Th e phe n ot ypic e x pre ssion of Pu r ple Body ( Pb) in dom e st ic gu ppy st r a in s of Poe cilia r e t icu la t a 1 Alan S. Bias, 2 Richard D. Squire 1 I ndependent Researcher and Sw ordt ail Guppy Breeder. Mailing address: P.O. Box 1508, Lew isburg, West Virginia 24901, USA. orcid.org/ 0000- 0002- 9093- 619X; 2 Biology Depart m ent ( retired) , University of Puert o Rico, Mayaguez cam pus, Mayaguez, Puert o Rico, USA. Mailing address: P. O. Box 3227, Mayaguez, P.R., USA 00681- 3227. orcid.org/ 0000- 0002- 3916- 0672. rickdsquire@gm ail.com . Corresponding aut hor: A. S. Bias, alansbias@aol.com Abst ract . Modification of wild- type carotenoid orange and pteridine red coloration and spotting of m ale ornam ents in m odern dom estic guppy strains ( Poecilia reticulata) by the naturally occurring Purple Body gene ( Pb) has been long incorporated into their strains by pedigree stock breeders. It is inherited as an autosom al incom pletely dominant trait. Its existence has allowed breeders to produce a vast array of Purple based phenotypes. Photographic evidence dem onstrates that Purple Body is a norm al polym orphism in dom estic guppies m odifying color pigm ented regions. When com bined with currently used m utant genes such as Albino, Blond, Golden, Asian Blau, Coral Red, Magenta, Grass, Moscow, Pink, Platinum, Red Mosaic, Multicolor, and Full Red, startling new phenotypes are created. The recently described Purple Body gene has long been overlooked in research articles and little understood in breeder publications. Ke y W or ds: guppy color and m odificat ion, dom est ic guppy st r ains, chr om at ophor e, violet ir idophor e, blue ir idophor e, violet - blue ir idophor e, xant hophor e, xant ho- er yt hr ophor e, Pur ple Guppy, Pur ple Body gene, Met al Gold ir idophor e, Pur ple Body gene, Vienna Em er ald Gr een ( VEG) , Albino, Blond, Golden, Asian Blau, Coral Red, Magenta, Grass, Moscow, Pink, Platinum, Red Mosaic, Multicolor, Full Red, Poecilia r et iculat a. I n t r odu ct ion. The Purple Body gene ( Figure 1) is locat ed on an aut osom e. Breeding t est s, involving t his m odification of orange spot t ing, reveal t his t rait t o have an incom plet ely dom inant m ode of inherit ance. As such a form al nam e and nom enclat ure of Purple Body ( Pb) has been suggest ed ( Bias & Squire 2017a) . [ Not e: Hereaft er Purple Gene, or Purple Body Gene used int erchangeably in reference.] The intent of t his brief paper is t o provide spect ral distinction, for ease in identification, bet w een Pb and non- Pb in m any of t he m ore com m only produced m odern dom est ic guppy st rains. The num ber of st rains and phenot ypes being produced for color and pat t ern cont inues t o increase, and is a t est am ent t o t he effort s of bot h pr ofessional and am at eur breeders around t he w orld. Em phasis is on t he prim ary color t rait s, and several com m on pat t ern t rait s. Groupings are present ed by Grey ( w ild- t ype) , aut osom al and sex- linked m odifiers. These should not be considered inclusive of all know n phenot ypes. Those included in t his paper are lim it ed t o Bias & Squire ( 2017a) Pb st udy st rains or t hose Pb st rains w hich have been w orked wit h in t he past by t he senior aut hor ( unpublished results) . Aut osom al incom plet ely dom inant Pb, sim ilar t o ident ified aut osom al recessives found in Poecilia ret iculat a, is a m odifier of t ot al existing body color and pat t ern pigm ent at ion ( involving xant ho- eryt hrophores, st ruct ural iridophores and m elanophores) in bot h m ales and fem ales. Purple Body is capable of m odifying ext ent color and pat t ern found in any dom est ic guppy st rain. When t he basic chrom at ophore int eractions of Pb are underst ood, visual identification of t he presence or absence of Pb in phenot ypes can be achieved t hrough skilled vision ( Grasseni 2004a, b, 2005, 2007, 2011) . Defined breeding t est s and resulting phenot ypic ratios are not alw ays necessary. Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 125 Figure 1. Purple Delt a ( Pb/ Pb) , phot o court esy of Terry Aley. Pb m odification is m ost noticeable in dom estic st rains as a m odifier of ornam ent s com prised of orange carot enoid color pigm ent , in bot h m ales and fem ales ( Figure 2) . Figur e 2. ( A) Hom ozygous Pb ( Pb/ Pb) m odif ied ornam ent s, expressing r em oval of xant hophores ( red circles) and incr eased v iolet - blue ir idophores ( r ed arrows) . ( B) Hom ozygous Pb ( Pb/ Pb) m odif ied ornam ent s, expr essing r educed xant hophores and increased v iolet - blue ir idophores. ( C- D) Non- Pb ornam ent s ( pb/ pb) ex pr essing no alt erat ion of xant ho- eryt hrophor es ( red arrows) . Visually, colorat ion is m odified from a highly reflective orange t o a “pinkish- purple” coloration in Grey ( “ w ild t ype” alleles A, B, G, R, and ab) corresponding t o aut osom al genes Albino ( a, Haskins & Haskins 1948) , Blond ( b, Goodrich et al 1944) , Golden ( g, Goodrich et al 1944) , European Blau ( r, Dzw illo 1959) , Asian Blau ( Ab, Undescribed - see Bias 2015) . I t is also m odified in various w ays w hen com bined wit h aut osom al genes Pink ( p, Luckm an 1990; Först er 1992, pi, Kem pkes 2007) , I vory ( I , Tsut sui 2006) , Magent a ( M, undescribed) , and Zebrinus ( Ze, Winge 1927) . I t com bines as w ell wit h sex-linked genes such as Coral Red ( Co, undescribed) Y-linked, Grass ( Gra, undescribed) X- and/ or Y- linked, Moscow (Mw, Y-linked, Kem pkes 2007] ) , Nigrocaudaut us, X and/ or Y-linked ( NiI , Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 126 Nybelin 1947] and NiI I , Dzwillo 1959) . Platinum (P, undescribed) X- and/ or Y- linked, Mosaic ( Mo, Khoo et al 1999) X- and/ or Y- linked, Multicolor ( no gene sym bol) X- linked, and Schim m elpennig Platinum ( Sc, described as Buxeus by Kem pkes 2007) , Y- linked. Exam ples provided in t his paper are prim arily lim it ed t o Delt a Tail and Sw ordt ail phenot ypes. I n het erozygous condit ion ( Pb/ pb) a distinct result is generat ed w hile in hom ozygous condition ( Pb/ Pb) t hese result s are furt her am plified. Pb is capable of pleiot ropic effect on all existing color and pat t ern elem ent s at m ultiple loci. The purple phenot ype has been present in hobbyist st ocks for decades, but has been largely unrecognized by m any breeders, except in t he case of pure- bred all- purple st rains. Pb m odification, zygosity dependent, rem oves cert ain classes of yellow- orange- red color pigm ent over silver iridophores or w hit e leucophores. Pb m odifies “ ot her existing” color in bot h body and fins, t hus suggestive of being a “ full body” m odifier, in hom ozygous condition. Dark red pt eridine color pigm ent does not seem t o be m odified by Purple Body in fins lacking an underlying silver iridophore or w hit e leucophore pat t ern. Modification by Pb seem s lim it ed predom inantly t o wild- t ype orange color pigm ent ; i.e. t hat w hich also cont ains yellow carot enoids in addition t o red pt