementary material, table S4), tree prior (electronic supplementary material, table S5) and fossil prior distribution (electronic supplementary material, table S7) are shown in figure 2, and more details are provided in electronic supplementary material, figure S3 and table S6. The genus Eranthis started to diversify at c. 41 Ma (95% highest posterior density (HPD): 25-54 Ma). Within group I, the east QTP taxa (I-1) diverged from the North Asian taxa at c. 26 Ma (95% HPD: 14-41 Ma). The split of the MP subclade and its Northeast Asian sister group occurred at c. 17 Ma (95% HPD: 8-29 Ma), and the MP subclade diversified around 6 Ma (95% HPD: 2-13 Ma).

Diversification of group II occurred at c. 12 Ma (95% HPD: 4-24 Ma). The split between southern European and Anatolian species occurred at c. 6 Ma (95% HPD: 2-13 Ma).

(c) Ancestral range reconstruction

The ancestral area reconstructions using the DEC and DIVALIKE models in BioGeoBEARS generated highly congruent results, although DIVALIKE was identified as the best-fitting model for our data (electronic supplementary material, table S9). Results

Discussion

Our study presents a phylogenetic tree for Eranthis with all 12 species (figure 1).

Divergence time estimates suggest a crown group age of 62 Ma (95%HPD: 57-68 Ma) for Cimicifugeae and a stem age of 57 Ma (95%HPD: 53-60 Ma) for Actaea, which are consistent with the estimates of Wang et al. [START_REF] Wang | The rise of angiosperm-dominated herbaceous floras: Insights from Ranunculaceae[END_REF] and Kadereit et al. [START_REF] Kadereit | Dual colonization of European high-altitude areas from Asia by Callianthemum (Ranunculaceae)[END_REF], yet they are much older than those of Zhai et al. [START_REF] Zhai | Chloroplast genomic data provide new and robust insights into the phylogeny and evolution of the Ranunculaceae[END_REF]. However, Zhai et al. [START_REF] Zhai | Chloroplast genomic data provide new and robust insights into the phylogeny and evolution of the Ranunculaceae[END_REF] only used the oldest known angiosperm tricolpate pollen as the fossil calibration point to constrain the crown group age of eudicots and estimated divergence dates within Ranunculaceae. Internal calibrations are regarded critical for obtaining accurate estimates [START_REF] Yang | Bayesian estimation of species divergence times under a molecular clock using multiple fossil calibrations with soft bounds[END_REF]. Detailed comparisons among divergence times estimated in this study and previous studies are shown in electronic supplementary material (Table S8).

(a) Spatiotemporal evolution of Eranthis

The results of our integrated molecular dating and ancestral range reconstruction analyses (figure 2) indicate that Eranthis originated in East Asia, and then dispersed to the west QTP and Mediterranean regions between 56 and 41 Ma. This period is prior to the interruption of the Turgai strait (an epicontinental sea from the Arctic Ocean to the Tethys Seaway) (c. 34 Ma) [START_REF] Tiffney | Perspectives on the origin of the floristic similarity between eastern Asia and eastern North America[END_REF] and the demise of the Tarim Sea (an epicontinental sea in the western Tarim Basin of China) (c. 34-37 Ma) [START_REF] Bosboom | Late Eocene sea retreat from the Tarim Basin (West China) and concomitant Asian paleoenvironmental change[END_REF]. Thus, the migration through north temperate regions of North Asia seems unlikely. A proto-Tibetan highland consisting of the Lhasa and Qiangtang terranes had been assumed to have attained an elevation of >4.5 km before the collision of India with Eurasia, but these two high terranes were at least separated by an east-west oriented elongate lowland along the Bangong-Nujiang suture, represented by a string of basins at present, such as the Lunpola basin [START_REF] Spicer | Why the 'Uplift of the Tibetan Plateau' is a myth[END_REF][START_REF] Spicer | Cenozoic topography, monsoons and biodiversity conservation within the Tibetan Region: An evolving story[END_REF][START_REF] Su | No high Tibetan Plateau until the Neogene[END_REF][START_REF] Su | A Middle Eocene lowland humid subtropical "Shangri-La" ecosystem in central Tibet[END_REF][START_REF] Fang | Revised chronology of central Tibet uplift (Lunpola Basin)[END_REF]. Fossil evidence suggests that a humid tropical or subtropical lowland flora existed in the central QTP until the latest Paleocene to late Oligocene [START_REF] Deng | Major turnover of biotas across the Oligocene/Miocene boundary on the Tibetan Plateau[END_REF][START_REF] Su | A Middle Eocene lowland humid subtropical "Shangri-La" ecosystem in central Tibet[END_REF][START_REF] Fang | Revised chronology of central Tibet uplift (Lunpola Basin)[END_REF]. Thus, it is more likely that ancestral populations of Eranthis would have migrated westward via this east-west lowland during the Eocene (figure 2a).

