Click Beetle Mitogenomics with the Definition of a New Subfamily Hapatesinae from Australasia (Coleoptera: Elateridae)

Electronic supplementary information

Click beetle mitogenomics with the definition of a new subfamily Hapatesinae from Australasia (Coleoptera: Elateridae)

Dominik Kusy, Michal Motyka & Ladislav Bocak

Supplementary Tables Table S1. The list of earlier reported taxa included in the present mitogenomic datasets with the number of genes included in the analysis. Table S2. Location of features in the newly sequenced mitogenomes. Table S3. Detailed overview of analysed datasets with partition schemes and results of the ModelFinder analysis.

Supplementary Illustrations Figure S1. Newly sequenced circularized mitogenomes. The grey circles inside each mitogenomes represent GC content. Figure S2. Gene order in newly sequenced linear mitogenomes. Figure S3. AliStat heat maps of pairwise completeness scores (Ca) in all produced datasets: (A) NUC123: 15 mitochondrial genes, (B) PCN12: 13 mitochondrial PCGs, (C) AA: amino acids of 13 mitochondrial PCGs, (D) MTallAS: dataset A analysed using AliScore. Figure S4. Heat maps calculated with SymTest showing p-values for the pairwise Bowker's tests in in all produced datasets: (A) NUC123: 15 mitochondrial genes, (B) PCN12: 13 mitochondrial PCGs, (C) AA: amino acids of 13 mitochondrial PCGs, (D) MTallAS: dataset A analysed using AliScore. Figure S5. Rectangular heat maps of heterogeneous sequence divergence within sequences in analyzed datasets: (A) NUC123: 15 mitochondrial genes, (B) PCN12: 13 mitochondrial PCGs, (C) AA: amino acids of 13 mitochondrial PCGs, (D) MTallAS: dataset A analysed using AliScore, (E) 13 PCGs third codon position. The mean similarity score between sequences is represented by a coloured square, based on AliGROOVE scores ranging from −1, indicating great difference in rates from the remainder of the data set, i.e. heterogeneity (red), to +1, indicating rates match all other comparisons (blue). Figure S6. Maximum likelihood trees from IQ-TREE analysis of datasets: A) NUC123: 15 mitochondrial genes partitioned by gene and PCGs further partitioned by codon position; B) PCN12: 13 mitochondrial PCGs partitioned by gene and by first and second codon positions with third codon position removed; C) AA: amino acids of 13 mitochondrial PCGs partitioned by gene; D) MTallAS: dataset A analysed using AliScore. The depicted branch support values represent SH-aLRT, abayes test, and ultrafast bootstrap. Figure S7. Bayesian trees inferred from unpartitioned datasets: (A) NUC123: 15 mitochondrial genes, (B) PCN12: 13 mitochondrial PCGs and (C) AA: amino acids of 13 mitochondrial PCGs in PhyloBayes under the siteheterogeneous mixture CAT+ GTR model. The values at nodes are Bayesian posterior probabilities. Table S1. The list of earlier reported taxa included in the present mitogenomic datasets with the number of genes included in the analysis.