eridines, over an iridophore pat t ern. Pb is alw ays found in all- purple fish, but is not by it self sufficient t o produce t he allpurple phenot ype in het erozygous expression. Hom ozygous Pb expression results in t he furt her rem oval of xant ho- eryt hrophores, in conj unct ion w it h bot h increased populations and/ or great er visibilit y of m odified m elanophores and nat urally occurring violet and blue iridophores. I t is required for t he production of the all- purple phenot ype ( Figure 1) . Pb causes a large reduct ion of yellow color pigm ent cell populations ( xant hophores) . I t t hus produces a m odified pinkish- purple expression from w hat w ould have been orange color pigm ent cells ( xant ho- eryt hrophores) . High resolution phot ography and m icroscopic st udy show s t he co- exist ence of varying populations of bot h violet and blue st ruct ural iridophores in all individuals, bot h m ale and fem ale ( Bias & Squire 2017a, b, c) . Violet and blue st ruct ural iridophores and m elanophores are alw ays found in close proxim ity wit h one anot her, form ing a type of chrom at ophore unit [ Not e: hereaft er referenced as violet - blue ( iridophores) for ease of discussion] . Violet - blue iridophores ( Figures 3A- B) are m ost visible along t he t opline and in bet w een regions lacking a clearly defined silver iridophore patt ern, oft en including t he caudal- peduncle base. By nat ure, yellow color pigm ent in guppies is highly m otile and m ood dependent w hile red color pigm ent is considered non- m ot ile. Red color pigm ent ( from eryt hrophores) is not altered by Pb, or at least alt ered t o a lesser degree, and a corresponding not iceable increase in t he visibility ( possibly increased population levels) of st ruct ural violet and blue iridophores is evident ( Figures 2A- B and Figure 3A) , resulting in t he increased reflective qualities of individuals. When not m asked by additional color and/ or pat t ern t rait s, t he identification of Purple Body ( Pb) in bot h wild- t ype and dom estic m ales can be easily accom plished t hrough visual phenot ypic observation. I n non- Purple Body ( pb/ pb) individuals carot enoid orange color pigm ent can be described as being vivid, bright orange spot s st ruct urally com prised of densely packed yellow and orange xant ho- eryt hrophores, norm ally extending t o t he very edge of t he spot. Though coverage over additional iridophore pat t erns m ay appear incom plet e. Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 127 Figur e 3. ( A) Pb/ Pb m odif ied caudal base expressin g increased v iolet ir idophores ( r ed circle) . ( B) Non- Pb pb/ pb caudal base expressin g balanced v iolet - blue ir idophores ( r ed circle) , phot os court esy of Christ ian Lukhaup. Het erozygous Purple Body ( Pb/ pb) alt ers orange spot s in select regions of t he body and in finnage t o “ pinkish- purple” . Thus, it m ay not act as a “ full body” m odifier in het erozygous form . Het erozygous Pb does not appear t o great ly reduce visible st ruct ural yellow color pigm ent cells over w hit e leucophore or reflective clust ered yellow cells, know n in breeder circles as Met al Gold ( Mg) ( Undescribed - Bias 2015) , in body and finnage. A slight increase in visibilit y of violet and blue iridophores is oft en det ected. Additionally not ed is an increase and m odification in existing m elanophore st ruct ure and possibly population num bers, as com pared t o het erozygous Pb. I n non- solid colored st rains, a reduct ion in t he num ber of yellow xant hophores result s in a corresponding reduct ion in overall size of individual spot ting ornam ent s. This reveals a “ circular ring” around rem aining color pigm ent produced by an underlying iridophore layer. This w elldefined layer of iridophores is an underlying precursor required for definition of shape over w hich color pigm ent cells populat e during m at urat ion. Hom ozygous Purple Body ( Pb/ Pb) alters all orange spot s found in t he body and in finnage t o “ pinkish- purple” , t hough m odification m ay not be so readily visible in regions of red solid color. I t t herefore should be considered a “ full body” m odifier. Hom ozygous Pb can also produce a purple guppy phenot ype. Hom ozygous Pb rem oves all visible yellow color pigm ent over w hit e leucophores, but not Mg in body and finnage. This in t urn, produces a dram atic increase in t he visibility of wild- type violet - blue iridophores. The num ber of m elanophores does not appear t o drastically increase in any given individual as com pared t o hom ozygous Pb, but t he size of t he m elanophores t hem selves w as great er. Het erozygous Pb exhibit s partial reduction in collect ed xant hophores, and hom ozygous Pb has a near com plet e rem oval of collect ed and clust ered xant hophores. How ever, yellow color cell populations consisting of isolat ed “ wild- type” single cell xant hophores rem ain int act. Furt her descriptions of guppy t rait s are available for dow nload in: Bias & Groenew egen ( 2016, w it h periodic updat es) Poecilia ret iculat a: Dom est ic Breeder Trait Mat rix Reference Guide. M a t e r ials See Bias & Squire ( 2017a) - Supplem ent al S2, S3 for det ail. Figure Figure Figure Figure 1. 2. 4. 5. Purple Delt a Vienna Low ersw ord ( A, C, D) and Purple Delt a Green Delt a x I FGA Purple Delt a ( A and B) Purple Delt a fem ale ( A) and Green Delt a fem ale ( B) Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 128 Figure Figure Figure Figure Figure Figure 6. Green Delt a ( A and B) 16. Vienna Low ersw ord ( A and B) 19. Panda Moscow Roundt ail ( A and B) 28. Golden Vienna Low ersw ord ( A and B) 29. Golden Panda Moscow Roundt ail ( A and B) 30. Vienna Low ersw ord ( A and B) Bias, prior breedings ( unpublished) and current observations. Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure 3. Sunburst Roundt ail ( A and B) 7. Blue Delt a ( A and B) 8. Purple Moscow Delt a ( A) , Blue Moscow Delt a ( B) , Green Moscow Delt a ( C) 9. Red Delt a ( A and B) 10. Half Black Red Delt a ( A and B) 11. Multi Delt a ( A and B) 12. Multi Delt a ( A and B) 13. Multi Delt a ( A and B) 14. Multi Delt a ( A and B) 15. Mosaic Delt a ( A and B) 17. Coral Red Doublesw ord ( A and B) 18. Magent a Delt a ( A and B) 20. Red Albino Delt a ( A and B) 21. Blond Albino Grass Delt a ( A) , Albino- Blond Low ersw ord ( B) 22. Blond Red Delt a ( A and B) 23. Blond Multi Delt a 24. Blond Multi Delt a ( A and B) 25. Vienna Ls ( A and B) 26. Blond Schim m elpennig Delt a ( A and B) 27. Blond Schim m elpennig Doublesw ord ( A and B) 31. Coral Red Ds ( A and B) 32. Grass Delt a ( A, B and C) 33. Grass Delt a ( A and B) 34. Grass Delt a ( A and B) 35. Grass Delt a 36. Blond Low ersw ord ( A and B) M e t h ods. St udy fish w ere raised in 5.75, 8.75 and 10- gallon all- glass aquaria dependent upon age. They received 16 hours of light and 8 hours of darkness per day. Tem perat ures ranged from 78°F t o 82°F. Fish w ere fe d a blend of com m ercially available veget able and algae