Ginga Sulphureus... ...Evolution of a new strain or variant upon existent?

© Alan S. Bias

Permission granted for nonprofit reproduction or duplication of photos and text with proper credit for learning purposes only.

August 9, 2012

Ginga Sulphureus   ...Evolution of a new strain or 

                                                      variant upon existent?

F2 Ginga Sulphureus

Introduction:

It seems most new strains come about from one of four scenarios:  1. Random hybridization. 2. Mutation.  3. Crossover.  4. Planned modification.  The latter should stem from a predetermined breed plan with well set goals working with a recognizable parental strain(s) and any additional genotype you wish to infuse.  In such instance one of the first questions that comes to mind is just how much modification is needed for distinction as a new strain from it's parent?  When is it a new strain and when it is just a variant of the parental phenotype?


Discussion:
So what do we consider a variant?  In a simple sense when limited sex-link or autosomal traits are introduced.  Such as blond, albino, another body or fin color.  Many breeders will often limit their discussion of variants to autosomal effect. In a strict sense whenever a breeder acquires a set of genetics, maintains it for several generations in his/her own fishroom under their own devices it becomes your strain.  Even when bred towards original well defined standards of the creator or a breed standard.  This results from a change in both environmental and genetic inputs that, while similar, are no longer under the same controls.

Ginga Rubra - photo K. Tanaka with permission

A couple of years ago I first saw a picture of Kenjiro Tanaka's Ginga Rubra.  Ken has been well known in Killifish circles for many years.  His 30 plus years of manipulating guppy genetics is far less known to breeders outside of his native Japan.  In email correspondence and review of his website:  http://www.ne.jp/asahi/medaka-ken/k.t/index.html it becomes clear Tanaka's interest in guppy breeding favors non delta strains that are small, highly active, with alot of coloration.  In variations of his Ginga you can find finnage that parallels caudal shape found in some killifish species.

Ginga Rubra - photo K. Tanaka with permission

For quite some time I pondered the Ginga Rubra phenotype and whether it would hold my interest.  A first step of any breeder when considering acquisition of new genetics should be an analysis of the potential makeup and it's value to your current breedings.   It was  apparent that a fair amount of the Ginga traits were autosomal and that the phenotype would always be fairly fluid in many respects.  Similar to how I maintain most of my Swordtails.  To those who appreciate non-delta type guppies Ginga are quite stunning to the eye, but in the end still a red dorsal guppy and they rarely hold my interest for long.  Which I have always found strange in that my primary focus in large animal livestock breeding has always been towards red:  Red Galloway cattle, Moorit Shetland sheep, Rhode Island Red chickens, and more.  So, after looking over limited photos of the strain for a number a months I felt I had a decent understanding of the genotype comprising this strain.  The honor of defining Ginga Rubra as a strain belongs to Tanaka as the breeder. In correspondence he indicates, "My Ginga strain is extremely variable and I'm working with many types to get (it) established."  I'll only list, describe and then discuss several visible components.

Ginga Rubra - photo K. Tanaka with permission

Today most informed breeders are inclined to view genotype based on gene complexes governed by regulation, rather than single genes resulting in specifics traits. The Overall Ginga phenotype is comprised in such a manner.  Where it differs from many modern strains is being a complex composition of recognizable traits described many years ago on a basic wild-type form.  Ginga genotype is not hidden like many modern strains if you know how to look.  You might describe it as a series of overlays and overlap.  Very similar to the Vienna type Swordtails I have worked with for nearly 30 years.

In total the Tanaka Ginga Rubra phenotype is bred for and contains at least the following visible traits:  Blond (IFGA gold), wild-type, possibly reduced Emerald Green Iridescence (EGI) from no visible peduncle spot, Iridescence, purple body mutation, barring overlayed in purple, roundtail (though some males exhibit sword-like extension), ocular spotting (in peduncle base into caudal), increased number of Wingean "orange spots" along the lateral line, red dorsal (sex linked), multiple genes identified by Winge and Kirpichnikov for caudal color patterns and shape.  

Winge, O. (1922b), male exhibiting Sulphureus and Rubra

O. Winge in his 1922b and 1923 papers describes both the sulphureus (su) and ruber (ru) genes and hints at the possibilities of crossover to explain phenotypes with only partial components of each.    In Winge's 1927 publication he goes one step further and not only illustrates this process of crossover, but reduces both su and ru from two complex genes into to four distinct genes.   The complex sulphureus becomes vitellinus (vi) and coccineus (co), while ruber becomes minutus (mi) and sanguineus (sa).  Why is this important?  Because it forms the basis of multiple components of Ginga Rubra and Ginga Sulphureus.  In 1922b he describes each:  Sulphureus;  1.  Sulphur yellow colour in the dorsal fin and a dark dot that only at times is visisble in the said fin.  2.  Sulphur yellow colour in the tial and in the caudal fins.  3.  Red colour in the lower edge of the caudal fin.  Rubra;  1.  Red colour proximally in the upper edge of the caudal fin.  2.  Large oblong red side spot, lying for the most part below and behind the dorsal fin.  3.  Dark side-dot in the tail at the base of the caudal fin.

