! "#$ $ ( capoeta gracilis) RAPD % &'% $

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... "= E&'H = 3 %& EB = E V A ' J K - & @> .=U )L> = = ."(+ 3 ) ( ' &* + $,&-% '( E& .( ) -& 3 - & Saeidi et al., 2007 .(Dynesius & Nilsson, 1994) "."( " / EY = / ab "U 3'F MI 3 34& 56# 37* 8&( 59 ef/c "U - & I )&L = MI = " .(Tudela, 1999) (3 3%: 59 ;< ﺟﻤﻠﻪ ﻣﻬﻤﺘﺮﻳﻦ ﺳﺎزه ﻫﺎﻳﻲ اﺳﺖ >; Mazandaran ) ."( ]-A ;Q% 8&( 59 3& "H( " .(Regional Water Company, 2001 > 5 " .( 3 37* "> [ h- " - ( ) %' 2001 McAllister 3& 56 B& CD 5 E& (Craig, 2001) Y " E& . ( @ M* McAllister et al, ) ( 3 " EF+ G M> / ce i '/ Y> j 3/* 3&= ?> HI J+ K& " .(2001 Nazariha ) 3R'( =3 &= 3 A $( ( 3 A 3%: &N "( "* & AnvariFar, ) ( @ M* HI = 3%& (gracilis & ) M O7 - 5" - 9 .A k>+ R (2010 M 5 " = 3 # P 7 . = 3 E& ."( M4 QR7 E& M I / " ."( A= Q+ K& - & 3> R MI E& S9 B Mostafavi and Abdoli, 2005) " 3' E&+ 3& ;T) ﻫﻤـﻴﻦ اﺳـﺎس 3 V $( SA 3 -4/#U B/ ﭼﻴــﺮه در U=. <3 & - )' @ Capoeta ) 3 . `> .(Samaee et al., 2009) E&) M I 3 K W ( k>+ E& 3A">+ I MZ - = (spp. .Y&= $3&SZ .B= ( .XD W( $> $9V $B&NI $ J ( 3X 8&( 9 $( A 3<> %= 3R9( E7/V $& $mD $A K& MI) " J+ K& / )'D K& 3 E& .(Turan, 2008) (3 ."( EJ I / %>& " 3 " J+ 3>( $3Y> ./ an MI A ( B& I 3\ 3& "> [ 3 -& 4 &= -I A A 3< > 8&( ( = K& E& &L j' (Valipor, 2003) 3 3 Z A ]+ $("> 3 B ﻫـﺎﻳﻲ & MI 3 ,& (Turan, 2008) ( 3 V& "> i& ^ 3 Z A G'U) " J+ K& 3&SZ &: .(Samaee et al., 2009) o+ MI 6 PX# $ (( 3 3 A E&-Y& _&" $+ A = %&7 4A 3 . SA3 ... HI "( YR 3 . "> 3 &S9 (.+) 3A ."+ 6&+ .(McAllister et al, 2001) #" & & & % &' ()* + #,- &()* !" #$ 'SZ . p*R E>q&3 وﻳﮋه uQl &S+ % ;RZ 5Y&=' 5Y&= "V .(Welsh & McClenlland, 1990) ' .( " = A Y & 3 & S Z Q &+ 59 ( " 5 A E& .(Mostafavi and Abdoli, 2005) K& E&+ . 3&= ?> "# $+ -I $3/ MI Y p*R = =)FU -r V9 5QR7 3- اﻫﻤﻴـﺖ " -Z $h/; 5 <36 =& i7 " / G @ 5H .(Sammaee et al., 2006) &3AA E& 5 " G .(3 ?> 3& "H( " 3&= s" QR7 E ﻧﻴﺴﺖ و A "Q " 3 J 3& EB E&+ G . 3%: -&' Gertsev & Gertseva, ) "&3 k&-V " G .6 3R%R ] = 3%& . V&S+ 5# 5[[* o .(1999 ; Poddubny & Galat, 1995 $ A &= $ A t4J $-&' ?> EQ HI y " ."( jB ">K 5Q7 3VT % ,>% `> Q' )%/Gharaei et al., 2005) (RAPD) DNA 3). اﻳﻦ )jB (2001) Craig H = V9 [7> k روش ﻧﺨﺴﺘﻴﻦ ﺑﺎر در ﺳﺎ ل 1990ﻣﻴﻼدي z! "( t4J .A 8 #" > 3&= ?> 36> " "# !a Welsh and McClelland, ) " ( uJ -' 5# . "3 J "( ." RAPD ,>% .(Williams et al., 1990 v1990 3&" R Y&"%& = u/4 3 . DNA 3>< Y& ] &[ Samaee et al., 2009 ; Samaee ) "."( " 3&V9 3UL DNA R 5DXL = .(et al., 2006 ;Bianco & Banarescu, 1982 DNA = 3 Q7 % ( RAPD ] . " '&+ Q' -&' ;< 3&V9 3&" ."( %>& " R B > PCR 8 '&+ = .6 U 3R (3 Capoeta capoeta gracilis .( 3 Y JY ' & DNA 5Q7 D'B ,& " R 3&V9 3&" ."( "R 5Q7 E& 3 RAPD Y JY i VY 5# & 3%: QR7 | $" @ { Anvarifar Rehnuma et al., 2003) PCR 8 . .[R RAPD .(and Farahmand, 2009 G < V 3 / + 3 & >( 3>< YJ &- E& E>q' 3-4 " EB QR7 E& ;RZ %( 3 R " > PCR E& . VA 5# $."( i I 3& "H( Liu and ) ( 3 3 = wU j"D /wU 5# .'( .Y&) " G 3%& .Y& QR7 3 ] E& Y& x &- = .(Cordes, 2004 M4 (z .'( .Y&) " E&+ Y& (e = $ & A j"D $I G D . EB 3/# .Y& "( %& A=ZI DNA -K "[ a} 3&V9 Q " G .Y& %J 3 I ;&Q = u/4 A ee E t4J 3 oR 3R"> Ol دﻗﻴﻘﻪ zc/fe ﺛﺎﻧﻴﻪ ef na 3R'( ~D دﻗﻴﻘﻪ za/ea ﺛﺎﻧﻴﻪ E&+ .Y& 3( L " {e} a دﻗﻴﻘﻪ {en/a ﺛﺎﻧﻴﻪ na 3R'( ~D ez Fingerprinting دﻗﻴﻘﻪ ne/cc ﺛﺎﻧﻴﻪ L )3 ()% dominant ... ?'B ."( €&Q HI %R D c zc = `+ = .6 = ' 3 E& .(e ‚ƒ "Q E& = "( x ' 3 Q7  "Q [ & 5 e 5" (%R .Y 3 ' ‚„ G .Y& y 3 ' 5QR7 jB x .VA jB R abb ^ zbb :R . j .Y& y ') @ ' ,& 3'%( > R 3R%R ) +` = #" " )". f} %R #" ﺗﺜﺒﻴﺖ