based flake foods and Ziegler Finfish St art er ( 50/ 50 m ix ratio) twice daily, and new ly hat ched live Art em ia nauplii twice daily. A high volum e feeding schedule w as m aint ained in an at t em pt t o produce t w o positive result s: 1. reduce t he tim e t o onset of initial sexual m at urit y and coloration, t hus reduce tim e between breedings; 2. increase m at ure size for ease of phenot ypic evaluat ion and relat ed m icroscopic st udy. Virgin fem ales w ere used in all crosses. Virgin fem ales w ere obt ained by rem oving all fem ales from t he brood t anks by “ gravid spot selection” prior t o m ale anal fins changing int o gonopodia. Crosses involved breeding groups of single m ales ( unknow n or assum ed genot ype) or 2- 3 m ales ( know n genot ype) and 1- 6 fem ales. Pregnant fem ales w ere isolat ed in breeding net s for collection of fry, and ret urned t o breeding groups once t hey had produced young. All broods from each fem ale w ere t ransferred t o individual t anks and reared independently. Thus t he genot ypes of parent al sires and dam s could be verified wit h analyses. All phot os by t he senior aut hor, unless ot herwise credit ed, w ere t aken w it h a Fuj ifilm FinePix HS25EXR; set t ings Macro, AF: cent er, Aut o Focus: cont inuous, varying Exposure Com pensat ion, I m age Size 16: 9, I m age Quality: Fine, I SO: 200, Film Sim ulation: Ast ia/ Soft , Whit e Balance: 0, Tone: STD, Dynam ic Range: 200, Sharpness: STD, Noise Reduction: High, I nt elligent Sharpness: On. Lens: Fuj inon 30x Opt ical Zoom . Flash: Ext ernal m ount ed EF- 42 Slave Flash; set tings at EV: 0.0, 35m m , PR1/ 1, Flash: - 2/ 3. Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 129 Phot os cropped or bright ness adj ust ed w hen needed w it h Microsoft Office 2010 Pict ure Manager and Adobe Phot oshop CS5. Re su lt s Ph e n ot ypic e x pre ssion of Pb an d n on - Pb m odifica t ion in Gr e y ( w ild- t ype) Figur e 4. ( A) Purple Delt a ( Pb/ pb) m ales. Resu lt s of a hom ozygous Green ( pb/ pb) m ale x hom ozygous Purple ( Pb/ Pb) fem ale breeding. ( B) Hom ozygous Purple ( Pb/ Pb) m ale x hom ozygous Green ( pb/ pb) fem ale breeding. Th is t ype m ale w ill ex pr ess as eit her blue or purple depending upon t he angle of light . Not e part ial m odif icat ion of orange ornam ent s t o pink ish- pur ple and increased v iolet iridophor es ( red circles) . Figur e 5. ( A) Purple Delt a Fem ale ( Pb/ Pb) and ( B) Green Delt a Fem ale ( pb/ pb) , phot os court esy of Bryan Chin. Not e t he dark violet - blu e color of t he caudal fin w it h Pb ( red circles) , gr een reduced t hrough xant hophore rem oval as com pared t o t he non- Pb ( red circle) . Figur e 6. ( A) Green Delt a expressin g Pb m odif ied ornam ent s ( Pb/ pb) . ( B) Green Delt a ( pb/ pb) , phot os court esy of Bryan Chin. Not e t he deepenin g of t he orange body spot s t o pink ish- purple w it h Pb t hrough xant hophore rem oval ( ar rows) . Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 130 Figur e 7. ( A) Blue Delt a expr essing Pb m odified or nam ent s ( Pb/ pb) . ( B) Blue Delt a ( pb/ pb) , phot os cour t esy of Br yan Chin. Not e t he deepening of t he or ange body spot t o pinkish- pur ple w it h Pb t hr ough xant hophor e r em oval ( ar r ow s) . Figur e 8. ( A) Pur ple Moscow delt a ( Y- link Moscow , Mw + X- link Moscow Blau Addit ional Gene, MBAG) , Delt a expr essing Pb m odified or nam ent s and over all pur ple sheen ( r ed cir cles) ( Pb/ Pb) . ( B) Blue- Gr een Moscow ( Mw + MBAG) Delt a expr essing Pb m odified or nam ent s ( Pb/ pb) . Not e v iolet - blue ir idophor e based pat t er n and r educt ion of xant hophor es in het er ozygous Pb condit ion. Unm odified ant er ior or ange body spot par t ially m ask ed by MBAG in peduncle ( ar r ow s) . ( C) Gr een Moscow ( Mw + MBAG) Delt a ( pb/ pb) . Not e absence of Pb effect s. Unm odified ant er ior and post er ior or ange body spot s par t ially m asked in peduncle by MBAG ( ar r ow s) , phot os cour t esy of I gor Dusanic. Figur e 9. ( A) Red Delt a expr essin g Pb m odif ied ornam ent s ( Pb/ pb) . ( B) Red Delt a ( pb/ pb) , phot os court esy of Kiddo Huang. Not e how Pb darkens deep orange t o dark red t hrough xant hophore rem oval ( yellow circles) , and m odifies body spot s from orange t o pink ish- pu rple ( gr een circle) w it h increase v iolet - blue ir idophore ex pr ession. Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 131 Figur e 10. ( A) Half Black ( NiI I ) Red Delt a expressin g Pb m odified ornam ent s ( Pb/ pb) . ( B) Half Black ( NiI I ) Red Delt a ( pb/ pb) , phot os court esy of Bryan Chin and Cheng- Hsien Yang. Not e t he dark er red r eplacing light er orange- red by Pb t hrough xant hophore rem oval. Peduncle and t opline expresses incr eased ir idophor es in Pb, and dorsal shows reduct ion of xant hophores revealing v iolet blu e ir idophores ( ar row) . Figur e 11. ( A) Red Mult i- Color ( Pb/ pb) Delt a ex pr essing part ial Pb m odif ied ornam ent s. Not e t he change of orange spot s in body and finnage t o pink ish- purple from xant hophore rem oval ( red circles) . ( B) Red Mu lt i- Color ( pb/ pb) delt a w it h unm odif ied spot s ( red circles) , phot os court esy of Bryan Chin. Figur e 12. ( A) Purple Mu lt i- Color ( Pb/ Pb) . ( B) Red Mu lt i- Color Delt a expressing Pb m odif ied ( r ed circles) ornam ent s ( Pb/ Pb) , phot os court esy of Bryan Chin. Not e t he ery t hrophores show part ial Pb m odif icat ion in bot h m ales’ finnage and t he violet colorat ion in t he fish t o t he left . Genes ot her t han Pb are responsible for som e of t hese differences. Som e orange spot s are m odif ied t o pink ishpurple. Pb r educes sex- link ed xant hophores in dorsal and caudal fins, revealing wh it e leucophores ( Le) in finnage. Ther e is a slight incr ease in v iolet - blue st ruct ural color in body. Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 132 Figur e 13. ( A) Purple Mu lt i- Color Delt a ( Pb/ Pb) . ( B) Red Mult i- Color Delt as expressin g Pb m odif ied ( red circles) ornam ent s ( Pb/ Pb) , phot os court esy of Bryan Chin. Again, addit ional genes are involved her e. Orange spot s are m odified t o pink ish- pur ple. Pb reduces sex- linked xant hophores in dorsal and caudal ( yellow circles) , revealin g whit e leucophores ( Le) in finnage. There is a slight increase in violet - blue st ruct ural color in body. Figur e 14. ( A) Purple Mu lt i- Color ( Pb/ Pb) . ( B) Red Mu lt i- Color Delt as expressin g Pb m odif ied ornam ent s ( Pb/ Pb) , phot os court esy of Bryan Chin. Orange spot s are m odif ied t o pink ish- purple, and increased v iolet - blue ir idophores ( r ed arrows) . Som e ant erior Met al Gold ( Mg) rem ains over blu e ir idophores and in t he VEG peduncle spot ( gr een ar rows) . Figur e 15. ( A) Red Mosaic Delt a ( Pb/ Pb or Pb/ pb) ex pr essing Pb m odif ied ornam ent s. ( B) Red Mosaic Delt a ( pb/ pb) , phot os court esy of Kevin and Karen Yang. Pb convert s light er carot enoid orange t o pink ish- pu rple on t he body ( ar rows) , but t he effect on t he darker pt er idine r ed in caudal fin is less pronounced. Pb reduces sex- link ed xant hophores in dorsal and caudal, revealin g whit e leucophor es ( Le) in finnage. Ther e is a slight incr ease in violet - blue st ruct ural color in body. Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 133 Figur e 16. ( A) Vienna Lowerswor d expressing Pb m odif ied ornam ent s ( Pb/ Pb) . ( B) Vienna Lowersword ( pb/ pb) . Green colorat ion is t he product of yellow xant hophores over blue ir idophores. Not e t hat when Pb reduces t he yellow, it reduces green as well. Post er ior orange spot s are m odified t o pink ish- pur ple ( ar rows) . Ther e is increased expression of violet - blue ir idophores while collect ed sex- linked xant hophores are rem ov ed and clust er ed Met al Gold ( Mg) ar e only r educed by Pb ( arrow) . Figur e 17. ( A) : Coral Red ( Co) Doublesword expressing Pb m odif ied ornam ent s ( Pb/ Pb or Pb/ pb) . ( B) Coral Red ( Co) Doublesword ( pb/ pb) , phot os court esy of Kriszt ián Medv eczk i and Gary Lee. Not e how Pb changes orange t o deep r ed. Pb r em oves sex- linked xant hophores in t he dorsal and caudal, leav ing only whit e leucophores ( Le) , as no sex- linked er yt hrophor es are pr esent . Som e orange spot s are m odif ied t o pink ish- pu rple, and t her e is proliferat ion of ir idophor es in t he body . Sex- lin ked collect ed yellow pigm ent is rem oved from bot h caudal and dorsal ( arrow) . Ther e is a heavy increase in violet - blu e st ruct ural color in body, but not finnage, as Pb m odificat ion of eryt hrophores is m inim al. Figur e 18. ( A) Magent a ( M) Delt a expressin g Pb m odif ied ornam ent s ( Pb/ pb) . ( B) Magent a ( M) Delt a ( pb/ pb) , phot os court esy of Kriszt ián Medv eczk i. Not e how Pb convert s orange t o red by reduct ion of xant hophores ( arrows) and deepens and expands t he violet - blue ir idophore colorat ion. Orange spot s are m odified t o pink ish- purple, and increased expression of violet - blu e ir idophores by xant hophore reduct ion ( arrows) . Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 134 Figur e 19. ( A) Purple Panda Moscow ( Mw + pp) Roundt ail expressin g Pb m odif ied ornam ent s ( red circles) ( Pb/ Pb) . ( B) Panda Moscow ( Mw + pp) Roundt ail ( pb/ pb) . Gr een is replaced by purple w it h Pb, by rem oval of xant hophores wit h incr eased ex pression of violet - blue ir idophor es. Ph e n ot ypic e x pre ssion of Pb an d n on - Pb m odifica t ion in Albin o ( a a ) Figur e 20. ( A) Full Red Albino ( aa) Delt a ex pressing Pb m odif ied ornam ent s ( Pb/ pb) . ( B) Full Red Albino ( aa) Delt a ( pb/ pb) , phot os court esy of Tseng Huang Chieh. Melanophores are rem oved by Albino. ( A) Orange r ed is changed t o dark red by Pb t hrough xant hophore rem oval, w it h prolif erat ion of violet - blue ir idophores v isibly m odif ied t o pink ish- pur ple ( arrows) . Figur e 21. ( A) Albino ( aa) Delt a expr essin g Pb m odif ied ornam ent s ( red circles) ( Pb/ Pb or Pb/ pb) , phot o court esy of Benson Liu. ( B) Albino ( aa) Lowersword ( pb/ pb) . Melanophores are rem ov ed by Albino, t hough dendr it ic pat t ern rem ains. Post er ior orange spot s ( red circles) rem ain unm odified wit h reduced expression of violet - blu e ir idophores in t he body. Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 135 Ph e n ot ypic e x pre ssion of Pb an d n on - Pb m odifica t ion in Blon d ( bb) Figur e 22. ( A) Blond ( bb) Red Delt a expressing Pb m odified ornam ent s ( Pb/ pb or Pb/ Pb) . ( B) Blond ( bb) Red Delt a ( pb/ pb) , phot os court esy of Bryan Chin and Kiddo Huang. Rem oval of xant hophores and a proliferat ion of violet - blue ir idophores m odifies orange t o a darker red and spot s are m odif ied t o pink ish- pur ple ( ar rows) . Figur e 23. Blond ( b) Red Mult i- Color Delt a expressin g Pb m odif ied ornam ent s ( Pb/ pb) , phot o court esy of Bryan Chin. Blond reduces m elanophore size. Post erior orange is convert ed t o pink ishpurple by Pb ( arrows) , w it h increased expression of violet - blu e ir idophore st ruct ural color. Pb reduces sex- link ed xant hophores in dorsal and caudal, revealing wh it e leucophores ( Le) in finnage. Figur e 24. ( A) Blond ( bb) Mult i- Color Delt a expr essing Pb m odif ied ornam ent s ( Pb/ pb) . Het erozygous Pb convert s som e orange spot s over Zebr inus ( Ze) t o pink ish- purple ( ar rows) w it h increased purple v iolet iridophor es, som e ant erior orange rem ains. Pb reduces sex- linked xant hophores in dorsal and caudal fins, revealing wh it e leucophor es ( Le) in finnage. Ther e is a slight incr ease in v iolet - blue st ruct ural color in body. ( B) Blond ( bb) Mult i- Color Delt a ( pb/ pb) , phot os court esy of Bryan Chin. Blond reduces m elanophore size. There is no m odificat ion t o orange spot t ing ornam ent s ( arrows) com pr ised of xant ho- eryt h rophores. Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 136 Figur e 25. ( A) Blond ( bb) Vienna Lowersword ex pr essin g Pb m odif ied ornam ent s ( Pb/ Pb) . ( B) Blond ( bb) Vienna Lowersword ( pb/ pb) . Blond reduces m elanophore size. Orange is convert ed t o pink ishpurple by xant hophore rem oval ( arrows) , gr een is alm ost elim inat ed by Pb. Figur e 26. ( A) Blond ( bb) Schim m elpenn ig Plat inum ( Sc) Delt a ex pr essing Pb m odified ornam ent s ( Pb/ Pb) . ( B) Blond ( bb) Schim m elpennig Plat inum ( Sc) Delt a ( pb/ pb) , phot os court esy of Bryan Chin. Blond reduces m elanophore size. Orange is convert ed t o pink ish- pu rple by xant hophore rem oval ( ar rows) , green is elim inat ed by Pb. Pb r educes sex- link ed xant hophores in dorsal and caudal fins, revealing whit e leucophores ( Le) in finnage. Met al Gold ( Mg) rem ains in body and finnage. Ther e is a slight incr ease in v iolet - blue st ruct ur al color in body. Figur e 27. ( A) Blond ( bb) Schim m elpenn ig Plat inum ( Sc) Doublesword ex pr essing Pb m odified ornam ent s ( Pb/ Pb) . ( B) Blond ( bb) Schim m elpenn ig Plat inum ( Sc) Doubleswor d ( pb/ pb) . Blond reduces m elanophore size. Orange is convert ed t o pink ish- purple by xant hophore rem oval ( arrows) , green is elim inat ed by Pb. Not e t hat t he lar ge plat inum shoulder area, com prised of clust er ed Met al Gold ( Mg) xant hophores, is st ill present in hom ozygous Pb. Pb reduces sex- lin ked xant hophores in dorsal and caudal, rev ealing wh it e leucophores ( Le) in finnage. Ther e is a slight increase in violet - blue st ruct ural color in body. Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 137 Ph e n ot ypic e x pre ssion of Pb an d n on - Pb m odifica t ion in Golden ( gg) Figur e 28. ( A) Golden ( gg) Vienna Lowerswor d expressing Pb m odified ornam ent s ( Pb/ Pb) . ( B) Golden ( gg) Vienna Lowersword ( pb/ pb) . Melanophores are reduced and collect ed by Golden. Orange is convert ed t o pink ish- pu rple by Pb by xant hophore rem oval ( arrows) , pale blu e is deepened t o violet , gr een is r educed. Pb reduces sex - lin ked xant hophores in dorsal and caudal, rev ealing wh it e leucophores ( Le) in finnage. Met al Gold ( Mg) r em ains in body and finnage. Slight increase in violet - blue st ruct ural color in body. Figur e 29. ( A) Golden ( gg) Purple Panda Moscow ( Mw + pp) Roundt ail ex pr essing Pb m odified ornam ent s ( Pb/ Pb) . ( B) Golden ( gg) Panda Moscow ( Mw + pp) Roundt ail ( pb/ pb) . Green is replaced by purple w it h Pb, t hrough xant hophore rem oval and pr oliferat ion of violet - blu e iridophor es ( arrow) . Reduct ion in m elanophore num bers, especially ect opic, reduces dark peduncle colorat ion ( arrow) . Ph e n ot ypic e x pre ssion of Pb an d n on - Pb m odifica t ion in Asia n Bla u ( Ab) Figur e 30. ( A) Asian Blau ( Ab) Vienna Lowersword ex pr essing Pb m odified ornam ent s ( Pb/ Pb. ( B) Asian Blau ( Ab) Vienna Lowerswords ( pb/ pb) . Orange is convert ed t o pink ish- pur ple by Pb xant hophore rem oval ( arrows) . Ery t hrophor es are furt her rem ov ed by Ab rev ealing m odif ied v iolet blu e ir idophores ( ar rows) . Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 138 Figur e 31. ( A) Asian Blau ( Ab) I vory ( ii) ; i. e. Lavender Coral Red ( Co) Doublesword expressin g Pb m odif ied ornam ent s ( Pb/ Pb or Pb/ pb) , phot o court esy of Taket oshi Sue. Orange is conver t ed t o pink ish- purple by Pb. Asian Blau rem oves orange and I vory rem ov es yellow- orange rev ealing m odif ied violet - blu e iridophor es and whit e leucophores ( red arrows) . ( B) Asian Blau ( Ab) Coral Red ( Co, pb/ pb) . Doublesword Orange convert ed t o pink ish- purple by Pb, and rem oved by Ab rev ealing under ly ing leucophores and m odif ied ir idophor es ( red arrows) . Figur e 32. ( A) Asian Blau ( Ab) I vory ( ii) ; i. e. Lavender Grass Delt a expr essing Pb m odified ornam ent s ( Pb/ pb) . Orange conver t ed t o pink ish- pu rple by Pb. Asian Blau rem oves orange and I vory rem oves yellow - orange r evealing m odified v iolet - blu e ir idophores and whit e leucophores ( r ed circles) . ( B) Red Grass Delt a expressin g orange ornam ent s ( red circles) ( pb/ pb) . Orange in peduncle and caudal convert ed t o pink ish- pur ple by Pb ( yellow circles) , phot os court esy of Taket oshi Sue. ( C) Pink Grass Delt a ex pr essing Pb m odified ornam ent s ( Pb/ pb) . Orange conver t ed t o pink ish- pur ple by Pb ( red circles) , phot o court esy of Gyula Pasarét i. Figur e 33. ( A) Asian Blau ( Ab) Purple Grass Delt a expr essing Pb m odif ied ornam ent s ( Pb/ pb) . Orange convert ed t o pink ish- pur ple by Pb ( red circles) . Asian Blau rem oves orange revealing m odif ied violet - blu e iridophor es. Phot o court esy of Leanne Shore. ( B) Asian Blau ( Ab) Blue Grass Delt a ( pb/ pb) . Asian Blau rem oves orange r ev ealing m odif ied blue ir idophor es ( red circles) . Phot o court esy of Kevin Yao. Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 139 Figur e 34. ( A) Asian Blau ( Ab) I vory ( i) ; i. e. Lavender Coral Red ( Co) Grass Delt a expr essing Pb m odif ied ornam ent s ( Pb/ pb) . Orange convert ed t o pink ish- purple by Pb. Asian Blau rem oves orange and I vory rem ov es yellow- orange revealin g m odified v iolet - blue ir idophores and whit e leucophor es ( red circles) . ( B) Asian Blau ( Ab) Coral Red ( Co) Blue Grass Delt a ( pb/ pb) . Asian Blau rem oves orange r ev ealing m odified blu e ir idophores and whit e leucophores ( r ed circles) , and m in im al Met al Gold ( Mg) . Phot os court esy of Taket oshi Sue. Figur e 35. Top: Asian Blau ( Ab) Albino ( aa) I vory ( ii) ; i. e. Albino Lavender Grass Delt a expressin g Pb m odif ied ornam ent s ( Pb/ Pb or Pb/ pb) . Albino rem ov es m elanophores. Orange convert ed t o pink ish- purple by Pb. Asian Blau rem oves orange and I vory rem ov es yellow- orange rev ealing m odif ied violet - blu e iridophor es ( red circles) . Bot t om : Asian Blau ( Ab) Albino ( aa) ; i.e. Albino Blue Grass Delt a, non- Pb ( pb/ pb) . Albino ( aa) rem oves m elanophores. Asian Blau ( Ab) rem oves orange rev ealing m odif ied blu e ir idophor es ( yellow circles) and Met al Gold ( Mg) . Phot o court esy of Taket oshi Sue. Figur e 36. ( A) Asian Blau ( Ab) Blond ( bb) Vienna Lower sword ex pr essing Pb m odified ornam ent s ( Pb/ Pb) . ( B) Asian Blau ( Ab) Blond ( bb) Vienna Lowersword ( Pb/ pb) . Blond r educes m elanophores. Orange convert ed t o pink ish- pur ple by Pb, by xant hophore rem oval ( arrows) . Asian Blau rem oves orange rev ealing m odif ied light v iolet - blue ir idophores. Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 140 D iscussion. The genes kit a, kitla, csf1ra, and csf1rb have been ident ified in Guppies ( Kot t ler et al 2013, 2015) . Kit a is a required precursor gene in t he developm ent of early form ing m otile m elanophores ( kit a- dependent ) in guppies. Kit a is not required for developm ent of genet ically distinct late form ing, possibly non- m ot ile, m elanophores ( kit a-independent ). Toget her, t hey produce t he ret iculated scale pat t ern found in bot h sexes; wild and dom estic. Csf1ra is a required precursor gene ( colony- stim ulating fact or 1 recept or) in t he developm ent and form ation of xant ho- eryt hrophores. I n guppies csf1ra is not required in developm ent of late form ing, possibly non- m ot ile, m elanophores ( kit a-independent ). Toget her, t hey cont ribut e t o non- defective ornam ent s of w ild- type m ales. I nt eract ions bet w een lat e form ing m elanophores in conj unct ion wit h xant hoeryt hrophores affect dom estic guppy st rain ornam ent s in bot h sexes. Thus, funct ional kit a and csf1ra are found in non- defect ive ornam ent s of dom est ic st rains in bot h sexes. Purple Bbody rem oves cert ain classes of yellow- orange- red color pigm ent over silver iridophores or white leucophores. Dark red pt eridine color pigm ent does not seem t o be m odified by Purple Body in fins lacking an underlying silver iridophore or w hit e leucophore pat t ern. Modification by Pb seem s lim it ed predom inantly t o wild- t ype orange color pigm ent ed x ant ho- eryt hrophores; i.e. t hose w hich also cont ain yellow- orange carot enoids in addition t o red pt eridines, over an iridophore pat t ern. Het erozygous Purple Body ( Pb/ pb) alt ers orange spot s in select regions of t he body and in finnage t o “pinkishpurple” . Het erozygous Pb does not appear t o greatly reduce visible st ruct ural yellow color pigm ent cells over w hit e leucophore or reflective clust ered yellow cells in body and finnage. A slight increase in visibilit y of violet and blue iridophores is oft en found. Hom ozygous Pb expression result s in a furt her rem oval of xant ho- eryt hrophores, in conj unct ion wit h bot h increased populations and/ or great er visibilit y of m odified m elanophores and nat urally occurring violet and blue iridophores. Pb/ Pb plus an unidentified additional genetic com ponent is required for product ion of t he all- purple phenot ype. When Pb is com bined wit h any of t he aut osom al or sex- linked m ut ant