Winge, O. (1922b)

Winge, O. (1927)

Winge, O. (1927)

Winge further describes in his 1927 publication two other genes that are potentials in the makeup of Ginga.  Those being Tigrinus (Ti); an X-link consisting of 1-2 black pigment stripes, and Zebrinus (Ze); a recessive autosome consisting of 2-5, generally 3 dark pigment stripes.  He indicates they resemble Ti, but are more pronounced.    As an autosome this trait can be both heterozygous or homozygous in form.  A heterozygous sire will statistically pass the trait to 1/2 of his sons, but not necessarily expression.  This is determined by the recessive or dominant nature of the autosome.

Winge, O. (1927)

Phang, V (1999)

There is a 3rd form of barring described by V. Phang, et al. (1999).  In conclusion she proposes:  "The designation of (bar) for an autosomal recessive gene, when in homozygous condition barbar and present with snakeskin body pattern (SSB) produces a vertical barred pattern".  The dominant and allele being bar⁺.  There still remains alot of speculation among breeders if bar and Ze are not one in the same.  Source fish listed were limited to a strain of Y-linked Singapore Yellow Snakeskins.  No where in Phang (1999) is there any reference to reciprocal outcross of bar⁺ Y to non snakeskin strains composed of barbar XX or barbar⁺ XX females.  Therefore, it is possible that bar and Ze are one in the same as her statistical analysis does not prove bar and SSB were found in complex.  Simply co-expressed on males as individual traits.

Iridescens (ir)  Winge 1922b, 1.  A characteristic mother of pearl sheen on the body.  2.  2-3 red smaller side spots.  3.  Black side-dot on the tial near the caudal fin.  4.  Black side-dot on the anterior body.

Winge, O. (1922b)

Ferrugineus (fe) Winge 1922b, 1.  Black rust-coloured part in the proximal part of the cuadal fin.  2.  Black side-dot in in the tail near the caudal find (viz. with ocular spotting in the caudal).

Winge, O. (1922b)

Purple Body Mutation - undescribed, is a very important Ginga trait that needs to me mentioned here.  Tanaka's Ginga Rubra are bred exclusively for Pb.  To the best of my knowledge there is no formal description of this trait in guppies, having found no published research or reliable documentation of it.  Discussion with other breeders and researchers has also provided minimal insight.  To my knowledge this trait is found in only two of three published members of the guppy family comprising the subgenus Acanthophacelus; Poecilia reticualata, Poecilia obscura, and Poecilia Wingei.  This trait does not seem to occur in any of the pure Poecilia Wingei (Enders) that I worked with, nor are there any photos online from reliably documented populations.  It is only expressed in hybrids.  In the past, having been able to breed for it reliably, it did not seem necessary to set up breeding groups, dedicate tanks and document results statistically.  My test crosses to decipher it, in several strains, to include Ginga, Vienna and Wild-type, are not yet conclusive.  

This trait I refer to as Purple Body Mutation (Pb).  It's counterpart is Green.  My test breedings to date with non Ginga strains point towards Pb being an autosomal recessive trait.  Green presumably could be a dominant wild-type allele or epistatic trait as evidenced by lack of Pb in Endlers.  Until it is proven whether the latter is a wild-type allele (Pb⁺⁾ or a distinct trait (Gb) I see no use in defining it as either.  To do so would require a larger number of free tanks than I have available.  Two strains; one homzygous for purple and another for green.  Documented analysis of reciprocal F1 crosses followed by F2 sib breedings.   

Vienna LS in both Pb (upper) and green (lower)

Purple Body Mutation as an autosomal recessive trait is a precursor in determining visible color resulting from pigment overlay.   Pb is what allows for creation of a true purple phenotype (viz. IFGA Purple).  Effect is limited to certain regions of the body.  Always in the peduncle in strains with reduced iridophores in this area.  In conversion of Wingean spots from orange to pink.  The expression can also extend along the shoulders and into the topline.  In most males, the belly region does not appear to be altered by the effect of Pb.  Green males express Wingean spots in orange.  Pb males express Wingean spots in pink.   While Pb is most noticeable in the peduncle region of male guppies, it is also expressed in females.  Along the lateral line and in the shoulders towards the topline, appearing as an iridescent purple sheen on the scales.  


Breedings:

In late summer of 2011 I arranged to obtain a trio of pure F1 Ginga Rubra from an individual who had imported directly from Tanaka.  My only requests were a male having a round tail and ocular spotting with no visible extensions, roundtail being a recessive not easily identifiable in my Vienna.  This request was fulfilled with the above below as he appeared after 4-5 months growth.

Ginga Rubra (P sire partially colored)

Ginga Rubra (P sire mature)

 As some of you have surmised based on yellow based breeding program my primary focus, at least initially, would be to remove red  in the dorsal and caudal via selective breeding.  If not feasible to at least mask it via epistasis from traits found in my Vienna LS.  To do this I would initiate breedings with a pure Ginga male and several pure Vienna females, blond and grey, to include both green and purple body mutation.  Then utilizing F1, F2 and F3 hybrid males exclusively bred the first 2-3 generations in backcross to pure Vienna females.  All hybrid filial sibling females were culled prior to onset of maturity.   Such a breed plan would avoid the pitfalls of segregation when using siblings.  In result:  (((Ginga * Vienna LS) * Vienna LS) * Vienna LS).  All initial Vienna females came from my Line1 as this line does not posses partially dominant autosomal Z-bar.  It does contain both Pb and green in composition.

One of the first things I wished to know about about Ginga had to do with barring.  Was this trait traditional autosomal Z-bar or the elusive recessive sex linked Tigrinus as documented by O. Winge early in the last century.