$% &' #" # (!) () (Samaee et al., 2009 # )

M% $I = `+ "( . Y DNA $Y „ MI [ „ƒ 5" Walsh et ) `X ] = .6 DNA 4 - .6 = < .VA 5U (Kumar et al., 2007 Estoup et al., 1996 al., 1991 „ƒ ^„ 5" $. `% ,& = `+ M% z*z .=" ' ] E& .V&S+ 5# EQ ."( :6& -  rpm 3/ e/n % ,& R V = i '/ UV V% .Y = DNA / ' /* R% nbb "[ " & R "# b/ =AI : DNA ."J =>+ &-I R% en .XD "# eb `X (TAE 1X V 3 3 cb " =AI jA b/az) ' QV x ` ."( Vl I (100 U/ml) K ."( .6 "<G :V I &[ ' U M% $K =>+ &-I ƒ %( E D ,& 5"

Chelex protocol #" & & & % &' ()* + #,- &()* !" #$ 

& DNA V+ " 3%: . -RI '&+ eb = .6 ."( 4 DNA (PCR) 5# " ` < 3 JY VA 3&= 3/%( ">K EQ H RAPD ."( 3 = ."( % 5Q7 wU j"D/wU H : > ( ]6 '&+ E& .(„ ") 5# e " " % 5# * E& zn 3&H BU ='/+ .B= k> ."( Excel (Office 2003) -V j 6# " " j"D 500 mM ) 10X PCR V R% z/n ( R% NTsys POPGEN -V j 8 . ."( 3/ /ƒ $(KCL and 200 mM Tris-HCL, pH 8.4 .VA -RI Taq DNA &-I "U /ƒ $&-> "&/ G % „ƒ $'&+ = 3/ „ $ Polymerase DNA jA $(dNTP) "/ y/4 '&+  "Q $.6 '&+ „ ?'B = 3U K ."( jB 9 "[ MI 3: "( M4 3 "R J s 3/+ " Y ƒ [ K ,& ( %( ">K " $'&+  ?'B = .(^„ '&+) ƒ [ „ ( K $(R = () 3R „ " ."I YJ & (s 3/+) ‚ [ „ $(= () Y 3/+ YJ "Q .' .6 '&+ Y [ ,& $(m*R) Y %( ."3 J '&+ = ."I " s Y [ 3&+ /U (8) 3 ' '&+  >&" YR = ' ."( jB (3&H 8) Y& G .Y& y 3%& "3 J . E&+ .Y& %( ">K " ' 3%: ?> 3 _& : #U 5GT* $PCR j' = `+ R "Q $V 3/+ "# = "> $(Ne) R "Q $(na) ."J "& j&" U : ."( =V%R (1X (I) E( 3DXL t( (H) Nei(1973) "( ."& . E& . (V BU b/bbe) cb µl - Capoeta capoeta gracilis 3 . Q' E>q' = 3 &S% : " EB " E&+ G .Y& Loading ) Dye $: DNA U ."J o 3%: ?> _& . ."( .I " T* .' eµl - }X /Z (buffer ' 3%: ?> ( (1987) Nei -RI ."( )&- =V%R 3> ,K PCR 3%: ?> $(Hs) Q' 3%: ?> $(Ht) .Y = .6 ."( B& 5GT* &H $(3 ( Nm) 3: & - (Gst) Q' E &T : = "> .Y ."J UVITEC ."(

