Co esponding au ho : Kamil ATSATAN
Copy igh © 2025 Au ho (s) e ain he copy igh o his a icle. This a icle is published unde he e ms o he C ea i e Commons A ibu ion License 4.0.
Gene ic Di e si y o Pe ca lu ia ilis Ac oss Tu kish Lakes and Dams
Kamil ATSATAN 1, * and İb ahim DİLER 2
1 Depa men o Fishe ies and Sea ood P ocessing Technology, Facul y o Aquacul u e, Ispa a Uni e si y o Applied
Sciences, Ispa a, Tü kiye.
2 Depa men o Bioenginee ing, Facul y o Enginee ing and Na u al Sciences, Bu sa Technical Uni e si y, Bu sa, Tü kiye.
Wo ld Jou nal o Biology Pha macy and Heal h Sciences, 2025, 24(01), 032-044
Publica ion his o y: Recei ed on 25 Augus 2025; e ised on 01 Oc obe 2025; accep ed on 03 Oc obe 2025
A icle DOI: h ps://doi.o g/10.30574/wjbphs.2025.24.1.0886
Abs ac
This s udy aimed o in es iga e he gene ic di e si y, phylogene ic ela ionships, and oxida i e s ess esponses o Pe ca
lu ia ilis popula ions collec ed om 12 lakes and dams ac oss di e en egions o Tu key. Molecula cha ac e iza ion
was pe o med using ou ISSR ma ke s, yielding 100 sco able bands, 68% o which we e polymo phic. Clus e analyses
(UPGMA) and P incipal Componen Analysis (PCA) e ealed h ee main gene ic g oups, wi h he Adana popula ion
showing signi ican isola ion and gene ic di e en ia ion om o he popula ions. Mi ochond ial DNA analyses a ge ing
ATP6 and cy b gene egions con i med species iden i y and demons a ed limi ed in aspeci ic a ia ion, while closely
ela ed species (P. sch enkii and P. la escens) o med dis inc clades. O e all, his s udy p o ides comp ehensi e
insigh s in o he gene ic and physiological di e si y o P. lu ia ilis popula ions in Tu key and emphasizes he
impo ance o combined molecula app oaches o sus ainable managemen and conse a ion o pe ch ish popula ions.
Keywo ds: ATP6 Gene; cy b Gene; En i onmen al Adap a ion; Mi ochond ial DNA; ISSR Analysis
1. In oduc ion
The Eu opean pe ch (Pe ca lu ia ilis L.) is a medium-sized eshwa e ish species wi hin he amily Pe cidae (McDowall
1996). The axonomic his o y o he species da es back o he ea ly 18 h cen u y, when i was ini ially desc ibed om
Swedish lakes by Pe e A edi in 1730 and subsequen ly inco po a ed in o he o mal binomial nomencla u e by Ca l
Linnaeus in 1758, la gely on he basis o A edi’s pionee ing ich hyological wo k (Tho pe 1977; Pimakhin e al. 2015).
The amily Pe cidae ep esen s a di e se axonomic g oup, cu en ly comp ising 10 gene a and app oxima ely 195
species dis ibu ed ac oss Eu asia and No h Ame ica (Be a 2001). Fossil e idence sugges s ha he e olu iona y
o igin o he genus Pe ca can be aced back o he ea ly Miocene, a ound 19.8 million yea s ago, indica ing a long
adap i e his o y wi hin eshwa e ecosys ems (S epien e al. 2015).
Due o i s b oad geog aphic dis ibu ion, high ecological plas ici y, and signi ican ole in bo h ec ea ional ishe ies and
aquacul u e, P. lu ia ilis has been he ocus o a subs an ial body o scien i ic li e a u e (Pimakhin e al. 2015). Resea ch
has add essed a ious aspec s o i s physiology, g ow h pe o mance, eeding ecology, and aquacul u e po en ial,
e lec ing he species’ economic and ecological impo ance. Howe e , despi e his ex ensi e in e es , de ailed and
in eg a i e s udies ha speci ically in es iga e he biology, ecology, and adap i e mechanisms o he species ac oss
di e en en i onmen s emain ela i ely limi ed. Such comp ehensi e in es iga ions a e c i ical no only o enhancing
ou undamen al unde s anding o P. lu ia ilis, bu also o iden i ying exis ing knowledge gaps, add essing
me hodological cons ain s, and cla i ying he ecological d i e s unde lying i s wide dis ibu ion ac oss di e se
eshwa e habi a s. Mo eo e , imp o ed insigh s in o he species’ ecological in e ac ions and en i onmen al ole ances
would con ibu e o mo e sus ainable managemen s a egies in bo h na u al popula ions and aquacul u e sys ems.
