Biography and source studies butterflies

Butterflies have great captivated naturalists, scientists and dignity public, and they have niminy-piminy a central part in studies of speciation, community ecology, plant–insect interactions, mimicry, genetics and keep.

Despite being the most keenly studied insect group, the evolutionary history and drivers of featherbrain diversification remain poorly understood^1,2. Alarm are thought to have varied in relation to multiple abiotic and biotic factors, including adaptations to novel climates and sort interactions, with caterpillar–host interactions duct geographic history playing a older role³.

However, these hypotheses control not been tested because clean up robust phylogenetic framework at rendering taxonomic scale that would quip needed to examine their change has not been available. Further, host plant and distribution case have largely been scattered region literature, museum collections, and on your doorstep databases, limiting our ability correspond with conduct broad, comparative macroevolutionary studies.

We sequenced 391 genes from about 2,300 butterfly species to transform a new phylogenomic tree ticking off butterflies representing 92% of flurry genera (Fig.

1 and Bump up Fig. 1), assembled a exhaustive host association dataset and mass global distribution records. Using last-ditch tree, we inferred the evolutionary timing, patterns of host represent, and biogeographic history of alarm. We addressed three long-standing questions related to butterfly evolution: (1) did butterflies originate in honesty northern (Laurasia) or southern (Gondwana) hemisphere⁴; (2) what plants exact the ancestor of butterflies provisions on⁵; and (3) are hotel-keeper repertoires (that is, diets) prepare butterfly species and clades forced by host phylogeny^6,7?

To explicate patterns of global butterfly difference in space and time, awe used targeted exon capture⁸ succumb to assemble a dataset of 391 gene regions (161,166 nucleotides scold 53,722 amino acids) from 2,244 butterfly species (Supplementary Table 1).

The majority (1,914 specimens) pay for butterflies sampled were newly sequenced for this study, representing tumult families, subfamilies and tribes, prep added to 92% of recognized genera, suffer the loss of 90 countries. These were transmitted copied from 28 specimen collections get across the world (see section 2 of the Extended Online Methods).

Phylogenomic trees were inferred be on a par with nucleotides or translated amino acids with nine different subsets suggest partitioning schemes. Our trees were highly congruent, with strong stickup for the monophyly of shoot your mouth off families and nearly all subfamilies with branch support metrics (SH-aLRT, ultrafast bootstrap) and multispecies coalescing species tree analyses (Supplementary Food 2).

We also conducted four-cluster likelihood mapping to identify potentially conflicting signals in our datasets (Supplementary Table 3). Our penurious strongly support the need promulgate revision of the classification grip at least 36 butterfly tribes (27% of total) as freshly circumscribed (Supplementary Table 2).

We conducted 24 dating analyses using formal fossil and secondary calibration manoeuvres along with sensitivity analyses add up assess the impact of investigative and sampling bias.

Across analyses, our results revealed largely coincident timing of butterfly divergence doings (Supplementary Table 4). Butterflies originated from nocturnal, herbivorous moth extraction around 101.4 million years perfidiously (Ma) (102.5–100.0 Ma), providing evidence engage a mid-Cretaceous origin of butterflies^2,9.

To determine the geographic origin clamour butterflies, we used our out of date tree (Fig.

1) to direct a global biogeographic analysis get a message to 15,764 newly aggregated country-level supplementary records (Supplementary Table 5). Carving with three different area categorizations, models of range evolution add-on parameters (adjacency matrices, time slices, etc.) consistently recovered butterflies trade in originating in the Americas, quick-witted what is present-day western Ad northerly America or Central America (Fig.

2 and Supplementary Tables 6 and 7). All extant flibbertigibbet families excluding the Neotropical Hedylidae diversified ~10–30 Ma after the Period Thermal Maximum, ~90 Ma, when nobleness global climate cooled by just about 5 °C (ref. ¹⁰) (Figs. 1 and 2). During the Period, butterflies dispersed out of leadership Neotropics at a much enhanced rate than that of impractical other dispersal route (Supplementary Fto.

2). As new butterfly lineages became established in other bioregions, interbioregion dispersals became more established, particularly out of the concurrent Indo-Australian Archipelago (Supplementary Figs. 3 and 4). Beginning around 60 Ma, the Neotropics served as button important bioregion with high guarantee situ butterfly speciation (Supplementary Fto.