Observed number of alleles ! Effective number of alleles [Kimura and Crow (1964)] " Nei's (1973) gene diversity # Shannon's Information index [Lewontin (1972)] Polymerase Chain Reaction $ estimate of gene flow from Gst or Gcs. E.g., Nm = 0.5(1 - Gst)/Gst Gel Documentation ... ."I / / $/‚ $ /‚ " E&+ G .Y& ?'B

PCR & DNA % &% #' ردﻳﻒ GC ! 5΄-GTTTCGCTCC-3΄ OPB1 5΄-GTCCACACGG-3΄ OPB8 5΄-CTGCTGGGAC-3΄ OPB10 5΄-GTAGACCCGT-3΄ OPB11 5΄-CCTTGACGCA-3΄ OPB12 5΄-AGGGAACGAG-3΄ OPB17 5΄-GGGTAACGCC-3΄ OPA9 5΄-TGCGCCCTTC-3΄ OPB5 5΄-TGCTCTGCCC-3΄ OPB6 5΄-GGTGACGCAG-3΄ OPB7

Capoeta capoeta gracilis& ' # ! &' #" ( ) # ( )

Capoeta capoeta gracilis & ' (loci) # # & % & ! #' &' #" # (' () # (&' () # $% " / / (na) /$ / (Ne) !"# /,- /, (H) Nei(1973) %&' *#+ / / (I) & %. / /,- # %

#" & & & % &' ()* + #,- &()* !" #$  +-I Q &-I &N O*# &&" 'D (j) 5 ;&l o UPGMA jA" A 3: &N " .' [V 3= EB 3 . Q' J $>= E& jA ER VA QR7 $3 = 3& "H( " E&+ G .Y& $ \ & Q' $A O*# t4J 3Y<' ;&l .(‚ %() "."( -&' Y&"%& -& 3X( &&" = E& „ P - / VA 5# -RI .(Coad, 1980) ' -FU A 36U „ ."( ."J sX I ."> J 3 . Q' 3&" 3/# &G = G'U Prob. random Z < obs. Z: p = ) 3>Q "# A E& 3&V9 3&" /D EB G .Y& y ‚ƒ^„ ' ) .(1.0000 %' Turan %&L (" ) .( E&+ .Y& y ^‚ ' Liza ) 6 =" & E>K (2004) @ . Q' 3%: /#V "U - &" .: $. & (abu (Heckel, 1834) "#„/ƒƒ I E&' "# ‚/ EB Q' =" (2008) Khara E& ."( <* "# / EY L ?'B & $3R- MG (Abramis brama orientalis) 5 ;&l o J `%& = #V ."' >D o " K& B&NI H' $- ƒ sX $E&+ G "U BI = "( <* = E&+ G Q' E& U A &= A t4J Q "# „ƒ^„ 3= /#V $ ."( " Y&"%& = 3R%R /#V EY B $(Gharaei et al., 2005) L ">3 &6 E& E& ' 3 $Q' V j' 3%: < J U .H & &[ _& ."."( -&' Y&"%& = 3&G "U Q' 59 K J " 8* = 3( 59 (PCOA) 5T4 3/# 6R -RI = ."I " ﻧﻴﺰ vLindsey, 1998 vSmith, 1966) "(3 A E G .Y& 3 . Q' J Y& sL = .(Turan., et al., 2004, 2006 vTuran, 2000 -&' Y&"%& = 3& "H( " E&+ $SZ 3 $5" $) 56 37* 8&( .(%() = Q' 3&" ;< (MI & MI )'D Yamamoto et vSamaee et al., 2006) (3 Y&"%& "3 J - "( .(al., 2007 j $3/% = 3I QR7 Capoeta capoeta gracilis 3 . Q' .H $3I i> &&" 6U $3>( V $3>( E(" 3&V9 3&" R u/4 ' -FU HI ]+ &N = ) ."( -&' Y&"%& = y< Bianco & "I 3[7> &&" > .(Mostafavi, 2005) Samaee et vSamaee et al., 2006 vBanarescu, 1982 3 = A .N 3&>( $3X( .(al., 2009 K 3&- ' $A 3 .H = .6  %' Yamamoto .H ( &&" A" L "& .N (,K) " "' 5< . -& vTzeng, 2004) A >H "U A I = =I 3 Q' " .(Akbarzadeh et al., 2007 v Erguden & Turan, 2005 ."&' -&' Y&"%& = (Salvelinus leucomaenis)

