The genetic basis of pheromone evolution in moths Astrid T. Groot, Teun Dekker, David G. Heckel Supplemental Table 1. Overview of the genetic analyses conducted to determine the heritability and/or mode of inheritance of variation in the female sex pheromone signal and the male preference response. # genes involved Species Type of analysis Type of cross Main findings ref (sex linked?) In heterozygous 2 genes: 1 major females much more gene (with two Z Crossing and selection F1 and variation than alleles) and 1 experiments with backcrosses (49) expected based on a modifying pheromone analysis phenotyped one locus-2 allele dominant locus in model E Sex pheromone is Crossing experiments F1, F2 and controlled by simple one gene, two with pheromone backcrosses Mendelian (27, 38) alleles analysis phenotyped inheritance involving a single pair of alleles Ostrinia one autosomal QTL analysis nubilalis E Not d11desat, but gene: pgFAR (backcrosses with vs Z Backcrosses FAR is the gene (unclear which pheromone analysis) (30, 31) phenotyped and underlying SNPs are and identifying genotyped pheromone involved, many candidate genes in differentiation SNPs between E QTL and Z Existence of all one gene (first Cloning and functional desats in both species gene duplication, analysis with (d11 in ACB, d14 in then desaturases NA ECB) without d11 or subfunctionalizati (39) (YEpOLEX for d9, d14 products suggest on, after which pYES2 for d11, gene duplication and neofunct to d14 pFastBac1 for d14) subfunctionalization happened) F1 produces 14:Ac one recessive O.nubilalis Crossing experiments F1, F2 and plus all pheromone gene, i.e. (40) vs O. with pheromone backcrosses components of both suppressor of furnacalis analysis phenotyped parents desaturase Genetic analyis one autosomal Ostrinia Inheritance of female sex pheromone signal pheromone sex female of Inheritance Crossing experiments F1, F2 and similar to O.nub, but locus with 2 scapulalis E (45) with pheromone backcrosses natural variation in alleles (plus vs O. analysis phenotyped EE, ZZ and EZ freqs modifiers for I furnacalis (in Japan) type) Ostrinia in F no Z9 produced, 1 two genes that are zealis, O. thus recessive. In Crossing experiments F1 and likely linked (one zaguliaevi, ZeaScaZ x Zea: 15/68 with pheromone backcrosses for EZ var and (44) and O. sp. females are ScaE-like analysis phenotyped one for the third near (Zea x ScaZ: F ScaI- 1 comp in O. zealis zaguliaevi like) Z9-14:OAc Ostrinia production controlled Crossing experiments scapulalis vs F1, F2 and through an autosomal One major with pheromone (43) backcrosses recessive gene, likely autosomal gene O. zealis analyses chain-shortening of fatty acid precursor Supplemental Material: Annu. Rev. Entomol. 2016. 61:99–117 doi: 10.1146/annurev-ento-010715-023638 The Genetic Basis of Pheromone Evolution in Moths Groot, Dekker, and Heckel Selection for 3 A few genes, Z11 and Z9 showed a generations presumably Ostrinia Selection on female heritability of 0.6-0.6. for E11-14:OAc, involved in (42) zaguliaevi pheromone E11 no heritable Z11-14:OAc and reduction or variation Z9-14:OAc acetylation Crosses were Planotortrix Crossing experiments F1, F2 and phenotyped, and two genes: most excessana, with pheromone backcrosses expression of desat5 likely desat5 and (2) analysis and qPCR of P. octo phenotyped was measured (also a repressor candidate gene sequenced) Co and Po use Z8- Ctenopseusti 14:Ac as extra sex s obliquana Desat expression & pheromone two genes: a (Co) and phenotyping in F1 and component; due to repressor of Planotortrix backcrosses; F1, F2 and changes in a trans- desat5 and a octo (Po) , sequencing, qPCR and backcrosses acting repressor and a (1) mutation in the vs C. herana functional analyses phenotyped cis-regulatory desat5 promoter (Ch) and P. with d9 and d11 mutation in an excessana desaturases activator binding (Pe) site within the desat5 promoter Different crosses yield different Ctenopseusti Crossing experiments F1, F2 and multiple genes, pheromone blends, s obliquana with pheromone backcrosses sex-linked and (14) suggesting >1 gene, vs C. herana analysis phenotyped autosomal sex-linked and autosomal Cloning and functional analysis of pheromone Identification of the (36) NA NA gland specific fatty first pg-FAR acyl reductase Bombyx Identification of mori desat1, a bifunctional Cloning and functional enzyme that catalyzes analysis of pheromone NA NA (35) both a Z11 specific desaturases desaturation and a d10,12 desaturation Helicoverpa Ratio of Z9-16:Ald to one autosomal Crossing experiments F1 and armigera Z11-16:Ald gene, two alleles, with pheromone backcrosses (47) and H. controlled by one with H.armigera analysis phenotyped assulta autosomal gene dominance Heliothis QTL analysis on Backcrosses Complex