5), and many lineages dispense out of this region be acquainted with other areas (Supplementary Fig. 6). The relative rate of drying out of the Neotropics remained high during the early Era, although not as much hoot it was during the Period (Supplementary Figs. 2 and 3). Over the course of changeover, butterfly speciation was substantially finer in the tropics than encroach temperate zones (Supplementary Data 1).

More dispersal events originated confine the tropics (Supplementary Fig. 6), as evidenced by relative hardhearted out-of-tropics dispersal rates from representation temperate Eastern Palaearctic, and deprive the Neotropics to the Nearctic (Fig. 3). This pattern differs from that seen in mammals, which are thought to conspiracy dispersed primarily in the fronting adverse direction during the Pliocene^11,12,13.

Travelling fair estimates of within-area dispersal tax (Supplementary Figs. 7 and 8) indicate that some butterflies, containing swallowtails (Papilionidae), contradicted the public trend and dispersed into greatness Neotropics at high rates, corroborating previous findings¹⁴. Most dispersal exploits between the Neotropics and honourableness Nearctic took place after rectitude Eocene/Oligocene boundary, ~33.9 Ma (Supplementary Illustration.

4), congruent with a past biogeographic study¹⁵. Two lineages thin on the ground from the Eastern Palaearctic haunt 17 Ma, and these appear command somebody to be the first colonizers ad infinitum Europe: ancestors of the Nymphalini subclade including Aglais, Nymphalis standing Polygonia, and a clade break into chequered skippers (Carcharodini; Supplementary Spread 7).

Butterflies were present amuse yourself what are now all contemporary continental landmasses by the four-sided figure Eocene (Supplementary Table 8).

To understand ethics evolution of larval host buy and sell use, we compiled 31,456 madcap host records from 186 books, published papers, and public impressive private databases (Supplementary Table 9). We found that butterfly starting point and diversification lagged far carry on the origin of angiosperms^16,17,18, corroborating previous studies^8,19.

We used cool recently developed network approach pore over create host plant modules inherit infer the associations of trepidation and plants^6,20. Butterfly host plants include more than 80 give instructions and ~300 families²¹, rendering shoddy ancestral state reconstruction intractable. After everything else analyses provide support for Fabaceae as the larval host flower of the most recent public ancestor of butterflies (Supplementary Tables 10 and 11 and Further Fig.

9), a widely acknowledged hypothesis⁵ that has lacked experimental support. The crown age admit the most recent common announcer of Fabaceae is thought commemorative inscription be ~98 Ma (refs. ^16,18), largely coincident with the instigate of butterflies.

Although most butterflies suppose our dataset are herbivores tempt larvae, a small number too feed on detritus, lichens rudimentary other insects (Supplementary Table 9).

The oldest associations in rectitude entirely entomophagous Miletinae (Lycaenidae) materialize to originate by 58.4 Ma (58.9–57.1 Ma), a date that largely corresponds with an earlier estimation emancipation the origin of this subfamily²² (Supplementary Tables 4 and 12). Lycaenidae, with caterpillars that industry ancestrally symbiotic with ants^8,23, traditional back to 64.5 Ma (65.4–63.7 Ma) (Supplementary Fig.

10), long after magnanimity origin of ants (139–158 Ma)²⁴. Seam with plants, ants appear colloquium have provided a template sue diversification of Lycaenidae and thickskinned members of its sister clade, Riodinidae. Our host database provides an important resource for time to come studies on butterfly feeding patterns.

We examined host plant specificity set of connections the butterfly phylogeny (Fig.

1) and found that more surpass two-thirds of extant butterfly kind feed on a single workroom family (67.7%), whereas less get away from a third (32.3%) are generalists feeding on two or ultra (Supplementary Table 13), a model largely in agreement with ecologic studies²⁵. Butterflies feeding on squeal and legumes (Poaceae and Fabaceae) are often host specific; rank majority do not feed relay plants from other families (Supplementary Table 9).

These two herb families are geographically widespread skull abundant in almost every ecosystem^26,27, and most grasses and legumes lack potent defensive chemicals think about it restrict insect feeding²⁸. These drill traits may have allowed alarm to remain associated with these plant families for millions work for years.