Ginga Sulphureus (F1 sire)

The F1 surprisingly resulted in at least two distinct phenotypes from several litters.  One was very Ginga in appearance to the sire with the addition of yellow finnage as hoped.  From the available males the one above was selected to sire the next generation.  You will notice a residual amount of red in the upper caudal.  While I did not do test crosses, this likely was the result of Y-link mi as it does not manifest in my Line1 Vienna stocks.  Dorsal size and shape was near identical between father and son at maturity.  Moderate in length and tapering to a well defined point.  Retention of overall body pattern still hinted at Y-link &/or autosomal traits.  In both F1 phenotypes barring was present in 100% of males from litters of four (4) females to varying degress  This without a doubt says autosomal Z-bar and not sex-linked Tirgrinus.  It also suggests an autosomal dominant version of Ze and not a recessive version as my Line1 Vienna LS traditionally lack Ze.  In F2 backcross Ze is still present nearly 100%, albeit weakened in some males.  This continues to suggest that the Ginga are comprised of a autosomal dominant version of Ze and not a recessive.  You will notice the appearance of a green peduncle spot in this F1 male.  It would be hopeful to suggest an autosomal input from his Vienna dam.  It is more likely visible from a reduction in barring.  Z-bar is often epistatic to peduncle spots when bred for posterior coverage toward the caudal base.  

F1 Ginga * Vienna hybrid

I mentioned a second distinct F1 phenotype that appeared in both blond & grey siblings.  A single grey female likely was the dam of all.   I have gradually culled or given away the blonds, but kept back a lone grey male who I am now breeding to both pure Vienna and F2 Ginga Sulphureus females.  His finnage is very reminiscent to the types of guppies I bred as a youngster back in the late 1960's and early 1970's.  An interesting Lyretail with little extension and in appearance mimics many Aphyosemion killifish species.  It will be interesting to see how much of this caudal type was passed, from his pure Vienna dam.  I have always suspected that much of either Y or X-link DS trait is carried by females both in sex link and autsomal, and is masked by epistatic Y-link LS.

F1 Ginga * Vienna hybrid

Outside of his caudal fin, other body color / pattern components are still very Ginga.  Note that all F1 males of this phenotype lacked dorsal color pigment.  Either being  clear or lightly colored  by metallic (Mg) and this is probably in complex with the unique caudal shape.  If so, a possible Y-link as my females will pass X-link yellow dorsal nearly 100% when linebred.  These males also lack a green peduncle spot.  In both F1 phenotypes the amount of orange / pink spotting seemed to increase and intensify with age.  Both in anterior regions and over posterior barring.

Segregation in the F2 has come about in the expected ratios.  While Ze has remained strong, the degree of intensity (viz. total number of bars) has decreased in some males from several generations of backcross.  A mixture of blond / grey in both green and purple body mutation.  For F3 breeding groups of each have been set up with sibling females to further this new strain at least one more generation.  With emphasis on increasing what I am sure Kenjiro Tanaka would consider not only a flaw, but one of his greatest challenges in breeding this unique strain.  Removing or at least masking the peduncle spot while increasing both ocular caudal coverage, also known as "peacock spots", and the degree / intensity of Wingean orange / pink spotting.

F2 Ginga Sulphureus

F2 Ginga Sulphureus

F2 Ginga Sulphureus

F2 Ginga Sulphureus

F2 Ginga Sulphureus

F2 Ginga Sulphureus

Conclusions:
I come across quite a few comments in correspondence or on the web hinting at Ginga's deriving much of their phenotype from possible Poecilia Wingei (Endlers Livebearer) influence.  Surprisingly, all of the individual traits I identified, less some finnage, were already existent in either the Line1 / Line2 or both of my combined Vienna Emerald Lowersword (LS) strain.  Those few that are not common today, such as ocular spotting, were often found in guppy strains of the past.  This has allowed me to utilize Vienna in conjunction with a new direction in breeding Ginga's.  That being a reduction of red finnage in favor of pure yellow.  The primary trait in found in Ginga Rubra lacking in my Vienna LS is the layering of purple striping over Ze.  I also found that one of the most striking traits on the Ginga to be a seeming ability to express both Pb and green simultaneously.  Again, this trait while rare is also found in P. Reticulata strains.   The male on the right has no Ginga genetics in his makeup, yet posses the same ability to express both orange anterior and pink posterior.  Note that I have never run across an inversion of this combination, pink in the anterior and orange in the posterior.

F1 Ginga Sulphureus (l) and Schimmelpennig Platinum DS crossover male (r).
 Both expressing Pb posterior in peduncle and orange anterior in shoulders.

Breedings continue to indicated Pb as an autosomal autosome.  Similar to Asian Blau (Ab).  Analysis of several prior non Ginga breeding test results, consisting of Vienna / Schim / JB albino, and lack of Pb in Endlers point to green being a wild-type allele Pb⁺.  As Tanaka line breeds for Pb in Ginga Rubra, it is assumed both males and females are homozygous for the Pb trait.  Yet, no green presented in F1 from a pure Ginga Rubra single sire mating to 2 females (Ginga * Vienna LS).  It is doubtful both of these females were homozygous for Pb as their sire had been green.

I'm looking forward to a series of reciprocal Vienna LS * Ginga Sulphureus crosses a couple generations from now.  It will be interesting to see what autosomal traits will be contained in the genotype of linebred  Ginga Sulphureus females.  In the long-run when I work with a new strain the end result is not only to see what traits I can stabilize, but identify those which I can feasibly incorporate into my Vienna LS.