genetic virescens multiple interspecific phenotyped and inheritance, 5-12 (18, 41) and H. autosomal genes pheromone variation genotyped QTL found subflexa Major QTL onto QTL analysis on Backcross Heliothis which d9-desaturases one autosomal intraspecific phenotyped and (19) virescens map, not d11- gene pheromone variation genotyped desaturase QTL analysis on Backcross Major QTL for Heliothis one autosomal intraspecific phenotyped and acetates overlaps with (20) subflexa gene pheromone variation genotyped interspecific QTL KPSE similar to other KPSE, Cloning and functional MsexAPTQThe Manduca characterization of NA possesses both NA (33) sexta desaturases KPSE and Z11 desaturase and APTQ 10,12-desaturase activities mutant pheromone Trichoplusia Crossing experiments F1, F2 and blend controlled by a one gene (24) ni with mutant line backcrosses single recessive autosomal gene Spodoptera sex pheromone Crossing experiments one major gene, descoinsi controlled by one with pheromone F1, F2 and with the Sl allele (Sd) and S. autosomal gene, (34) analysis and calling backcrosses semi-dominant latisfascia onset time of calling time over the Sd allele (Sl) likely polygenic QTL analysis on F1 and backcross Spodoptera multiple QTL with multiple pheromone variation in phenotyped and (21) frugiperda small effects autosomal genes corn and rice strain genotyped Selection (6 Response to Selection differentials generations) selection shows (sd) High line: 3.4- Cadra Selection on female for high and low additive genetic 4.2, (3, 4) cautella pheromone ratios of Z9,E12– variance to titers sd Low lines: 1.9 – 14:Ac to Z9– of components, 2.6 14:Ac not to ratio Autosomal Likely one major Crossing experiments inheritance with no F1, F2 and autosomal gene, Agrotis with Scandinavian and dominance for Z5- backcrosses with partial (29) segetum Zimbabwean 10/Z7-12 ratio, but phenotyped dominace for Z5- populations more complicated for 10:Ac Z9-14/Z7-12 ratio Genetic correlation F1 (heritability between the two Heritability of analysis, F2 isomers: 0.989 Not assessed (7) intraspecific variation (inbreeding Inbreeding: no effect analysis) on ratio, but total amt Pectinophor sign. lower a Selection for 12 gossypiella generations to change Heritability of variation Bidirectional the ratio of the binary in pheromone selection NA (9) blend showed production experiments heritability in one direction only Male response is Crossing experiments F1, F2 and heritable, NY (E) with male behavioral backcrosses males have broader probably one gene (27) response phenotyped response than P (Z) males Crossing experiments F1, F2, and SSRs segregate in one gene, with single sensillum backcrosses (23) Mendelian fashion autosomal recordings (SSR) phenotyped two genes: one Crossing experiments F1, F2 and SSR response autosomal for the with male backcrosses autosomal, behavioral periphery and one (10, 38) physiological and phenotyped response sex-linked sex-linked for the Ostrinia behavioral response behavior nubilalis E Crossing and vs Z Z chromosome behavioral analysis and ExZ strain F1 and origin(s) determines sex-linked (16) allozyme tracking of Z F2 crosses behavioral repsonse chromosome F1 and QTL analysis with backcrosses Behavioral response one gene, sex- male behavioral (11) phenotyped and sex-linked linked response Inheritance of male response and preference response and ofInheritance male genotyped Crossing experiments Antennal response, with antennal lobe F1 and antennal lobe recordings, backcrosses neuroanatomy and sex-linked (26) neuroanatomy and phenotyped volume of glomeruli antennal response are sex linked at least two genes, Crossing experiments F1 and SSR response both one autosomal with single sensillum backcrosses autosomal and sex- and one E- (37) recordings (SSR) phenotyped linked dominant sex- linked Gradual increase in Artificial selection for response breadth to broader response in selection (and gradual NA multiple genes (12) ECB Z males to ACB decrease in absence pheromone of selection) suggests a quantitative trait Hybrids have Crossing experiments swapped neuronal with sensory F1 phenotyped identity across all sex-linked (28) physiological analysis three neurons in the (SSR) same sensillum Comparison of A single nucleotide Ostrinia genetics and shift in one OR One gene, nubilalis and Pheromone receptor response profile underlies swap from pheromone (32) O. analyses of candidate E11-14:OAc to E12- receptor, OR3 furnacalis pheromone 14:OAc receptor Phenotype distribution suggests single-sex linked one gene, sex- locus with C. herana- Crossing experiments F1, F2 and linked, although type dominance. with single sensillum backcrosses additional genes (22) However, variability recordings phenotyped seem to be in the responses of involved too the