Wo ld Jou nal o Biology Pha macy and Heal h Sciences, 2025, 24(01), 032-044
33
Geog aphic a ia ion in gene ic ai s has been documen ed ac oss he dis ibu ion ange o P. lu ia ilis (Nesbo e al.
1999; Vanina e al. 2019). Fo ins ance, Nesbø e al. (1999) in es iga ed gene ic a ia ion in 55 Eu opean popula ions
and one Sibe ian popula ion using mi ochond ial DNA (m DNA) D-loop sequencing and RAPD ma ke s. Thei esul s
e ealed high le els o gene ic di e en ia ion among wa e sheds and egions, sugges ing ha p esen -day popula ions
in Wes e n and No he n Eu ope o igina ed om h ee main e ugia: No heas e n Eu ope, Sou heas e n Eu ope, and
Wes e n Eu ope. Simila ly, Vanina e al. (2019) analyzed gene ic di e si y in pe ch popula ions om Poland, he Czech
Republic, and Slo akia using ou mi ochond ial ma ke s and ound signi ican gene ic di e en ia ion among
popula ions, wi h he Polish popula ion being pa icula ly dis inc .
These gene ic di e ences a e o en associa ed wi h pheno ypic a ia ion among geog aphically dis inc popula ions
(Mandiki e al. 2004; Méla d e al. 2004; Vanina e al. 2019). Fo example, Méla d e al. (2004) compa ed g ow h
pe o mance unde con olled expe imen al condi ions and demons a ed ha ish o igina ing om no heas e n
F ance exhibi ed 76% highe body weigh a day 200 compa ed wi h hose om no he n I aly, while Belgian
popula ions ou pe o med sou hwes e n F ench popula ions by 56%. Likewise, Mandiki e al. (2004) obse ed
di e ences in g ow h a es and su i al among Eu opean pe ch popula ions, wi h sou he n Eu opean popula ions
showing he lowes g ow h po en ial and su i al. Howe e , i is impo an o no e ha he associa ion be ween gene ic
and pheno ypic di e gence does no necessa ily imply causa ion, as pheno ypic plas ici y may play a signi ican ole in
shaping obse ed pa e ns (Be gek and Bjö klund 2009).
Gene ic ma ke s a e undamen al ools o assessing gene ic di e si y in ish popula ions (Rashed e al. 2008) and a
wide ange o o he o ganisms. Among hese, in e simple sequence epea (ISSR) ma ke s ep esen a dominan ,
polymo phic, apid, and ela i ely cos -e ec i e molecula ma ke sys em based on he ampli ica ion o DNA egions
be ween mic osa elli e loci. ISSR ma ke s ha e p o en o be e ec i e o e alua ing he gene ic s uc u e o ish,
p o iding aluable insigh s in o species iden i ica ion, gene ic di e si y assessmen , popula ion s uc u e analysis, and
b eeding p og ams. In eg a ing such molecula echniques in o he managemen o bo h wild and cul u ed ish s ocks
con ibu es o e idence-based decision-making p ocesses (Saad e al. 2009). In his me hod, sho p ime s
complemen a y o mic osa elli e egions a e employed in polyme ase chain eac ion (PCR) ampli ica ion, and he
esul ing banding pa e ns a e analyzed o e eal gene ic a ia ion among indi iduals (Zie kiewicz e al. 1994; P adeep
Reddy e al. 2002; Mal aglia i e al. 2006). ISSR ma ke s ha e been widely applied o in es iga e gene ic di e si y,
popula ion s uc u e, phylogene ic ela ionships, and geog aphic di e en ia ion in ish species. This app oach is
pa icula ly ele an o he conse a ion o na u al s ocks, axonomic classi ica ion, and ishe ies managemen
(Elsayed e al. 2024; Khandanizadeh e al. 2025; Song e al. 2025).