References:

Winge, O., Genetic Laboratory of the Royal Veterinary and Agricultural College, Copenhaen

(1922b).  "ONE-SIDED MASCULINE AND SEX-LINKED INHERITANCE IN LEBISTES RETICULATUS"

(1923).  "CROSSING-OVER BETWEEN THE X- AND THE Y-CHROMOSOME IN LEBISTES"

(1927).  "THE LOCATION OF EIGTHEEN GENES IN LEBISTES RETICULATUS"

Phang, V. et al., Dept. of Biological Sciences, Faculty of Science, Nat. Univ. of Singapore (1999). "Interaction between the Autosomal Recessive bar Gene and the Y-linked Snakeskin Body (Ssb) Pattern Gene in the Guppy, Poecilia reticulata"

Tanaka, K & Bias, A. S. (2012).  Email correspondences on Ginga Rubra

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Which do you find of more challenge as a breeder, creation and stabilization of a new phenotype or continued propagation and improvement of and existing one?

  Each a unique challenge in their own right.   

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Asian Blau Mutation As A Tool For Yellow Breeders

Asian Blau Mutation As A Tool For Yellow Breeders

© Alan S. Bias

Permission granted for nonprofit reproduction or duplication of photos and text with proper credit for learning purposes only.

Some of you may have noticed I've been on a crusade for a Yellow Swordtail phenotype over the last decade.  But not in a traditional sense of trying to create a fish with yellow pigment in the body.  I raised IFGA type Yellow Delta's for quite a few years back in the 80's and know such a phenotype does not exist.  These fish were pretty much a dull yellow finnage and body color a multi pattern in either green or purple body mutation.  Today the better Yellow Deltas are still blond and often incorporate Half Black, Platinum, Micariff or Metal (Mg) combinations.  So for the most part I've been content identifying and adding color patterns to the body in both my Lower Swords & Schimmelpennig Platinum DS / Lyretails with yellow dorsal / caudal in a study of population genetics.  

Blond Vienna LS

Blond Schimmelpennig Platinum DS

That is, up until a couple years ago.  Since then I've been focusing on crossover Platinum genetics, concentrating gold iridophores and metal (Mg) into my Lower Swords.  For some reason I have not infused Stoerzbach into my lines, although I have just obtained these genetics.  The Platinum crossover has come about from the several Schimmelpennig Platinum Double Sword lines I maintain via common usage of females across all sword lines.  In addition to the normal green peduncle, which results from Emerald Green Iridescence (EGI), many breeders are familiar with "blue peduncle" in Vienna Swords as an autosome.  A third variant I been able to identify is "yellow peduncle".  Unfortunately,  it has been in a rather rigid complex for with a blunt sword shape even after numerous outcrosses.  Only now am I starting to see some improvement in mature males. 

Yellow Peduncle Blond Vienna LS w/purple body mutation

Yellow Peduncle Blond Vienna LS w/purple body mutation

Most breeders of swordtails still prefer clear caudals and dorsals in females as an indicator of her ability to enhance swordshape.  As a rule I still adhere to this principle, though find modern swords can successfully bend this old adage.  I do select for expression of such yellow genes as Flavus or presence of metal in female finnage.   These females will in turn pass the traits as visible expression in sons.  Increase your knowledge of both X-link and autosomes.  By doing so you can reduce your reliance on Y-link genetics.

So, when recent publications and research indicate creation of a yellow pigmented swordtail guppy as unlikely why continue? Yellow simply appeals to my eye.  In addition, we have a host of new genetic tools that allow us to bypass yellow pigment in the body.  Thus we can mimic what appears to be a yellow guppy.    You may have noticed my increasing interest in the Asian Blau (Ab) mutation for the last couple years.  Yeah, I can hear the rumblings in the background already, likely for the same reasons this particular mutation never held much attraction for myself.  From preconceived notions it just seems to be a blue / green pattern that would "washout" most of the color in any given strain.  Resulting in similarities found in a combined Red / Blue Grass strain.  Yet, as I wrote about in my last article on the influence of Asian Blau on Japan Blue Swords, this is not necessarily the case.  Especially if one's focus is on incorporating both X and Y-link yellow pigment and autosomal iridescence from metal (Mg) into your strains.  I have managed to breed out most of the genetics for red fins in my lines, and in where I haven't yellow is normally epistatic to red. 

Asian Blau Vienna Emerald Lower Sword

From planned mating's and analyzing the results it has become clear that Ab does not just simply remove red from a phenotype.  A goal oriented breeder should always keep in mind that color in a guppy is regulated in multiple zones within both the body and finnage.  Various zones can often seem larger than they actually are when a gene complex combines areas of independent regulation into seemingly larger ones.  Not only are each of the zones in the body often under independent control, but also the caudal and dorsal.  To go one step further various portions of the caudal and dorsal can be controlled as distinct regions from a host of identified genes.   

Asian Blau Vienna Emerald Lower Sword

What I realized early on working with Vienna pattern swordtail, which are pretty much a glorified multi pattern, is that color in each zone can be amplified depending upon how they come about.  Do they result from simple color pigment or the presence of Mg or a combination of both to create iridescence found strains.  To produce brilliant colored guppies you must concentrate on iridescence over pigment.  Depending upon your strain it may result in some gaps in color coverage, but the results can be phenomenal.  To create a guppy that glows almost "neon" an additional input is needed.  One that highlights iridophores and Mg.  This is what I have found to be the secret of Ab when you view it as a mutation that adds to effect and not simply as one which removes a portion of existing phenotype.  Ab has the ability to mask dull pigment and enhance the effect of Mg and iridophores.