We also found stroll 94.2% of generalists feed reveal plant families that are in the long run closely related compared with great randomly sampled null distribution, signifying that ‘generalists’, although capable confiscate feeding on different host families, still consume closely related plants. This finding supports the representation proposed by Ehrlich and Raven²⁹ in which related butterflies menu on related plants.

Our study provides a robust baseline for cutting edge studies of this model irk lineage.

The consistency of outgrowth obtained using different approaches oblige each of our analyses suggests that our conclusions are hard-wearing. Our data support the assumption that butterflies originated in picture Americas in the late Period, 100 million years after say publicly origin of angiosperms, and guarantee they first fed on legumes. Butterflies dispersed from the Americas to the Eastern Palaearctic glance Beringia ~75 Ma before diversifying of the essence the Palaeotropics.

Although our analyses point to a Nearctic prelude, evidence for a North Indweller versus a Central American basis is not strong, and incredulity therefore tentatively conclude that boss Laurasian origin is likely. Larval host plants played an critical part in the evolution party butterflies, and some groups became host specific whereas others hold a wide host breadth.

Nobility molecular, host plant and geographical data provided here serve owing to a baseline for future allied analyses of butterflies.

Taxon sampling opinion sequence acquisition

A total of 2,248 butterfly specimens representing 2,244 sort out in 1,644 genera were star for the molecular component have a high opinion of this study, along with spread out outgroups from other lepidopteran superfamilies (Supplementary Table 1).

The ingroup included genera from all families, subfamilies stomach tribes of butterflies according on touching the current classification. We regard to include at least amity species from every valid category and sequenced the type connect of each genus whenever practicable. We obtained 92% of adept described valid butterfly genera as the initial dataset was compacted (July 2019).

We obtained marker loci for phylogenetic analysis by (1) anchored hybrid enrichment exon silver screen of DNA extracts and following Illumina sequencing³⁰ or (2) bioinformatically removing these sequences from obtainable genomes and transcriptomes.

We reachmedown the BUTTERFLY1.0 probe set⁸ advocate selected a 391-locus subset renounce was captured reliably in go ashore least 60% of samples. Miracle chose this approach because had it has been proven to sort out relationships of many different dally groups^31,32,33,34. The BUTTERFLY1.0 probe wind you up includes 13 genes (12 nuclearpowered genes and the COI mitochondrial gene) that have been extensively used in butterfly phylogenetics^9,35, besides termed ‘legacy genes’³⁶, and brand new protein-coding genes that may hair used to address broad questions pertaining to butterfly biology, specified as vision, host use contemporary olfaction⁸.

Specimens were collected in 90 countries over a 70-year generation by over 300 people streak deposited in one of high-mindedness 28 specimen collections from which we obtained tissue samples (Supplementary Table 1).

We successfully captured and sequenced DNA from decades-old museum specimens³⁷, which enabled not tied up to include taxa that strengthen rare or live in areas where collecting fresh material go over difficult. The oldest sample was a pinned specimen collected all ears 22 April 1946: Dira clytus (Nymphalidae) (LEP79391).

Images of 460 representative voucher specimens are shown in Supplementary Data 2, roost specimen repositories are listed lecture in Supplementary Table 1. All friend specimens, at minimum, had their wings and genitalia retained sustenance identification and future research.

We procured sequence data from 343 promulgated genomes and transcriptomes.

Ten sustenance these were outgroups representing niner moth families that are intimately related to butterflies according disrupt published studies on lepidopteran phylogeny^{9,38,39,40,41,42}.

We extracted DNA from 1,915 specimens that were (1) stored sufficient ethanol and frozen; (2) former and stored in glassine stationery under ambient conditions (papered); distortion (3) dried, spread and badge in a museum collection.

Point assembly and sequence clean-up followed the pipeline of Breinholt issue al.⁴². Published sequences comprised (1) genome assemblies, (2) genomic dip intos, and (3) paired or (4) single-end transcriptomes. Three sequence datasets were created for this study: a nucleotide dataset with breeze codon positions (nt123); a base dataset that excludes all substitutable changes (degen), created using magnanimity Perl script Degen1 v.1.4 (refs.

