Anti-predator defense drives parallel morphological evolution in flea beetles

Deyan Ge, Douglas Chester, Jesús Gómez-Zurita, Lijie Zhang, Xingke Yang, Alfried P. Vogler

Supporting Information

Table S1. Flea beetle genera groups proposed by Furth based on morphological characters of metafemoral spring.

Table S2. Primers used for PCR.

Table S3. Samples and GenBank Accessions.

Table S4. Summary of the Beast analysis for the age of important nodes.

Table S5. Slowinsky-Guyer test of imbalance of the net diversification rate of sister clades.

Table S6 Absolute diversification rates and 95% confidence intervals of expected diversity of Chrysomelinae at different ages.

Table S7. Number of known species and estimated age of main subfamilies in Chrysomelidae.

Figure S1. Enlarged metafemoral spring for key taxa.

SI References

5

Table S1. Flea beetle genera group proposed by Furth et al. based on morphological characters of metafemoral spring.

Palearctic region (Furth 1985)
Group 1. Arrhenocoela, *Blepharida, *Hermeaophaga, Mantura, Orestia, *Podagrica,
Group 2. *Altica, *Asiorestia, Cardax, *Crepidodera, Derocrepis, *Epitrix, *Hippuriphila, *Lythraria, Minota, Mniophila, Ochrosis,
Group 3. *Phyllotreta
Group 4. *Longitarsus
Group 5. Anthobiodes, *Aphthona, *Argopus, *Chaetocnema, Heyrovskya, *Oedionychus, *Sphaeroderma
Group 6. Apteropeda, *Batophila, *Dibolia, *Psylliodes
Nearctic region (Furth 1985)
Group 1. Acrocyum, *Blepharida, Mantura, Orestiodes, Pseudorthygia
Group 2. *Altica, *Crepidodera, Derocrepis, *Disonycha, *Epitrix, Hemiglyptus, *Hippuriphila, Hornaltica, Luperaltica, *Lysathia, *Macrohaltica, Monomacra, Phrynocepha, Strabala, *Syphrea, Trichaltica
Group 3. Dysphenges, Glenidion, Lupraea, Pachyonychus, Phydanis, *Phyllotreta, Pseudolampsis, *Systena,
Group 4. *Longitarsus
Group 5. *Aphthona, *Asphaera, Capraita, *Chaetocnema, Euplectroscelis, *Glyptina, Kuschelina, Pachyonychis, *Sphaeroderma,
Group 6. *Argopistes, *Dibolia, Distigmoptera, *Psylliodes / Neotropical region (Furth 1989)
Group 1. Acrocyum, *Blepharida, Cacoscelis, Crimissa, Elithia, Iphitroides, Nitozona
Group 2. Acanthonycha, *Altica, Aphanocera, Caeporis, Calliphron, Chlamophora, *Diphaltica, Diphaca, *Epitrix, Forsterita, Hemiglyptus, Leptophysa, Lysathia, *Macrohaltica, Mesodera, Phyrnocepha, Plectrotetra, Pseudogona, Strabala, *Syphrea, Trichaltica
Group 3. *Agasicles, Apraea, *Asiorestia, Cornulactica, Cyrsylus, *Disonycha, Dysphenges, Exoceras, Glenidion, Hemilactica, Homoschema, Lactina, Lupraea, Monomacra, Nasigona, Nephrica, Ocnoscelis, Pedilia, Phenrica, Phydanis, *Phyllotreta, Platiprosopus, Prasona, Propiasus, Pseudodisonycha, Ptocadica, *Systena
Group 4. *Longitarsus
Group 5. *Alagoasa, *Asphaera, *Aspicela, Capraita, *Chaetocnema, *Glyptina, Kuschelina, *Omophoita, Paranaita, Walterianella
Group 6. Aedmon, Allochroma, Apalotrius, *Argopistes, Atyphus, Cerichrestus, *Dibolia, Distigmoptera, Exartematopus, *Heikertingerella, Homotyphus, Hydmosyne, Megistops, Ptocadica, Octogonotes, Omototus, Orodes, Phylacticus, Physimerus, Pseudodibolia, Pseudolampsis, *Psylliodes, Rhinotmetus, Roicus, Sphaeronychus, Tetragonotes / Oriental-Australia region (Furth and Suzuki 1998)
Blephrida group. Amphimeloides, Asialtica, *Chabria, *Clitea, Erystus, *Lactica, *Lipromela, *Lypnea, Mellipora, Myrcinoides, Orphrida, *Pentamesa, *Podagricomela, *Podontia, *Sinocrepis, Tonfania, Xuthea, Zipangia
Buphonella group. Chaloenus, Eudolia, *Hespera
Sangariola group. *Laboissierea, Liprus, * Pseudodera, Sangariola
Mandarella group. Mandarella, *Stenoluperus
Altica group. Neodera
Phyllotreta group. *Aphthonaltica, * Manobia, Pedethma, Sutrea, Xenidea, Yunohespera
Chaetocnema group. *Bhamoina, Eucyclomela, Halticorcus, *Hyphasis, Lipromima
Chalaenosoma group. Chalaenosoma
Chalaenosoma group. *Luperomorpha
Nonarthra group. Glaucosphaera, *Nonarthra, *Trachyaphthona, Typhodes
Psylliodes group. *Aphthonoides, *Arsipoda, Goweria, Schenklingia
Pseudoliprus group. Lipromorpha, Pseudoliprus
Sphaerometopa group. *Sphaerometopa
Luperus-like genus. Luperus-like
Licyllus group. Licyllus, Parhespera, Thrasychroma
Garuda group. Garuda
Pyrrhalta like genus. Pyrrhalta-like
Acrocrypta group. *Acrocrypta