NCBI Voucher Species Subfamily Family # of genes Outgroup MH065615 Sinopyrophorus schimmeli Sinopyrophorinae Sinopyrophoridae 15 Ingroup KT876879 Agriotes obscurus Elaterinae Elateridae 14 JX412737 Agriotes ustulatus Elaterinae Elateridae 12 KX087232 Adrastus rachifer Elaterinae Elateridae 15 MN306531 Ludioschema vittiger Elaterinae Elateridae 15 MH789726 Elateridae sp. Elaterinae Elateridae 15 KT876904 Melanotus villosus Elaterinae Elateridae 15 KJ922150 Pyrearinus termitilluminans Agrypninae Elateridae 15 EF398270 Pyrophorus divergens Agrypninae Elateridae 13 MG242621 Ignelater luminosus Agrypninae Elateridae 15 KJ922149 Hapsodrilus ignifer Agrypninae Elateridae 15 MK524933 Cryptalaus yamato Agrypninae Elateridae 15 MT118665 Cryptalaus larvatus Agrypninae Elateridae 15 MN370897 Agrypnus sp. Agrypninae Elateridae 15 HQ232815 Drilus ﬂavescens Agrypninae Elateridae 13 KX087237 Anostirus castaneus Dendrometrinae Elateridae 15 KX087306 Limonius minutus Dendrometrinae Elateridae 15 KT852377 Limonius californicus Dendrometrinae Elateridae 15 KT876881 Athous haemorrhoidalis Dendrometrinae Elateridae 15 MK692585 Cardiophorus signatus Cardiphorinae Elateridae 15 KX087283 Dicronychus cinereus Cardiphorinae Elateridae 15 JX412848 Dicronychus sp. Cardiphorinae Elateridae 12 KJ938491 Teslasena femoralis Cardiphorinae Elateridae 15 Table S2. Location of features in the newly sequenced mitogenomes. Taxon Name Type Minimum Maximum Length Direction Taxon Name Type Minimum Maximum Length Direc�on D19007 Eudicronychus rufus rrnL rRNA 12,547 13,832 1,286 reverse G20007 Hapatesus tropicus rrnL rRNA 12,504 13,828 1,325 reverse D19007 Eudicronychus rufus rrnS rRNA 13,903 14,659 757 reverse G20007 Hapatesus tropicus rrnS rRNA 13,884 14,647 764 reverse D19007 Eudicronychus rufus nad5 gene 6,254 7,990 1,737 reverse G20007 Hapatesus tropicus nad5 gene 6,256 7,974 1,719 reverse D19007 Eudicronychus rufus cox1 gene 1,399 2,961 1,563 forward G20007 Hapatesus tropicus cox1 gene 1,399 2,961 1,563 forward D19007 Eudicronychus rufus nad4 gene 8,056 9,387 1,332 reverse G20007 Hapatesus tropicus nad4 gene 8,042 9,373 1,332 reverse D19007 Eudicronychus rufus cob gene 10,310 11,443 1,134 forward G20007 Hapatesus tropicus cob gene 10,294 11,427 1,134 forward D19007 Eudicronychus rufus nad2 gene 201 1,229 1,029 forward G20007 Hapatesus tropicus nad2 gene 199 1,224 1,026 forward D19007 Eudicronychus rufus nad1 gene 11,531 12,481 951 reverse G20007 Hapatesus tropicus nad1 gene 11,512 12,462 951 reverse D19007 Eudicronychus rufus cox3 gene 4,672 5,520 849 forward G20007 Hapatesus tropicus cox3 gene 4,666 5,453 788 forward D19007 Eudicronychus rufus cox2 gene 3,033 3,746 714 forward G20007 Hapatesus tropicus cox2 gene 3,029 3,742 714 forward D19007 Eudicronychus rufus atp6 gene 3,998 4,672 675 forward G20007 Hapatesus tropicus atp6 gene 3,992 4,666 675 forward D19007 Eudicronychus rufus nad6 gene 9,804 10,310 507 forward G20007 Hapatesus tropicus nad6 gene 9,788 10,294 507 forward D19007 Eudicronychus rufus nad3 gene 5,521 5,877 357 forward G20007 Hapatesus tropicus nad3 gene 5,517 5,870 354 forward D19007 Eudicronychus rufus nad4l gene 9,381 9,671 291 reverse G20007 Hapatesus tropicus nad4l gene 9,367 9,657 291 reverse D19007 Eudicronychus rufus atp8 gene 3,849 4,004 156 forward G20007 Hapatesus tropicus atp8 gene 3,843 3,998 156 forward D19007 Eudicronychus rufus trnW(tca) tRNA 1,235 1,306 72 forward G20007 Hapatesus tropicus trnK(c�) tRNA 3,708 3,778 71 forward D19007 Eudicronychus rufus trnS2(tga) tRNA 11,442 11,513 72 forward G20007 Hapatesus tropicus trnV(tac) tRNA 13,813 13,883 71 reverse D19007 Eudicronychus rufus trnK(c�) tRNA 3,712 3,782 71 forward G20007 Hapatesus tropicus trnQ(�g) tRNA 64 132 69 reverse D19007 Eudicronychus rufus trnV(tac) tRNA 13,833 13,902 70 reverse G20007 Hapatesus tropicus trnC(gca) tRNA 1,291 1,358 68 reverse D19007 Eudicronychus rufus trnQ(�g) tRNA 65 133 69 reverse G20007 Hapatesus tropicus trnM(cat) tRNA 132 198 67 forward D19007 Eudicronychus rufus trnM(cat) tRNA 133 200 68 forward G20007 Hapatesus tropicus trnS1(tct) tRNA 6,063 6,129 67 forward D19007 Eudicronychus rufus trnA(tgc) tRNA 5,876 5,943 68 forward G20007 Hapatesus tropicus trnI(gat) tRNA 1 66 66 forward D19007 Eudicronychus rufus trnI(gat) tRNA 1 67 67 forward G20007 Hapatesus tropicus trnN(g�) tRNA 5,997 6,062 66 forward D19007 Eudicronychus rufus trnY(gta) tRNA 1,361 1,427 67 reverse G20007 Hapatesus tropicus trnH(gtg) tRNA 7,975 8,040 66 reverse D19007 Eudicronychus rufus trnD(gtc) tRNA 3,782 3,848 67 forward G20007 Hapatesus tropicus trnS2(tga) tRNA 11,426 11,491 66 forward D19007 Eudicronychus rufus trnS1(tct) tRNA 6,074 6,140 67 forward G20007 Hapatesus tropicus trnY(gta) tRNA 1,363 1,427 65 reverse D19007 Eudicronychus rufus trnE(�c) tRNA 6,142 6,208 67 forward G20007 Hapatesus tropicus trnD(gtc) tRNA 3,778 3,842 65 forward D19007 Eudicronychus rufus trnF(gaa) tRNA 6,207 6,273 67 reverse G20007 Hapatesus tropicus trnA(tgc) tRNA 5,869 5,933 65 forward D19007 Eudicronychus rufus trnR(tcg) tRNA 5,943 6,008 66 forward G20007 Hapatesus tropicus trnR(tcg) tRNA 5,932 5,996 65 forward D19007 Eudicronychus rufus trnN(g�) tRNA 6,008 6,073 66 forward G20007 Hapatesus tropicus trnE(�c) tRNA 6,131 6,195 65 forward D19007 Eudicronychus rufus trnH(gtg) tRNA 7,991 8,056 66 reverse G20007 Hapatesus tropicus trnP(tgg) tRNA 9,722 9,786 65 reverse D19007 Eudicronychus rufus trnP(tgg) tRNA 9,738 9,802 65 reverse G20007 Hapatesus tropicus trnL2(taa) tRNA 2,964 3,027 64 forward D19007 Eudicronychus rufus trnL2(taa) tRNA 2,965 3,028 64 forward G20007 Hapatesus tropicus trnG(tcc) tRNA 5,453 5,516 64 forward D19007 Eudicronychus rufus trnT(tgt) tRNA 9,674 9,737 64 forward G20007 Hapatesus tropicus trnT(tgt) tRNA 9,659 9,722 64 forward D19007 Eudicronychus rufus trnL1(tag) tRNA 12,483 12,545 63 reverse G20007 Hapatesus tropicus trnF(gaa) tRNA 6,194 6,256 63 reverse D19007 Eudicronychus rufus trnC(gca) tRNA 1,299 1,360 62 reverse G20007 Hapatesus tropicus trnL1(tag) tRNA 12,464 12,526 63 reverse G19011 Diplophoenicus sp. rrnL rRNA 12,521 13,802 1,282 reverse G20007 Hapatesus tropicus trnW(tca) tRNA 1,230 1,283 54 forward G19011 Diplophoenicus sp. rrnS rRNA 13,875 14,630 756 reverse G19006 Parablax sp. rrnL rRNA 12,517 13,844 1,328 reverse G19011 Diplophoenicus sp. nad5 gene 6,263 7,978 1,716 reverse G19006 Parablax sp. rrnS rRNA 13,893 14,653 761 reverse G19011 Diplophoenicus sp. cox1 gene 1,417 3,000 1,584 forward G19006 Parablax sp. nad5 gene 6,256 7,929 1,674 reverse G19011 Diplophoenicus sp. nad4 gene 7,978 9,324 1,347 reverse G19006 Parablax sp. cox1 gene 1,439 2,980 1,542 forward G19011 Diplophoenicus sp. cob gene 10,292 11,425 1,134 forward G19006 Parablax sp. nad4 gene 8,052 9,387 1,336 reverse G19011 Diplophoenicus sp. nad2 gene 196 1,224 1,029 forward G19006 Parablax sp. cob gene 10,310 11,443 1,134 forward G19011 Diplophoenicus sp. nad1 gene 11,507 12,457 951 reverse G19006 Parablax sp. nad2 gene 222 1,250 1,029 forward G19011 Diplophoenicus sp. cox3 gene 4,668 5,516 849 forward G19006 Parablax sp. nad1 gene 11,525 12,475 951 reverse G19011 Diplophoenicus sp. cox2 gene 3,032 3,745 714 forward G19006 Parablax sp. cox3 gene 4,683 5,470 788 forward G19011 Diplophoenicus sp. atp6 gene 3,994 4,668 675 forward G19006 Parablax sp. cox2 gene 3,046 3,759 714 forward G19011 Diplophoenicus sp. nad6 gene 9,777 10,292 516 forward G19006 Parablax sp. atp6 gene 4,009 4,683 675 forward G19011 Diplophoenicus sp. nad3 gene 5,517 5,873 357 forward G19006 Parablax sp. nad6 gene 9,804 10,310 507 forward G19011 Diplophoenicus sp. nad4l gene 9,366 9,656 291 reverse G19006 Parablax sp. nad3 gene 5,534 5,887 354 forward G19011 Diplophoenicus sp. atp8 gene 3,845 4,000 156 forward G19006 Parablax sp. nad4l gene 9,381 9,671 291 reverse G19011 Diplophoenicus sp. trnV(tac) tRNA 13,803 13,874 72 reverse G19006 Parablax sp. atp8 gene 3,860 4,015 156 forward G19011 Diplophoenicus sp. trnK(c�) tRNA 3,711 3,781 71 forward G19006 Parablax sp. trnW(tca) tRNA 1,257 1,327 71 forward G19011 Diplophoenicus sp. trnQ(�g) tRNA 61 129 69 reverse G19006 Parablax sp. trnK(c�) tRNA 3,725 3,795 71 forward G19011 Diplophoenicus sp. trnS1(tct) tRNA 6,064 6,131 68 forward G19006 Parablax sp. trnV(tac) tRNA 13,823 13,892 70 reverse G19011 Diplophoenicus sp. trnM(cat) tRNA 129 195 67 forward G19006 Parablax sp. trnQ(�g) tRNA 64 132 69 reverse G19011 Diplophoenicus sp. trnW(tca) tRNA 1,227 1,293 67 forward G19006 Parablax sp. trnR(tcg) tRNA 5,950 6,016 67 forward G19011 Diplophoenicus sp. trnF(gaa) tRNA 6,197 6,263 67 reverse G19006 Parablax sp. trnS1(tct) tRNA 6,086 6,152 67 forward G19011 Diplophoenicus sp. trnA(tgc) tRNA 5,872 5,937 66 forward G19006 Parablax sp. trnI(gat) tRNA 1 66 66 forward G19011 Diplophoenicus sp. trnE(�c) tRNA 6,133 6,198 66 forward G19006 Parablax sp. trnM(cat) tRNA 156 221 66 forward G19011 Diplophoenicus sp. trnS2(tga) tRNA 11,424 11,489 66 forward G19006 Parablax sp. trnN(g�) tRNA 6,020 6,085 66 forward G19011 Diplophoenicus sp. trnN(g�) tRNA 5,999 6,063 65 forward G19006 Parablax sp. trnE(�c) tRNA 6,155 6,220 66 forward G19011 Diplophoenicus sp. trnY(gta) tRNA 1,349 1,412 64 reverse G19006 Parablax sp. trnP(tgg) tRNA 9,737 9,802 66 reverse G19011 Diplophoenicus sp. trnD(gtc) tRNA 3,781 3,844 64 forward G19006 Parablax sp. trnS2(tga) tRNA 11,442 11,507 66 forward G19011 Diplophoenicus sp. trnP(tgg) tRNA 9,721 9,784 64 reverse G19006 Parablax sp. trnY(gta) tRNA 1,382 1,446 65 reverse G19011 Diplophoenicus sp. trnI(gat) tRNA 1 63 63 forward G19006 Parablax sp. trnD(gtc) tRNA 3,795 3,859 65 forward G19011 Diplophoenicus sp. trnC(gca) tRNA 1,286 1,348 63 reverse G19006 Parablax sp. trnA(tgc) tRNA 5,886 5,950 65 forward G19011 Diplophoenicus sp. trnL2(taa) tRNA 2,969 3,031 63 forward G19006 Parablax sp. trnH(gtg) tRNA 7,999 8,063 65 reverse G19011 Diplophoenicus