 Ab effect is "overall" on any particular phenotype, or to state in another way, "it modifies all color / pattern on the body evenly".  At first I thought just the opposite as Ab appeared to have zones of effect.  Further breedings revealed these apparent differences in any given male to result from the following reasons:  1.  Absence or presence of yellow color pigment (or any color pigment);  2.  Absence or presence of Mg;  3.  Color of visible iridophores (silver / blue / gold);  and 4.   Independent control of color in body and fins. 

Asian Blau Vienna Emerald Lower Sword

In contrast to European Blau (Eb), which is considered autosomal recessive, Ab has been thought of in simple terms as autosomal partially dominant.   Disregarding homozgous Ab individuals, what is particularly interesting about Ab are the presence of males, often in F1 from blau * non-blau matings, who seem not to fit into two basic catagories of blau or non-blau.  Such males exhibit two orange Wingean spots (sometimes a bit lighter than normal), but the body color just seems "not what you would expect".  I refrain from using the term "washed out or diluted" as they are not.  They also seem to lack yellow pigment &/or show reduced Mg in finnage.  Resulting in a blue / green appearance.  These variants hint to Ab being more than "simply partially dominant" as subsequent offspring from this type male when backcrossed to non-blau females (with no blau in the background) has produced at least one regular blau offspring.  This surprised me.   I think if one could produce similar results, while adhering to strict scientific method and analysis of breeding results, it would possibly show a "plus effect" to Asian Blau (Ab+) dominance.

Grey Ab Vienna Emerald Lower Sword variant

I know that there are those breeders who in a quest to produce a yellow phenotype, or any solid (or predominant) color,  would say that it must be in a grey body to be meaningful.  That in a blond (gold) form is just masking imperfections, such as visible removal of black which is always found in complex with yellow pigment.   While a valid argument from one perspective, it is also flawed for a serious breeder should use any non lethel genetic trait as a tool in a quest to produce desired results.   After all, the more traits we identify & combinations we create, the more we will find that some combinations or results are just not scientifically feasible.  So use your genetic tools to their maximum potential.

When all is said and done my goals have led me to focus in two distinct directions in a quest for a Yellow phenotype.  The first has resulted in Asian Blau Blond Vienna Lower Swords and the second Asian Blau Blond Schimmelpennig Platinum DS.  Please note:  These photos have not been color enhanced in any form or manner.

Asian Blau Blond Vienna Emerald Lower Sword

Asian Blau Blond Vienna Emerald Lower Sword

Asian Blau Blond Schimmelpennig Platinum Veil Tail

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Random exploration of genetics through test breedings can often prove fruitful in the short-term.

  Yet,   for any long-term breeding program to be successful and endure it must first have defined goals and objectives to preserve not only core genetics, but breeder interest.

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Asian Blau Japan Blue Swordtails

Asian Blau Japan Blue Swordtails

© Alan S. Bias

Permission granted for nonprofit reproduction or duplication of photos and text with proper credit for learning purposes only.

April 9, 2012

Mixed tank of pure JB and  JB * Vienna outcross with X-link yellow

Introduction:

When a new strain of guppies or a variation upon one makes an appearance it can seem natural to credit its existence on newly discovered genetics.  Occasionally this is the case.  More often than not after careful test breeding and analysis the truth is revealed to be new combinations of already identified genes.

A year and a half ago I once again gave in to the appeal of Japan Blue (JB) and a chance to work with this alluring phenotype.  My first hands on experience with JB came back in 2004.  I started with a typical grey wild-type strain with a clear roundtail lacking extension or caudal color.  Combined with females from an X-link Lace Snakeskin Double Sword strain it was easy to produce, and then fix a nice line of Variegated (Var) JB Double Swords after 2-3 generations.

  Variegated (Var) JB Double Swords

Over the last couple years a line of Japan Blue Swordtails has been making the rounds on Aquabid.  Inquiries with several sellers as to origins were met with the same vague responses that hinted to importation from Asian Breeders.  What drew me to this strain were two particular traits:  1. First and foremost they were swordtails to varying degrees.  2.  Second intense iridescence and what appeared to be more than normal blue coloration into the anterior portion of the body.

Discussion:  

Accepted theory states that JB was initially identified as a Y-link mutation found in a feral population of wild-type guppies in Japan.  Infrequent crossover to X-link has since been identified in a few strains.  Blue patterns in guppies is not limited to expression as a result of JB.  Recently, in Venezuela, French breeder Frederic Nicole collected a stunning new blue phenotype in a population of Endlers Livebearer.  Blue covers the upper quadrant along the entire length of the peduncle and then some.  It seems to have more in common with Lazuli than Japan Blue.  In my fishroom there has been only one suspected instance of X-link JB in the last 10 years.  Interestingly, the incident involved ½ a dozen males in a single drop of about 12 males in total from a Schimmelpennig Platinum Double Sword * Lace Snakeskin Top Sword breeding, and not an isolate individual.   I had culled the dam prior to sons coloring up and worse failed to capitalize on the potentially X-linked JB type pattern for lack of tank space at the time, or so I decided.  This was a very poor decision in hindsight.

