An integrative approach to unravel the Ceratitis FAR (Diptera, Tephritidae) cryptic species complex: a review

This is an elec onic ep in o he o iginal a icle. This ep in may di e om he o iginal in pagina ion and ypog aphic de ail. Au ho (s): Ti le: Yea : Ve sion: Please ci e he o iginal e sion: All ma e ial supplied ia JYX is p o ec ed by copy igh and o he in ellec ual p ope y igh s, and duplica ion o sale o all o pa o any o he eposi o y collec ions is no pe mi ed, excep ha ma e ial may be duplica ed by you o you esea ch use o educa ional pu poses in elec onic o p in o m. You mus ob ain pe mission o any o he use. Elec onic o p in copies may no be o e ed, whe he o sale o o he wise o anyone who is no an au ho ised use . An in eg a i e app oach o un a el he Ce a i is FAR (Dip e a, Teph i idae) c yp ic species complex: a e iew De Meye , Ma c; Dela e, Hélène; Ekesi, Sunday; Jo daens, Ku ; Kalino a, Blanka; Man akhan, A una; Mwa awala, Maulid; S eck, Ga y; Van Cann, Joannes; Vanciko a, Lucie; B izo a, Radka; Vi gilio, Massimiliano De Meye , M., Dela e, H., Ekesi, S., Jo daens, K., Kalino a, B., Man akhan, A., Mwa awala, M., S eck, G., Van Cann, J., Vanciko a, L., B izo a, R., & Vi gilio, M. (2015). An in eg a i e app oach o un a el he Ce a i is FAR (Dip e a, Teph i idae) c yp ic species complex: a e iew. ZooKeys, 540, 405-427. h ps://doi.o g/10.3897/zookeys.540.10046 2015 An in eg a i e app oach o un a el he Ce a i is FAR (Dip e a, Teph i idae)... 405 An in eg a i e app oach o un a el he Ce a i is FAR (Dip e a, Teph i idae) c yp ic species complex: a e iew Ma c De Meye 1, Hélène Dela e2, Sunday Ekesi3, Ku Jo daens1,8, BlankaKalino á4, A una Man akhan5, Maulid Mwa awala6, Ga y S eck7, JoannesVanCann8,9, Lucie Vaníčko á4,10, Radka Břízo á11, Massimiliano Vi gilio1 1 Royal Museum o Cen al A ica, In e eb a es Sec ion and JEMU, Leu enses eenweg 13, B3080 Te u en, Belgium 2 CIRAD, UMR PVBMT, 7 ch de l’IRAT, 97410 Sain -Pie e, La Réunion, F ance 3 In e na ional Cen e o Insec Physiology and Ecology (icipe), P.O. Box 30772 – 00100, Nai obi, Kenya 4 Ins i u o O ganic Chemis y and Biochemis y, Academy o Sciences o he Czech Republic, P ague 5 Ci us Resea ch In e na io- nal, PO Box 28, Nelsp ui 1200, Sou h A ica 6 Sokoine Uni e si y o Ag icul u e, Dep . o C op Science and P oduc ion, Mo ogo o, Tanzania 7 Flo ida Depa men o Ag icul u e and Consume Se ices, Gaines ille FL, USA 8 Uni e si y o An we p, G oenenbo ge laan 171, B-2020 An we p, Belgium 9 Uni e si y o Jy äskylä, PO Box 35, 40014 Finland 10 Labo a ó io de Ecologia Química, Ins i u o de Química e Bio ecnologia, Uni e sidade Fede al de Alagoas, A . Lou i al de Melo Mo a, s/n, Tabulei o, 57072-970, Maceió, AL, B azil 11Ins i u e o Chemical Technology in P ague, Technická 5, CZ-166 28 P ague 6, Czech Republic Co esponding au ho : Ma c De Meye ([email p o ec ed]) Academic edi o : J. Hend ichs|Recei ed 26 May 2015|Accep ed 5 Augus 2015|Published 26 No embe 2015 h p://zoobank.o g/C072905C-89EE-4BE4-B5B7-33819CD86A3F Ci a ion: De Meye M, Dela e H, Ekesi S, Jo daens K, Kalino á B, Man akhan A, Mwa awala M, S eck G, Van Cann J, Vaníčko á L, Břízo á R, Vi gilio M (2015) An in eg a i e app oach o un a el he Ce a i is FAR (Dip e a, Teph i idae) c yp ic species complex: a e iew. In: De Meye M, Cla ke AR, Ve a MT, Hend ichs J (Eds) Resolu ion o C yp ic Species Complexes o Teph i id Pes s o Enhance SIT Applica ion and Facili a e In e na ional T ade. ZooKeys 540: 405–427. doi: 10.3897/zookeys.540.10046 Abs ac This pape e iews all in o ma ion ga he ed om di e en disciplines and s udies o esol e he species s a us wi hin he Ce a i is FAR (C. asci en is, C. anonae, C. osa) complex, a g oup o polyphagous ui ly pes species (Dip e a, Teph i idae) om A ica. I includes in o ma ion on la al and adul mo phol- ogy, wing mo phome ics, cu icula hyd oca bons, phe omones, mic osa elli es, de elopmen al physiol- ogy and geog aphic dis ibu ion. The gene al consensus is ha he FAR complex comp ises C. anonae, wo species wi hin C. osa (so-called R1 and R2) and wo pu a i e species unde C. asci en is. The in o ma- ion ega ding he la e is, howe e , oo limi ed o d aw inal conclusions on speci ic s a us. E idence o his ecogni ion is discussed wi h e e ence o publica ions p o iding u he de ails. ZooKeys 540: 405–427 (2015) doi: 10.3897/zookeys.540.10046 h p://zookeys.penso .ne Copy igh Ma c De Meye e al. This is an open access a icle dis ibu ed unde he e ms o he C ea i e Commons A ibu ion License (CC BY 4.0), which pe mi s un es ic ed use, dis ibu ion, and ep oduc ion in any medium, p o ided he o iginal au ho and sou ce a e c edi ed. RESEARCH ARTICLE Launched o accele a e biodi e si y esea ch A pee - e iewed open-access jou nal Ma c De Meye e al. / ZooKeys 540: 405–427 (2015) 