sp. trnR(tcg) tRNA 5,937 5,999 63 forward G19006 Parablax sp. trnL2(taa) tRNA 2,982 3,045 64 forward G19011 Diplophoenicus sp. trnT(tgt) tRNA 9,658 9,720 63 forward G19006 Parablax sp. trnG(tcc) tRNA 5,470 5,533 64 forward G19011 Diplophoenicus sp. trnL1(tag) tRNA 12,459 12,520 62 reverse G19006 Parablax sp. trnF(gaa) tRNA 6,219 6,281 63 reverse G18004 Drilus mauritanicus rrnL rRNA 12,422 13,708 1,287 reverse G19006 Parablax sp. trnL1(tag) tRNA 12,477 12,539 63 reverse G18004 Drilus mauritanicus rrnS rRNA 13,761 14,507 747 reverse G19006 Parablax sp. trnC(gca) tRNA 1,320 1,381 62 reverse G18004 Drilus mauritanicus nad5 gene 6,207 7,927 1,721 reverse G19006 Parablax sp. trnT(tgt) tRNA 9,675 9,736 62 forward G18004 Drilus mauritanicus cox1 gene 1,376 2,937 1,562 forward A01544 Plastocerus angulosus rrnL rRNA 12,532 13,840 1,309 reverse G18004 Drilus mauritanicus nad4 gene 7,924 9,306 1,383 reverse A01544 Plastocerus angulosus rrnS rRNA 13,892 14,659 768 reverse G18004 Drilus mauritanicus cob gene 10,206 11,336 1,131 forward A01544 Plastocerus angulosus nad5 gene 6,242 7,987 1,746 reverse G18004 Drilus mauritanicus nad2 gene 197 1,216 1,020 forward A01544 Plastocerus angulosus cox1 gene 1,418 2,957 1,540 forward G18004 Drilus mauritanicus nad1 gene 11,415 12,365 951 reverse A01544 Plastocerus angulosus nad4 gene 8,047 9,376 1,330 reverse G18004 Drilus mauritanicus cox3 gene 4,629 5,474 846 forward A01544 Plastocerus angulosus cob gene 10,298 11,431 1,134 forward G18004 Drilus mauritanicus cox2 gene 3,001 3,685 685 forward A01544 Plastocerus angulosus nad2 gene 200 1,228 1,029 forward G18004 Drilus mauritanicus atp6 gene 3,955 4,626 672 forward A01544 Plastocerus angulosus nad1 gene 11,515 12,465 951 reverse G18004 Drilus mauritanicus nad6 gene 9,706 10,206 501 forward A01544 Plastocerus angulosus cox3 gene 4,663 5,448 786 forward G18004 Drilus mauritanicus nad3 gene 5,475 5,831 357 forward A01544 Plastocerus angulosus cox2 gene 3,025 3,703 679 forward G18004 Drilus mauritanicus nad4l gene 9,300 9,587 288 reverse A01544 Plastocerus angulosus atp6 gene 3,989 4,663 675 forward G18004 Drilus mauritanicus atp8 gene 3,806 3,961 156 forward A01544 Plastocerus angulosus nad6 gene 9,783 10,298 516 forward G18004 Drilus mauritanicus trnK(c�) tRNA 3,674 3,743 70 forward A01544 Plastocerus angulosus nad3 gene 5,515 5,868 354 forward G18004 Drilus mauritanicus trnQ(�g) tRNA 63 131 69 reverse A01544 Plastocerus angulosus nad4l gene 9,370 9,660 291 reverse G18004 Drilus mauritanicus trnV(tac) tRNA 13,693 13,761 69 reverse A01544 Plastocerus angulosus atp8 gene 3,840 3,995 156 forward G18004 Drilus mauritanicus trnW(tca) tRNA 1,222 1,289 68 forward A01544 Plastocerus angulosus trnW(tca) tRNA 1,235 1,307 73 forward G18004 Drilus mauritanicus trnM(cat) tRNA 131 196 66 forward A01544 Plastocerus angulosus trnK(c�) tRNA 3,704 3,774 71 forward G18004 Drilus mauritanicus trnE(�c) tRNA 6,082 6,147 66 forward A01544 Plastocerus angulosus trnR(tcg) tRNA 5,932 6,002 71 forward G18004 Drilus mauritanicus trnI(gat) tRNA 1 65 65 forward A01544 Plastocerus angulosus trnV(tac) tRNA 13,822 13,891 70 reverse G18004 Drilus mauritanicus trnS1(tct) tRNA 6,017 6,081 65 forward A01544 Plastocerus angulosus trnQ(�g) tRNA 65 133 69 reverse G18004 Drilus mauritanicus trnL2(taa) tRNA 2,938 3,001 64 forward A01544 Plastocerus angulosus trnX(tct) tRNA 6,068 6,135 68 forward G18004 Drilus mauritanicus trnN(g�) tRNA 5,953 6,016 64 forward A01544 Plastocerus angulosus trnS2(tga) tRNA 11,430 11,497 68 forward G18004 Drilus mauritanicus trnD(gtc) tRNA 3,743 3,805 63 forward A01544 Plastocerus angulosus trnI(gat) tRNA 1 67 67 forward G18004 Drilus mauritanicus trnA(tgc) tRNA 5,830 5,892 63 forward A01544 Plastocerus angulosus trnM(cat) tRNA 133 199 67 forward G18004 Drilus mauritanicus trnT(tgt) tRNA 9,589 9,651 63 forward A01544 Plastocerus angulosus trnD(gtc) tRNA 3,774 3,839 66 forward G18004 Drilus mauritanicus trnY(gta) tRNA 1,343 1,404 62 reverse A01544 Plastocerus angulosus trnA(tgc) tRNA 5,867 5,932 66 forward G18004 Drilus mauritanicus trnF(gaa) tRNA 6,146 6,207 62 reverse A01544 Plastocerus angulosus trnN(g�) tRNA 6,002 6,067 66 forward G18004 Drilus mauritanicus trnP(tgg) tRNA 9,652 9,713 62 reverse A01544 Plastocerus angulosus trnE(�c) tRNA 6,139 6,204 66 forward G18004 Drilus mauritanicus trnS2(tga) tRNA 11,335 11,396 62 forward A01544 Plastocerus angulosus trnY(gta) tRNA 1,361 1,425 65 reverse G18004 Drilus mauritanicus trnC(gca) tRNA 1,282 1,342 61 reverse A01544 Plastocerus angulosus trnG(tcc) tRNA 5,450 5,514 65 forward G18004 Drilus mauritanicus trnR(tcg) tRNA 5,892 5,952 61 forward A01544 Plastocerus angulosus trnF(gaa) tRNA 6,203 6,267 65 reverse G18004 Drilus mauritanicus trnL1(tag) tRNA 12,367 12,427 61 reverse A01544 Plastocerus angulosus trnH(---) tRNA 7,985 8,049 65 reverse G20004 Eudicronychus sp. rrnL rRNA 12,548 13,832 1,285 reverse A01544 Plastocerus angulosus trnL1(tag) tRNA 12,467 12,531 65 reverse G20004 Eudicronychus sp. rrnS rRNA 13,903 14,659 757 reverse A01544 Plastocerus angulosus trnT(tgt) tRNA 9,663 9,726 64 forward G20004 Eudicronychus sp. nad5 gene 6,256 7,995 1,740 reverse A01544 Plastocerus angulosus trnP(tgg) tRNA 9,727 9,790 64 reverse G20004 Eudicronychus sp. cox1 gene 1,402 2,964 1,563 forward A01544 Plastocerus angulosus trnL2(taa) tRNA 2,958 3,020 63 forward G20004 Eudicronychus sp. nad4 gene 8,058 9,389 1,332 reverse A01544 Plastocerus angulosus trnC(gca) tRNA 1,300 1,360 61 reverse G20004 Eudicronychus sp. cob gene 10,312 11,445 1,134 forward G20012 Tibionema abdominalis rrnL rRNA 12,518 13,798 1,281 reverse G20004 Eudicronychus sp. nad2 gene 202 1,230 1,029 forward G20012 Tibionema abdominalis rrnS rRNA 13,869 14,624 756 reverse G20004 Eudicronychus sp. nad1 gene 11,532 12,482 951 reverse G20012 Tibionema abdominalis nad5 gene 6,231 7,970 1,740 reverse G20004 Eudicronychus sp. cox3 gene 4,675 5,523 849 forward G20012 Tibionema abdominalis cox1 gene 1,413 2,954 1,542 forward G20004 Eudicronychus sp. cox2 gene 3,036 3,749 714 forward G20012 Tibionema abdominalis nad4 gene 7,967 9,358 1,392 reverse G20004 Eudicronychus sp. atp6 gene 4,001 4,675 675 forward G20012 Tibionema abdominalis cob gene 10,287 11,420 1,134 forward G20004 Eudicronychus sp. nad6 gene 9,806 10,312 507 forward G20012 Tibionema abdominalis nad2 gene 201 1,226 1,026 forward G20004 Eudicronychus sp. nad3 gene 5,524 5,880 357 forward G20012 Tibionema abdominalis nad1 gene 11,504 12,454 951 reverse G20004 Eudicronychus sp. nad4l gene 9,383 9,673 291 reverse G20012 Tibionema abdominalis cox3 gene 4,666 5,453 788 forward G20004 Eudicronychus sp. atp8 gene 3,852 4,007 156 forward G20012 Tibionema abdominalis cox2 gene 3,021 3,737 717 forward G20004 Eudicronychus sp. trnW(tca) tRNA 1,236 1,308 73 forward G20012 Tibionema abdominalis atp6 gene 3,992 4,666 675 forward G20004 Eudicronychus sp. trnK(c�) tRNA 3,715 3,785 71 forward G20012 Tibionema abdominalis nad6 gene 9,778 10,287 510 forward G20004 Eudicronychus sp. trnS2(tga) tRNA 11,444 11,514 71 forward G20012 Tibionema abdominalis nad3 gene 5,516 5,869 354 forward G20004 Eudicronychus sp. trnV(tac) tRNA 13,833 13,902 70 reverse G20012 Tibionema abdominalis nad4l gene 9,355 9,645 291 reverse G20004 Eudicronychus sp. trnQ(�g) tRNA 65 133 69 reverse G20012 Tibionema abdominalis atp8 gene 3,843 3,998 156 forward G20004 Eudicronychus sp. trnM(cat) tRNA 134 201 68 forward G20012 Tibionema abdominalis trnK(c�) tRNA 3,703 3,773 71 forward G20004 Eudicronychus sp. trnA(tgc) tRNA 5,879 5,946 68 forward G20012 Tibionema abdominalis trnS1(tct) tRNA 6,062 6,131 70 forward G20004 Eudicronychus sp. trnI(gat) tRNA 1 67 67 forward G20012 Tibionema abdominalis trnV(tac) tRNA 13,799 13,868 70 reverse G20004 Eudicronychus sp. trnY(gta) tRNA 1,364 1,430 67 reverse G20012 Tibionema abdominalis trnQ(�g) tRNA 65 133 69 reverse G20004 Eudicronychus sp. trnD(gtc) tRNA 3,785 3,851 67 forward G20012 Tibionema abdominalis trnW(tca) tRNA 1,234 1,302 69 forward G20004 Eudicronychus sp. trnS1(tct) tRNA 6,077 6,143 67 forward G20012 Tibionema abdominalis trnS2(tga) tRNA 11,419 11,486 68 forward G20004 Eudicronychus sp. trnE(�c) tRNA 6,145 6,211 67 forward G20012 Tibionema abdominalis trnI(gat) tRNA 1 67 67 forward G20004 Eudicronychus sp. trnR(tcg) tRNA 5,946 6,011 66 forward G20012 Tibionema abdominalis trnM(cat) tRNA 135 200 66 forward G20004 Eudicronychus sp. trnN(g�) tRNA 6,011 6,076 66 forward G20012 Tibionema abdominalis trnD(gtc) tRNA 3,777 3,842 66 forward G20004 Eudicronychus sp. trnF(gaa) tRNA 6,210 6,275 66 reverse G20012 Tibionema abdominalis trnP(tgg) tRNA 9,711 9,776 66 reverse G20004 Eudicronychus sp. trnH(gtg) tRNA 7,993 8,058 66 reverse G20012 Tibionema abdominalis trnA(tgc) tRNA 5,868 5,932 65 forward G20004 Eudicronychus sp. trnP(tgg) tRNA 9,740 9,804 65 reverse G20012 Tibionema abdominalis trnN(g�) tRNA 5,997 6,061 65 forward G20004 Eudicronychus sp. trnL2(taa) tRNA 2,968 3,031 64 forward G20012 Tibionema abdominalis trnY(gta) tRNA 1,357 1,420 64 reverse G20004 Eudicronychus sp. trnT(tgt) tRNA 9,676 9,739 64 forward G20012 Tibionema abdominalis trnF(gaa) tRNA 6,193 6,256 64 reverse G20004 Eudicronychus sp. trnC(gca) tRNA 1,301 1,363 63 reverse G20012 Tibionema abdominalis trnT(tgt) tRNA 9,647 9,710 64 forward G20004 Eudicronychus sp. trnL1(tag) tRNA 12,484 12,546 63 reverse G20012 Tibionema abdominalis trnC(gca) tRNA 1,295 1,357 63 reverse G20012 Tibionema abdominalis trnL2(taa) tRNA 2,959 3,021 63 forward G20012 Tibionema abdominalis trnG(tcc) tRNA 5,453 5,515 63 forward G20012 Tibionema abdominalis trnE(�c) tRNA 6,132 6,194 63 forward G20012 Tibionema abdominalis trnR(tcg) tRNA 5,932 5,993 62 forward G20012 Tibionema abdominalis trnL1(tag) tRNA 12,456 12,517 62 reverse Table S3. Detailed overview of analysed datasets with partition schemes and results of the ModelFinder analysis.

