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Lu (Ling) Yang @edenling3 Princeton University

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2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is prohibited] . 2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is prohibited.

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A adaptation B

A B

or How often do cases of convergent evolution share a common 2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction evenmolecular in part is prohibited. mechanism? A B How predictable is evolution on the molecular level?

2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is 3prohibited. 2018 © 48th European Contact Lens Society Of Ophthalmologists. All rights reserved - Any reproduction even in part is prohibited] .

2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is 4prohibited. 2018 © 48th European Contact Lens Society Of Ophthalmologists. All rights reserved - Any reproduction even in part is prohibited] .

2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is prohibited] . Q111V Q111L Q111L/V/T Q111T A119S A119S A119N A119S/N A119S N122H N122H N122H N122H Target site insensitivity

Extracellular

Butterflies Flies Wasps Beetles True bugs COOH moths Diptera Hymenoptera Coleoptera Hemiptera NH 2 Cytoplasmic Lepidoptera References F. Holzinger, et al., 1992.

2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is prohibited. E. Labeyrie, et al., 2004. Y. Zhen, et al., 2012. S. Dobler, et al., 2012. S. Dobler, et al., 2015. G. Petschenka, et al., 2017.

(Pictures from the Internet)

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Grasshoppers Orthoptera

Butterflies Flies Wasps Beetles True bugs moths Diptera Hymenoptera Coleoptera Hemiptera Lepidoptera

Photo credits: Top, Middle: Ricardo Mariño-Pérez. Bottom: Krushnamegh Kunte.

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Grasshoppers Orthoptera

Butterflies Flies Wasps Beetles True bugs moths Diptera Hymenoptera Coleoptera Hemiptera Lepidoptera ? 2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction evenIf in partpredictable is prohibited. … Amino acid substitutions? Duplications? Photo credits: Top, Middle: Ricardo Mariño-Pérez. Bottom: Krushnamegh Kunte.

2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is 7prohibited. 2018 © 48th European Contact Lens Society Of Ophthalmologists. All rights reserved - Any reproduction even in part is prohibited] .

Gather Pyrgomorphidae grasshopper samples from collaborators (Krushnamegh Kunte, Hojun Song, Ricardo Mariño-Pérez.)

Grasshoppers Orthoptera RNA-seq library preparation.

Hi-Seq sequencing

Sequences of ATP1α

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Outgroup Mantodea:Mantidae Acrididae Orthoptera Pyrgomorphidae (Yang et al. unpublished)

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2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is prohibited] . Color codes: • Outgroups sensitive to cardenolides Orthoptera • Resistant single copy of ATP1α or the more conserved copy in duplicates • The duplicated copy of ATP1α that Hemiptera harbors more mutations

Coleoptera

Diptera

Lepidoptera

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Coleoptera Parallelism #1 Most of the non-synonymous substitutions are also present in 2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is prohibited.

Diptera other insect orders.

Lepidoptera

Coleoptera Parallelism #2 Most of the unique non-synonymous substitutions occurred on ATP1α 2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is prohibited.

Diptera duplicates.

Lepidoptera

2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is prohibited] . 1 1 1 1 1 1 1 1 1 1 3 3 7 7 8 8 9 0 0 1 1 1 1 1 2 2 2 0 1 8 9 7 8 7 4 9 1 2 5 8 9 0 1 2 9 5 6 7 4 0 2

reference C S Q A V P A D D N H I F T L R R L. migratoria · · L · · · S · · · · · · · · · · A. miliaris · · L · · · S · · · · · · · · · · a

T. calliparea · · L · · · S · · · · · · · M · · r e t

Z. elegans · · L · · · S · · · · · · · · · · p o

· · L · · · S · · · · · · · · · · h P. pictus B t r

P. leprosus B · · L · · · S · · · · · · · · · · O P. pictus A · · L · T S A D S · · H · V · · · · ·

2018 © II Joint Congress on Evolutionary Biology 2018. AllP rights. l ereservedpro - Anysu reproductions A even in part is prohibited.· · L · T S T D S E N H · V · · M · · B. trivittata · · · · · · S · · · Y · · · T · · a r

O. fasciatus D · · · · · · S · · · Y · · · V · · e # of non-synonymous substitutions t p O. fasciatus C · · · · · · S · · H Y · · S M · · i m