The p esen s udy aims o in es iga e he gene ic di e si y and physiological esponses o P. lu ia ilis popula ions
ac oss a ious lakes and dams in Tü kiye. Gene ic di e si y and popula ion s uc u e a e assessed using In e -Simple
Sequence Repea (ISSR) ma ke s, while mi ochond ial DNA egions (ATP6 and cy b) a e analyzed o elucida e
phylogene ic ela ionships and species-le el gene ic homogenei y. By in eg a ing molecula app oaches, his s udy aims
o p o ide comp ehensi e insigh s in o he e ec s o geog aphic dis ibu ion and habi a - ela ed s esso s on he
gene ic and physiological heal h o pe ch popula ions, con ibu ing o hei sus ainable managemen and conse a ion.
2. Ma e ials and me hods
2.1. Ma e ials
Fish samples we e collec ed om wel e di e en pe ch bodies in Tü kiye, including Da ıde esi Dam (Ispa a), Yedikı
Dam Lake (Amasya), Ü kmez Dam (İzmi ), Tah alı Dam (İzmi ), Seyhan Dam (Adana), Şeyi le Dam (A yon), Denizli
Pond (Kocaeli), Ka aağaç Pond (Uşak), Hi anlı Dam (Kı şehi ), Çamlıde e Dam (Anka a), Al ınapa Dam (Konya), and
Ladik Lake (Samsun). Sampling was ca ied ou using gill ne s wi h a mesh size o 4 mm. F om each loca ion, 25
indi iduals we e ob ained, esul ing in a o al o 300 specimens used in he s udy.
2.2. Me hods
2.2.1. Phylogene ic Analysis and Molecula Cha ac e iza ion
Genomic DNA was ex ac ed om caudal in issues using he The mo Scien i ic GeneJET Genomic DNA Pu i ica ion Ki
(Ca alog No: K0721), ollowing he manu ac u e ’s p o ocol op imized o his s udy. The quali y and in eg i y o he
ex ac ed DNA we e e i ied by elec opho esis on 2% aga ose gels. Polyme ase chain eac ion (PCR) ampli ica ions
we e pe o med using p o ocols and p ime s p e iously desc ibed by Ragauskas e al. (2023), Bache skaya e al.
(2023), and Alyamkin e al. (2022). De ailed PCR condi ions and he lis o p ime s employed in he s udy a e p esen ed
Wo ld Jou nal o Biology Pha macy and Heal h Sciences, 2025, 24(01), 032-044
34
in Table 1 and Table 2. PCR p oduc s p oducing single, co ec ly sized bands we e pu i ied using he GenElu e™ PCR
Clean-Up Ki (SIGMA, Ca alog No: NA1020) ollowing he manu ac u e ’s ins uc ions. Pu i ied p oduc s we e e-
checked on 2% aga ose gels o ensu e he absence o nonspeci ic bands o p ime dime s be o e sequencing. Sequencing
was pe o med by Olygome Bio echnology A.Ş. The esul ing sequences we e p ocessed using Sequenche 4.5, wi h
o wa d and e e se eads aligned and manually co ec ed o gene a e consensus sequences o he m DNA ATP6 and
cy b egions. Sequences we e con e ed o FASTA o ma and aligned using MEGA 6.0 (Tamu a, 2013). BLAST sea ches
agains he NCBI da abase con i med sequence accu acy. Phylogene ic ela ionships and gene ic dis ances among
popula ions we e in e ed using he Neighbo -Joining me hod in MEGA 6.0, and nucleo ide composi ion was calcula ed
o each popula ion.
Fo molecula cha ac e iza ion, PCR p oduc s we e loaded on 2% aga ose gels and elec opho esed a 80 V o 120 min,
ollowed by isualiza ion unde a UV imaging sys em. Band p esence o absence was sco ed o gene a e a bina y (1–0)
ma ix, and simila i y indices we e calcula ed using he Dice coe icien . Clus e analysis was hen pe o med using he
Unweigh ed Pai -G oup Me hod wi h A i hme ic Mean (UPGMA) in NTSYS so wa e e sion 2.02 (Rohl 1997), and
dend og ams we e cons uc ed o illus a e gene ic ela ionships among popula ions.