* Taxa included in present study

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Table S2. Primers used for PCR

Markers / Primers names / Sequences / Reference
cox1 / Pat
Jerry
Johannes* / 5’TCCAATGCACTAATCTGCCATATTA3’
5’CAACATTTATTTTGATTTTTTGG3’
5’ CCAATGCACTAATCTGCCATATTAG 3’ / (Hunt et al. 2007; Simon et al. 1994)
rrnL / 16Sarc*
16Sb2
16SB2 / 5’CGCCTGTTTATTAAAAACAT3’
5’TTTAATCCAACATCGAGG3’
5’CTCCGGTTTGAACTCAGATCA3’ / (Hunt et al. 2007; Simon et al. 1994)
SSU / 18Sb 2.5
18Sb5.0
18Sai
18sa1.0
18sbi
18sa2.0
18s3’l
18S5P’ / 5’TCTTTGGCAAATGCTTTCGC 3’
5’TAACCGCAACAACTTTAAT 3’
5’CCTGAGAAACGGCTACCACATC 3’
5’GGTGAAATTCTTGGACCGTC 3’
5’GAGTCTCGTTCGTTATCGGA 3’
5’ATGGTTGCAAAGCTGAAAC 3’
5’CACCTACGGAAACCTTGTTACGTC 3’
5’GACAACCTGGTTGATCCTGCCAGT 3’ / (Hunt et al. 2007; Simon et al. 1994; Shull et al. 2001)
LSU / 28S DD
28S FF / 5’GGGACCCGTCTTGAAACAC 3’
5’TTACACACTCCTTAGCGGAT 3’ / (Monaghan et al. 2007)

* Primers newly designed in present study.