406 Keywo ds Taxonomy, Ce a i is osa, Ce a i is asci en is, Ce a i is anonae, A ica, ui ly In oduc ion His o ical backg ound and sys ema ic posi ion Ce a i is MacLeay is an A o opical genus o eph i id ui lies comp ising close o 100 species ound in Sub-Saha an A ica and he islands o he Wes e n Indian Ocean. Phylogene ically i belongs o he sub ibe Ce a i idina wi hin he ibe Dacini. The la e ibe includes all main pes gene a occu ing na u ally in A ica, i.e. Bac oce a Macqua , Cappa imyia Bezzi, Ce a i is, Dacus Fab icius, Neoce a i is Hendel and T i hi h um Bezzi. Howe e , he monophyly o he genus is no suppo ed and some species appea phylo- gene ically mo e closely ela ed o T i hi h um, while o he Ce a i is species p obably do no belong o he genus (Ba and McPhe on 2006, Vi gilio e al. 2015). Con a y o o he indigenous A ican ui ly pes g oups ha ha e a pa icula hos niche wid h a - acking pa icula plan amilies, species o he genus Ce a i is demons a e a e y a iable hos speci ici y. Some species a e la gely monophagous, while o he s o m monophyle ic s enophagous clus e s specialized on pa icula hos gene a. Ye , o he s a e polyphagous species a acking a wide a ie y o un ela ed plan s (De Meye e al. 2002, E bou e al. 2011). Fo example, he Medi e anean ui ly, Ce a i is capi a a (Wiedemann), is he mos ex eme case o polyphagy, wi h 304 di e en hos plan s belonging o 154 gene a in 57 amilies conside ed o be sui able hos plan s (Liquido e al. 2014). The Ce a i is FAR complex is a g oup o polyphagous species comp ising h ee mo phologically simila species: C. asci en is (Bezzi), C. anonae G aham and C. osa Ka sch (Figu e 1). Al hough he FAR complex is ecognized as a g oup o closely ela - ed species, hei monophyly is no always suppo ed in phylogene ic s udies. Ba and Wiegmann (2009) p esen ed a molecula analysis o a numbe o Ce a i is (P e an- d us) species including one specimen o each Ce a i is FAR complex species. They s a e ha CAD1 and ND6 ma ke s suppo ed he monophyly o he complex, while ango and pe iod ma ke s placed he h ee species in di e en clus e s wi hin he so- called ‘P e and us A’ g oup. A majo i y ule Bayesian ee o he combined molecula da a suppo ed he monophyly o he complex. Ye , a cladis ics analysis based on adul mo phological cha ac e s could no ecognize hem as a monophyle ic clus e wi hin he subgenus P e and us (De Meye 2005). Cu en axonomic s a us o ecognized species and sexual dimo phism The Na al ui ly, C. osa, was o iginally desc ibed om ’Delagoabai’ (nowadays Baia de Mapu o, Mozambique and p obably e e ing o a loca ion nea Mapu o) by Ka sch in 1887, based on a single male specimen (Ka sch 1887). Bezzi (1920) desc ibed as- An in eg a i e app oach o un a el he Ce a i is FAR (Dip e a, Teph i idae)... 407 c b a Figu e 1. Habi us image o a Ce a i is asci en is b C. anonae c C. osa (a, c pho os R.S. Copeland, b pho o G. Goe gen). Ma c De Meye e al. / ZooKeys 540: 405–427 (2015) 408 ci en is as a a ie y o P e and us osa, based upon ma e ial om Uganda (unknown locali y). De Meye (2001) conside ed i o be a sepa a e en i y wi h speci ic s a us. Ce a i is anonae was desc ibed by G aham om ma e ial ea ed om an Annona ui in Ashan i, Ghana (G aham 1908). All h ee species show ema kable sexual dimo phism, wi h he males ha ing leg o namen a ion ha is absen in emales. Ce a i is anonae males ha e he mid leg (Fig- u e2) wi h a ow o long da k, la ened se ae (so-called ‘ ea he ing’) en ally along he en i e leng h o he mid emu . The mid ibia is b oadened wi h ea he ing do sally along dis al 0.9 and en ally along he dis al 0.8. The mid leg is la gely b ownish o b ownish black in colou . In C. osa males (Figu e 3), he en al ea he ing on mid emu is absen (a mos he e a e a ew hin and dispe sed se ulae en ally). The mid ibiae is mode a ely b oadened, an e io ly black wi h a conspicuous sil e y e lec ion seen when kep unde a ce ain angle and black ea he ing do sally along dis al 0.75 and en ally along dis al 0.66–0.75. O he wise, he leg is yellow. Ce a i is asci en is males ha e he mid leg shaped simila ly o C. osa, excep ha he mid ibia is no dis- inc ly b oadened and he black ea he ing is es ic ed o he dis al 0.5 a mos (Figu e 4). The leg is colou ed uni o mly yellow, excep in some specimens whe e he an e io pa is pa ially b ownish in he dis al 0.3 (De Meye 2001). P elimina y cou ship beha io s udies by Quilici e al. (2002) gi e some indica ion ha , o a leas in C. osa, he mid legs a e used in p ecopula o y beha io . Figu e 2. Mid leg o male Ce a i is anonae, an e io iew o emu and ibia (pho o I.M. Whi e, NHM). An in eg a i e app oach o un a el he Ce a i is FAR (Dip e a, Teph i idae)... 