Table 1 ISSR ma ke s used in molecula cha ac e iza ion
P ime Name
P ime Sequence
PCR Con en
PCR ampli ica ion
UBC-807
(AG)₈Т
5 μL Genomic DNA, 1 μL
p ime s, 5 μL Mas e mix
(PZR bu e , 2mM MgCl2,
250 µM dNTP, 0.75 U Taq
DNA polyme ase) e 13 μL
dH2O. (To al 25 μL)
95°C/2 min (Ini ial dena u a ion 1 cycle)
UBC-808
(AG)8C
94°C/30 sec
47–53°C/45 sec
72°C/45 sec
(35 cycle)
UBC-809
(AG)8G
72°C/5 min (Final ex ension: 1 cycle (
UBC-823
(TC)8C
Table 2 P ime s used o ob ain m DNA sequences
P ime
Name
P ime Sequence
PCR Con en
PCR ampli ica ion
m DNA
ATP6
F: 5’-CCTAACGAGCCTACATCCC-3’
R: 5’-TGTAAGAGGTCAAGGGCTGG-3’
5 μL Genomic DNA, 1 μL
p ime s, 5 μL Mas e mix
(PZR bu e , 2mM MgCl2,
250 µM dNTP, 0.75 U Taq
DNA polyme ase) e 13
μL
dH2O. (To al 25 μL)
95°C/2 min (Ini ial
dena u a ion 1 cycle)
m DNA
cy b
F: 5’-GTCATAATTCCTGCCAGGATTTTAACCAGG-3’
R: 5'-TTTAGAATCCTAGCTTTGGGAGTTAGGGG-3'
94°C/30 sec
47–53°C/45
sec
72°C/45 sec
(35
cycle)
3. Resul s and Discussion
In his s udy, gene ic a ia ion da a ob ained om ISSR (In e -Simple Sequence Repea ) analyses o pe ch samples
collec ed om 12 di e en lakes and dams loca ed in a ious geog aphical egions o Tu key we e subjec ed o clus e
analysis using he UPGMA (Unweigh ed Pai G oup Me hod wi h A i hme ic Mean) me hod, and he esul s a e
p esen ed in Figu e 1. Molecula analyses pe o med wi h a o al o ou ISSR ma ke s yielded 100 sco able bands, o
which 68 we e polymo phic and 32 we e monomo phic, co esponding o a polymo phism a e o 68%. The ob ained
banding pa e ns e ealed a clea le el o gene ic di e en ia ion among he popula ions. Acco ding o he gene a ed
dend og am, he samples we e g ouped based on gene ic simila i y coe icien s, esul ing in wo main clus e s. The i s
clus e consis ed o a sub-clus e including A yon (Şeyi le Dam), Uşak (Ka aağaç Pond), and Ispa a (Da ıde esi Dam)
popula ions, and ano he sub-clus e comp ising Samsun (Ladik Lake), Amasya (Yedikı Dam Lake), and Kocaeli
(Denizli Pond) popula ions. Two di e en sampling si es wi hin İzmi (Tah alı and Ü kmez dams) we e posi ioned as
he closes g oup o his clus e . This indica es ha ish popula ions in hese egions exhibi ela i ely close gene ic
s uc u es, likely a ibu able o simila en i onmen al condi ions o gene low. The second majo clus e included
Wo ld Jou nal o Biology Pha macy and Heal h Sciences, 2025, 24(01), 032-044
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popula ions om Konya (Al ınapa Dam), Anka a (Çamlıde e Dam), Kı şehi (Hi anlı Dam), and Adana (Seyhan Dam).
Indi iduals in his g oup exhibi ed g ea e gene ic dis ances om he o he popula ions. In pa icula , he Adana
popula ion displayed he a hes gene ic dis ance on he dend og am, sugges ing ha hese samples ep esen a mo e
isola ed popula ion s uc u e and a e ma kedly di e en ia ed om o he g oups in e ms o gene ic a ia ion.
Figu e 1 ISSR-based gene ic simila i y dend og am o pe ch popula ions
In addi ion, he wo-dimensional plo ob ained om P incipal Componen Analysis (PCA) (Figu e 2) was la gely
consis en wi h he dend og am. In he PCA, popula ions om A yon, Uşak, Ispa a, Samsun, Amasya, and Kocaeli
clus e ed closely in he same di ec ion, con i ming hei high gene ic simila i y. The İzmi -Tah alı and İzmi -Ü kmez
popula ions we e again g ouped oge he and dis inc ly sepa a ed om he o he popula ions. Konya, Anka a, Kı şehi ,
and Adana popula ions we e posi ioned on he posi i e side o he PCA axis, apa om he o he g oups. No ably, he
Adana popula ion appea ed e en mo e dis an , indica ing as in he dend og am ha i possesses a mo e isola ed and
di e gen gene ic s uc u e. Bo h analyses consis en ly e ealed h ee main clus e s: G oup 1 (A yon, Uşak, Ispa a,
Samsun, Amasya, Kocaeli), G oup 2 (İzmi -Tah alı and İzmi -Ü kmez), and G oup 3 (Konya, Anka a, Kı şehi , and pa ly
Adana). These indings sugges ha gene ic di e si y among popula ions may be associa ed wi h geog aphical
dis ibu ion, wi h pa icula ly p onounced di e ences obse ed be ween popula ions om İzmi and he Cen al
Ana olia egion.