Dataset A) 15 mitochondrial genes partitioned by gene and PCGs further partitioned by codon position. Input data: 31 taxa with 41 partitions and 13341 total sites (2.29296% missing data)

Gene Seq Site Unique Infor Invar Const ModelFinder 12S 28 820 486 372 347 347 TPM3+F+I+G4 16S 28 1331 825 669 511 510 TPM3+F+R4 atp6 30 224 110 93 116 116 TIM3+F+I+G4 atp6b 30 224 57 36 171 171 TVM+F+G4 atp6c 30 224 217 204 9 9 TN+F+G4 atp8 29 51 39 32 15 15 K3Pu+F+G4 atp8b 29 51 34 24 21 21 TPM3+F+G4 atp8c 29 51 48 46 4 4 TIM3+F+G4 cox1 31 520 173 118 374 374 SYM+I+G4 cox1b 31 520 73 29 474 474 TVM+F+R2 cox1c 31 520 513 485 15 15 TPM3+F+R4 cox2 31 226 111 87 119 119 GTR+F+I+G4 cox2b 31 226 68 44 164 164 TPM3+F+I+G4 cox2c 31 226 224 220 4 4 TPM3+F+I+G4 cox3 31 262 113 89 150 150 TIM2+F+G4 cox3b 31 262 60 38 204 204 TVM+F+I+G4 Legend: cox3c 31 262 250 242 15 15 TPM3+F+I+G4 Unique: Number of unique site patterns cytb 31 381 176 136 210 210 GTR+F+I+G4 Infor: Number of parsimony-informative sites cytbb 31 381 109 62 281 281 TIM3+F+I+G4 Invar: Number of invariant sites cytbc 31 381 375 349 10 10 HKY+F+I+G4 Const: Number of constant sites (can be subset nad1 31 316 169 134 151 151 GTR+F+I+G4 of invariant sites) nad1b 31 316 92 53 226 226 TVM+F+G4 nad1c 31 316 304 282 9 9 HKY+F+I+G4 nad2 29 342 237 208 113 113 GTR+F+I+G4 nad2b 29 342 149 113 188 188 GTR+F+I+G4 nad2c 29 342 339 325 5 5 HKY+F+R4 nad3 31 118 72 55 52 52 TIM3+F+I+G4 nad3b 31 118 53 34 71 71 TVM+F+G4 nad3c 31 118 116 113 4 4 HKY+F+G4 nad4 31 443 271 224 179 179 GTR+F+I+G4 nad4b 31 443 169 120 281 281 GTR+F+I+G4 nad4c 31 443 429 395 16 15 TPM2+F+R4 nad4l 31 100 65 51 40 40 TPM3+F+G4 nad4lb 31 100 49 26 60 60 TVM+F+G4 nad4lc 31 100 94 80 9 9 HKY+F+G4 nad5 31 575 317 268 256 256 GTR+F+I+G4 nad5b 31 575 206 150 375 375 GTR+F+I+G4 nad5c 31 575 545 506 25 25 HKY+F+G4 nad6 31 172 132 110 47 47 GTR+F+I+G4 nad6b 31 172 107 83 73 73 TVM+F+I+G4 nad6c 31 172 172 158 3 3 HKY+F+I+G4