409 Figu e 3. Mid leg o male Ce a i is osa, an e io iew (pho o I.M. Whi e, NHM). Figu e 4. Mid leg o male Ce a i is asci en is, an e io iew (pho o I.M. Whi e, NHM). Ma c De Meye e al. / ZooKeys 540: 405–427 (2015) 410 Unlike he males, emales, a e mo phologically almos indis inguishable. Ce a i is anonae emales di e om he o he wo axa in he pilosi y o he anepis e num and o e emu . In C. anonae he anepis e num has some ew da k se ulae medio en ally, and he o e emu has dispe sed sho da k se ulae be ween he en al se ae and pos- e io ow o se ae. In C. osa and C. asci en is, he anepis e nal pilosi y is comple ely pale and he o e emu usually only has pale se ulae p esen be ween he en al se ae and pos e io ow o se ae (De Meye and F eidbe g 2006). Females o C. osa and C. asci en is emales canno be eliably di e en ia ed on mo phological cha ac e s. Dis ibu ion pa e ns h oughou A ica The occu ence o he h ee species h oughou A ica shows di e en dis ibu ion pa - e ns wi h only a pa ial o e lap be ween some o he species. Al hough some species do occu sympa ically o pa apa ically, nowhe e do all h ee species co-occu . Ce a i is osa is ound h oughou Sou h ( om wes e n Cape onwa ds) o Eas A ica wi h he no he nmos eco ds om he Cen al Highlands in Kenya (Figu e 5). The la e oc- cu ence seems o be a ecen expansion (Copeland and Wha on 2006) wi h i s p e i- ous ex ension eaching ill he Kenyan coas only. I has been in oduced o he Indian Ocean Islands o La Réunion and Mau i ius in he 20 h Cen u y (Whi e e al. 2000). The single eco d o C. osa (one male) om Came oon in Wes A ica is p obably due o mislabeling, while he p esence o he species in I o y Coas (N’dépo e al. 2010, 2013) is an e oneous eco d. No eliable eco ds om wes e n A ica ha e been ound. Ce a i is asci en is has a wide dis ibu ion and is ound h oughou wes e n A ica, wi h isola ed eco ds om Cen al A ica, and ex ensi e dis ibu ion along he Albe - ine and G ego y Ri s in eas e n A ica, as a no h as E hiopia (Figu e 5). Ce a i is anonae has a p edominan ly equa o ial bel dis ibu ion, being widesp ead h ough Wes ( om Senegal onwa ds) and Cen al A ica (Figu e 6). I s eas e nmos dis ibu- ion seems o be con ined o he wes e n side o he G ego y Ri in Kenya (Copeland e al. 2006) ( eco ds u he o he eas need con i ma ion). Sympa ic and allopa ic occu ence is ound be ween C. anonae and C. asci en is in wes e n A ica (Vayssiè es e al. 2004), and wes e n Kenya (Copeland and Wha on 2006), while C. osa and C. asci en is occu oge he in pa s o Kenya and Tanzania (Figu e 5) (Copeland e al. 2006, Mwa awala e al. 2006). Ce a i is anonae and C. osa we e no ound oge he . Indica ions o inconsis encies Despi e he mo phological di e ences in adul males and he pa ially disjunc dis i- bu ion, he e a e indica ions ha c yp ic specia ion occu s wi hin he h ee cu en ly ecognized mo phospecies. An in eg a i e app oach o un a el he Ce a i is FAR (Dip e a, Teph i idae)... 411 Figu e 5. Geog aphical dis ibu ion o Ce a i is osa (blue iangles) and C. asci en is (yellow ci cles). Figu e 6. Geog aphical dis ibu ion o Ce a i is anonae. Ma c De Meye e al. / ZooKeys 540: 405–427 (2015) 412 When compa ing sequences o mi ochond ial and nuclea ma ke s, Vi gilio e al. (2008) could no eco e he h ee mo phospecies as monophyle ic g oups, al hough di e en ma ke s eco e ed well-suppo ed clades wi hin he ep esen a i es o C. as- ci en is om Wes and Eas A ica. So, al hough he molecula da a did no con adic o suppo he mo phological sepa a ion, i did sugges ha C. asci en is is i sel a complex o c yp ic species. This s udy, as well as Douglas and Hayme (2001) and Ba e al. (2006) indica ed mo eo e he exis ence o o he sepa a e clus e s wi hin he complex (al hough no always wi h high suppo ). Finally, co ela i e ecological niche modeling showed ha C. osa p e e s clima ic condi ions wi h lowe empe a u es when compa ed o C. capi a a (De Meye e al. 2008). This was co obo a ed by indings on he island o La Réunion whe e C. osa occupies a colde and mo e humid clima e niche han C. capi a a (Duyck e al. 2006). Howe e , hese esul s we e con adic ed by G ou and S ol z (2007) who s udied he de elopmen al h esholds o a numbe o Ce a i is species, based on Sou h A ican popula ions and concluded ha he si ua ion in Sou h A ica is di e en om ha in La Réunion. This led o he sugges ion ha C. osa could comp ise en i ies wi h di e - en biological equi emen s. Objec i e The inclusion o he Ce a i is FAR complex in he Coo dina ed Resea ch P ojec (CRP) on c yp ic species, he e o e, was o es ablish whe he he h ee mo phological en i ies ac ually ep esen h ee dis inc species, o i he ela ionship can be esol ed in a di e en manne . The au ho s op ed o an in eg a i e app oach, using di e - en me hodologies, including popula ion gene ics, la al mo phology, wing mo pho- me ics, cu icula hyd oca bons, de elopmen al physiology and p e- and pos