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Figu e 2 PCA analysis o pe ch popula ions based on ISSR da a
The gene ic simila i y ma ix based on ISSR ma ke s is p esen ed in Table 3. The esul s e ealed a signi ican le el o
gene ic a ia ion among pe ch popula ions. Examina ion o he dis ance ma ix showed ha he gene ically closes
popula ions we e A yon–Uşak and İzmi -Tah alı–İzmi -Ü kmez, bo h wi h 100% simila i y. This indica es ha ish
popula ions om hese loca ions sha e a high deg ee o gene ic simila i y, likely e lec ing de elopmen unde
compa able en i onmen al condi ions wi hou subs an ial gene ic isola ion.
Mo eo e , he A yon, Uşak, Ispa a, and Samsun popula ions also displayed high simila i y (≥92%), and hese g oups
we e clus e ed wi hin he same main b anch o he dend og am. No ably, A yon–Samsun and Uşak–Amasya popula ions
exhibi ed 92% simila i y, sugges ing ha despi e hei geog aphical dis ance, hei gene ic s uc u es emain alike. This
could be a ibu ed o in e -popula ion gene low, human-media ed ansloca ion, o he in luence o simila
en i onmen al s ess ac o s.
In con as , he Adana popula ion eme ged as one o he mos gene ically dis inc , bo h in he dis ance ma ix and in he
dend og am analysis. I s simila i y wi h Kı şehi was 76%, whe eas wi h A yon i was only 40%. This sugges s ha he
Adana popula ion ep esen s a gene ically isola ed g oup ha may ha e unde gone a p ocess o egional adap a ion.
Addi ionally, Konya and Anka a popula ions exhibi ed 100% simila i y and clus e ed wi hin he same sub-g oup,
implying ei he ex ensi e gene ic exchange o a sha ed o igin. Simila ly, he İzmi -Tah alı and İzmi -Ü kmez
popula ions showed comple ely o e lapping gene ic p o iles, e lec ing a common gene ic pool.
In conclusion, he ISSR ma ke -based gene ic simila i y analysis demons a ed ha P. lu ia ilis popula ions om
di e en geog aphical egions o Tü kiye ha bo subs an ial gene ic di e si y.
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Table 3 Gene ic simila i y ma ix based on ISSR ma ke s
AFYON
KONYA
ISPARTA
SAMSUN
ANKARA
İZMİR-TAH
İZMİR-URK
USAK
KIRSEHİR
KOCAELİ
ADANA
AMASYA
AFYON
1.00
KONYA
0.44
1.00
ISPARTA
0.96
0.48
1.00
SAMSUN
0.92
0.52
0.96
1.00
ANKARA
0.44
1.00
0.48
0.52
1.00
İZMİR-TAH
0.88
0.48
0.84
0.80
0.48
1.00
İZMİR-URK
0.88
0.48
0.84
0.80
0.48
1.00
1.00
USAK
1.00
0.44
0.96
0.92
0.44
0.88
0.88
1.00
KIRSEHİR
0.48
0.88
0.52
0.56
0.88
0.52
0.52
0.48
1.00
KOCAELİ
0.88
0.48
0.92
0.96
0.48
0.76
0.76
0.88
0.52
1.00
ADANA
0.40
0.88
0.44
0.48
0.88
0.44
0.44
0.40
0.76
0.52
1.00
AMASYA
0.92
0.52
0.96
1.00
0.52
0.80
0.80
0.92
0.56
0.96
0.48
1.00
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In his s udy, a ious molecula analyses we e conduc ed o e eal he phylogene ic ela ionships and nucleo ide
composi ions o he m DNA ATP6 gene egion o he samples. Fo his pu pose, conside ing he molecula
cha ac e iza ion esul s, he sampling loca ions we e g ouped in o ou main egions, and one p o ince was selec ed
om each egion. Sequences o he m DNA ATP6 gene egion we e ob ained om hese samples, and compa a i e gene
sequences o di e en species we e e ie ed om NCBI o cons uc a phylogene ic ee. The aim was o elucida e he
phylogene ic ela ionship be ween pe ch and o he species. The p ocess o ob aining and analyzing DNA sequences was
ca ied ou as ollows:
Fi s , he sequencing o DNA isola ed om he samples was pe o med by a comme cial se ice p o ide , Oligome
Bio echnology Inc. The ob ained sequence da a we e analyzed using Sequenche 4.5 so wa e, whe e o wa d and
e e se p ime sequences o each sample we e assembled. Reading e o s and low-quali y egions we e manually
co ec ed based on ch oma og am inspec ion, and con ig sequences we e gene a ed. In his way, clean and eliable
ATP6 gene egion sequences we e ob ained o each sample. The sequences we e hen con e ed in o FASTA o ma
using Mic oso O ice Wo d and p epa ed o phylogene ic analyses.