Dataset B) 13 mitochondrial PCGs partitioned by gene and by first and second codon positions with third codon position removed. Input data: 31 taxa with 26 partitions and 7460 total sites (0.873476% missing data)

Gene Seq Site Unique Infor Invar Const ModelFinder atp6 30 224 110 93 116 116 TIM3+F+I+G4 atp6b 30 224 57 36 171 171 TVM+F+G4 atp8 29 51 39 32 15 15 K3Pu+F+G4 atp8b 29 51 34 24 21 21 TPM3+F+G4 cox1 31 520 173 118 374 374 SYM+I+G4 cox1b 31 520 73 29 474 474 TVM+F+R2 cox2 31 226 111 87 119 119 TIM3+F+I+G4 Legend: cox2b 31 226 68 44 164 164 TPM3+F+I+G4 Unique: Number of unique site patterns cox3 31 262 113 89 150 150 TIM2+F+G4 Infor: Number of parsimony-informative sites cox3b 31 262 60 38 204 204 TVM+F+I+G4 Invar: Number of invariant sites cytb 31 381 176 136 210 210 GTR+F+I+G4 Const: Number of constant sites (can be subset cytbb 31 381 109 62 281 281 TIM3+F+I+G4 of invariant sites) nad1 31 316 169 134 151 151 TIM2+F+I+G4 nad1b 31 316 92 53 226 226 TVM+F+G4 nad2 29 342 237 208 113 113 GTR+F+I+G4 nad2b 29 342 149 113 188 188 GTR+F+I+G4 nad3 31 118 72 55 52 52 TIM3+F+I+G4 nad3b 31 118 53 34 71 71 TVM+F+G4 nad4 31 443 271 224 179 179 TIM3+F+I+G4 nad4b 31 443 169 120 281 281 GTR+F+I+G4 nad4l 31 100 65 51 40 40 TPM3+F+G4 nad4lb 31 100 49 26 60 60 TVM+F+G4 nad5 31 575 317 268 256 256 GTR+F+I+G4 nad5b 31 575 206 150 375 375 GTR+F+I+G4 nad6 31 172 132 110 47 47 GTR+F+I+G4 nad6b 31 172 107 83 73 73 TVM+F+I+G4

Dataset C) Amino acids of 13 mitochondrial PCGs partitioned by gene. Input data: 31 taxa with 13 partitions and 3730 total sites (0.873476% missing data)

Gene Seq Site Unique Infor Invar Const ModelFinder atp6 30 224 118 78 126 126 mtART+I+G4 atp8 29 51 46 34 14 14 mtMet+G4 cox1 31 520 180 78 409 409 mtZOA+I+G4 cox2 31 226 136 88 113 113 mtMet+R3 cox3 31 262 131 87 150 150 mtZOA+I+G4 Legend: cytb 31 381 211 131 197 197 mtZOA+G4 Unique: Number of unique site patterns nad1 31 316 181 124 154 154 mtZOA+F+I+G4 Infor: Number of parsimony-informative sites nad2 29 342 263 212 101 101 mtZOA+F+R4 Invar: Number of invariant sites nad3 31 118 81 52 56 56 mtMet+G4 Const: Number of constant sites (can be subset nad4 31 443 303 214 180 180 mtZOA+F+R4 of invariant sites) nad4l 31 100 82 47 40 40 mtInv+G4 nad5 31 575 347 260 254 254 mtInv+F+R4 nad6 31 172 151 115 39 39 mtMet+F+G4

Dataset D) Dataset A analysed using AliScore and AliCut software to remove possibly ambiguously aligned regions. Input data: 31 taxa with 15 partitions and 12613 total sites (2.24397% missing data)

Gene Seq Site Unique Infor Invar Const ModelFinder 12S 28 795 461 349 346 346 TPM3+F+I+G4 cox3 31 768 397 351 369 369 GTR+F+I+G4 cytb 31 1126 620 530 501 501 GTR+F+I+G4 nad2 29 910 600 534 305 305 TPM3+F+R4 nad4l 31 287 182 145 109 109 K3Pu+F+G4 atp6 30 654 358 316 295 295 TIM+F+I+G4 nad4 31 1259 776 671 475 475 TIM+F+I+G4 Legend: nad5 31 1631 945 836 653 653 TIM+F+I+G4 Unique: Number of unique site patterns nad1 31 927 526 448 386 386 TIM+F+I+G4 Infor: Number of parsimony-informative sites nad3 31 315 196 164 127 127 GTR+F+I+G4 Invar: Number of invariant sites cox2 31 653 364 326 287 287 GTR+F+I+G4 Const: Number of constant sites (can be subset nad6 31 388 272 223 123 123 TN+F+I+G4 of invariant sites) 16S 28 1229 724 572 510 509 GTR+F+I+G4 cox1 31 1557 719 629 863 863 GTR+F+I+G4 atp8 29 114 78 63 40 40 HKY+F+I+G4 atp6

atp6

atp8 trnD(gtc) trnK(ctt) 6 cox3 cox3 cox3 trnG(tcc) p

t atp8 cox2 trnD(gtc) trnK(ctt) a trnG(tcc) trnG(tcc) atp8 trnD(gtc) trnK(ctt) cox2 trnL2(taa) trnA(tgc) nad3 trnR(tcg) cox2 nad3 trnN(gtt) nad3 trnA(tgc) trnL2(taa) trnX(tct) trnR(tcg) trnE(ttc) trnA(tgc) trnR(tcg) trnN(gtt) trnL2(taa) trnS1(tct) trnN(gtt) trnS1(tct) trnE(ttc) trnE(ttc) cox1

cox1

trnF(gaa) cox1 trnF(gaa) trnF(gaa)

nad5 trnW(tca) trnW(tca) nad5 trnW(tca) nad5 trnY(gta) trnC(gca) trnY(gta) trnC(gca) trnY(gta) nad2 trnC(gca) nad2 trnH(---) nad2

trnH(gtg)

trnM(cat) trnQ(ttg) trnM(cat) trnI(gat) nad4 trnM(cat) trnQ(ttg) Plastocerus angulosus trnQ(ttg) Tibionema abdominals Pararablacinae sp. trnI(gat) trnI(gat)

nad4

17,780 bp nad4 16,451 bp 17,057 bp

nad4l

trnP(tgg) nad4l trnT(tgt)

nad4l trnP(tgg)

nad6 trnP(tgg) trnT(tgt)

trnT(tgt) nad6 rrnS

trnV(tac) nad6

cob rrnS trnV(tac)

trnL1(tag)

rrnL

trnV(tac)

nad1 rrnS cob

rrnL

nad1 nad1

trnL1(tag) cob

trnL1(tag)

rrnL trnS2(tga)

trnS2(tga)

atp6

8 atp6 atp6

a trnD(gtc) trnK(ctt) atp8 trnD(gtc) trnK(ctt) atp8 trnD(gtc) trnK(ctt) cox2 cox2 cox3 cox3 cox2 cox3

trnL2(taa) trnL2(taa) trnG(tcc) trnL2(taa)

nad3 nad3 trnA(tgc) nad3 trnR(tcg) trnA(tgc) trnA(tgc) trnN(gtt) trnR(tcg) trnS1(tct) trnR(tcg) cox1 cox1 trnN(gtt) trnN(gtt) trnS1(tct) trnE(ttc) cox1 trnS1(tct) trnE(ttc) trnE(ttc)

trnF(gaa) trnF(gaa) trnF(gaa)

trnW(tca) trnW(tca) trnW(tca)

trnY(gta) trnY(gta) trnY(gta) trnC(gca) trnC(gca) trnC(gca) nad5 nad5 nad5 nad2 nad2 nad2

trnM(cat) trnM(cat) trnM(cat) trnH(gtg) trnQ(ttg) trnQ(ttg) trnQ(ttg) trnI(gat) Diplophoenicus sp. trnI(gat) trnI(gat) Hapatesus tropicus trnH(gtg) Eudicronychus sp. 15,995 bp 16,290 bp 15,916 bp

nad4 nad4 nad4

nad4l

nad4l nad4l rrnS rrnS trnP(tgg) trnP(tgg) trnP(tgg) trnV(tac) trnV(tac) rrnS trnT(tgt) trnV(tac)

trnT(tgt) trnT(tgt) nad6 rrnL rrnL trnL1(tag) trnL1(tag)

nad6 trnL1(tag) rrnL nad6

nad1

n nad1

atp6 cob 8

atp6

a trnD(gtc) cob trnK(ctt) cob

cox2

atp8 trnD(gtc) trnK(ctt) cox3

cox2 cox3 trnL2(taa)

trnL2(taa) trnS2(tga)

trnS2(tga) trnS2(tga)

nad3 nad3 trnA(tgc) trnA(tgc) trnR(tcg) trnR(tcg) cox1 trnN(gtt) trnN(gtt) trnS1(tct) trnS1(tct) cox1 trnE(ttc) trnE(ttc)

trnF(gaa) trnF(gaa)

trnW(tca) trnW(tca) trnY(gta) trnC(gca)

trnY(gta) trnC(gca) nad5 nad5 nad2 nad2

trnM(cat) trnM(cat) trnQ(ttg) Eudicronychus rufus trnI(gat) trnQ(ttg) Drilus mauritanicus trnI(gat) trnH(gtg) 16,014 bp 15,914 bp

nad4 nad4 complex I (NADH dehydrogenase)

rrnS complex III (ubichinol cytochrome c reductase) nad4l nad4l

trnV(tac) trnP(tgg) trnP(tgg) complex IV (cytochrome c oxidase) rrnS

ATP synthase trnV(tac) trnT(tgt) trnT(tgt) transfer RNAs rrnL

trnL1(tag) nad6 nad6 ribosomal RNAs trnL1(tag) rrnL

nad1

cob cob

trnS2(tga)