zygo ic ma ing compa ibili y. The esul s o hese di e en app oaches a e p esen ed in his olume o ha e been published elsewhe e (Vi gilio e al. 2013, Vaníčko á e al. 2014). We he ewi h summa ize he main esul s ob ained by each o hese app oaches and p o ide a syn hesis on he cu en knowledge and s a us o he di e en en i ies wi hin he Ce a i is FAR complex. Resul s Mic osa elli e analysis Using mic osa elli es, Bali aine e al. (2004) e ealed signi ican di e en ia ion wi h espec o geno ypic equencies be ween C. osa specimens om he A ican main- land and he Indian Ocean islands, and be ween C. asci en is om Kenya and om Uganda. Dela e e al. (2013) isola ed and cha ac e ized 16 mic osa elli e ma ke s us- An in eg a i e app oach o un a el he Ce a i is FAR (Dip e a, Teph i idae)... 419 Many ola iles iden i ied in Ce a i is FAR male emana ion ha e been p e iously iden i ied in o he eph i id phe omones (C uz-López e al. 2006, Hea h e al. 1991, Lu and Teal 2001, Mile -Pinhei o e al. 2014, Robacke 1988, Rocca e al. 1993, Vaníčko á 2012, Vaníčko á e al. 2012). O he s, speci ically isop enoid ge anyl ac- e one, alipha ic ola ile (E)-non-2-enal, and me hyl (2E,6E)- a nesoa e, ha e no been epo ed be o e. In e es ingly, he la e is a c us acean ep oduc i e ho mone, s uc- u ally simila o insec ju enile ho mone, which is esponsible o enhancing ep o- duc i e ma u a ion, main aining ju enile mo phology, and in luencing male sex de e - mina ion (Olms ead and LeBlanc 2007, Naga aju and Bo s 2008). In insec s, me hyl (2E,6E)- a nesoa e ep esen s he immedia e p ecu so o insec ju enile ho mone III (Teal e al. 2014). As a semiochemical, me hyl (2E,6E)- a nesoa e was epo ed in pen a omid bug phe omones (Milla e al. 2002). The phe omone composi ion as well as elec oan ennog aphy may be used o spe- cies iden i ica ion. Simila ly as o he composi ion o CHCs (Vaníčko á e al. 2014), he phe omone composi ion and an ennal speci ici y sugges ha he h ee nominal species o he Ce a i is FAR complex co espond o axonomically well-de ined en i ies. I is ecom- mended, howe e , ha his wo k should be expanded by including phe omone s udies o F1 and R1 and u he sampling and analysis o di e en popula ions o all ypes, o de e mine in e -popula ion a iabili y. De elopmen al physiology De elopmen al physiology s udies can assis in de ec ing di e ences be ween species wi h ega d o biological equi emen s. Only he de elopmen al physiology o he wo C. osa ypes was s udied in de ail (Tanga e al. 2015), because o he lack o es ablished colonies o he o he species and because i was he di e ences obse ed be ween de- elopmen al s udies conduc ed in La Réunion (Duyck and Quilici 2002) and Sou h A ica (G ou and S olz 2007) ha ini ia ed he idea ha C. osa may consis o di e en bio ypes wi h di e en clima ic equi emen s. R1 and R2 popula ions we e, he e o e, s udied simul aneously in Kenya and Sou h A ica (Tanga e al. 2015). Depending on locali y and empe a u e, ma ked di e ences we e obse ed be ween R1 and R2 popula- ions in he de elopmen al du a ion o imma u e li e s ages. In bo h Kenyan and Sou h A ican popula ions, R2 appea s o be less adap ed o ho e en i onmen s han R1. In Kenya, R2 appea ed o be be e adap ed o colde en i onmen s, while in Sou h A - ica, bo h C. osa ypes we e able o ole a e lowe empe a u es. Despi e disc epancies in empe a u e ela ed de elopmen al physiology o he wo ypes in he wo locali ies, esul s om Kenya and Sou h A ica clea ly demons a e and suppo he exis ence o wo gene ically dis inc popula ions o C. osa ha a e di e gen in hei physiological esponse o empe a u e. Disc epancies be ween he locali ies wi h ega ds o obse ed and es ima ed de elopmen al ime pa ame e s o he wo C. osa ypes can be due o ge- og aphic a ia ion (popula ions om Sou h A ica o igina ed om empe a e clima es Ma c De Meye e al. / ZooKeys 540: 405–427 (2015) 420 and popula ions om Kenya o igina ed om opical clima es). O he ac o s such as ood quan i y and quali y, ea ing condi ions, acclima iza ion and gene a ion age a e also men ioned as ac o s in luencing he de elopmen al physiology. Geog aphical dis ibu ion and al i udinal ansec The dis ibu ion o C. asci en is and C. osa was e-analyzed, aking in o accoun he exis ence o wo ypes o each o hese species. As he known dis ibu ion was la gely based upon museum specimens collec ed o e a pe iod o 130 yea s and he DNA e ie al om olde specimens (i.e. >10y s) is cumbe some and wi h low success a e, i was decided o e-assign specimens based only on mo phological cha ac e s which excluded emale specimens. In o al, specimens om 218 locali ies we e e-examined and assigned o one o he ou ypes (F1, F2, R1, R2). The obse ed dis ibu ions a e gi en in Figu es 10–11. F1 is mainly ep esen ed in wes e n A ica bu ex ends i s dis ibu