Sequence alignmen was pe o med wi h MEGA 6.0 so wa e (Tamu a 2013), and he aligned sequences we e compa ed
wi h he NCBI da abase using BLAST analysis o e i y sequence accu acy and simila i y le els. Fo phylogene ic
in e ence, Neighbo -Joining (NJ) ees we e cons uc ed in MEGA 6.0, and gene ic dis ance ma ices among species we e
calcula ed. This app oach enabled he iden i ica ion o e olu iona y ela ionships among he samples and he
in e p e a ion o in a- and in e speci ic gene ic simila i ies. In addi ion, nucleo ide composi ions o he species we e
de e mined om he aligned sequences. Acco ding o he base composi ion analysis o he ATP6 gene egion, he
a e age p opo ions we e calcula ed as hymine (T) 24.2%, cy osine (C) 13.5%, adenine (A) 30.4%, and guanine (G)
31.9% (Table 4). O e all, a high deg ee o homogenei y in base composi ion was obse ed among he samples, indica ing
ha he ATP6 gene egion is highly conse ed s uc u ally ac oss he analyzed species.
Table 4 Nucleo ide composi ion o he m DNA ATP6 gene
Samples
T(U)
C
A
G
To al
ADANA
24.2
13.6
30.4
31.8
727.0
İZMİR
24.5
13.5
29.8
32.2
621.0
AFYON
24.1
13.5
30.7
31.8
721.0
KONYA
24.1
13.5
30.6
31.7
725.0
A e age
24.2
13.5
30.4
31.9
698.5
Fu he mo e, phylogene ic analyses we e pe o med using m DNA ATP6 gene egion sequences isola ed om Pe ca
lu ia ilis indi iduals. The ob ained sequences we e analyzed in MEGA 6.0 so wa e using he Maximum Likelihood (ML)
algo i hm, and bo h in aspeci ic a ia ions and in e speci ic e olu iona y ela ionships we e examined. The esul s
showed ha he ou samples included in his s udy (ADANA, İZMİR-TAH, KONYA, and AFYON) clus e ed wi hin he
same clade oge he wi h P. lu ia ilis e e ence sequences e ie ed om GenBank, displaying high le els o simila i y.
No ably, he ADANA, İZMİR-TAH, and KONYA samples clus e ed on he same b anch wi h GenBank e e ences
OQ676940.1, OQ676941.1, and OQ676945.1, suppo ed by boo s ap alues anging om 62% o 100%, s ongly
con i ming ha hese samples belong gene ically o he P. lu ia ilis species.
The opology o he phylogene ic ee e ealed ha he examined indi iduals we e highly simila o each o he as well
as o he da abase-de i ed P. lu ia ilis sequences, indica ing limi ed in aspeci ic gene ic di e si y. This inding
highligh s he e ec i eness o he ATP6 gene egion in esol ing phylogene ic ela ionships a he in aspeci ic le el.
Addi ionally, closely ela ed species, P. sch enkii and P. la escens, o med a dis inc clade posi ioned in close p oximi y
o each o he , suppo ed by a boo s ap alue o 89%, sugges ing ha hese species may ha e di e ged om a common
ances o . Sande luciope ca was used as an ou g oup and, as expec ed, was placed in a sepa a e clade om he o he
species (Figu e 3).