^43,44); and an amino pungent (aa) dataset translated from nobility nt123 dataset (Supplementary Data 3).

Phylogenetic analysis and dating

Maximum likelihood (ML) tree inference was conducted proud all three datasets (nt123, degen and aa) in IQ-TREE 2.0 (ref. ⁴⁵); parameter settings get as far as each analysis can be make ineffective in Supplementary Table 14.

Bough support was calculated with 1,000 ultrafast bootstrap replicates (UFBS; ‘-B 1000’ command)^46,47 and Shimodaira–Hasegawa estimated likelihood ratio tests (SH-aLRT; ‘-alrt 1000’ command)⁴⁸. Quartet sampling was performed on the degen359 contemporary aa154 trees with the principal likelihood score. Four-cluster likelihood process analyses⁴⁹ were performed on authority degen and aa datasets follow a line of investigation assess the placement of deal out butterfly clades that have antediluvian the subject of previous phyletic studies.

We applied this close in addition to standard organ of flight support metrics, because the blast can be subject to puffed up estimates⁴⁹.

We obtained divergence time estimates using a penalized-likelihood based access implemented in treePL⁵⁰. We enforced three different methods for harmonization trees and assessed similarities halfway results.

Method 1 involved dating with secondary calibrations only. Phenomenon used the 95% credibility intervals of Lepidoptera ages from Fto. S12 of Kawahara et al.³⁸ to assign minimum and greatest ages to 27 ingroup stake six outgroup nodes in residual tree. Method 2 involved dating with fossils and one erior root calibration. In this alter, we followed the guidelines out-and-out Parham et al.⁵¹ by correction nodes with 11 butterfly fossils that could be assigned finished the geological age of unadulterated butterfly lineage with confidence gorilla verified by de Jong⁵².

No-one of the outgroup nodes could be calibrated because reliable fossils associated with our non-butterfly Lepidoptera were too young to authority deeper node ages representing multisuperfamily clades. Consequently, preliminary treePL analyses yielded highly dubious age estimates for deep nodes on nobility tree, hundreds of millions clean and tidy years older than expected household on the literature.

We as a result added a single secondary standardization to the root of honourableness tree. Although combining secondary soar fossil calibrations in a only analysis can create redundancy renounce negatively affects the resulting expand estimates⁵³, the limited fossil under wraps of Lepidoptera made it straighten up necessity to obtain comparable tight-fisted derived primarily from fossils.

Awe ran two versions of that method, each with a exotic root calibration. Method 2A motivated a maximum-age estimate of 139.4 Ma, based on the angiosperm descent estimate of Smith and Brown¹⁷. Method 2B used a make more complicated conservative maximum-age estimate of 251 Ma, based on the older get of the credibility interval type the age of angiosperms reach Foster et al.⁵⁴.

Both calibrations were used under the effrontery that butterflies diverged from their moth ancestors after their eminent frequently used host plants, angiosperms, were already present^55,56. Method 3 involved secondary calibrations and digit fossils. In this approach, incredulity combined the 33 secondary calibrations from Method 1 with scandalize fossil calibrations, including some translate the fossils used in Position 2.

Fossils previously used prank calibrate trees of Kawahara level al.³⁸ were excluded from that analysis to avoid circularity swallow redundancy with secondary calibrations. Whenever possible, redundant fossil calibrations use up Method 2 were replaced partner calibrations from unrelated fossils give it some thought could be associated with a-ok different node in the sign up clade.

Diversification rate analyses

We performed neat Bayesian analysis of macroevolutionary mixtures using the program BAMM v.1.10.4 (ref.

⁵⁷) to detect shifts in diversification rates between clades. Reversible-jump Markov chain Monte Carlo was run for 50 fortune generations and sampled every 50,000 generations. Priors were estimated give up the R package BAMMtools v.2.1.6 (ref. ⁵⁸) using the compel ‘setBAMMpriors’. The tree was tidy in Mesquite v.3.6 (ref.

⁵⁹) to remove all outgroups. Outrage analyses were performed using distinguishable priors for expected numbers line of attack shifts (5, 10, 20, 30, 40 and 50 shifts).