The spatiotemporal evolution of Eranthis may have resulted from the uplifts of the three great Asian plateaus and associated mountain belts, as well as associated environmental changes. The uplift of the QTP region induced by the India-Eurasia collision might have prevented the migration of Eranthis through the east-west lowland and thereby resulted in the deep divergence between groups I and II at c. 41 Ma (95% HPD: 25-54 Ma).

Later, the QTP grew continuously upward and outward and caused the formation of Lüliang and Qinling orogens in the northeast [START_REF] Li | The tectonic-sedimentary evolution of the Lüliang mountains since the Miocene[END_REF][START_REF] Wang | The tectonic uplift and its environmental effects of the Qinling Mountains during the Cenozoic Era: Progress and problems[END_REF] and northern Chinese desertification [START_REF] Qiang | New eolian red clay sequence on the western Chinese Loess Plateau linked to onset of Asian desertification about 25 Ma ago[END_REF]. These events could have caused the extinction of Eranthis in the northern and central China and thereby resulted in the split between clades I-1 and I-2 around 26 Ma (95% HPD: 14-41 Ma). Contrary to the Northeast Asian taxa, the east QTP taxa are distributed at higher elevations (figure 3). Our analyses recovered one dispersal from Northeast Asia to the MP, suggesting Northeast Asia as the source for the MP biodiversity. The split between the MP subclade and its Northeast Asian sister group at c. 17 Ma (95% HPD: 8-29 Ma) is likely related to the uplift of the MP region, which was suggested to be the consequence of the far-field effect of the India-Eurasia and Arabia-Eurasia collisions [START_REF] Yin | Cenozoic tectonic evolution of Asia: A preliminary synthesis[END_REF]. The divergence of the west QTP clade (II-1) from the Mediterranean taxa (II-2) around 12 Ma (95% HPD: 4-24 Ma) is probably linked to the uplift of the IP region and the extrusion of the Anatolia block resulting from the Arabia-Eurasia collision.

(b) Biogeographic implications for the geological evolution of Asia

Because of its conserved habitat, poor dispersal ability, and scattered geographic distribution across Eurasia, Eranthis possesses unique genetic imprints that reflect the history of geological and environmental changes in Asia during the Cenozoic.

Following the initial India-Eurasia collision in the early Cenozoic (~ 61 Ma) [START_REF] Yuan | Rapid drift of the Tethyan Himalaya terrane before two-stage India-Asia collision[END_REF], the uplift and outward growth of the QTP occurred and continued well into the Miocene [START_REF] Spicer | Cenozoic topography, monsoons and biodiversity conservation within the Tibetan Region: An evolving story[END_REF].