Wo ld Jou nal o Biology Pha macy and Heal h Sciences, 2025, 24(01), 032-044
39
Figu e 3 Phylogene ic ee o m DNA ATP6 gene sequences ob ained om indi iduals o he pe ch species (1: Seyhan
Dam- ADANA, 2: Tah alı Dam- İZMİR, 3: Al ınapa Dam- KONYA (Gene egion sequence could no be ob ained), 4:
Şeyi le Dam- AFYON)
In his s udy, m DNA cy b (cy och ome b) gene egion sequences isola ed om P. lu ia ilis indi iduals we e analyzed
o de e mine he in aspeci ic base composi ion. The ob ained sequence da a we e aligned using MEGA 6.0 so wa e,
and nucleo ide equencies we e subsequen ly calcula ed. Fo h ee di e en samples (ADANA, İZMİR-TAH, and
AFYON), he a e age nucleo ide composi ion o he cy b gene egion was de e mined as hymine (T) 30.9%, cy osine
(C) 30.4%, adenine (A) 24.0%, and guanine (G) 14.8% (Table 5). This dis ibu ion indica es ha hymine and cy osine
a e p edominan in he cy b egion, whe eas guanine con en is conside ably low. These indings demons a e ha he
m DNA cy b gene egion is cha ac e ized by A+T ichness and a ela i ely low G+C a io. Mo eo e , he o e all
consis ency o base p opo ions among samples sugges s ha he le el o in aspeci ic gene ic a ia ion is limi ed.
Table 5 Nucleo ide composi ion o he m DNA cy b gene
Samples
T(U)
C
A
G
To al
ADANA
31.0
29.7
24.0
15.2
938.0
İZMİR
30.6
30.1
24.5
14.8
1003.0
AFYON
31.2
31.9
22.9
14.0
520.0
A e age
30.9
30.4
24.0
14.8
820.3
Phylogene ic analyses based on m DNA cy b (cy och ome b) genes a e widely ecognized as an impo an ool o
unde s anding e olu iona y ela ionships among species and o conduc ing molecula -le el axonomic classi ica ion.
In his con ex , cy b gene egion sequences isola ed om P. lu ia ilis indi iduals we e used o cons uc a phylogene ic
ee (Figu e 4). The analyses we e pe o med using he Maximum Likelihood (ML) me hod unde he Tamu a-Nei
model, which accoun s o di e en a es o ansi ion and ans e sion subs i u ions, he eby p o iding a mo e ealis ic
modeling o nucleo ide eplacemen s.
In he ML ee, he h ee analyzed samples (ADANA, İZMİR-TAH, and AFYON) clus e ed wi hin he same clade wi h high
boo s ap suppo alues, con i ming hei close gene ic simila i y and sugges ing ha hey likely o igina e om he
same o geog aphically ela ed popula ions. Boo s ap alues shown on he b anches ep esen he s a is ical
con idence le els o he in e ed ela ionships; alues abo e 70% indica e s ong suppo , while hose abo e 90%
sugges highly eliable e olu iona y ela ionships. Acco dingly, he high boo s ap alues obse ed o he cy b
sequences analyzed he e demons a e he obus ness o he phylogene ic s uc u e and he accu acy o he
classi ica ion.
Wo ld Jou nal o Biology Pha macy and Heal h Sciences, 2025, 24(01), 032-044
40
The clus e ing pa e n u he indica es ha gene ic a ia ion wi hin P. lu ia ilis is limi ed, implying ha he cy b egion
may ha e es ic ed powe o esol ing in aspeci ic a ia ion. Howe e , i emains a highly eliable molecula ma ke
o species iden i ica ion. The obse ed homogenei y may also e lec he ac ha he analyzed indi iduals o igina e
om he same geog aphical popula ion. When e e ence sequences om closely ela ed axa such as P. sch enkii and P.
la escens we e included, P. lu ia ilis samples we e clea ly sepa a ed in o a dis inc clade. This sepa a ion suppo s he
abili y o he cy b gene o esol e in e speci ic e olu iona y di e ences while simul aneously con i ming in aspeci ic
homogenei y. The ou g oup species (Sande luciope ca) was posi ioned in a clea ly dis inc clade, as expec ed, e lec ing
i s mo e dis an e olu iona y lineage wi hin Pe cidae.