Figure S1. Newly sequenced circularized mitogenomes. The grey circles inside each mitogenomes represent GC content. trnK(ctt) trnS1(tct) trnD(gtc) trnA(tgc) trnR(tcg) trnN(gtt) trnE(ttc) trnI(gat) atp8 Eudicronychus rufus trnM(cat) nad2 trnW(tca) cox1 trnL2(taa) cox2 atp6 cox3 nad3 trnT(tgt) nad6 cob trnS2(tga) 15,914 bp trnQ(ttg) trnF(gaa) nad5 trnH(gtg) nad4 trnP(tgg) nad1 trnL1(tag) rrnL trnV(tac) rrnS nad4l trnR(tcg) trnN(gtt) trnS1(tct) trnK(ctt) trnD(gtc) trnA(tgc) trnE(ttc) trnY(gta) trnC(gca) atp8 Eudicronychus sp. trnI(gat) trnM(cat) nad2 trnW(tca) cox1 trnL2(taa) cox2 atp6 cox3 nad3 trnT(tgt) nad6 cob trnS2(tga) 15,916 bp trnQ(ttg) trnF(gaa) nad5 trnH(gtg) nad4 trnP(tgg) nad1 trnL1(tag) rrnL trnV(tac) rrnS nad4l trnK(ctt) trnD(gtc) trnA(tgc) trnN(gtt) trnS1(tct) trnE(ttc) trnR(tcg) trnY(gta) trnC(gca) trnM(cat) atp8 Diplophoenicus sp. trnI(gat) nad2 trnW(tca) cox1 trnL2(taa) cox2 atp6 cox3 nad3 trnT(tgt) nad6 cob trnS2(tga) 15,995 bp trnQ(ttg) trnF(gaa) nad5 nad4 trnP(tgg) nad1 trnL1(tag) rrnL trnV(tac) rrnS nad4l trnK(ctt) trnR(tcg) trnN(gtt) trnS1(tct) trnD(gtc) trnA(tgc) trnE(ttc) trnY(gta) trnC(gca) trnI(gat) trnM(cat) atp8 trnG(tcc) Parablax sp. nad2 trnW(tca) cox1 trnL2(taa) cox2 atp6 cox3 nad3 trnT(tgt) nad6 cob trnS2(tga) 17,057 bp trnQ(ttg) trnF(gaa) nad5 trnH(gtg) nad4 trnP(tgg) nad1 trnL1(tag) rrnL trnV(tac) rrnS nad4l trnD(gtc) trnS1(tct) trnK(ctt) trnA(tgc) trnR(tcg) trnN(gtt) trnE(ttc) trnY(gta) trnC(gca)

Hapatesus tropicus trnI(gat) trnM(cat) nad2 trnW(tca) cox1 trnL2(taa) cox2 atp8 atp6 cox3 trnG(tcc) nad3 trnT(tgt) nad6 cob trnS2(tga) 16,290 bp trnQ(ttg) trnF(gaa) nad5 trnH(gtg) nad4 trnP(tgg) nad1 trnL1(tag) rrnL trnV(tac) rrnS nad4l atp8 trnA(tgc) trnX(tct) trnK(ctt) trnD(gtc) trnR(tcg) trnN(gtt) trnE(ttc) trnY(gta) trnC(gca) trnI(gat) Plastocerus angulosus trnM(cat) nad2 trnW(tca) cox1 trnL2(taa) cox2 atp6 cox3 trnG(tcc) nad3 trnT(tgt) nad6 cob trnS2(tga) 17,780 bp trnQ(ttg) trnF(gaa) nad5 trnH(---) nad4 nad1 trnL1(tag) rrnL trnV(tac) rrnS nad4l trnP(tgg) trnA(tgc) trnE(ttc) trnK(ctt) trnD(gtc) trnR(tcg) trnN(gtt) trnS1(tct) trnY(gta) trnC(gca) trnM(cat) Tibionema abdominalis trnI(gat) nad2 trnW(tca) cox1 trnL2(taa) cox2 atp8 atp6 cox3 trnG(tcc) nad3 trnT(tgt) nad6 cob trnS2(tga) 16,451 bp trnQ(ttg) trnF(gaa) nad5 nad4 trnP(tgg) nad1 trnL1(tag) rrnL trnV(tac) rrnS nad4l trnD(gtc) trnN(gtt) trnK(ctt) trnA(tgc) trnR(tcg) trnS1(tct) trnE(ttc) trnY(gta) trnC(gca)

Drilus mauritanicus trnI(gat) trnM(cat) nad2 trnW(tca) cox1 trnL2(taa) cox2 atp8 atp6 cox3 nad3 trnT(tgt) nad6 cob trnS2(tga) 16,014 bp trnQ(ttg) trnF(gaa) nad5 nad4 trnP(tgg) nad1 trnL1(tag) rrnL trnV(tac) rrnS nad4l

complex I (NADH dehydrogenase) complex IV (cytochrome c oxidase) transfer RNAs trnY(gta) trnC(gca) complex III (ubichinol cytochrome c reductase) ATP synthase ribosomal RNAs

Figure S2. Gene order in newly sequenced linear mitogenomes. Ca: 0.97; (C) AA: Ca: Parablax sp. Parablax sp. Agrypnus sp. Agrypnus sp. Elateridae sp. Elateridae sp. Elateridae Dicronychus sp. Dicronychus sp. Drilus flavescens Adrastus rachifer Drilus flavescens Adrastus rachifer Adrastus Limonius minutus Eudicronychus sp. Agriotes ustulatus Limonius minutus Agriotes obscurus Eudicronychus sp. Melanotus villosus Agriotes ustulatus Agriotes Diplophoenicus sp. Agriotes obscurus Agriotes Hapsodrilus ignifer Cryptalaus yamato Melanotus villosus Melanotus Diplophoenicus sp. Drilus mauritanicus Hapsodrilus ignifer Cryptalaus larvatus Cryptalaus yamato Hapatesus tropicus Ignelater luminosus Drilus mauritanicus Cryptalaus larvatus Hapatesus tropicus Teslasena femoralis Teslasena Ignelater luminosus Anostirus castaneus Teslasena femoralis Teslasena Eudicronychus rufus Ludioschema vittiger Anostirus castaneus Limonius californicus Eudicronychus rufus Ludioschema vittiger Ludioschema Limonius californicus Dicronychus cinereus Pyrophorus divergens Dicronychus cinereus Pyrophorus divergens Cardiophorus signatus Plastocerus angulosus Cardiophorus signatus Tibionema abdominalis Tibionema Plastocerus angulosus Tibionema abdominalis Tibionema Athous haemorrhoidalis Athous haemorrhoidalis Sinopyrophorus shimmeli Sinopyrophorus shimmeli Pyrearinus termitilluminans Pyrearinus termitilluminans , Ca: 0.95; (B) PCN12: 13 mitochondrial PCGs, , Ca: Ca: 0.95. AliScore, Ca: using analysed A dataset 0.97; (D) MTallAS: Ca:

= 0.0 < 0.1 < 0.2 < 0.3 < 0.4 < 0.5 < 0.6 < 0.7 < 0.8 < 0.9 ≤ 1.0 AliStat heat maps of pairwise completeness scores (Ca) in all produced datasets: (A) NUC123: AliStat heat maps of pairwise completeness scores (Ca) in all produced Parablax sp. Parablax sp. Agrypnus sp. Agrypnus sp. Elateridae sp. Elateridae Elateridae sp. Dicronychus sp. Dicronychus sp. Drilus flavescens Adrastus rachifer Adrastus Drilus flavescens Adrastus rachifer Limonius minutus