s w ell know n t o breeders, Albino ( aa) , Blond ( bb) , Golden ( gg) , and Asian Blau ( Ab) , furt her com bined effect s occur. The m odifications of each of t hese genes on “ w ild- type” grey and resulting co- expressions wit h t he Purple Body ( Pb) gene are briefly discussed as follow s: Au t osom a l Ge n e s w it h Re fe re nce s Albin o ( aa, Haskins & Haskins 1948) . Recessive. Also know n as Real Red Eye Albino ( RREA) or ( Type B) . There is an inability t o produce black m elanophores in t he body and finnage. I t elim inates all classes of m elanophores; dendritic, corolla and punct at e. Therefore m elanin is absent as w ell. Albino is epist atic t o bot h t he Blond and Golden genes; t hus m ut ant alleles of each m ay be found in t he albino genot ype (e.g., aa Bb, aa bb, aa Gg, aa gg, etc.) ( Haskins & Haskins 1948; Goodrich et al 1944; Bias 2017, The albino- blond double recessive, fort hcom ing) . I n Albino, t he result w hen com bined wit h Pb is sim ilar t o t hat wit h Blond, except t hat t he m elanin is com pletely absent and t he colors appear even paler ( except w hen a different pigm ent cell is over expressed as in full red) . Red can appear deepened and “ darker” w hen Pb is present . Blon d ( bb, Goodrich et al 1944; Kot t ler et al 2015) . Recessive. I t has been ident ified as a defective gene m ut ation in adenylate ( adenylyl) cyclase 5 ( adcy5; ac5) , and m apped t o Linkage Group ( LG) 2 ( Kot tler et al 2015) . I t is also know n t o pedigree breeders as I FGA Gold, Asian Gold, European Blond. This m ut ation produces a near norm al num ber of black m elanophores of all t ypes; Dendritic, Corolla and Punct at e. How ever, t he size of each is greatly reduced and t he st ruct ure m odified as com pared t o “ wild- type” grey. According t o published result s Blond is not linked t o Golden. This gene should be viewed not as a suppressor of m elanophores, but rat her one t hat alt ers m elanophore size and shape. When Pb is com bined wit h hom ozygous blond ( bb) , a violet - blue sheen co- expresses w it h oft en paler and less int ense xant ho- eryt hrophores, resulting from t he reduction in Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 141 m elanophore size. The reflective qualities of xant ho- eryt hrophores in Blond are det erm ined by t he com position of underlying st ruct ural colors (iridophores are m ore reflective and leucophores are less reflective) and angles at w hich cryst alline plat elet s reside. Golde n ( gg, Haskins & Druzba 1938; Goodrich et al 1944; Winge & Ditlevsen 1947) . Recessive. The Golden gene is a defective ort holog of kit a ( Kot tler et al 2013) . Kit a has been m apped on Guppy aut osom al LG 4 ( Tripat hi et al 2008) . I t is also know n t o pedigree breeders as European Gold, I FGA Bronze or Asian Tiger. This gene produces a reduced am ount of black m elanophores ( approxim at ely 50% ) of all t ypes; dendritic, corolla and punct at e. How ever, t he size of dendrit ic and corolla m elanophores is greatly increased and t he collection of corolla m elanophores is concent rat ed int o "clum ps" or "islands" of m elanophores along t he scale edges. Males and fem ales lack skin m elanophores at birt h, but t hey develop w it h m at urit y. Scale edging will becom e lighter w it h a higher inbreeding co- efficient ; i.e. long- term Golden x Golden breeding’s. This suggest s t hat t here are non- allelic m odifier genes affect ing t he Golden phenot ype. According t o published result s (Kot tler et al 2013, 2015; Tripat hi et al 2008) Golden and Blond are in different linkage groups and assum edly different chrom osom es and t hus can not be alleles of each ot her or linked t o each ot her. This gene should be view ed as a suppressor of m elanophore population num bers. I n Pb plus Golden ( gg) , t he effect is sim ilar t o Blond, except t hat m elanophores are present at higher frequencies and wit h a m odified dist ribution. As a result , t he colors w hile paler t han in grey are not as pale as in Blond. Asia n Blau ( Ab, Bias 2015; Bias & Squire 2017b) . I ncom plet ely Dom inant . Also know n t o pedigree breeders as ( r2) Europe and ( Rr) Asia. [ Not e: The use of low er case “ r” violat es t he accept ed genetic use of sym bols since t his is not a recessive gene, t his usage cam e about prior t o identification of Ab as a second eryt hrophore defect] . I n het erozygous condition red color pigm ent is rem oved, w hile collect ed yellow color pigm ent and clust ered Met al Gold ( Mg) is lit tle affect ed. This produces an iridophore based phenot ype. Snakeskin pat t erns degrade in bot h het erozygous and hom ozygous expression, as a result of disruption of m elanophore st ruct ure or m elanin cont ent . The Purple ( Violet ) sheen found above t he lat eral line of bot h m ales and fem ales is rem oved. I n hom ozygous condit ion cert ain black m elanophores are rem oved along w it h red and yellow color pigm ent s. I n hom ozygous condit ion finnage m ay be reduced in size, but t he genes are st ill present in t he genot ype for norm al finnage. An out cross of hom ozygous Ab w ill produce t he expect ed finnage in F1 offspring. [ Not e: As t here are distinct t ypes of red color pigm ent ( carot enoid and pteridine) present in bot h body and finnage, rem oval m ay not be com plet e, as in a red “ Old Fashioned” shoulder st ripe. A very faint “ red shoulder st ripe” is som etim es visible] . The result w hen Asian Blau is com bined wit h Pb can range from highly reflective violet - blue t o non- reflective violet - blue in com binat ion wit h additional genes t hat rem ove and/ or reduce iridophores or alt er angles of cryst alline plat elet s. Eu r ope a n Bla u ( r or r1, Dzwillo 1959; Kot t ler et al 2013) ; also ( eb) . Recessive. Bot h csf1ra and csf1rb genes have been identified in guppies, and are t he result of an ancest ral genom e duplication event t hat produced four copies of each gene rat her t han t w o ( t he guppy is an ancest ral t et raploid) . I n m any fish species one or t he ot her pair of som e genes has been lost t o reduce t he t ot al gene dosage back t o a “ diploid level” of tw o rat her t han four copies. I n som e ot her cases, t he t w o genes diverge from each ot her and assum e different functions. The European Blau gene is a defective ort holog of csf1ra. Expression levels of csf1rb w ere not upregulat ed t o com pensat e for t he deficiency in csf1ra ( Kot t ler et al 2013) , w hich suggests t hat Csf1ra and Csf1rb have functionally diverged from each ot her in t he guppy. Csf1ra has been m apped on Guppy aut osom al LG 10 ( Tripat hi et al 2008) . European Blau is also know n as Dunkel in Asia. Csf1ra act ivity is required for t he dispersal or different iation of m ale- specific xant hophores ( Kot t ler et al 2013) . I n hom ozygous condition it is epist atic t o w ild type genes for red and yellow ; m aj or red and yellow color pigm ent s are rem oved from t he body . Cert ain red color pigm ent s m ay be present in finnage, and t o a lesser degree in t he body. Reflective qualities are reduced. Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 142 Ecot opic m elanophores m ay be rem oved, w hile basal level m elanophores such as are found in Half Black ( NiI I ) are only slightly reduced. “ The salient feat ure of t he csf1ra m ut ant m ales w as t he absence of all orange t rait s, wit h concom it ant severe changes in black ornam ent s” ( Kot t ler et al 2013) . Snakeskin pat terns degrade in hom ozygous expression. The purple ( violet ) sheen found above t he lateral line of bot h m ales and fem ales is rem oved. There is m inim al finnage reduct ion. Addit ion a l Au t osom a l Ge ne s Re fe r re d To Pin k ( p, Luckm an 1990; Först er 1992; pi Kem pkes 2007) Recessive. Rem oval of orange eryt hrophores in body result ing in a “ yellow - orange” cast in finnage. Hom ozygous reduct ion of NiI I m elanophores and increase in MBAG. Rem oves blue iridophores. Reduces size of finnage. Pb m odification: orange spot t ing is convert ed t o pinkish- purple. Collected yellow pigm ent cells are rem oved, but not Clust ered Mg. Body color m ay be m odified t o violet - blue. I vor y ( I , Tsut sui 2006) Aut osom al Dom inant . Het erozygous suppression of eryt hrophores ( red color) . Hom ozygous suppression of xant ho- eryt hrophores ( yellow- red color) , wit h reduction in fin size. Possible differences in m elanophore m odifications in het erozygous vs hom ozygous st at es. Resulting in a "w hit e" appearance.I I ( hom ozygous) , I i ( het erozygous) and ii ( non- I vory) . Pb m odification: previous orange spot ting is rem oved by I vory. Underlying iridophores and leucophores convert ed t o light pinkishpurple. M a ge n t a ( M, undescribed, Bias, unpublished breeding result s) Aut osom al Dom inant . Proliferation of red color pigm ent w hen present and an increase of violet - blue iridophore st ruct ural color. Converts yellow color pigm ent cells ( xant hophores) t o red ( eryt hrophores) , t hough Met al Gold ( Mg) m ay rem ain. Concent rat es black m elanophores. There is a reduction in fin size. Pb m odification: orange spot t ing is convert ed t o pinkishpurple. Collect ed yellow pigm ent cells are rem oved, but not Clust ered Mg. Convert s orange t o red and deepens violet blue coloration. Ze br in us ( Ze, Winge 1927) Aut osom al Dom inant . Color Charact er; Barred pat t ern of vertical st ripes on t he peduncle, viz. 2- 5, generally 3 dark pigm ent st ripes. Effect resem bles t hat of Tigrinus gene, but is as a rule m ore pronounced. ZeZe ( hom ozygous) , Zeze ( het erozygous) and zeze ( non- Zebrinus) . Pb m odification: no direct effect on barring pat t ern. Overlaying orange spot t ing is convert ed t o pinkish- purple. Collect ed yellow pigm ent cells are rem oved, but not Clust ered Mg. M a j or Se x - link e d Tra it s Refe r re d To Cor a l Re d ( Co, undescribed; Turesson 2005; Bias, unpublished breeding result s) Ylinked. Red color pigm ent shoulder patt ern. Linked in com plex wit h Ds. Probably a Full Body m odifier. I t originat ed out of Vienna Em erald Green Ds. Pb m odification: proliferation of violet st ruct ural color. Orange spot t ing is convert ed t o pinkish- purple. Collected yellow pigm ent cells are rem oved, but not clust ered Mg. Gr a ss ( Gra, Tsut sui 2006; I w asaki 1989) X and/ or Y- linked dom inant . The Grass phenot ype is a highly variable random “ fine dot " circular m elanophore pat t ern in finnage. Prim arily lim it ed t o caudal ornam ent at ion, wit h lim ited dorsal influence. Oft en associat ed w it h “ Nike Melanophore St ripe” body pat t ern. Variegation shape is dependent upon inbreeding co- efficient . Color pigm ent s can be added. “ Glass Grass” genot ype is sim ilar t o Multi wit h a t ranslucent background and color pigm ent s. “ Grass Grass” genot ype is oft en linked in com plex wit h sex -linked xant ho- eryt hrophore color pigm ent spot s. Pb m odification: som e orange spot ting is converted t o pinkish- purple. Collect ed yellow pigm ent cells are rem oved, but not Clustered Mg. Xant hophore rem oval m ay reveal w hit e leucophores if present . M oscow Bla u Addit ion al Ge ne ( MBAG, undescribed) X- linked dom inant. Half body pat t ern expressing m otile black m ediating m oderat e & t ranslucent m elanin developm ent over ent ire body area post erior t o dorsal fin, and in caudal peduncle. Ot her post erior peduncle color pat t erns m ay be nearly or w holly obscured. Early Russian MBAG st rains, in addition t o NiI I , m ay have been identified as “ Tuxedo; i.e. HalfBlack” ( I w asaki 1989) . Pb m odification: orange spot ting is converted t o pinkish- purple. Collect ed yellow pigm ent Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 143 cells are rem oved, but not Clust ered Mg. Peduncle m ay t ake on violet - blue reflective coloration. M osa ic ( Mo, Khoo et al 1999) X- linked dom inant . The Mosaic phenot ype is a highly variable random “ large spot ” crescent shaped m elanophore pat t ern in finnage. Prim arily lim ited t o caudal ornam ent at ion, wit h lim it ed dorsal influence. Variegation shape is dependent upon in- breeding co- efficient . Norm ally associat ed wit h eryt hrophore color pigm ent ( carot enoid and/ or pt eridine) . Pb m odificat ion: som e orange spot t ing in t he body is convert ed t o pinkish- purple. Collect ed yellow pigm ent cells are rem oved, but not Clust ered Mg. Xant hophore rem oval m ay reveal w hit e leucophores if present. Dark Red Caudal pigm ent is generally not m odified. M u lt i ( - - , undescribed) X-linked dom inant . The Multi phenot ype is a highly variable random “ fine dot " circular m elanophore pat t ern in finnage. Prim arily lim it ed t o caudal ornam ent at ion, wit h lim it ed dorsal influence. Variegation shape is dependent upon inbreeding co- efficient. Color pigm ent s can be added. Not linked wit h eryt hrophore color pigm ent ( carot enoid and/ or pt eridine) spot s. This m ust be added t hrough out crossing. Lit tle or no effect on existing body color or pat t ern. Pb m odification: som e orange spot t ing in t he body is converted t o pinkish- purple. Collect ed yellow pigm ent cells are rem oved, but not Clust ered Mg. Xant hophore rem oval m ay reveal w hit e leucophores if present . Dark Red Caudal pigm ent is generally not m odified. M oscow ( Mw , Kem pkes 2007) Y- linked. Blue iridophore shoulder pat t ern. Likely a Full Body m odifier. Color variation w it h addition or rem oval of xant ho- eryt hrophores. Pb m odification: orange spot ting is converted t o pinkish- purple. Collect ed yellow pigm ent cells are rem oved, but not Clust ered Mg. Body color is m odified t o violet - blue. V a r ie ga t ion ( Var, Khoo et al 1999) . ( See Gr ass, Mosaic, Multi) X and/ or Y- linked dom inant gene. I nherit ance of variegated t ail pat t erns appears t o be det erm ined by a single locus on t he X and Y chrom osom es. The gene st udy of Variegat ion focused on variable random “ large spot ” shaped m elanophore pat