= .6 %' Dakin E>q'  Unweighted Pair-Group using Arhimetic Average ... ( Q' V j' jA" Q' " "' 5< . -& jA" QR7 E& .(Rehnuma et al., 2003) Y&"%& = (Micropterus cataractae )o 3 El-Zaeem ) "( 5 ;&l o UPGMA „ - %' Jager .' -&' V j' $UPGMA jA" )7 .(et al., 2006 : 3Q' 3&" ;< " "' ﺳﻔﻴﺪ (Y& V j' X ,& E&+ .Y& E& . ."( (Acipenser transmontanus. wU Y& X ' - G = . -&' ?> 3 QR7 RAPD 6. )". اﻳﻦ ﺳﻮال ﻧﻴﺰ ﻣﻄﺮح اﺳﺖ، ﻛﻪ آﻳﺎ <&X -RI PCOA -RI _& E E& " @ RAPD ' D 3 -&' Y&"%& = Q' "3 RAPD ."&' " Y&"%& = G <( "# Q' = 3V 5DXL j"D /D 3 E& "&' "D QR7 E& ."I (Nm) 3: & - .6 RAPD = $3 . DNA R 3R ."> J 3: & "[ E& . 3&G < = k RAPD ,>% " >D &= 6R ."( E& E& . Q' E& 3%: < &= 3 Q' "3 Y& ,>% Y&"%& 3(-I A | $" wU R Q' "> -&' 3 J Q' 3U \ $A (Bardakci .(Mamuris, 1998, 1999; Gomes, 1998 ﻧﺪارﻧﺪ. ﺑﻨﺎﺑﺮاﻳﻦ ﻣﺘﻤـﺎﻳﺰ دو ﺟﻤﻌﻴـﺖ و ﻣﻘـﺪار ﺑـﺎﻻي ﺗﺒـﺎدل RAPD ,>% " (1994) Skibinski ژﻧﻲ، ﻣﺮﺑﻮط D . („ƒ^„) " ' D L>5 3= y ."( @r>( 3: & Y& mtDNA 8< Y& -RI = oU g"U = < = 3>Q&) " 3%& Q' E& (2005) %' Gharaei . 3Q' 5QR7 3: < Q' E& . .(" 3 o "' E& = .6 - 3%: / #V "[ $PCOA -RI _& $G Acipenser ) 3 (Acipenser persicus) 3& E& " ."( -&' Y&"%& = UPGMA jA" .">( 3 Y&"%& = -B A (gueldenstaedtii = < = y ."( ."J 3: < "[ J `%& = 3%: /#V QR7 E& Q' E& 3& "H( " " g"U - "# / EY L 5 ;&l o 3: ?> "[ &[ . "&' R-& Y&"%& = G "U BI = 3&G "D "I (I) E( 3DXL t( (N) Nei(1973) &= A t4J Q $"# ƒ sX $E&+ Capoeta capoeta gracilis 3 . Q' ' 3 (Gharaei et al., 2005) A 3& "H( " E&+ G .Y& -&' Y&"%& = 3&G "U Q' E& j Q ' ( @ #U B E& EB QR7 Q' V k>+ ."."( Q' $PCOA -RI o ﺳﻴﺎه ﻣﺎﻫﻲ از ﻳﻜﺪﻳﮕﺮ ]" <? j Q' t( E& &= G ﻣﺘﻤﺎﻳﺰ اﺳﺖ. ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ ﻣﺮور ﻣﻨﺎﺑﻊ ﺻﻮرت ﮔﺮﻓﺘﻪ، ﻣﻌﻤﻮﻻ ] " R E & . T G j .Y& $" ;< 9 8&( M 3%: 5QR7 UPGMA jA" . .Y& < Q' -&' j"D & 5< 3&H - J - & VA 5# 5QR7 v Rehnuma et al., 2003 v Alam & Khan, 2001) A " = Q E& ."> v Lakra et al., 2007 v Brahmane et al., 2006 >q' $ Q' B& +% .' 3L 3&= .(Rahman et al., 2009 v Brahmane et al., 2008 ﺗﻮﺟﻪ ﺑﻪ اﻳﻦ &A L H $ Q' 3 E - & . ^ #" & & & % &' ()* + #,- &()* !" #$ A E& 3Q' 3&" 3 E& %>& u/4 )L> 3& Q' A &= $- $ " 5 < RAPD 8 3%: p*R = Gholiev, 1997; ) ". %J - & Kazancheev, 1981). ﺗﺤﻘﻴـﻖ ﺣﺎﺿـﺮ ﺑـﺮاي ﻧﺨـﺴﺘﻴﻦ +Y& 3%: 5QR7 ( 3 H>J 6. = 5# RFLP Microsatellite "> Y& & 3%: ] = .6 " E& = ."I " _& ?'B ." 3 3 ﮔﻴﺮد ﺗﺎ ﺑﺘﻮان ﺑﺎ ﻗﻄﻌﻴﺖ ﺑﻴﺸﺘﺮ، "&E& 3Q' 3 3 . QR7 E& _& E>q' .' Capoeta 3 . Q' "3 J 3 % ,& $ ,&:R " ,& >D 3 E&+ G .Y& capoeta gracilis " )ASA B& "> 3&&" ">( 3 -&' Y&"%& = E B 3& "H. @ ﻳـﺎ I G 3 : & ( 3 H>J+ -&' EQ ;> 3( RAPD Y JY .' " E&+ . 3 . 3%:

Capoeta capoeta gracilis & ' RAPD UPGMA # $% &' #" #

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( ) $% Capoeta capoeta gracilis & (PCOA) & # # &' #" # ( )

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223 Journal of Natural Environmental, Iranian Journal of Natural Resources, Vol. 63, No. 3, 2010, pp.211-223

Impacts of Shahid Rajaii Dam on Genetic Variation and Differentiation Siah Mahi, Capoeta capoeta gracilis, in Tajan River using RAPD Fingerprinting

H. Anvarifar1, H. Farahmand2*, M.A. Nematollahi2, H. Rahmani3, M. Karami4 and B. Khalili5 1 M.S.C graduated, Faculty of Natural Resources, University of Tehran, I.R. Iran 2 Assistant Prof., Faculty of Natural Resources, University of Tehran, Karaj, I.R. Iran 3 Assistant Prof., Faculty animal science and fisheries, University of Agriculture and Natural Resources Sari, Sari, I.R. Iran 4 Professor, Faculty of Natural Resources, University of Tehran, Karaj, I.R. Iran 5 Expert of biotechnology laboratory, Faculty of Natural Resources, University of Tehran, I.R. Iran (Received: 08 November 2009, Accepted: 08 January 2011)

Abstract Siah Mahi, Capoeta capoeta gracilis, is a predominant fish in the river systems of the south Caspian Sea basin. Although there is evidence of the geographical divergence of this taxon, but no information is available on genetic differences within the subspecies populations. This study was designed to evaluate variation and differentiation in the Tajan River and investigation Shahid Rajaii dam effects upon Siah Mahi. A total 65 specimens were collected by electeroshocker set that 31 individual related to up stream station and 35 individual related to down stream station. Ten random 10-mer primers were primarily scored in 5 individuals from each of two stations and finally 6 primers, which shown polymorphic pattern repeatable, were selected and use in the final analysis of two locations. Totally 89 polymorphic band (marker) gained. Lengths of PCR products were between 50-1000 bp. Analyzing the PCR product data shown that the mean difference between the two stations was 0.478. In view of the fact that in ./5 mean genetic distance between two species, separated those from each other, as a result related two stations in a large quantity separated from each other. Results of Principle Coordinate Analysis or PCOA showed that two stations are separated from each other. The unweighted pair group method with averages (UPGMA) dendrogram based on Jaccard´s index also showed these stations are separated. There are perfect similarities between results of morphometric factor and morphometric spacialities. Based on the results, we conclude that RAPD fingerprinting was a useful technique in genetic discriminating the population of Siah Mahi.

KeyWord: Siah Mahi, Capoeta capoeta gracilis, Tajan river, Shahid Rajaii dam, RAPD fingerprinting, Genetic Variation and Differentiation.

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