Limonius minutus Eudicronychus sp. Agriotes ustulatus Agriotes Eudicronychus sp. Agriotes obscurus Agriotes Agriotes ustulatus Agriotes obscurus

Melanotus villosus Melanotus Diplophoenicus sp. Melanotus villosus

Diplophoenicus sp. Hapsodrilus ignifer Cryptalaus yamato Hapsodrilus ignifer Cryptalaus yamato

Drilus mauritanicus Cryptalaus larvatus Hapatesus tropicus Drilus mauritanicus Cryptalaus larvatus Hapatesus tropicus Ignelater luminosus

Ignelater luminosus

Teslasena femoralis Teslasena Teslasena femoralis Teslasena Anostirus castaneus Eudicronychus rufus Anostirus castaneus

Eudicronychus rufus Ludioschema vittiger Ludioschema

Ludioschema vittiger Limonius californicus Limonius californicus

Dicronychus cinereus Dicronychus cinereus Pyrophorus divergens Pyrophorus divergens

Cardiophorus signatus

Plastocerus angulosus

Cardiophorus signatus C) D) C)

Plastocerus angulosus Tibionema abdominalis Tibionema

Tibionema abdominalis Tibionema A) B) A)

Athous haemorrhoidalis Athous haemorrhoidalis

Sinopyrophorus shimmeli 15 mitochondrial genes (13 PCGs and 2 rRNA) amino acids of 13 mitochondrial PCGs,

Figure S3. Sinopyrophorus shimmeli

Pyrearinus termitilluminans Pyrearinus termitilluminans Agriotes obscurus Agriotes obscurus A) Agriotes ustulatus B) Agriotes ustulatus Adrastus rachifer Adrastus rachifer Ludioschema vittiger Ludioschema vittiger Elateridae sp. Elateridae sp. Melanotus villosus Melanotus villosus Pyrearinus termitilluminans Pyrearinus termitilluminans Pyrophorus divergens Pyrophorus divergens Ignelater luminosus Ignelater luminosus Hapsodrilus ignifer Hapsodrilus ignifer Cryptalaus yamato Cryptalaus yamato Cryptalaus larvatus Cryptalaus larvatus Agrypnus sp. Agrypnus sp. Drilus flavescens Drilus flavescens Drilus mauritanicus Drilus mauritanicus Anostirus castaneus Anostirus castaneus Limonius minutus Limonius minutus Limonius californicus Limonius californicus Athous haemorrhoidalis Athous haemorrhoidalis Cardiophorus signatus Cardiophorus signatus Dicronychus cinereus Dicronychus cinereus Dicronychus sp. Dicronychus sp. Teslasena femoralis Teslasena femoralis Diplophoenicus sp. Diplophoenicus sp. Tibionema abdominalis Tibionema abdominalis Parablax sp. Parablax sp. Eudicronychus rufus Eudicronychus rufus Eudicronychus sp. Eudicronychus sp. Hapatesus tropicus Hapatesus tropicus Plastocerus angulosus Plastocerus angulosus Sinopyrophorus shimmeli Sinopyrophorus shimmeli

Agriotes obscurus Agriotes obscurus C) Agriotes ustulatus D) Agriotes ustulatus Adrastus rachifer Adrastus rachifer Ludioschema vittiger Ludioschema vittiger Elateridae sp. Elateridae sp. Melanotus villosus Melanotus villosus Pyrearinus termitilluminans Pyrearinus termitilluminans Pyrophorus divergens Pyrophorus divergens Ignelater luminosus Ignelater luminosus Hapsodrilus ignifer Hapsodrilus ignifer Cryptalaus yamato Cryptalaus yamato Cryptalaus larvatus Cryptalaus larvatus Agrypnus sp. Agrypnus sp. Drilus flavescens Drilus flavescens Drilus mauritanicus Drilus mauritanicus Anostirus castaneus Anostirus castaneus Limonius minutus Limonius minutus Limonius californicus Limonius californicus Athous haemorrhoidalis Athous haemorrhoidalis Cardiophorus signatus Cardiophorus signatus Dicronychus cinereus Dicronychus cinereus Dicronychus sp. Dicronychus sp. Teslasena femoralis Teslasena femoralis Diplophoenicus sp. Diplophoenicus sp. Tibionema abdominalis Tibionema abdominalis Parablax sp. Parablax sp. Eudicronychus rufus Eudicronychus rufus Eudicronychus sp. Eudicronychus sp. Hapatesus tropicus Hapatesus tropicus Plastocerus angulosus Plastocerus angulosus Sinopyrophorus shimmeli Sinopyrophorus shimmeli

< 5e-11< 5e-10< 5e-9< 5e-8< 5e-7< 5e-6< 5e-5< 5e-4< 5e-3< 5e-2≥ 5e-2

Figure S4. Heat maps calculated with SymTest showing p-values for the pairwise Bowker's tests in in all produced datasets: (A) NUC123: 15 mitochondrial genes (13 PCGs an 2 rRNA), (B) PCN12: 13 mitochondrial PCGs, (C) AA: amino acids of 13 mitochondrial PCGs, (D) MTallAS: dataset A analysed using AliScore. A) B)

Adrastus rachifer Adrastus rachifer Agriotes obscurus Agriotes obscurus Agriotes ustulatus Agriotes ustulatus Elateridae sp. Elateridae sp. Eudicronychus rufus Eudicronychus rufus Eudicronychus sp. Eudicronychus sp. Ludioschema vittiger Ludioschema vittiger Melanotus villosus Melanotus villosus Agrypnus sp. Agrypnus sp. Cryptalaus larvatus Cryptalaus larvatus Cryptalaus yamato Cryptalaus yamato Drilus flavescens Drilus flavescens Drilus mauritanicus Drilus mauritanicus Hapsodrilus ignifer Hapsodrilus ignifer Ignelater luminosus Ignelater luminosus Pyrearinus termitilluminans Pyrearinus termitilluminans Pyrophorus divergens Pyrophorus divergens Anostirus castaneus Anostirus castaneus Athous haemorrhoidalis Athous haemorrhoidalis Limonius californicus Limonius californicus Limonius minutus Limonius minutus Plastocerus angulosus Plastocerus angulosus Cardiophorus signatus Cardiophorus signatus Dicronychus cinereus Dicronychus cinereus Dicronychus sp. Dicronychus sp. Teslasena femoralis Teslasena femoralis Diplophoenicus sp. Diplophoenicus sp. Tibionema abdominalis Tibionema abdominalis Parablax sp. Parablax sp. Sinopyrophorus shimmeli Sinopyrophorus shimmeli Hapatesus tropicus Hapatesus tropicus

C) D)

E) Adrastus rachifer Agriotes obscurus Agriotes ustulatus Elateridae sp. Eudicronychus rufus Eudicronychus sp. Ludioschema vittiger Melanotus villosus Agrypnus sp. Cryptalaus yamato Drilus flavescens Drilus mauritanicus Hapsodrilus ignifer Ignelater luminosus Pyrearinus termitilluminans Pyrophorus divergens Cryptalaus larvatus Anostirus castaneus Athous haemorrhoidalis Limonius californicus Limonius minutus Plastocerus angulosus Cardiophorus signatus Dicronychus cinereus Dicronychus sp. Teslasena femoralis Diplophoenicus sp. Tibionema abdominalis Parablax sp. Sinopyrophorus shimmeli Hapatesus tropicus

Figure S5. Rectangular heat maps of heterogeneous sequence divergence within sequences in analyzed datasets: (A) NUC123: 15 mitochondrial genes, (B) PCN12: 13 mitochondrial PCGs, (C) AA: amino acids of 13 mitochondrial PCGs, (D) MTallAS: dataset A analysed using AliScore, (E) 13 PCGs third codon position. The mean similarity score between sequences is represented by a coloured square, based on AliGROOVE scores ranging from −1, indicating great difference in rates from the remainder of the data set, i.e. heterogeneity (red), to +1, indicating rates match all other comparisons (blue). Sinopyrophorus shimmeli Sinopyrophorus shimmeli A B Adrastus rachifer Adrastus rachifer 100/1/100 100/1/100 99.8/1/100 Agriotes obscurus 100/1/100 Agriotes obscurus

81.3/0.991/77 Agriotes ustulatus 86.7/1/92 Agriotes ustulatus 100/1/100 Eudicronychus sp 100/1/100 Ludioschema vittiger

99.9/1/84 Eudicronychus rufus 100/1/100 Elateridae sp. Melanotus villosus Melanotus villosus 59.1/0.965/70 75.3/0.999/86 100/1/100 Elateridae sp. 100/1/100 Eudicronychus sp. Eudicronychus rufus Ludioschema vittiger Hapatesus tropicus Hapatesus tropicus 97.6/1/75 98.8/1/78 99.9/1/100 Parablax sp. 100/1/100 Tibionema abdominalis Parablax sp. Tibionema abdominalis 99.3/1/52 74.2/0.997/83Anostirus castaneus Diplophoenicus sp. 99.9/1/66 39.7/0.884/77 Plastocerus angulosus 99.9/1/99 Plastocerus angulosus Athous haemorrhoidalis 99.2/1/96 Anostirus castaneus 97.1/1/96 93.7/0.998/47 Athous haemorrhoidalis 99.8/1/100 Limonius californicus 98.1/1/98 Limonius minutus 71.5/0.999/51 Limonius minutus 95.2/1/98 Diplophoenicus sp. Limonius californicus 66.9/0.951/40 Teslasena femoralis Teslasena femoralis 100/1/100 100/1/100 94.2/1/56 Cardiophorus signatus Cardiophorus signatus 100/1/100 100/1/100 100/1/100 Dicronychus cinereus 100/1/100 Dicronychus sp.