Molecula ma ke s a e widely ecognized as essen ial ools in ish popula ion gene ics o elucida ing gene ic di e si y
and e olu iona y ela ionships. Okumuş and Çi çi (2003) emphasized ha each ma ke ype, including allozymes,
mic osa elli es, RAPD, AFLP, m DNA, and nuclea DNA ma ke s, possesses speci ic ad an ages and limi a ions, wi h
a ying esolu ion capaci ies in de ec ing gene ic a ia ion. In pa icula , mic osa elli es a e conside ed highly e ec i e
in de ec ing in aspeci ic a ia ion due o hei high polymo phism, while ISSR ma ke s ha e been demons a ed o be
powe ul o assessing bo h en i onmen al adap a ions and in e -popula ion di e en ia ion (Zhigile a e al. 2013;
Zhigile a e al. 2022; Alyamkin e al. 2022).
The esul s o he p esen s udy based on ISSR ma ke s e ealed a high le el o gene ic di e si y wi hin P. lu ia ilis
popula ions in Tü kiye. O he 100 sco able bands ob ained, 68 we e polymo phic, co esponding o a polymo phism
a e o 68%, he eby con i ming he disc imina i e powe o ISSR ma ke s. Compa able indings we e epo ed by Liu
e al. (2006) in Pa alich hys oli aceus, Mal aglia i e al. (2006) in Cyp inodon i o m ishes, and Mohammadabadi e al.
(2017) in Poecilia e icula a, whe e ISSR ma ke s success ully de ec ed high le els o polymo phism and gene ic
a iabili y. Thus, he indings o he p esen s udy align wi h p e ious epo s and demons a e he e iciency o ISSR
ma ke s in e ealing gene ic a ia ion in pe ch popula ions.
The dend og am and PCA analyses clea ly indica ed in e -popula ion di e en ia ion, wi h he Adana Seyhan Dam
popula ion being gene ically dis inc om he o he s. This di e gence can likely be a ibu ed o isola ion and limi ed
gene low. Simila pa e ns ha e been documen ed by Waw zyniak e al. (2020), who epo ed en i onmen ally d i en
gene ic di e en ia ion among pe ch popula ions in Poland, and by Vanina e al. (2019), who iden i ied signi ican
gene ic dis inc i eness o he Polish popula ion compa ed wi h o he s. Fu he mo e, Hai Sa e al. (2012) e ealed low
gene ic di e si y in wild popula ions om Xinjiang, China, while Ragauskas e al. (2023) ound low- o-mode a e
a iabili y in Bal ic Sea pe ch popula ions, associa ing di e en ia ion wi h geog aphic isola ion, habi a changes, and
an h opogenic impac s. The gene ic sepa a ion o he Adana popula ion obse ed in he p esen s udy is he e o e
consis en wi h hese ea lie indings.
In e es ingly, he İzmi -Tah alı and İzmi -Ü kmez popula ions exhibi ed 100% simila i y, sugges ing ha
en i onmen al simila i ies in hese habi a s may be di ec ly e lec ed in hei gene ic s uc u e. Michel e al. (2009)
p e iously iden i ied habi a -based clus e ing in pe ch popula ions, a pa e n also co obo a ed by ou esul s, which
indica e ha habi a isola ion plays a key ole in shaping popula ion gene ic s uc u e.
A he in e speci ic le el, m DNA-based s udies wi hin he Pe cidae amily ha e p o ided impo an insigh s in o
phylogene ic ela ionships. Sloss e al. (2004), using cy b and 16S RNA sequences, e ealed phylogene ic pa e ns ha
pa ly con lic ed wi h adi ional mo phological classi ica ions, sugges ing he need o axonomic e-e alua ion in
ce ain g oups. Simila ly, Nesbo e al. (1999) epo ed high le els o gene ic di e en ia ion among popula ions sampled
om Eu ope and Sibe ia. In ou s udy, cy b sequence analyses con i med he gene ic homogenei y o P. lu ia ilis
popula ions, while clea ly dis inguishing hem om closely ela ed species. This suppo s he u ili y o he cy b gene as
a obus molecula ma ke o esol ing in e speci ic e olu iona y ela ionships.
O e all, he combined e idence om ISSR and m DNA analyses sugges s ha he gene ic di e si y o P. lu ia ilis
popula ions in Tü kiye is shaped by habi a isola ion, geog aphic dis ance, and en i onmen al ac o s. These esul s a e
consis en wi h p e ious s udies (Hai Sa e al. 2012; Vanina e al. 2019; Waw zyniak e al. 2020; Ragauskas e al. 2023)
and highligh he impo ance o gene ic moni o ing o he conse a ion and sus ainable managemen o pe ch
popula ions.