X-Link Japan Blue type pattern

In any JB strain the pattern is almost exclusively ½ body with a clear line of demarcation starting just below the base of the dorsal fin, or slightly before, and running the length of the peduncle.  The exact location often seems relevant to the level of improvement found in a strain.  There also seems to be a direct correlation between Emerald Green Iridescence (EGI) determining how much the pattern bleeds into the caudal.     

Other factors can influence the intensity and shade of blue coloration found in a Japan Blue male.  The addition of yellow pigment in the body from genes such as vitellinus (vi) can modify appearance from blue to green or even yellow.  The addition of red pigment can dilute from blue to lavender.  

 Blond JB * Vienna Lower with X-Link yellow and metal

Based in part on these facts the perceived increase in blue coloration into the anterior portion of the body in this JB strain should result from additional additive effect or other genes.  Not the JB itself. 

Test Breedings:

In brief email correspondence I requested only males exhibiting a very long lower sword and if possible dorsal color of any type.  The seller obliged on the first request, but was unable to provide any males with dorsal color as the strain did not have any.    My initial approach would be several generations of sib breeding to identify existing traits in pure form.  This revealed a percentage of males expressing “blue lips”.  Colored lips are often found in metal / platinum strains.  These males also tend to express the most intense blue and increased level of coverage.  Males exhibiting a lower sword would generally pass the trait to sons to varying degrees in shape and length.

Japan Blue male with hyaline dorsal

In multiple purebred litters over 3 generations a portion of males would posses lower swords and a portion fairly mismatched double swords.  The reminder being clear roundtails.  This indicated not all the JB females were homozygous for producing or enhancing sword shape.  Many of the roundtails and lower swords also showed oculatus (oc) spotting.   Sib breeding also produced about 25% males with a red shoulder stripe.  The presence of Zebrinus Bar (Ze) was evident as secondary sex link colors started to manifest in young males.  This was quickly covered over as JB made its appearance.  At the time, while suspecting it to be autosomal dominant Zebrinus Bar (Ze), I could not rule out autosomal recessive Tigrinus (Ti).

Autosomal Zebrinus Bar, as later identified

Less than 1 in 10 males would exhibit some patchy black dorsal color.  Purebred sib breeding also told me the hyaline (clear) dorsal resulted from a lack of X-link dorsal color or it was epistatic to any existing genes for dorsal color.  Therefore, likely near impossible to eliminate without outcross.  Hence, making an outcross with my Vienna lowers a logical choice as they posses both Y and X-link for yellow pigment and gold iridophores in caudal / dorsal and to some degree.  At this point I discontinued pure breeding the strain with better lower males resembling the individual below.

 Asian Blau Japan Blue Lower Sword with hyaline dorsal

Reciprocal outcrosses to my Line 1Vienna Lower lines helped substantiate several developing theories about this JB strain.  It showed JB females were actually contributing not only to caudal fin length, but the overall shape itself.  Results hinted at fin extension being mostly X-link for swords, regardless of type.  Notice that I say extension and not the sword itself.  Sib breeding of F1 Vienna Lower * JB revealed in the F2 and F3 via recombination, that the foundation JB females often only carried extension on a single X-link or non at all.  I note that not a single incidence of bar has occurred in my Line 1 Vienna Lowers in over a decade, though I breed for it regularly in my Line 2.  Reciprocal crosses between JB and Line 1Vienna Lower showed the barring to be autosomal dominant Ze as it appeared in both directions of F1 offspring.  So this should eliminate further consideration of Ti as the source of barring.

Lacking tanks space for breeding the double swords I have culled them for the most part.  While I have done a few test breedings to evaluate, outcross with Vienna Lower females and recombination in the F2 / F3 hinted at two forms of doubles.  The best matched possibly resulting from a single X-link and another resulting from a combination of X-link top sword and X or Y-link lower sword.  The latter version likely in complex as no pure Top Swords have ever presented.

Segregation and recombination in F2 / F3 crosses (from both reciprocal crosses) in addition to F1 * Vienna backcross (BC) revealed two concerns.  First and foremost the JB Y-link lower sword does not appear completely dominant to either type of double sword or recurring clear roundtails, though the latter phenotype has been reduced in occurrence.  I would not be surprised if better defined test breedings of doubles found them to be epistatic to lowers as is common with Vienna Swordtails.  Second, optimal lower sword shape is not exclusively controlled by Y-link alone.  Suggesting JB X-links &/or autosomal input for sword shape and length.

The continued use of Line 1 Vienna lower females for BC creates an issue all to itself.  It produces JB males that are a definite shade of Apple Green or a blue with distinct regions of yellow pigment or gold iridophores.

 F1 Grey JB * Vienna Lower with X-Link yellow influence

Two of the first traits I wished to test for / rule out were Stoerzbach and Lazuli.  I see no evidence of Stoerzbach to date.    While I rule out Lazuli as unlikely, I do see similarities to this trait;  1.  Blue coloration in the upper shoulders of some males.  2.  Variation in shades of blue on individual males in distinct regions.

Reciprocal outcrossing to my Line 1 Vienna Lowers has revealed the true secret to this strain.  This being the presence of autosomal Asian Blau (Ab), which is partially dominant.  It not only helps add to the additional anterior color, but the overall color itself.

            I.    Abab (heterozygous) Asian Blau

            II.   AbAb (homozygous) Asian Blau

            III.  abab (non Asian Blau)

I.  Unlike European Blau (Eb), Abab heterozygotes can express yellow pigment and possibly gold iridophores in body and finnage when the genes are present. 