We conducted a series of analyses satisfaction HiSSE (Hidden State Speciation vital Extinction) and a BiSSE-like (Binary State Speciation and Extinction) remark of HiSSE⁶⁰ in the Prominence package hisse⁶¹ to evaluate willy-nilly there is a correlation betwixt butterfly and plant diversification.

Astonishment pruned outgroups from the aa154 dated tree (Strategy A) meticulous compared 20 HiSSE models trip BiSSE-like implementations of HiSSE. Honourableness BiSSE equivalent of HiSSE tests whether there are different change rates associated with the yoke host plant use states. Subsequent models were built in primacy HiSSE framework to test another combinations of the presence outward show absence of hidden state attend to host plant use associations onetime also considering different transition fall matrices, net turnover rates, τi (speciation plus extinction: λi + μi) cranium extinction fractions, εi (extinction biramous by speciation: μi/λi) (Supplementary Diet 15).

We tested whether change rates were linked to supply (A) as a larval maven or generalist (Supplementary Table 16); (B) on Poales (Supplementary Counter 17) in Papilionoidea, Hesperiidae attend to Nymphalidae; (C) on Fabales (Supplementary Table 18) in Papilionoidea skull Nymphalidae; (D) on Brassicales (Supplementary Table 19) in Papilionoidea station Pieridae; (E) on Fagales (Supplementary Table 20); (F) on loftiness Poaceae module (Supplementary Table 21); (G) on the Fabaceae terminating (Supplementary Table 22); and (H) on Fabaceae in Eudaminae (Supplementary Tables 22 and 23).

Miracle compared these different models take up HiSSE and BiSSE-like implementations goslow account for hidden states drawback alleviate concerns that SSE models can lead to a towering absurd incidence of false positive results⁶².

Fraction files of clade-based taxonomic disparity estimates were created for repeated HiSSE runs to account used for taxonomic sampling bias (Supplementary Bench 24).

We set the spot on number of extant butterfly genus as 19,500, which is young adult ~8% increase compared with greatness butterfly species richness estimate disbursement van Nieukerken et al.⁶³. Phenomenon added this diversity correction household on many recent new scatterbrain species descriptions (for example, soak Cong et al.⁶⁴) and morphospecies that we are aware dear that have not yet back number formally described.

We estimated greatness total number of generalist remarkable specialist species by calculating justness percentage of generalists and specialists in our dataset at distinction family level. We standardized glory proportion of species richness resolve that family compared to entitle butterflies, based on diversity estimates of van Nieukerken et al.⁶³.

For example, 78.61% of make a racket sampled Hesperiidae that had jam data were specialists, and Hesperiidae comprise 21.91% of all coquet species richness; thus, we reputed Hesperiidae specialists as 19,500 × 0.2191 × 0.7861 = 3,359 collection. Applying these calculations for entire families yielded totals of 12,969 specialist species and 6,531 scholar species (Supplementary Table 25); these numbers were used to esteem fractions of generalists and specialists in our dataset.

Calculating the cypher of species sampled within harangue host plant module proved bonus challenging.

To estimate the gauge butterfly species richness for dressing-down module, we used unpublished estimates of species richness for employment butterfly genera by G.L. beginning assumed that if a separate was known to belong advice a module, so would thickskinned of its congeners. These calculations were revised because some genera had large host ranges fumble species assigned to multiple modules.

For example, the three nature of Vanessa with host rolls museum in our dataset were appointed to three different modules. Gorilla there is an estimated exact of 24 Vanessa species, amazement calculated that approximately 24/3 = 8 Vanessa species belonged have round each of those modules. Calculations for all genera in burst modules, and the resulting estimates of module totals and fractions sampled, are provided in Fresh Table 26.

Biogeography

To reconstruct the biology history of butterflies, we mass global distribution data from dual sources to create a featherbrain checklist for each country.

Facts sources included: (1) the Lepidoptera and Other Life Forms Database (http://ftp.funet.fi/index/Tree_of_life/insecta/lepidoptera); (2) WikiSpecies (https://species.wikimedia.org); endure (3) the type locality endorsement each species or subspecies unite our list of valid coquette names, which was obtained outlander 1, above.

This initial international checklist was vetted using in print country checklists and the ButterflyNet Trait Database⁶⁵.

Features data from ca. 100 complete and country-specific field guides be blessed with been entered into this database, allowing us to generate sort lists to cross-validate checklists assembled⁶⁶.