ion h oughou sou he n and eas e n pa s o he con inen wi h eco ds om Angola, Zambia, Malawi and Tanzania (Figu e 10). F2 is con ined o he Ri a eas o E hiopia, Kenya and eas e n pa o he Democ a ic Republic o Congo, wi h sou he n expansion in o Ka anga egion o he Democ a ic Republic o Congo and Zambia. So a , no sympa ic o pa apa ic occu ence o he wo ypes has been obse ed. R1 and R2 on he o he hand do no show a clea geog aphic isola ion (Figu e 11). Only in he Cape and cen al pa s o Sou h A ica is a single ype (R2) p esen , as well as in he ad en i e popula ions on he Indian Ocean islands. In he no he n pa o Sou h A ica and no hwa ds he e a e eco ds showing o e lap o dis ibu ions. A de ailed s udy was conduc ed in he Ulugu u Moun ains nea Mo ogo o (Tan- zania) whe e bo h ypes a e known o occu (Mwa awala e al. 2015). Along an al- i udinal ansec , anging om 540 o 1650 masl he occu ence o bo h ypes was moni o ed using aps wi h EGO-lu e. A g adual shi was obse ed wi h bo h ypes occu ing a lowe al i udes (wi h p edominance o R1) while only R2 was obse ed a he highes ele a ions. Geu s e al. (2012) obse ed a empe a u e shi o 7–8 °C along his ansec . When looking a he de elopmen al di e ences obse ed o , a leas he Kenyan popula ions, empe a u e could well play a majo ole in he obse ed pa e n, al hough i emains o be seen whe he o he aspec s such as di e en ial hos ange and a ailabili y also ha e an in luence ega ding he di e ences obse ed. P e- and pos zygo ic incompa ibili y Al hough no de ailed da a a e p esen ed in his olume, p elimina y da a by S. Ekesi (pe s. communica ion), indica e ha in ield cage s udies he e is a signi ican p e- and pos -zygo ic incompa ibili y bo h be ween Ce a i is osa and C. asci en is (F2), as well as be ween he wo geno ypic clus e s o C. osa: R1 and R2. None o he o he geno ypic clus e s o mo phospecies we e included. An in eg a i e app oach o un a el he Ce a i is FAR (Dip e a, Teph i idae)... 421 Figu e 10. Geog aphical dis ibu ion o Ce a i is asci en is, F1 (b own ci cles), F2 (yellow ci cles), un- assigned (whi e ci cles). Conclusion The majo i y o he esea ch app oaches discussed he e indica e ha he Ce a i is FAR complex consis s o a leas he h ee ecognized mo phospecies, bu possibly o i e di e en species. All me hodologies (excep la al mo phology) used con i m ha Ce - a i is anonae, C. asci en is and C. osa a e well ecognized g oups. Ce a i is anonae is a uni o m g oup showing no appa en mo phological o gene ic a iabili y and has a well-de ined dis ibu ion ange. The wo o he en i ies consis each o wo sepa a e g oups. Fo C. osa, he wo en i ies (called ‘R1’, ‘lowland’ o ‘ho osa’ on one hand, and ‘R2’, ‘highland’ o ‘cold osa’ on he o he hand) can be dis inguished mo pho- logically in males, as well as by o he means and demons a e a di e en de elopmen- al physiology. They occu sympa ically in some egions, bu also show a disjunc dis ibu ion ha appea s o be co ela ed wi h ambien empe a u e. I is concluded ha bo h ypes should be conside ed as wo di e en species. Taxonomically, he ype Ma c De Meye e al. / ZooKeys 540: 405–427 (2015) 422 ma e ial o C. osa belongs o he R1 ype, which means ha he R2 ype should be conside ed as a new species, and a o mal desc ip ion will be published in he nea u u e. Simila ly, he e iew indica es ha also C. asci en is ends o be composed o wo en i ies. Ye , because he da a a e cu en ly insu icien o es ablish clea ly whe he hey should also be conside ed wo di e en species o no , we cu en ly sugges o main ain he wo ypes unde one and he same species. Acknowledgemen s We would like o hank he Join FAO/IAEA P og amme o ini ia ing he Coo di- na ed Resea ch P ojec on “Resolu ion o c yp ic species complexes o eph i id pes s o o e come cons ain s o SIT applica ion and in e na ional ade”, and o p o iding suppo o he au ho s o conduc ing pa o he esea ch and o a end he esea ch Figu e 11. Geog aphical dis ibu ion o Ce a i is osa, R1 ( ed ci cles), R2 (blue ci cles), sympa ic occu ence (b own ci cles), un-assigned (whi e ci cles). An in eg a i e app oach o un a el he Ce a i is FAR (Dip e a, Teph i idae)... 