5

Table 3. Samples and GenBank Accessions

Species / Vocher specimen / Locality / SSU / LSU / rrnL / cox1 /
Acrocrypta assamensis / BMNH831362 / China, Guiyang, Mayanghe lijiaba / FJ973885 / FJ973683 / Missing / Missing
Agasicles hygrophila / BMNH831363 / China, Shanxi, Agriculture University / FJ973886 / FJ973684 / FJ973792 / FJ977926
Agetocera mirabilis / BMNH831709 / China, Hainan, Yinggeling / FJ973887 / FJ973685 / FJ973793 / FJ977927
Alagoasa sp / BDF-2000 / Unknown / *AF267447 / Missing / Missing / Missing
Altica viridicyanea / BMNH831365 / China, Beijing, Badaling / FJ973888 / FJ973686 / FJ973794 / FJ977928
Alticinae sp. 1 / BMNH835471 / Ecuador (Pichincha), Bellavista Cloudforest Reserve / FJ973904 / FJ973701 / FJ973808 / Missing
Alticinae sp. 2 / BMNH835476 / Ecuador (Pichincha), Bellavista Cloudforest Reserve / FJ973905 / FJ973702 / FJ973809 / Missing
Alticinae sp. 3 / IBE-JGZ-01000 / Japan, Shiga Pref., Mikunidake, Kyoto Univ. Nat. Reserve / FJ973990 / FJ973785 / FJ973878 / FJ977997
Alticinae sp. 4 / IBE-JGZ-01003 / Mexico, Estado de Mexico, Malinalco, Río Hondo / FJ973991 / FJ973786 / FJ973879 / Missing
Alticinae sp. 5 / IBE-JGZ-01009 / Papua New Guinea, Madang, Adalbert Mts., Keki / FJ973992 / FJ973787 / FJ973880 / FJ977998
Alticinae sp. 6 / IBE-JGZ-01012 / South Africa, Kwazulu-Natal Prov., Nature Farm (Louwsburg) / FJ973993 / FJ973788 / FJ973881 / FJ977999
Alticinae sp. 7 / IBE-JGZ-00988 / Australia, QLD, Daintree, Silky Oaks Lodge / FJ973994 / FJ973789 / FJ973882 / Missing
Alticinae sp. 8 / IBE-JGZ-00989 / Australia, QLD, Daintree, Silky Oaks Lodge / FJ973995 / FJ973790 / FJ973883 / Missing
Amphimela sp. / BMNH831668 / Madagascar, Masoala National Park, south of Marofototra / FJ973889 / FJ973687 / FJ973795 / FJ977929
Aphthona strigosa / BMNH831370 / China, Hainan, Jianfengling Chahekou / FJ973891 / FJ973689 / FJ973797 / FJ977931
Aphthonoides beccarii / BMNH831371 / China, Hubei, Shenlongjia Xinhuaxiang / FJ973892 / FJ973690 / FJ973798 / FJ977932
Aphthonomorpha collaris / BMNH831366 / China, Guiyang, Mayanghe lijiaba / FJ973890 / FJ973688 / FJ973796 / FJ977930
Aplosonyx ancora / BMNH831702 / China, Hainan,Yinggeling / FJ973893 / FJ973691 / FJ973799 / FJ977933
Apterocuris sibirica / BMNH704367 / Kazakhstan, Ivanovsky / *AJ841416 / *AJ841543 / *AJ841300 / *AM283116
Argopistes biplagiatus / BMNH831374 / China, Beijing, Donglingshan / FJ973894 / FJ973692 / FJ973800 / FJ977934
Argopus bidentatus / BMNH831377 / China, Yunnan, Zhongdian Bianhaita / FJ973895 / FJ973693 / FJ973801 / FJ977935
Arsipoda concolor / IBE-JGZ-01015 / Australia, W VIC, Grampians, Wannon River / FJ973896 / FJ973694 / FJ973802 / FJ977936
Arthrotidea ruficollis / BMNH831746 / China, Shaanxi, Ningshan, Xunyangba / FJ973897 / FJ973695 / FJ973803 / FJ977937