100/1/100 Dicronychus sp. 100/1/97 Dicronychus cinereus

100/1/100 Drilus mauritanicus 100/1/100 Drilus mauritanicus Drilus flavescens Drilus flavescens 96.5/1/94 92.8/1/91 100/1/100 Cryptalaus yamato 100/1/100 Cryptalaus larvatus Cryptalaus yamato 100/1/100 Cryptalaus larvatus 100/1/100 Agrypnus sp. Agrypnus sp. 98.4/1/98 88.9/1/88 Pyrearinus termitilluminans Pyrearinus termitilluminans 100/1/100 99.9/1/100 0.6 Pyrophorus divergens Pyrophorus divergens 24.4/0.809/66 71.1/0.986/77 99.5/1/98 Hapsodrilus ignifer 0.07 94.9/1/91 Hapsodrilus ignifer Ignelater luminosus Ignelater luminosus C D Sinopyrophorus shimmeli Sinopyrophorus shimmeli Teslasena femoralis Adrastus rachifer 100/1/100 100/1/100 Cardiophorus signatus 99.9/1/100 100/1/100 Agriotes obscurus 100/1/100 Dicronychus sp 95.6/1/88 Agriotes ustulatus

56/0.993/70 Dicronychus cinereus 100/1/100 Ludioschema vittiger

100/1/100 Drilus flavescens 100/1/98 Elateridae sp. Drilus mauritanicus Melanotus villosus 77.4/1/85 94.8/1/93 100/1/100 Cryptalaus larvatus 100/1/100 Eudicronychus rufus Cryptalaus yamato Eudicronychus sp 100/1/100 Agrypnus sp. Hapatesus tropicus 99/1/87 98.6/1/99 Ignelater luminosus Parablax sp. 10.5/0.694/43 100/1/100 99.9/1/100 Pyrophorus divergens 58.7/0.997/60 Tibionema abdominalis 82/0.999/75 Hapsodrilus ignifer Plastocerus angulosus 98/1/95 Pyrearinus termitilluminans Athous haemorrhoidalis 51.1/0.931/48 100/1/100 Tibionema abdominalis Anostirus castaneus 41.9/0.983/53 98.9/1/91 Parablax sp. 95/1/95 Limonius californicus Plastocerus angulosus Limonius minutus 95.1/1/83 97.9/1/96 Anostirus castaneus Diplophoenicus sp. 27.8/0.883/69 Athous haemorrhoidalis 99.4/1/99 Teslasena femoralis 100/1/100 Limonius californicus 97.9/1/79 Cardiophorus signatus 98.5/1/100 100/1/100 89.9/1/65 Limonius minutus 100/1/100 Dicronychus cinereus

95.5/1/67 Hapatesus tropicus 99.9/1/99 Dicronychus sp.

Diplophoenicus sp. 100/1/100 Drilus mauritanicus Adrastus rachifer 74.8/0.997/44 100/1/100 Drilus flavescens 92.1/1/89 Agriotes obscurus 100/1/100 Cryptalaus yamato 99.9/1/100 100/1/100 Agriotes ustulatus 100/1/100 Cryptalaus larvatus 100/1/100 Eudicronychus rufus Agrypnus sp. 95.5/1/96 99.9/1/100 Eudicronychus sp. Pyrophorus divergens 0.08 100/1/100 Melanotus villosus Pyrearinus termitilluminans 5.52/0.559/59 43.5/0.966/75 100/1/100 Ludioschema vittiger 97.5/1/98 Hapsodrilus ignifer 0.2 Elateridae sp. Ignelater luminosus

Figure S6. Maximum likelihood trees from IQ-TREE analysis of datasets: A) NUC123: 15 mitochondrial genes partitioned by gene and PCGs further partitioned by codon position; B) PCN12: 13 mitochondrial PCGs partitioned by gene and by first and second codon positions with third codon position removed; C) AA: amino acids of 13 mitochondrial PCGs partitioned by gene; D) MTallAS: dataset A analysed using AliScore. The depicted branch support values represent SH-aLRT, abayes test, and ultrafast bootstrap. A B Sinopyrophorus shimmeli Sinopyrophorus shimmeli

Adrastus rachifer 0.97 Ludioschema vittiger 1

1 Agriotes obscurus 0.61 Elateridae sp. Agriotes ustulatus Melanotus villosus 0.69

1 Ludioschema vittiger 1 Eudicronychus sp.

1 Elateridae sp. Eudicronychus rufus Melanotus villosus Adrastus rachifer 0.86 1

1 Eudicronychus sp. 1 Agriotes obscurus Eudicronychus rufus Agriotes ustulatus

Hapatesus tropicus 0.81 Hapatesus tropicus 1 1

1 Tibionema abdominalis 1 Tibionema abdominalis Parablax sp. Parablax sp.

1 Diplophoenicus sp 1 Diplophoenicus sp.

0.99 Plastocerus angulosus 0.98 Plastocerus angulosus

0.98 Anostirus castaneus 0.7 Anostirus castaneus 0.97 0.87 Athous haemorrhoidalis Athous haemorrhoidalis 0.99 0.95

1 Limonius minutus 0.94 Limonius minutus Limonius californicus Limonius californicus 0.71 0.99 Teslasena femoralis Teslasena femoralis 1 1 Cardiophorus signatus Cardiophorus signatus 1 0.99

1 Dicronychus cinereus 1 Dicronychus cinereus Dicronychus sp. 1 Dicronychus sp. 0.99

1 Drilus flavescens 1 Cryptalaus larvatus Drilus mauritanicus Cryptalaus yamato 1 1 Cryptalaus larvatus 1 1 Drilus flavescens Cryptalaus yamato Drilus mauritanicus 0.86 Agrypnus sp. Agrypnus sp. 0.9 0.59 Pyrophorus divergens Pyrophorus divergens 0.3 0.99 0.94 Pyrearinus termitilluminans Pyrearinus termitilluminans 0.98 0.57

0.99 Ignelater luminosus 0.97 Ignelater luminosus 0.5 Hapsodrilus ignifer Hapsodrilus ignifer

C Sinopyrophorus shimmeli Adrastus rachifer 1

1 Agriotes obscurus

0.6 Agriotes ustulatus Melanotus villosus

0.99 1 Ludioschema vittiger

0.68 Elateridae sp.

1 Eudicronychus sp. Eudicronychus rufus Hapatesus tropicus 0.79

1 Tibionema abdominalis Parablax sp.

0.93 Diplophoenicus sp

0.98 Plastocerus angulosus

0.99 Anostirus castaneus 0.64 Athous haemorrhoidalis 1

1 Limonius minutus Limonius californicus 0.71 Teslasena femoralis 1 Cardiophorus signatus 1

1 Dicronychus cinereus

0.99 Dicronychus sp

1 Drilus flavescens Drilus mauritanicus 1 1 Cryptalaus larvatus Cryptalaus yamato 0.91 Agrypnus sp. 0.95 Ignelater luminosus 1 Pyrophorus divergens 0.3 0.86 Pyrearinus termitilluminans Hapsodrilus ignifer

Figure S7. Bayesian trees inferred from unpartitioned datasets: (A) NUC123: 15 mitochondrial genes, (B) PCN12: 13 mitochondrial PCGs and (C) AA: amino acids of 13 mitochondrial PCGs in PhyloBayes under the siteheterogeneous mixture CAT+ GTR model. The values at nodes are Bayesian posterior probabilities.