423 coo dina ion mee ings. Some o he da a p esen ed he e a e also he esul o inancial suppo by he Belgian De elopmen Coope a ion ( h ough he F amewo k Ag ee- men wi h he Royal Museum o Cen al A ica, RMCA) o Sokoine Uni e si y o Ag icul u e, and by he Belgian Science Policy o he Join Expe imen al Molecula Uni (JEMU) o he RMCA and he Royal Belgian Ins i u e o Na u al Sciences. The de elopmen al physiology wo k on he wo C. osa ypes in Sou h A ica was unded by Ci us Resea ch In e na ional. Re e ences Bali aine FN, Bonizzoni M, Guglielmino CR, Osi EO, Lux SA, Mulaa FJ, Gomulski LM, Zheng L, Quilici S, Gaspe i G, Malac ida AR (2004) Popula ion gene ics o he po en ially in asi e A ican ui ly species, Ce a i is osa and Ce a i is asci en is (Dip e a: Teph i idae). Molecula Ecology 13: 683–695. doi: 10.1046/j.1365-294X.2004.02105.x Ba NB, McPhe on BA (2006) Molecula phylogene ics o he genus Ce a i is (Dip e a: Teph i idae). Molecula Phylogene ics and E olu ion 38: 216–230. doi: 10.1016/j. ympe .2005.10.013 Ba NB, Copeland RS, De Meye M, Masiga D, Kibogo HG, Billah MK, Osi E, Wha on RA, McPhe on BA (2006) Molecula diagnos ics o economically impo an ui ly spe- cies (Dip e a: Teph i idae) in A ica using PCR and RFLP analyses. Bulle in o En omo- logical Resea ch 96: 505–521. Ba NB, Wiegmann BM (2009) Phylogene ic ela ionships o Ce a i is ui lies in e ed om nuclea CAD and ango/ARNT gene agmen s: es ing monophyly o he subgene a Ce a- i is (Ce a i is) and C. (P e and us). Molecula Phylogene ics and E olu ion 53: 412–424. doi: 10.1016/j.ympe .2009.07.008 Bezzi M (1920) No es on he E hiopian ui lies o he amily T ypaneidae, o he han Dacus. – III. Bulle in o En omological Resea ch 10: 211–272. doi: 10.1017/S000748530004414X Blomquis GJ, Bagnè es AG (2010) Insec hyd oca bons: biology, biochemis y, and chemi- cal ecology. Camb idge Uni e si y P ess, New Yo k. doi: 10.1017/CBO9780511711909 Blomquis GJ, Jackson LL (1979) Chemis y and biochemis y o insec waxes. P og ess in Lipid Resea ch 17: 319–345. doi: 10.1016/0079-6832(79)90011-9 Břízo á R, Vaníčko á L, Faťa o á M, Ekesi S, Hosko ec M, Kalino á B (2015) Analyses o ola iles p oduced by he A ican ui ly species complex (Dip e a, Teph i idae). In: De Meye M, Cla ke AR, Ve a MT, Hend ichs J (Eds) Resolu ion o C yp ic Species Com- plexes o Teph i id Pes s o Enhance SIT Applica ion and Facili a e In e na ional T ade. ZooKeys 540: 385–404. doi: 10.3897/zookeys.540.9630 Ca oll LE (1998) Desc ip ion o he hi d ins a la a o Ce a i is osa Ka sch (Dip e a: Teph- i idae). P oceedings o he En omological Socie y o Washing on 100(1): 88–94. Copeland RS, Wha on RA (2006) Yea - ound p oduc ion o pes Ce a i is species (Dip e a: Te- ph i idae) in ui o he in asi e species Solanum mau i ianum in Kenya. Annals o he En- omological Socie y o Ame ica 99: 530–535. doi: 10.1603/0013-8746(2006)99[530:YP OPCS]2.0.CO;2 Ma c De Meye e al. / ZooKeys 540: 405–427 (2015) 424 Copeland RS, Wha on RA, Luke Q, De Meye M, Lux S, Zenz N, Mache a P, Okumu M (2006) Geog aphic dis ibu ion, hos ui s, and pa asi oids o A ican ui ly pes s Ce a i is anonae, Ce a i is cosy a, Ce a i is asci en is, and Ce a i is osa (Dip e a: Teph- i idae) in Kenya. Annals o he En omological Socie y o Ame ica 99: 261–278. doi: 10.1603/0013-8746(2006)099[0261:GDHFAP]2.0.CO;2 C uz-López L, Malo EA, Toledo J, Vi gen A, Del Mazo A, Rojas JC (2006) A new po en ial a ac an o Anas epha obliqua om Spondias mombin ui s. Jou nal o Chemical Ecol- ogy 32: 351–365. doi: 10.1007/s10886-005-9006-7 De Meye M (2001) On he iden i y o he Na al ui ly Ce a i is osa Ka sch (Dip e a, Teph i idae). Bulle in de l’Ins i u Royal des Sciences Na u elles de Belgique En omologie 71: 55–62. De Meye M (2005) Phylogeny o he ui ly genus Ce a i is MacLeay (Dip e a: Teph i idae). Insec Sys ema ics and E olu ion 36: 459–480. doi: 10.1163/187631205794761012 De Meye M, Copeland RS, Lux S, Mansell M, Wha on R, Whi e IM, Zenz N (2002) Anno- a ed check lis o hos plan s o A o opical ui lies (Dip e a: Teph i idae) o he genus Ce a i is. Zoologische Documen a ie Koninklijk Museum oo Midden A ika 27: 1–92. De Meye M, F eidbe g A (2006) Re ision o he Subgenus Ce a i is (P e and us) Bezzi (Dip- e a: Teph i idae). Is ael Jou nal o En omology 36: 197–315. De Meye M, Robe son M, Pe e son T, Mansell M (2008) Clima ic modeling o he med ly and Na al ui ly. Jou nal o Biogeog aphy 35: 270–281. Dela e H, Vi gilio M, Simiand C, Quilici S, De Meye M (2013) Isola ion and cha ac e isa ion o six een mic osa elli e ma ke s c oss-ampli ying in a complex o h ee A ican ag icul u al pes s (Ce a i is osa, C. anonae and C. asci en is, Dip e a: Teph i idae). Conse a ion Gene ic Resou ces 5: 31–34. doi: 10.1007/s12686-012-9722-6 Douglas L, Hayme DS (2001) Ribosomal ITS1 polymo phism in Ce a i is capi a a and Ce a i- is osa (Dip e a: Teph i idae). Annals o he En omological Socie y o Ame ica 9: 726–731. doi: 10.1603/0013-8746(2001)094[0726:RIPICC]2.0.CO;2 Duyck PF, Quilici S (2002) Su i al and de elopmen o di e en li e s ages o h ee Ce a i is spp. (Dip e a: Teph i idae) ea ed a i e cons an empe a u es. Bulle in o En omological Resea ch 92: 461–469. doi: 10.1079/BER2002188 Duyck PF, Da id P, Quilici S (2006) Clima ic niche pa i ioning ollowing successi e in asions by ui lies in La Réunion. Jou nal o Animal Ecology 75: 518–526. doi: 10.1111/j.1365- 2656.2006.01072.x E bou N, Vi gilio M, Lens L, Ba N, De Meye M (2011) Disc epancies be ween subgene ic classi ica ion and molecula phylogeny o Ce a i is (Dip e a: Teph i idae), can he e olu- ion o hos use p o ide some clues? Molecula Phylogeny and E olu ion 60: 259–264. doi: 10.1016/j.ympe .2011.04.004 Geu s K, Mwa awala M, De Meye M (2012) Indigenous and in asi e ui ly (Dip e a: Teph i idae) di e si y along an al i udinal ansec in Cen al Tanzania. Jou nal o Insec Science 12: 1–18. doi: 10.1673/031.012.1201 G aham WM (1908) Some new and undesc ibed insec pes s a ec ing cocoa in Wes A ica. Jou nal o Economic Biology 3: 113–117. An in eg a i e app oach o un a el he Ce a i is FAR (Dip e a, Teph i idae)... 