The deep divergence between the east and west groups (I and II) took place in the central part of the QTP around 41 Ma, associated with the first uplift of the QTP (figure 2a). This timeframe is in concert with the results of geological studies, which indicate that the east and central QTP and its northern margin were elevated around 40 Ma [START_REF] Chung | Diachronous uplift of the Tibetan plateau starting 40 Myr ago[END_REF][START_REF] Wang | Constraints on the early uplift history of the Tibetan Plateau[END_REF][START_REF] Shi | A prolonged Cenozoic erosional period in East Kunlun (Western China): Constraints of detrital apatite (U-Th)/He ages on the onset of mountain building along the northern margin of the Tibetan Plateau[END_REF]. The origin of alpine ginger [START_REF] Zhao | Evolutionary diversification of alpine ginger reflects the early uplift of the Himalayan-Tibetan Plateau and rapid extrusion of Indochina[END_REF] and the divergence of west Chinese salamanders [START_REF] Zhang | Phylogeny, evolution, and biogeography of Asiatic Salamanders (Hynobiidae)[END_REF] were estimated to have occurred during the same period, both of which were also suggested to be related to this uplift of the QTP region. The paleoelevational estimate for the hinterland of the QTP, i.e., a string of basins between the Lhasa and Qiangtang terranes, is crucial for understanding Tibetan landscape evolution, but has been intensely debated [START_REF] Yin | Cenozoic tectonic evolution of Asia: A preliminary synthesis[END_REF][START_REF] Deng | Major turnover of biotas across the Oligocene/Miocene boundary on the Tibetan Plateau[END_REF][START_REF] Su | No high Tibetan Plateau until the Neogene[END_REF]. Our elevation reconstructions suggest that the most recent common ancestor of Eranthis likely occurred up to the highest altitude of about 1.8 km (figure 3). Accordingly, we hypothesize that after the first uplift (c. 40 Ma), the central QTP likely had an elevation of <2 km, likely about 1.8 km (figure 2a). This differs from the hypothesis that during the first uplift the QTP reached an altitude of at least 2.5 km [START_REF] Zhang | Phylogeny, evolution, and biogeography of Asiatic Salamanders (Hynobiidae)[END_REF]. Our estimate is compatible with the result of the magnetostratigraphic and radiochronologic dating that indicates that the elevations of central Tibet were generally low (<2.3 km) at 39.5 Ma [START_REF] Fang | Revised chronology of central Tibet uplift (Lunpola Basin)[END_REF]. Also, the paleobotanical evidence suggests an elevation of <2.3 km for the Lunpola basin of central QTP at c. 25 Ma [START_REF] Su | No high Tibetan Plateau until the Neogene[END_REF] and an elevation of 1.5-2.9 km for the Kailas Basin in the western part of the Lhasa terrane during the latest Oligocene [START_REF] Ai | The uppermost Oligocene Kailas flora from southern Tibetan Plateau and its implications for the uplift history of the southern Lhasa terrane[END_REF].

The split between the east QTP clade (I-1) and the North Asian clade (I-2) (figure 2b; c. 26 Ma) suggests a second uplift of the QTP region and the resulting paleoclimatic changes. This age fell in line with the deep splits of lineages in alpine ginger (c. 32 Ma, 95% HPD: 18-50 Ma) [START_REF] Zhao | Evolutionary diversification of alpine ginger reflects the early uplift of the Himalayan-Tibetan Plateau and rapid extrusion of Indochina[END_REF] and in spiny frogs (c. 27 Ma, 95% HPD: 19-36 Ma) [START_REF] Che | Spiny frogs (Paini) illuminate the history of the Himalayan region and southeast Asia[END_REF], both of which were postulated to have been triggered by this rapid uplift. A few endemic plant genera of the QTP, such as Nannoglottis (Asteraceae) [START_REF] Liu | Molecular phylogeny and biogeography of the Qinghai-Tibet Plateau endemic Nannoglottis (Asteraceae)[END_REF] and Gymnaconitum (Ranunculaceae) [START_REF] Wang | Gymnaconitum, a new genus of Ranunculaceae endemic to the Qinghai-Tibetan Plateau[END_REF] also originated during this period. This uplift has not only significantly modified Asian physiognomy, but has also greatly affected the regional and even the global climate and biotic evolution [START_REF] Deng | Major turnover of biotas across the Oligocene/Miocene boundary on the Tibetan Plateau[END_REF][START_REF] Chung | Diachronous uplift of the Tibetan plateau starting 40 Myr ago[END_REF]. During this period, the QTP extended outward and consequently gave rise to adjacent mountain buildings by a far-field effect, such as the Emei mountain in the east [START_REF] Meng | Uplift of the Emei Shan, western Sichuan Basin: Implication for eastward propagation of the Tibetan Plateau in Early Miocene[END_REF] and Lüliang and Qinling mountains in the northeast [START_REF] Li | The tectonic-sedimentary evolution of the Lüliang mountains since the Miocene[END_REF][START_REF] Wang | The tectonic uplift and its environmental effects of the Qinling Mountains during the Cenozoic Era: Progress and problems[END_REF], the onset of Asian interior desertification (c. 25 Ma) [START_REF] Qiang | New eolian red clay sequence on the western Chinese Loess Plateau linked to onset of Asian desertification about 25 Ma ago[END_REF], and the establishment of East Asian monsoon climate [START_REF] Ding | Palaeovegetation variation in response to the late Oligocene-early Miocene East Asian summer monsoon in the Ying-Qiong Basin, South China Sea[END_REF]. Paleontological evidence shows that radical changes in vegetation and biotic components occurred in the TP and adjacent regions around the Oligocene-Miocene transition [START_REF] Deng | Major turnover of biotas across the Oligocene/Miocene boundary on the Tibetan Plateau[END_REF][START_REF] Sun | How old is the Asian monsoon system? Palaeobotanical records from China[END_REF].