 Asian Blau Japan Blue with X-link yellow

II.  AbAb homozygotes remove / inhibit red pigment to reveal silver (or white) or blue depending upon the type of irridophore lying beneath the color pigment.  As Japan Blue is a mutation of blue irridophores it seems there should be little or no silver in the peduncle.  As seen below in grey form, AbAb fish appear blue/grey and in blond double recessive white.  Epistasis likely prevents formation of swords and colors are limited to blue iridophores, possibly a theoretical gold as evidenced by the presence of green in the body.  

Homozygous Asian Blau Japan Blue (Grey on left & Blond on right)

III.  abab Japan Blue males (those lacking the Asian Blau gene) may show a red stripe on the shoulders or caudal outside the region of influence in traditional JB.

Japan Blue lacking Asian Blau

Reciprocal crosses with Vienna Lower * Abab JB females show they also pass extension genetics to the F1 males.  With selection this enhancement has been easily retained to a high degree in F2 and F3 sib-breedings.  Having worked with Y-link lower swords for many years I am of the opinion that the best length and shape is only possible in combination with the additive effect of X-link &/or autosomal genes.  I believe this is often taken for granted by many swordtail breeders working predominantly with X-link double or top swords in which the majority of extension genetics are also passed in a tight linked gene complex.    

Few if any colors / phenotypes are the result of a single gene in guppies, more likely multiple genes by regulation(s).  The initial reciprocal breeding involved my Line 1 using a Blond Vienna Lower * JB.  It produced predominantly Abab males in both green and purple body mutation.  This high degree of autosomal dominance has maintained in subsequent Vienna breedings.  It likely expresses similar in JB.  You will notice below in these Asian Blau Vienna Lower normally “Orange” Wingean spots are still visible, with red pigment removed, solely by remaining iridophores which are the basis for patterns.  Prior to outcross my Line 1 Lowers did not contain Ze, yet barring is present in these F1 males.  Thus, further evidencing the trait was passed by JB females as an autosome.

Another visible trait found in both my Line 1 and the JB, is a very blue belly patch.  Resulting from blue iridophores it has been little modified by the presence of Ab.  Passed by males and females, this trait helps ad to the additional anterior color of the JB strain in combination with Ab.  A similar related effect can manifest above the lateral line on the shoulder.  However, it is more often evident in conjunction with a blue belly patch.

Asian Blau Vienna Lowers w/blue belly

The male below is multi generation pure from my Vienna Lower Line 1.  There seems to be a correlation between expression of the blue belly trait and EGI.  You will notice in this individual how the blue is also found in the lower peduncle.

Normal non Asian Blau Blond Vienna Lower w/blue belly

One of the hypotheses I wished to test for was the presence of Wild-Type Wingean Super gene traits being masked by Japan Blue.  To do so I employed a simple “fright test” and moved males directly from their tank into my photo tank without a period of acclimation.  You will notice that these males immediately revealed an orange spot in mid body.  While not visible in this photo a second spot was also present in the peduncle.   

 Japan Blue males without period of acclimation

Conclustions:

Within this JB strain in any given litter, both purebred and outcross with Vienna Lower, exists a broad range of phenotypic expression.  The overall basis for both the increased level of coloration and its intensity in this strain results from Asian Blau (Ab).  Additional color enhancement comes from a “blue belly” created by blue iridophores in conjunction with EGI.  A lack of yellow pigment or gold iridophores in the body further enhances coloration.  Each trait is independent of traditional Japan Blue (JB) and results from either X / Y sex link or autosomal genes.

  

 Asian Blau Japan Blue with X-link yellow

 Asian Blau Japan Blue with X-link yellow, including blond on the right

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Enjoy the strains you have labored to create?  Pass them around so others can do the same and maybe further your efforts.  Never know when you might want some back …

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Science as A Tool and Benefit to Guppy Breeders…

Science as A Tool and Benefit to Guppy Breeders…

© Alan S. Bias

Permission granted for non profit reproduction or duplication of photos and text with proper credit for learning purposes only.

January 11,  2012

Grey Vienna Emerald Lower with Blue Peduncle

Outcrossing is often offered as a cure all for a declining strain showing the effects of inbreeding. However, this is not the whole story. Just as the negative aspect of inbreeding can be described as inbreeding depression, outcrossing can result in outcrossing depression. What is “out-breeding depression?”  The answer can be found in one of the many tools available to the successful guppy breeder.  Science… Out-breeding depression is a loss of robustness due to reproduction among very dissimilar individuals; characterized as hybrid breakdown.

Out-breeding to a substantially different strain can break up the gene complexes needed to produce the initial traits a breeder is selecting for.  Science is not the cure all for a guppy breeder.   It is but one of many tools that can be used to balance out a successful breeding program for show, research, preservation or plain eye appeal.

Grey Asian Blau Lower with Z-Bar and Blue Peduncle

Understanding of traits we now take for granted such as blond (gold), albino, snakeskin, basic caudal extension all resulted from breeding experiments, analysis and testing of hypothesis.  Breeding guppies is already a science for those who take it seriously and are willing to admit it.

As a breeder I view guppies as a genetic puzzle to be decoded and expanded upon through phenotype.  Over the last 100 years our knowledge as grown by leaps and bounds.  Try to imagine the entire potential of the guppy genome.  You cannot.  Each of us is bound by our narrow focus on not only how we describe a quality guppy, but the breeding technique(s) we use to obtain them.  If a breeder is not careful self imposed limitations may hinder our interpretation of accrued knowledge from all sources and its possible uses.  To further compound events as breeders we often do not document results of our crosses very well. 