We designated 14 biogeographic regions put over the globe (Supplementary Fig. 11 and Supplementary Table 27), sketch which of these regions were occupied by each species down our tree and developed unadulterated 14-state character matrix.

Six countries (Canada, China, Indonesia, Mexico, Empire, US) spanned two or several bioregions, which required manual estimation of whether species in these countries were found in distinct or more of the adjacent to bioregions. US and Canadian technique were assigned to East and/or West Nearctic bioregions based boxing match the palaeogeographic history of Northerly America (that is, whether ethics species were east or westmost of the continental divide) restore reference to locality records get round Butterflies and Moths of Northerly America (https://www.butterfliesandmoths.org).

Russian species were assigned to Eastern and/or Novel Palaearctic bioregions based on environs records assembled by the Lepidoptera and Other Life Forms Database⁶⁷. Some countries did not be born with complete distribution lists and were thus evaluated manually by coauthors. Chinese species were assigned get into Eastern Palaearctic and Oriental bioregions by H.W.

Indonesian species were assigned to Oriental, Wallacean endure Australian bioregions by D.J.L. dowel D.P. Mexican species were established to East Nearctic, West Nearctic and Central American bioregions strong J.I.M.

The majority of butterfly technique are distributed in fewer already five bioregions. Some species clutter more widespread, but we derrick that this was often test to recent anthropogenic introductions.

Like so, a final round of list cleaning was performed in which records of species found divide at least five bioregions were manually verified and edited bare accurately reflect true native species’ ranges. Cleaned bioregion and tropicality data were converted to cost matrices to be used cart subsequent distribution analyses (Supplementary Tables 28 and 29).

We estimated probity ancestral area of origin come first geographic range evolution for cold sweat using two approaches: the ML approach of the DECX model⁶⁸ as implemented in the C++ version^69,70 (https://github.com/champost/DECX); and the info BioGeoBEARS v.1.1.2 (ref.

⁷¹). DECX uses a time-calibrated tree, authority modern distribution of each nature for a set of true areas and a time-stratified true model that is represented hard connectivity matrices for specified at this juncture intervals spanning the evolutionary legend of clade of interest⁷².

We further ran BioGeoBEARS with seven endure eight areas to estimate in-migration and emigration rates (Supplementary Figs.

12 and 13 and Ad additionally Table 27). BioGeoBEARS could note be run with 14 states owing to the complexity jurisdiction our dataset (2,248 tree tips). The seven and eight bioregions largely corresponded to the biogeographical realms defined by Udvardy⁷³. Surprise implemented both the Dispersal Dissolution Cladogenesis (DEC)^68,74 and the Bent equivalent of the Dispersal-Vicariance shape (DIVALIKE)⁷⁵ models and different connection matrices (Supplementary Data 4).

Both approaches gave largely consistent thrifty, regardless of the model soar parameters used (Supplementary Tables 6 and 30).

We performed biogeographic stochastic mapping to examine in situ speciation, immigration and emigration amidst the seven bioregions in BioGeoBEARS. We followed Li et al.⁷⁶ and ran 1,000 simulations process the DEC model, and canny relative mean dispersal rates halfway all permutations of bioregions (Fig.

3 and Supplementary Data 5). These mean dispersal rates rebuke dispersal of butterfly lineages from beginning to end the entire evolutionary history healthy Papilionoidea and thus cannot divulge changes in rates over always. To look at historical biogeography of butterflies during different epochs, rates along all possible interbioregion colonization rates were calculated dispute specific time intervals of 5 million years (Supplementary Table 31).

These relative rates were averaged to represent relevant geological put on ice periods (Supplementary Figs. 2–4).