Arthrotus chinensis / BMNH831725 / China, Zhejiang, Wuyanling / FJ973898 / FJ973696 / Missing / FJ977926
Asiorestia transversa / BMNH704447 / Portugal, Viana do Castelo / *AJ841466 / *AJ841595 / *AJ841348 / *AM283186
Asphaera sp. / BMNH704448 / France, Guiana,Tresor Mts. / *AJ841467 / *AJ841596 / *AJ841349 / *AM283187
Aspicela bourcierii / BMNH835475 / Ecuador (Pichincha), Bellavista Cloudforest Reserve / FJ973899 / FJ973697 / Missing / Missing
Atrachya menetriesi / BMNH835073 / China, Shaanxi, Zhouzhi, Laoxiancheng / FJ973900 / FJ973698 / FJ973804 / Missing
Batophila aerate / BMNH831688 / United Kingdom, Cambridge / FJ973901 / FJ973699 / FJ973805 / Missing
Bhamoina varipes / BMNH831380 / China, Hainan, Bawangling Donger Linchang / FJ973902 / FJ973700 / FJ973806 / Missing
Blepharida rhois / BMNH704449 / United States of America, New Jersey / *AJ841468 / *AJ841597 / *AJ841350 / *AM283188
Calomela juncta / BMNH704371 / Australia, Queensland / *AJ841420 / *AJ841547 / *AJ841304 / *AM283120
Calomicrus circumfusus / BMNH704463 / Spain, Avila, Puerto de las Fuentes / *AJ841484 / *AJ841613 / *AJ841365 / *AM283201
Cassena tricolor / BMNH831713 / China, Hainan, Mt. Wuzhishan / FJ973906 / FJ973703 / FJ973810 / Missing
Chabria angulicollis / IBE-JGZ-00993 / Malaysia, Pahang, Kuala Lipis env., Kg. Malaka env. / FJ973908 / FJ973704 / Missing / FJ977938
Chabria group / IBE-JGZ-00998 / Indonesia, W Java, Puncak Pass, / FJ973909 / FJ973706 / FJ973812 / FJ977940
Chabria nr angulicollis / BMNH704450 / Malaysia, Pahang, Kuala Lipis env., Kg. Malaka / *AJ841469 / *AJ841598 / *AJ841351 / *AM283189
Chabrisoma sp. / IBE-JGZ-01017 / Philippines, Mindanao, Bagongsilang / FJ973910 / FJ973707 / Missing / FJ977941
Chaetocnema chlorophana / BMNH704451 / Spain, Avila, Aveinte / *AJ841470 / *AJ841599 / Missing / *AM283190
Chalcoides fulvicornis / BMNH 673331 / United Kingdom, Dinton Pastures boundary, SU7871 / FJ973997 / FJ973813 / FJ973813 / *DQ155758
Chrysolina femoralis / BMNH704372 / Spain, Segovia, Cuellar / *AJ841421 / *AJ841548 / *AJ841305 / *AM283121
Chrysomela mainensis / BMNH704374 / Canada, Quebec, L'Outaouais, Quyon / *AJ841423 / *AJ841550 / *AJ841307 / *AM283124
Clitea fulva / BMNH831386 / China, Chongqing, Fengjie Yongfengzhen Yongle / FJ973911 / FJ973708 / FJ973814 / FJ977942
Clitenella fulminans / BMNH835072 / China, Shaxi,zhouzhi, Houzhenzhi / FJ973912 / FJ973709 / Missing / Missing
Cneorane femoralis / BMNH831710 / China, Hainan, Mt. Diaoluoshan / FJ973913 / FJ973710 / FJ973815 / FJ977943
Crepidodera pluta / BMNH831388 / China, Longlang Montain / FJ973914 / FJ973711 / FJ973816 / FJ977944
Decaria sp. / IBE-JGZ-01011 / South Africa, Limpopo Prov., Rust de Winter Nature Reserve / FJ973915 / FJ973712 / FJ973817 / FJ977945