425 G ou TG, S ol z KC (2007) De elopmen al a es a cons an empe a u e o h ee economically impo an Ce a i is spp. (Dip e a: Teph i idae) om Sou he n A ica. En i onmen al En- omology 36: 1310–1317. doi: 10.1603/0046-225X(2007)36[1310:DRACTO]2.0.CO;2 Hea h RR, Landol PJ, Tumlinson JH, Chambe s DL, Mu phy RE, Dooli le RE, Dueben BD, Si inski J, Calkins CO (1991) Analysis, syn hesis, o mula ion, and ield es ing o 3 majo componen s o male Medi e anean ui ly phe omone. Jou nal o Chemical Ecology 17: 1925−1940. doi: 10.1007/BF00993739 Jennings JH, E ges WJ, Schmi T, Hoikkala A (2014) Cu icula hyd oca bons o D osophila mon ana: Geog aphic a ia ion, sexual dimo phism and po en ial oles as phe omones. Jou nal o Insec Physiology 61: 16–24. doi: 10.1016/j.jinsphys.2013.12.004 Ka sch F (1887) Dip e ologisches on de Delagoabai. En omologische Nach ich en 13: 22–26. Ka he R, Ma in SJ (2012) Cu icula hyd oca bon p o iles as a axonomic ool: ad an ages, lim- i a ions and echnical aspec s. Physiological En omology 37: 25–32. doi: 10.1111/j.1365- 3032.2011.00826.x Liquido NJ, McQua e GT, Sui e KA (2014) MEDHOST: An encyclopedic bibliog aphy o he hos plan s o he Medi e anean F ui Fly, Ce a i is capi a a (Wiedemann), Ve sion 2.0. Uni ed S a es Depa men o Ag icul u e, Cen e o Plan Heal h Science and Tech- nology, Raleigh. h ps://medhos 2.cphs .o g/ Lu F, Teal PEA (2001) Sex phe omone componen s in o al sec e ions and c op o male Ca ib- bean ui lies Anas epha suspensa (Loew). A chi es Insec Biochemis y and Physiology 48: 144−154. doi: 10.1002/a ch.1067 Mile -Pinhei o P, Na a o DMA, De Aquino NC, Fe ei a LL, Ta a es RF, Da Sil a RCC, Li- ma-Mendonça A, Vaníčko á L, Mendonça AL, Do Nascimen o RR (2014) Iden i ica ion o male-bo ne a ac an s in Anas epha a e culus (Dip e a: Teph i idae). Chemoecology 25(3): 115–122. doi: 10.1007/s00049-014-0180-3 Milla JG, McB ien HL, Ho H-Y, Rice RE, Cullen E, Zalom FG, Üokl A (2002) Pen a omid bug phe omones in IPM: possible applica ions and limi a ions. IOBC Bulle in 25: 1–11 Mwa awala MW, De Meye M, Makundi RH, Mae e e AP (2006) Biodi e si y o ui lies (Dip e a, Teph i idae) a o cha ds in di e en ag o-ecological zones o he Mo ogo o e- gion, Tanzania. F ui s 61: 321–332. doi: 10.1051/ ui s:2006031 Mwa awala M, Vi gilio M, Joseph J, De Meye M (2015) Niche pa i ioning among wo Ce a i- is osa mo pho ypes and o he Ce a i is pes species (Dip e a, Teph i idae) along an al i udi- nal ansec in Cen al Tanzania. In: De Meye M, Cla ke AR, Ve a MT, Hend ichs J (Eds) Resolu ion o C yp ic Species Complexes o Teph i id Pes s o Enhance SIT Applica ion and Facili a e In e na ional T ade. ZooKeys 540: 429–442. doi: 10.3897/zookeys.540.6016 Naga aju GPC, Bo s DW (2008) Me hyl a nesoa e couples en i onmen al changes o es icu- la de elopmen in he c ab Ca cinus maenas. Jou nal o Expe imen al Biology 211(17): 2773–2778. doi: 10.1242/jeb.019133 N’Dépo OR, Hala N, Gnago A, Allou K, Kouassi KP, Vayssiè es J-F, De Meye M (2010) In en ai e des mouches des ui s de ois egions ag o-écologiques e des plan es-hô es associées à l’espèce nou elle, Bac oce a (Bac oce a) in adens D ew e al. (Dip e a: Teph i- idae) en Cô e-d’I oi e. Eu opean Jou nal o Scien i ic Resea ch 46: 62–72. Ma c De Meye e al. / ZooKeys 540: 405–427 (2015) 426 N’Dépo OR, Hala N, N’Da Adopo A, Coulibaly F, Kouassi KP, Vayssiè es J-F, De Meye M (2013) F ui lies (Dip e a: Teph i idae) popula ions dynamic in mangoes p oduc ion zone o Cô e-d’I oi e. Ag icul u al Science Resea ch Jou nal 3: 352–363. Olms ead AW, LeBlanc GA (2007) The en i onmen al-endoc ine basis o gynand omo phism (in e sex) in a c us acean. In e na ional Jou nal o Biological Sciences 3(2): 77–84. doi: 10.7150/ijbs.3.77 P i cha d JK, S ephens M, Donnelly P (2000) In e ence o popula ion s uc u e using mul ilo- cus geno ype da a. Gene ics 155: 945–959. Quilici S, F anck A, Peppuy A, Dos Reis Co eia E, Mouniama C, Bla d F (2002) Compa a i e s udies o cou ship beha io o Ce a i is spp. (Dip e a: Teph i idae) in Reunion island. Flo ida En omologis 85: 138–142. doi: 10.1653/0015-4040(2002)085[0138:CSOCBO ]2.0.CO;2 Robacke DC (1988) Beha io al esponses o emale