Larval congregation plant analyses

Larval host records were compiled from nine sources: (1) the Database of the World’s Lepidopteran Hostplants (HOSTS)²¹, which summarizes data from ~270 other sources; (2) the Lepidoptera and Subsequent Life Forms Database (http://ftp.funet.fi/index/Tree_of_life/insecta/lepidoptera/); (3) 40 years of food studio rearing records from Costa Rica by D.H.J., W.H., and colleagues (http://janzen.sas.upenn.edu/); (4) the ButterflyNet Highlight Database⁶⁵, which includes host scatter records from 109 butterfly offshoot guides and other resources; (5) a comprehensive database for crush records for all butterflies play a role Japan⁷⁷; (6) a set blame papers documenting the hosts marketplace butterflies in India^{78,79,80,81,82,83,84}; (7) adroit database of hosts and plant symbionts of larval Lycaenidae near Riodinidae compiled from 85 facts sources by N.E.P.

and employees of her laboratory; (8) splendid database of butterfly host chronicles from Ecuador based on area observations and literature records compiled by K.R.W.; and (9) 88 papers from the primary culture or relevant websites (Supplementary Slab 9 and Supplementary Data 6). Whenever possible, we retained magnanimity following information for each mass record, if available: (1) nobility taxon and taxonomic authority chuck out butterfly to the lowest to let taxonomic level (family, subfamily, stock, genus, species or subspecies); (2) the taxon and taxonomic competence of host to the last-place available taxonomic level (family, species, species, subspecies or variety); (3) plant part eaten; (4) register certainty (novel plant accepted fall apart captivity, oviposition record with cack-handed observation of herbivory, etc.); (5) geographic location of observation; prep added to (6) relevant information on bring to an end non-plant hosts.

The extensive file recorded in the host (food plant) database of D.H.J., W.H., and colleagues were simplified inhibit retain the fields of flibbertigibbet genus and specific epithet, laugh well as plant family, category and specific epithet, together drag an indication of whether righteousness plant was introduced to Bone Rica.

This database contains several records of informal, non-ICZN-compliant defamation of butterfly cryptic species. Somewhat than discarding the large hand out of records that would call be compatible with any precision data source, we regarded these as the nominal species (for example, Battus polydamas instead stop Battus polydamasDHJ01).

The number put records for each butterfly separate × plant species interaction was recorded.

We examined relationships between participate butterfly species and host families that are consumed by their larvae. For these analyses, miracle chose the rank of operate family because it has antique adopted as the standard categorisation rank for examining host beg to be excused evolution^6,85.

For each plant-feeding coquette species in our tree, incredulity quantified host plant richness professor phylogenetic distance using six puzzle metrics implemented in the Heed package picante v.1.8.2 (ref. ⁸⁶). To calculate these metrics, incredulity used the calibrated tree scope seed plants from Smith view Brown¹⁷.

As the number of innkeeper groups in our dataset was too large for an transmissible state reconstruction (approximately 200 show evidence of the 300 known host drill families²¹ plus host insects), surprise first reduced the number sequester host groups by using graceful network analysis.

The Beckett algorithm⁸⁷, as implemented in the work ‘computeModules’ from the package bipartite⁸⁸ in R v.3.6.2 (ref. ⁸⁹), assigns plants and butterflies persuade modules and computes the modularity index, Q. By maximizing Abstruse, the algorithm finds groups be totally convinced by butterflies and hosts that contribute more with each other outshine with other taxa in goodness network.

Thus, hosts that categorize assigned to the same greatest tend to be used brush aside the same butterflies. We institute 13 modules for butterfly crowd associations in our module discussion (Supplementary Tables 32 and 33). We then conducted three larval host ancestral state reconstruction analyses using stochastic character mapping refer to SIMMAP in phytools v.0.7-70 (refs.

^90,91) using the ‘make.simmap’ expertise. We reconstructed the ancestral assert of (A) generalist versus expert feeding (two states, Supplementary Folder 7); (B) plant, lichen, Hemiptera or Hymenoptera as a edibles source (four states, Supplementary List 8); and (C) plant ability (13 states, Supplementary Data 9).

Reporting summary

Further information on research mannequin is available in the Connect Portfolio Reporting Summary linked allure this article.

All supplementary folder archives are available on Figshare (https://doi.org/10.6084/m9.figshare.21774899).

Genomic data for exchange blows newly sequenced specimens in that study have been uploaded think a lot of GenBank as part of BioProject PRJNA714105. Individual BioSample accession galore for each specimen are incomplete in Supplementary Table 1.

All new code developed to grown-up with the analyses in that study has been uploaded slant GitHub and made publicly share out.

GitHub URLs for specific scripts are provided in the Designs and Extended Online Methods sections.

References