The split between the west QTP clade (II-1) and the Mediterranean clade (II-2) (figure 2c; c. 12 Ma) suggests a rapid uplift of the IP region triggered by the Arabia-Eurasia collision, which served slab detachment in eastern Turkey and northwestern Zagros [START_REF] Mouthereau | Building the Zagros collisional orogen: Timing, strain distribution and the dynamics of Arabia/Eurasia plate convergence[END_REF].

Our dating is highly consistent with the geological hypothesis that uplift, exhumation and shortening in the Zagros mountains and the IP region accelerated at about 15-12 Ma [START_REF] Mouthereau | Building the Zagros collisional orogen: Timing, strain distribution and the dynamics of Arabia/Eurasia plate convergence[END_REF].

The Arabia-Eurasia collision displaced the Anatolia block westward relative to the IP, which occurred at about or 7-5 Ma [START_REF] Westaway | Kinematic consistency between the Dead Sea Fault Zone and the Neogene and Quaternary left-lateral faulting in SE Turkey[END_REF][START_REF] Hubert-Ferrari | Morphology, displacement, and slip rates along the North Anatolian Fault, Turkey[END_REF]. Contrary to the eastward lateral extrusion of Indochina that has been widely investigated [START_REF] Zhao | Evolutionary diversification of alpine ginger reflects the early uplift of the Himalayan-Tibetan Plateau and rapid extrusion of Indochina[END_REF][START_REF] Che | Spiny frogs (Paini) illuminate the history of the Himalayan region and southeast Asia[END_REF][START_REF] Luo | Caver Stedocys spitting spiders illuminate the history of the Himalayas and southeast Asia[END_REF], the westward lateral extrusion of the Anatolia block has yet to be examined using modern phylogenetic and biogeographic methods. Our data suggest that the uplift and tectonic extrusion should have occurred synchronously to generate the observed distribution pattern in Eranthis around 12 Ma (figure 2c). The split between the Anatolian and southern European species was estimated at c. 6 Ma, associated with the progressive uplift and expansion of the IP region. This is supported by geological studies, indicating an about 7-8 Ma uplift of the southern margin of the Anatolia block [START_REF] Meijers | Rapid late Miocene surface uplift of the Central Anatolian Plateau margin[END_REF] and an about 5 Ma active mountain building of the Zagros mountains [START_REF] Lacombe | Late Cenozoic and modern stress fields in the western Fars (Iran): Implications for the tectonic and kinematic evolution of central Zagros[END_REF].

The split of the MP subclade and its Northeast Asian sister group occurred at about 17 Ma. The species in the MP are distributed at relatively higher elevations than the species in the Northeast Asia (figure 3). We hypothesize that the split was associated with geological events that separated the MP and Northeast Asia. Our estimated age is compatible with the geological hypothesis that the rapid uplift of the MP region occurred around 20-10 Ma [START_REF] Smith | Paleotopography and erosion rates in the central Hangay Dome, Mongolia: Landscape evolution since the mid-Miocene[END_REF][START_REF] Feng | Thrusting and exhumation of the southern Mongolian Plateau: Joint thermochronological constraints from the Langshan Mountains, western Inner Mongolia, China[END_REF]. The diversification of the MP subclade arose at about 6 Ma (figure 2d), which implies a second uplift and expansion of the MP region. This hypothesis is consistent with the geological hypotheses that the summit of the MP emerged at about 5 to 10 Ma [START_REF] Kurushin | The surface rupture of the 1957 Gobi-Altay, Mongolia, earthquake[END_REF] and that the uplift of the Gobi-Altai mountain range of Mongolia started at 5 ± 3 Ma [START_REF] Vassallo | Uplift age and rates of the Gurvan Bogd system (Gobi-Altay) by apatite fission track analysis[END_REF]. The two discrete intervals of uplift of the MP region that our analyses revealed are concurrent with those of the rapid uplift and expansion of the QTP and IP regions (figures 2b, c) [START_REF] Mouthereau | Building the Zagros collisional orogen: Timing, strain distribution and the dynamics of Arabia/Eurasia plate convergence[END_REF][START_REF] Kurushin | The surface rupture of the 1957 Gobi-Altay, Mongolia, earthquake[END_REF][START_REF] Hodell | Variations in the strontium isotopic composition of seawater during the Neogene[END_REF], and with those of the Tian Shan and Altai Shan [START_REF] Caves | Late Miocene uplift of the Tian Shan and Altai and reorganization of Central Asia climate[END_REF][START_REF] Wang | The uplift history of south-western Tian Shan: Implications from AFT analysis of detrital samples[END_REF]. The MP, Tian Shan, and Altai Shan are all located in the Central Asian right-slip fault zone (electronic supplementary material, figure S1) [START_REF] Yin | Cenozoic tectonic evolution of Asia: A preliminary synthesis[END_REF]. Thus, we suggest that the two uplifts of the MP region in the Miocene likely represent two rapid responses to the interaction of the India-Eurasia and Arabia-Eurasia collisions.