Mexican ui lies Anas epha ludens o componen s o male-p oduced sex phe omone. Jou nal o Chemical Ecology 14: 1715– 1726. doi: 10.1007/BF01014639 Rocca JR, Na ion JL, S ekowski L, Ba is e MA (1993) Compa ison o ola iles emi ed by male Ca ibbean and Mexican ui lies. Jou nal o Chemical Ecology 18: 223−244. doi: 10.1007/BF00993755 S eck G, Ekesi S (2015) Desc ip ion o hi d ins a la ae o Ce a i is asci en is, C. anonae, C. osa (FAR complex) and C. capi a a (Dip e a, Teph i idae). In: De Meye M, Cla ke AR, Ve a MT, Hend ichs J (Eds) Resolu ion o C yp ic Species Complexes o Teph i id Pes s o Enhance SIT Applica ion and Facili a e In e na ional T ade. ZooKeys 540: 443–466. doi: 10.3897/zookeys.540.10061 Tanga CM, Man akhan A, Daneel JH, Mohamed SA, Khamis FM, Ekesi S (2015) Compa a- i e analysis o de elopmen and su i al o wo Na al ui ly Ce a i is osa Ka sch (Dip- e a, Teph i idae) popula ions om Kenya and Sou h A ica. In: De Meye M, Cla ke AR, Ve a MT, Hend ichs J (Eds) Resolu ion o C yp ic Species Complexes o Teph i id Pes s o Enhance SIT Applica ion and Facili a e In e na ional T ade. ZooKeys 540: 467–487. doi: 10.3897/zookeys.540.9906 Teal PEA, Jones DJG, To o B, Nyasembe V, Bo gemeis e C, Albo n HT, Kaplan F, Boucias D, Lie ze VU (2014) Iden i ica ion o me hyl a nesoa e om he hemolymph o insec s. Jou nal o Na u al P oduc s 77(2): 402–405. doi: 10.1021/np400807 Van Cann J, Vi gilio M, Jo daens K, De Meye M (2015) Wing mo phome ics as a possible ool o he diagnosis o he Ce a i is asci en is, C. anonae, C. osa complex (Dip e a, Teph i idae). In: De Meye M, Cla ke AR, Ve a MT, Hend ichs J (Eds) Resolu ion o C yp ic Species Complexes o Teph i id Pes s o Enhance SIT Applica ion and Facili a e In e na ional T ade. ZooKeys 540: 489–506. doi: 10.3897/zookeys.540.9724 Vaníčko á L (2012) Chemical ecology o ui lies: gene a Ce a i is and Anas epha. PhD hesis, Ins i u e o Chemical Technology and Ins i u e o O ganic Chemis y and Biochemis y, P ague. Vaníčko á L, do Nascimen o RR, Hosko ec M, Ježko á Z, Břízo á R, Tomčala A, Kalino á B (2012) A e he wild and labo a o y insec popula ions di e en in semiochemical emis- An in eg a i e app oach o un a el he Ce a i is FAR (Dip e a, Teph i idae)... 427 sion? The case o med ly sex phe omone. Jou nal o Ag icul u al and Food Chemis y 60: 7168−7176. doi: 10.1021/j 301474d Vaníčko á L, Vi gilio M, Tomčala A, Břízo á R, Ekesi S, Hosko ec M, Kalino á B, Do Nas- cimen o RR, De Meye M (2014) Resolu ion o h ee c yp ic ag icul u al pes s (Ce a i is asci en is, C. anonae, C. osa, Dip e a: Teph i idae) using cu icula hyd oca bon p o il- ing. Bulle in o En omological Resea ch 104: 631–638. Vaníčko á L, Břízo á B, Mondonça AL, Pompeiano A, Do Nascimen o RR (2015a) In aspe- ci ic a ia ion o cu icula hyd oca bon p o iles in he Anas epha a e culus (Dip e a: Te- ph i idae) species complex. Jou nal o Applied En omology. doi: 10.1111/jen.12204 Vaníčko á L, Břízo á R, Pomeiano A, Ekesi S, De Meye M (2015b) Cu icula hyd oca bons co obo a e he dis inc ion be ween lowland and highland Na al ui ly (Teph i idae, Ce a i is osa) popula ions. In: De Meye M, Cla ke AR, Ve a MT, Hend ichs J (Eds) Reso- lu ion o C yp ic Species Complexes o Teph i id Pes s o Enhance SIT Applica ion and Facili a e In e na ional T ade. ZooKeys 540: 507–524. doi: 10.3897/zookeys.540.9619 Vayssiè es J-F, Sanogo F, Noussou ou M (2004) In en ai e des espèces de mouches des ui s (Dip e a:Teph i idae) in éodées au manguie au Mali e essais de lu e aisonnée. F ui s 59: 1–14. Vi gilio M, Backeljau T, Ba N, De Meye M (2008) Molecula e alua ion o nominal species in he Ce a i is asci en is, C. anonae, C. osa complex (Dip e a: Teph i idae). Molecula Phylogene ics and E olu ion 48: 270–280. doi: 10.1016/j.ympe .2008.04.018 Vi gilio M, Dela e H, Quilici S, Backeljau T, De Meye M (2013) C yp ic di e si y and gene low among h ee A ican ag icul u al pes s: Ce a i is osa, Ce a i is asci en is and Ce a i is anonae (Dip e a, Teph i idae). Molecula Ecology 22: 2526–2539. doi: 10.1111/ mec.12278 Vi gilio M, Jo daens K, Ve wimp C, De Meye M (2015) Highe phylogeny o ugi o ous ui lies (Dip e a, Teph i idae, Dacini) as in e ed om mi ochond ial and nuclea gene agmen s. Molecula Phylogene ics and E olu ion doi: 10.1016/j.ympe .2015.01.007 Whi e IM, Elson-Ha is MM (1992) F ui lies o economic signi icance: hei iden i ica ion and bionomics. CAB In e na ional, Walling o d, 599 pp. Whi e IM, De Meye M, S onehouse J (2000) A e iew o he na i e and in oduced ui lies (Dip e a, Teph i idae) in he Indian Ocean Islands o Mau i ius, Réunion, Rod igues and Seychelles. In: P ice NS, Seewoo u hun I (Eds) P oceedings o he Indian Ocean Commis- sion Regional F ui Fly Symposium, Mau i ius, 5–9 h June 2000. Indian Ocean Commis- sion, Mau i ius, 15–21.