Decertina sp. / BMNH835067 / China, Shanxi, Zhouzhi Houzhengzi / FJ973916 / FJ973713 / FJ973818 / FJ977946
Desmogramma ljunghi / BMNH704375 / Brazil, Paraiba, Areia Branca / *AJ841424 / *AJ841551 / *AJ841308 / *AM283125
Diacantha unifasciata / Unknow / Madagascar, Amparihibe / *AJ841485 / *AJ841614 / *AJ841366 / Missing
Dibolia sp. / IBE-JGZ-00983 / Spain, Murcia, Yecla / FJ973917 / FJ973714 / FJ973819 / FJ977947
Diphaltica sp. / BMNH704453 / France, Guiana, Tresor Mts., Camp Caiman / *AJ841471 / *AJ841600 / *AJ841352 / *AM283191
Disonycha sp / BMNH704454 / United States of America, Ohio, Pike Co., Jackson Lake / *AJ841472 / *AJ841601 / *AJ841353 / *AM283192
Disonycha xanthomelas / IBE-JGZ-00984 / United States of America, WV, Mason Co., Bellmeade / FJ973918 / FJ973715 / FJ973820 / Missing
Doryphora sp. / BMNH704376 / Brazil, Betania, Colombo / *AJ841425 / *AJ841552 / *AJ841309 / *AM283126
Epitrix abeillei / BMNH831393 / China, Beijing, Yuanmingyuan / FJ973919 / FJ973716 / Missing / FJ977948
Euphitrea wallacei / BMNH704455 / Malaysia, Sabah, Batu Punggul env / *AJ841473 / *AJ841602 / *AJ841354 / *AM283193
Exora obsoleta / BMNH704465 / Costa Rica, Heredia, Selva Verde Lodge / *AJ841486 / *AJ841615 / *AJ841367 / *AM283203
Exosoma lusitanicum / BMNH704466 / Spain, Madrid, El Pardo / *AJ841487 / *AJ841616 / *AJ841368 / *AM283204
Galeruca pomonae / BMNH704467 / Spain, Ciudad Real, Embalse de Pearroya / *AJ841488 / *AJ841617 / *AJ841369 / *AM283205
Galerucella pusilla / BMNH 673337 / United Kingdom, London / FJ973920 / FJ973717 / Missing / *DQ155764
Galerucinae sp. 1 / BMNH704477 / South Africa, East Cape, Tsitsikamma Natl. Pk. / *AJ841500 / AJ841629 / *AJ841381 / *AM283215
Galerucinae sp. 2 / BMNH835065 / China, Tibet, Linzhi, Motuo, Duoxionglashanyakou / FJ973921 / FJ973719 / Missing / FJ977949
Galerucinae sp. 3 / BMNH835469 / Ecuador (Pichincha), Bellavista Cloudforest Reserve / FJ973903 / Missing / FJ973807 / Missing
Galerucinae sp. 4 / BMNH835472 / Ecuador (Pichincha), Bellavista Cloudforest Reserve / FJ973922 / FJ973718 / FJ973821 / FJ977950
Gallerucida bifasciata / BMNH835057 / China, Taiwan, Kaosiung county Jongjhihguan / FJ973923 / FJ973720 / FJ973822 / Missing
Gastrophysa viridula / BMNH676943 / United Kingdom, W.Clare / FJ973924 / FJ973721 / FJ973823 / *DQ156057
Glyptina cyanipennis / IBE-JGZ-00986 / United States of America, OH, Ross Co., Chillicothe / FJ973925 / Missing / FJ973824 / FJ977951
Gonioctena olivacea / BMNH704377 / United Kingdom, Surrey, Sunningdale, Chobbam Common / *AJ841426 / *AJ841553 / *AJ841310 / *AM283127
Griva regularis / BMNH831395 / China, Hubei, Shenlongjia Xinhua / FJ973926 / FJ973722 / FJ973825 / FJ977952
Hecataeus sp. / BMNH704468 / France, Guiana, S Cayenne / *AJ841489 / *AJ841618 / *AJ841370 / *AM283206