In conclusion, our study provides the first case, to our knowledge, of an organismal group reconciling and illuminating the geological history of the QTP, IP and MP. The biogeography of Eranthis may have been shaped by multiple uplifts of these three Asian plateaus and the extrusion of the Anatolia block. Considering the wide credibility intervals of the estimated times of divergence, we urge caution interpreting these results and the geological hypotheses that we put forward should be further tested by studying other taxa that exhibit similar habitat requirements and dispersal abilities in Eurasia. (c) The map shows sampling locations, detailed sampling information is presented in electronic supplementary material, table S3. Photographs by Andrey S. Erst. 2d shows the interacted zone between the India-Eurasia and Arabia-Eurasia collisions [START_REF] Yin | Cenozoic tectonic evolution of Asia: A preliminary synthesis[END_REF]. A series of palaeo-maps were reconstructed from http:// www.odsn.de/odsn/index.html (link "Plate Tectonic Reconstructions"), and tectonic structures are modified from Yin [START_REF] Yin | Cenozoic tectonic evolution of Asia: A preliminary synthesis[END_REF]. AB, Anatolia block; AS, Altai Shan; IP, Iranian Plateau; LB, Lunpola basin; LM, Lüliang mountains; LT, Lhasa terrane; MP, Mongolian Plateau; QL, Qinling; QT, Qiangtang terrane; QTP, Qinghai-Tibetan Plateau; TS, Tian Shan; ZM, Zagros mountains.

  under the DIVALIKE model are shown in figure 2. The genus Eranthis originated in East Asia, and its most recent common ancestor had likely spread to the west QTP and Mediterranean regions. A subsequent vicariance event led to group I in East Asia and group II in the west QTP, Anatolia, and South Europe. Within group I, a vicariance event separated the east QTP clade (I-1) from the North Asian clade (I-2), and a dispersal event from Northeast Asia to the MP within clade I-2 took place. Within group II, a vicariance event isolated the west QTP clade (II-1) and the Mediterranean clade (II-2).

Figure 1 .

 1 Figure 1. Phylogenetic relationships of Eranthis inferred from the combined plastid and

Figure 2 .

 2 Figure 2. Evolution of Eranthis with associated tectonic events. Timetree is modified from

Figure 3 .

 3 Figure 3. Ancestral reconstruction of the mean and boundaries of the 95% elevational

Figure 1 .E

 1 Figure 1.

Figure 2 .

 2 Figure 2.

Figure 3 .

 3 Figure 3.

Acknowledgements. We thank the associated editor and two anonymous reviewers for constructive comments which greatly improved this manuscript. We thank Cynthia Hong Wa for reading an early draft of the manuscript and Xiao-Guo Xiang for critical comments on the manuscript.

Funding. This work was supported by the Strategic Priority Research Program of Chinese Academy of Sciences (XDB31000000), the National Natural Science Foundation of China (31770231, 31770233 and 32011530072), K.C.Wong Education Foundation (GJTD-2020-05), and the Russian Science Foundation (19-74-10082).

Data accessibility. The sequences reported in this paper have been deposited in the GenBank database (accession nos. MW716473-MW716501; MW722251-MW722357).

The datasets supporting this article can be accessed from Dryad (doi:10.5061/dryad.0zpc866wz).

Authors' contributions. W.W. conceived the research. K.L.X., A.S.E., H.W.P, R.C.O., F.J. and TVE carried out taxon sampling. K.L.X., A.S.E. and J.Y. generated all the data. K.L.X., A.S.E., J.Y., H.W.P. and W.W. performed the data analyses. W.W. led the writing with contributions from all co-authors.

Competing interests. The authors declare no competing interests.