t ern in t he caudal, t hough specim ens exhibited sim ilar dorsal patt ern. Variegat ion shape is dependent upon inbreeding co- efficient . Color pigm ents ( xant ho- eryt hrophores) w ere not linked. Pb m odification: som e orange spot ting in t he body is convert ed t o pinkish- purple. Collect ed yellow pigm ent cells are rem oved, but not Clust ered Mg. Xant hophore rem oval m ay reveal w hit e leucophores if present . Dark Red Caudal pigm ent is generally not m odified. N igr oca u da u t us ( NiI , Nybelin 1947 and NiI I , Dzw illo 1959) X and/ or Y- linked dom inant gene. Full body m odifier, epist atic t o m any ot her genes in out crosses. Pb m odification: orange spot t ing is convert ed t o pinkish- purple. Collected yellow pigm ent cells are rem oved, but not Clust ered Mg. Sch im m e lpen nig Pla tinu m ( Sc) ; Bu x e us ( Kem pkes 2007) . Y- linked dom inant gene. Silver- Blue iridophore shoulder patt ern wit h Metal Gold ( Mg) overlay. Probably a Full Body m odifier. Linked in com plex wit h Ds. Originat ed out of Vienna Em erald Green Ds. Pb m odification: orange spot t ing is convert ed t o pinkish- purple. Collect ed yellow pigm ent cells are rem oved, but not Clust ered Mg. Con clusion s. The newly described gene Purple Body and previously described genes Albino, Blond, Golden, Asian Blau, and European Blau each lim it s or ot herwise reduces t he norm al expression of chr om at ophores found in “ w ild- type” Grey. I n general, visual distinction bet w een Pb and non- Pb phenot ypes is easily accom plished. When t hey are com bined t oget her and wit h ot her frequent ly used color genes, new phenot ypes are produced w hich are useful t o pedigree guppy breeders and com m ercial farm ers alike. Their basic effect s are of int erest t o geneticist s, biochem ist s and m olecular biologist s. Et h ics St a t e m e n t . This st udy adhered t o established et hical practices under AVMA Guidelines for t he Eut hanasia of Anim als: 2013 Edition, S6.2.2 Physical Met hods ( 6) . No specim ens w ere eut hanized or harm ed in t his st udy. Com pe t in g I n t e re st s an d Fun din g. The aut hors declare t hat t hey have no com peting int erest s. Senior aut hor is a m em ber of t he Edit orial Board for Poeciliid Research; Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 144 I nt ernat ional Journal of t he Bioflux Societ y, and request ed non- affiliat ed independent peer review volunt eers. The aut hors received no funding for t his w ork. N ot e s. This publication is num ber four ( 4) of four ( 4) by Bias and Squire in t he st udy of Purple Body ( Pb) in Poecilia ret iculat a: 1. The Cellular Expression and Genet ics of an Est ablished Polym orphism in Poecilia ret iculat a; “ Purple Body, ( Pb) ” is an Aut osom al Dom inant Gene, 2. The Cellular Expression and Genetics of Purple Body ( Pb) in Poecilia ret iculat a, and it s I nt eract ions w it h Asian Blau ( Ab) and Blond ( bb) under Reflect ed and Transm it t ed Light , 3. The Cellular Expression and Genetics of Purple Body (Pb) in t he Ocular Media of t he Guppy Poecilia ret iculat a, 4. The Phenot ypic Expression of Purple Body ( Pb) in Dom estic Guppy St rains of Poecilia ret iculat a. Ack n ow le dgm en t s. To m y best friend and wife Deana Bias, for her support and persist ence over t he last several years in t his four part st udy... To m y co- aut hor and dear friend Rick Squire for his patience as a m entor… To t hose Dom est ic Breeders w ho w illingly provided additionally needed pedigree st rains and st udy populations for com pletion of t his paper… Re fe r en ce s Bias A. S., 2015 Working wit h aut osom al genes for color and pat t ern: a dom estic guppy breeder’s best friend and often w orst night m are. Present ed Sept . 5, 2015 t o at t endees of t he 18 t h World Guppy Cont est held in Tam pa, Florida, USA. Available at : ht t ps: / / www.academ ia.edu/ 15488221/ Wor king_Wit h _Aut osom al_Genes_for_Color_and_P at t ern_A_Dom est ic_Guppy_Breeders_best _friend_and_oft en_worst _night m are. Accessed: February, 2017. Bias A. S., Groenewegen C. A., 2016 Poecilia ret iculat a: Dom estic Breeder Trait Mat rix Reference Guide. 1 st Edition, wit h periodic updates. Available at : ht t ps: / / w w w .academ ia.edu/ 29928596/ Poecilia_reticulat a_Dom estic_Breeder_Trait _M at rix_Reference_Guide. Accessed: January, 2017. Bias A. S., Squire R. D., 2017a The cellular expression and genetics of an est ablished polym orphism in Poecilia ret iculat a; “ Purple Body ( Pb) ” is an aut osom al dom inant gene. Poec Res 7( 1) : 1- 32. Bias A. S., Squire R. D., 2017b The cellular expression and genetics of Purple Body (Pb) in Poecilia ret iculat a, and it s int eractions wit h Asian Blau ( Ab) and Blond ( bb) under reflect ed and t ransm it t ed light. Poec Res 7( 1) : 59- 85. Bias A. S., Squire R. D., 2017c The cellular expression and genetics of Purple Body (Pb) in t he ocular m edia of t he guppy Poecilia reticulata. Poec Res 7( 1) : 93- 119. 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Nybelin O., 1947 Et t fall av X- bunden nedärvning hos Lebist es ret iculat us ( Pet ers) . Zoologiska Bidrag fran Uppsala 25: 448- 454. Tripat hi N., Hoffm ann M., Dreyer C., 2008 Nat ural v ariation of m ale ornam ent al t rait s of t he guppy, Poecilia ret iculat a. Zebrafish 5( 4) : 265- 278. Tsut sui Y., 2006 [ The t rail of t he guppy] . Aqualife m ont hly, pp. 244, 268- 269, 287, 308309. I SBN 4- 89512- 544- 0. [ in Japanese] Turesson H., 2005 Coral red doublesw ords. Guppy Labs e- Bulletin, vol. 8. Available at : ht t ps: / / w eb.archive.org/ w eb/ 20060516012637/ ht t p: / / w w w .guppylabs.info: 80/ oct obe r2005/ hakan.ht m . Accessed: July, 2017. Winge Ö., 1927 The location of eight een genes in Lebist es ret iculat us. Journal of Genet ics 18( 1) : 1- 43. Winge Ö., Ditlevsen E., 1947 Colour inherit ance and sex det erm ination in Lebist es. Heredit y 1( 1) 65- 83. Received: 28 May 2017. Accept ed: 13 July 2017. Published online: 20 July 2017. Aut hor s: Alan S. Bias, I ndependent Resear cher and Sw or dt ail Guppy Br eeder . Mailing addr ess: P.O. Box 1508, Lewisbur g, West Vir ginia 24901, USA. or cid.or g/ 0000- 0002- 9093- 619X. alansbias@aol.com Squir e Richar d D., Biology Depar t m ent ( r et ir ed) , Univer sit y of Puer t o Rico, Mayaguez cam pus, Mayaguez, Puer t o Rico, USA. Mailing addr ess: P. O. Box 3227, Mayaguez, P.R., USA 00681- 3227. or cid.or g/ 0000- 00023916- 0672. r ickdsquir e@gm ail.com This is an open- access ar t icle distr ibut ed under t he t er m s of t he Cr eat ive Com m ons At t r ibut ion License, which per m it s unr estr ict ed use, dist r ibution and r epr oduct ion in any m edium , pr ovided t he or iginal aut hor and sour ce ar e cr edit ed. How t o cit e t his ar t icle: Bias A. S., Squir e R. D., 2017 The phenot ypic expr ession of Pur ple Body ( Pb) in dom est ic guppy st r ains of Poecilia r et iculat a. Poec Res 7( 1) : 125- 146. Poeciliid Research, 2017, Volum e 7, I ssue 1. ht t p: / / www.pr.bi oflux .com .ro/ 146