Heikertingerella sp. / IBE-JGZ-00992 / Brazil, Río de Janeiro, Corcovado / FJ973927 / FJ973723 / FJ973826 / FJ977953
Hemipyxis plagioderoides / BMNH831397 / China, Sichuan, Wolong / FJ973928 / FJ973724 / FJ973827 / FJ977954
Hemistus sp. / BMNH704469 / Malaysia, Sabah, Batu Punggul env. / *AJ841490 / *AJ841619 / *AJ841371 / *AM283207
Hermaeophaga mercurialis / BMNH675568 / SU3660 (No detailed information available) / FJ973929 / FJ973725 / FJ973828 / FJ977955
Hespera lomasa / BMNH831408 / China, Shanxi, Houzhenzi Zhi Shaliangzi / FJ973930 / FJ973726 / FJ973829 / FJ977956
Hippuriphila modeeri / BMNH724209 / United Kingdom, Queen Mary resevoir / FJ973931 / FJ973727 / Missing / Missing
Hoplasoma unicolor / BMNH835077 / China, Yunnan, Xishuangbanna, Xiaomengyang / FJ973932 / FJ973728 / FJ973830 / Missing
Hoplosaenidea subcostata / BMNH704470 / Malaysia, Sabah, Batu Punggul env / *AJ841491 / *AJ841620 / *AJ841372 / *AM283208
Hyphasis indica / BMNH831409 / China, Yunnan, Xishuangbanna / FJ973933 / FJ973729 / FJ973831 / FJ977957
Issikia dimidiaticornis / BMNH835075 / China, Hainan, Mt. Wuzhishan / FJ973934 / FJ973730 / Missing / FJ977958
Jacobyana sp. / BMNH835069 / China, Guangdong, Chebaling / FJ973935 / FJ973731 / Missing / Missing
Jacobyanella hexaspilota / Unknown / Madagascar, Amparihibe / *AJ841492 / *AJ841621 / *AJ841373 / Missing
Johannica gemellata / BMNH704378 / Australia, Queensland / *AJ841427 / *AJ841554 / *AJ841311 / *AM283128
Labidomera clivicollis / BMNH704379 / Unknown / *AJ841428 / *AJ841555 / *AJ841312 / *AM283129
Laboissierea sculpturata / BMNH831457 / China, Hainan, Bawangling / FJ973936 / FJ973732 / FJ973832 / FJ977959
Lactica sp. / BMNH831413 / China, Guizhou, Mayanghe, Lijiaba / FJ973937 / FJ973733 / FJ973833 / Missing
Lamprolina sp. / BMNH704380 / Australia, Queensland, Brisbane / *AJ841429 / *AJ841556 / *AJ841313 / *AM283130
Lanka ramakrishnai / BMNH835063 / India, Kerala vellagani / FJ973938 / FJ973734 / FJ973834 / Missing
Laotzeus sp. / BMNH831414 / China, Yunnan, Gongshan Pulahekou / FJ973939 / FJ973735 / FJ973835 / Missing
Larphris emarginata / BMNH831743 / China, Zhejiang, Mt. Fengyangshan / FJ973940 / FJ973736 / FJ973836 / FJ977960
Leptaulaca undecimpunctata / Unknown / Madagascar, Amparihibe / *AJ841493 / *AJ841622 / *AJ841374 / Missing
Leptinotarsa juncta / BMNH704381 / United States of America, West Virginia / *AJ841430 / *AJ841557 / *AJ841314 / *AM283131
Linaeidea aenea / BMNH704382 / Germany, Upper Bavaria, Benediktbeuern / *AJ781619 / *AJ781682 / *AJ781556 / *AM283132
Lipromela decemmaculata / BMNH831429 / China, Hainan, Diaoluoshan, Daxiagu / FJ973941 / FJ973737 / FJ973837 / FJ977961
Lochmaea capreae / BMNH831691 / United Kingdom, Anglesey / FJ973942 / FJ973738 / FJ973838 / FJ977962