O. fasciatus B · F T S E S S · · H F V N · M · · e

=> Relaxed selection on the duplicated copies of ATP1α H O. fasciatus A · F T S · A S · N H F V N A E · Q

T. castaneum 2018 ©· II Joint· Congress· · on ·Evolutionary· Biology 2018.· ·All rights· reserved· · - Any· reproduction· · evenT in· part· is prohibited. R. lineaticollis A · G T · · · S · · · · · · · M · · 2018 R©. lin e48thaticollis B EuropeanY G · · · · S · ·ContactY · · · · M · · Lens Society Of Ophthalmologists. All rights reserved - Any reproduction even in part is prohibited] . a

T. tetraophthalmus · · L · · · S · · · · V · · R · · r e t

P. versicolora · · · · · · · · · · · · · · M · · p o e

L. clivicolis · G V · A · S · · H · · · · M · · l o

C. cobalitinus A · · L · · · S · · · · · · · M · · C C. auratus A · · L · · · S · · · · · · · M · · C. cobalitinus B · A V · · A S · · H Y L · · M · · C. auratus B · A V · · A S · · H Y L · · M S · 2018 © II Joint Congress on EvolutionaryL. eup atBiologyorii · · · 2018.· S · S ·All· · · rights· · · - - - reserved - Any reproduction even in part is prohibited] . L. sp. · · L · T · S · · · · · · · - - - N. lateralis · A L · E · S · · · · · · · - - - a r

N. scrophulariae · A L · E · S · · · · · · · - - - e t p P. ilicis · · · · S · S · · · · · · · - - - i D P. fallaciosa · · · · S · S · · · · · · · - - - P. hellebori B · · · · S · S · · · · · · · - - - P. hellebori A · · H · S · S · · H · · · · - - - 2018 © IIB. mJointori Congress· G · · · · · ·on· · · Evolutionary· · · M · · Biology 2018. All rights reserved - Any reproduction even in part is prohibited] . T. legitima · G L · · · · · · · · · · · M · · a C. tenera · G L · · · S · · · · · · · M · · r e t

L. caryae · G L · · · · · · · · · · · · · · p o d L. archippus · G · · · · · · · · · · · · · · · i p e

L. halia · G L · · · S · · · · · · · T · · L D. gilippus · G V · · · S · · · · · · · T · · D. plexippus · G V · · · S · · H · · · · T · ·

2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is prohibited] . Parallelism #3 The duplicated copies that harbor more non-synonymous substitutions all experienced relaxed selection.

Whereas, some amino acid sites were positively selected. (Yang’s branch-site analysis)

Poekilocerus pictus

High Toxin concentration Dogbane beetle Weevil

2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in partParallelism is prohibited. #4 Higher expression of the presumably more resistant copy in gut than in brain.

Y. Zhen, et al., 2012. 2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is prohibited. 2018 © 48th European Contact Lens Society Of Ophthalmologists. All rights reserved - Any reproduction even in part is prohibited] .

2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is prohibited] . Q111V Q111L Q111L/V/T Q111T Q111L A119S A119S A119N A119S/N A119S A119S N122H N122H N122H N122H N122H

Butterflies Flies Wasps Beetles True bugs Grasshoppers moths Diptera Hymenoptera Coleoptera Hemiptera Orthoptera Lepidoptera Amino acid substitutions ✔️

2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is prohibited. Duplication ✔️ RelaxedPREDICTABLE& positive selection✔️ Differential expression ✔️

(Yang et al. unpublished)

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Krushnamegh Kunte, Nitin R., Riddhi Deshmukh National Center of Biological Sciences, India.

Funding

Hojun Song, Ricardo Mariño-Pérez Texas, A&T. 2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is18 prohibited. 2018 © 48th European Contact Lens Society Of Ophthalmologists. All rights reserved - Any reproduction even in part is prohibited] .

2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is19 prohibited. 2018 © 48th European Contact Lens Society Of Ophthalmologists. All rights reserved - Any reproduction even in part is prohibited] .

[email protected] @edenling3 www.luyangevolution.com

2018 © II Joint Congress on Evolutionary Biology 2018. All rights reserved - Any reproduction even in part is20 prohibited. 2018 © 48th European Contact Lens Society Of Ophthalmologists. All rights reserved - Any reproduction even in part is prohibited] .