Breeders should consider the possibilities as they present themselves.  In example; a cull is not just an individual who fails in expectations.  You can still learn from it.  Did it come about from crossover or simple recombination of alleles?  Worse yet, will a gene complex inhibit your desired goals or is a form of gene regulation preventing it?

2.5 month old Asian Blau Blond & Grey Vienna Lower 

There are many elements that compose the whole of breeding domesticated guppies.  They include:  purebred, hybrid, research, enhancement, sale, and show to name a few.  Value of each is determined by personal interests.  Therefore, no single component or the resulting fish is of greater value than another in genotype if healthy and viable.  Breeder preference should not be misconstrued to suggest otherwise.  In each case the primary criteria for selection of potential stock should be limited to:  “Has it been bred toward goals that reflect my intended use and will it reproduce to type?”

Preservation, Improvement, and Promotion are considered the primary functions of a breed association. Showing is normally considered a by-product of purebred stock breeding, and is prohibited by some tasked w/preservation of gene pools to avoid judgment that results in inflated value.

Show stock is an expression of a particular phenotype and not always a reflection of breeding value.  Only a very small percentage of any purebred (registered) population is considered to be of show quality.  It takes the use of diverse breeding techniques by individual breeders to promote overall population health.  The value of purebred strains is to produce predictable results within itself and for infusion into that part of a population which does not.

Many authors continue to profess the benefits of out-breeding as a cure all for a declining strain showing the effects of inbreeding.  Normally I refrain from directly critiquing others, even when positive in intent.  Last month on one of my regular web rambles involving “Guppies” as part of the search criteria I ran across an article posted to the Malaysia Guppy Forum.  Originally published in the IFGA eBulletin Volume 7 # 7 July 2011, and titled “Follow the Best Genetics by Jim Alderson.”  It was originally a short article written by a prominent IFGA officer and breeder that has had me pondering ever sense.  Though vague in use of terms, it was not so much the content of the article that created issue for me as the perceived intent of summation.

As a linebreeder, in general, I could agree with much the author stated if his implication was his successful hybrid crosses are limited to highly inbred IFGA strains.  Broad based statements lacking clarity of definition are subject to a wide range of interpretation.  Crosses involving non IFGA strains or even dissimilar IFGA strains have the potential to be subject to out-breeding depression.  As a result, there are several conclusions offered by Alderson where I would disagree based not only on personal experiences as a breeder, but backed by scientific knowledge.

An example is: “Selecting more dissimilar fish will give you a wider range of variability and more hybrid vigor.”  Unless the intent was to introduce traits not already found in my strain, in this instance one could state that, “such an approach is just as likely to result in out-breeding depression in a fixed strain.”

In another quote, “A more sustainable approach is to use out-crossing to instill new genes into your pure lines and make rapid improvement in your stock. Remember, every time you out-cross you are improving the hardiness and disease resistance of the the fish as well.”  One could easily state that, “such an approach is just as likely to result in out-breeding depression in a fixed strain by reducing or eliminating positive benefit derived from already existing co-adaptive gene complexes for disease resistance.”

Grey Schimmelpennig Platinum Lyretail

While we lack corresponding “registration papers,” as found in animal breed associations, with line-bred Guppy strains an old adage in the Pedigree Livestock World is very applicable.  It simply states, “A pedigree is only as good as the breeder who produced it.”   This is a personal preference based on not being interested in creating hybrids for show.  As a result I seek out methods that will reliably produce sound pedigreed breeding stock and promote the results.  I would not take a route that derives short-term result from long-term failure.  Genetic dead-ends are of little value in a breeding program.

Some years back I wrote a couple articles for the sheep world that were published by several breed groups:   Breeding Strategies and Genetic Manipulation ~and~ Breeding Strategies and Genetic Consequences.  Recently, after review, retired biology professor Richard “Rick” D. Squire, Ph. D., and now active guppy breeder felt I should rework them for the benefit of other breeders.  Here are the links to each as rewritten:    

                                                                                                                                      

Breeding Strategies and Genetic Manipulation in Guppies

~ and~

Breeding Strategies and Genetic Consequences in Guppies 

There has always been a view in show circles that claims many new breeders are successful until they step out of the box, break from established practice and try to do things on their own.  This seems to indicate much initial success results from starting off with reliable strains produced by those breeders who utilize all tools available.  Tools which can provide definition to the more static results we often observe as breeders to provide forethought in planning matings.  Science is one of them. 

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In the end what was in Alderson’s summation that was so perplexing it led me to spend several weeks procrastinating a response?  It stated:  Guppy genetics is not a science. Guppies are a tapestry that we can learn to paint with a few genetic principles, trial and error, and records. Those who claim to study guppy genetics do so by making the crosses and dissecting the results, not by predicting the outcome in advance, claiming to have discovered the inheritance of a new trait, until the next cross is made that does not follow the rules. Breeding guppies may someday be a science, but I will be pushing up daisies by then.

Having thought about this statement for some time, it dawned on me what I found so disconcerting is that a prominent IFGA officer and breeder would not only appear to frown upon, but downplay one of the organizations founding principles as stated: 

IFGA Constitution

ARTICLE 2 – PURPOSE AND OBJECTIVES

Section 2.1 – Purpose

            2.1.1    To promote interest in, and scientific knowledge about guppies.

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