AFL P. Journal of Xiamen University (Natural Science) ( Scombroidei), ( Scombridae), , ( ) cm, DNA, 1. 3 A FL P, RA PD.

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1 ( ) Journal of Xiamen University (Natural Science) Vol. 48 No. 6 Nov AFL P,,,, 3 (, ) : (Amplified fragment length polymorphism,a FL P), ( Thunnus thy nnus) ( T hunnus albacares). : ( bp), Shannon % %, AMOVA :, %, %.,2,.. : ; ;A FL P ; ; :Q 75 :A : (2009) ( T hunnus) ( Perciformes), ( Scombroidei),( Scombridae),,, [ 1 ]., ( T hunnus t hy nnus) ; ( T hunnus albacares) ( Katsuw onus pel a2 mis), [ 2 ]. [3-4 ].,., [5-10 ].,, [ ],. RA PD [14 ]. A FL P, : :(2004NZ03) 3 edu. cn , ,( ) cm, ( ) kg ; 44,2006 9,( ) cm, ( ) kg DNA 30,[ 15 ], K + / (90 % - 20 ) DNA, 1 % DNA, A FL P, bp, Vos [ 16 ], DNA, DNA. EcoR I / Mse I,,3 1, EcoR I + A (5 2GACT GCGTACCAA T TC + A23 ) Mse I + C (5 2GA T GA GTCCT GA GTAA + C23 ),, 16 9 A FL P. 3

2 6 :AFL P Tab. 1 Selective amplification primers and their sequences used in A FL P analysis EcoR I (5-3 ) Mse I (5-3 ) E1 GACT GCGTACCAA T TC AAC M1 GA T GA GTCCT GA GTAA CAC E2 GACT GCGTACCAA TTC A GC M2 GA T GA GTCCT GA GTAA CT G E3 GACT GCGTACCAA T TC ACG M3 GA T GA GTCCT GA GTAA CCA E4 GACT GCGTACCAA T TC A T G 3, %,. 1. 4,, 1, 1 ( ),0. POP GEN E Shannon ( Gst ). AMOVA ( Fst ), A FL P 9 30, bp, 35 54, % %, A FL P. ( 2) , , Shannon % % ( p > 0. 05),Shannon, ,94, Tab. 2 Number of fragments,percent polymorphic loci,polymorphic rate of yellowfin tuna ( Thunnus albacare) and bluefin tuna ( Thunnus thy nnus) using 9 pairs of primers / % E1/ M E1/ M E1/ M E2/ M E2/ M E2/ M E3/ M E3/ M E4 / M

3 892 ( ) Tab. 3 Parameters of genetic diversity for two tested tuna populations Nei Shannon Gst : 43, 51 A FL P. 3, Gst Fst = , %, %. AMOVA, %, %. Fst Gst, A FL P, DNA SSR [17 ] RA PD [ 18 ] A FL P [ 19 ]. Han A FL P 3,A FL P, [20 ]. 9 A FL P, 675, 75, 35 54, % %,, A FL P Shan2 non s % , % Han [20 ] A FL P, Han 3 %, , 180, 57 %,,.,,. RA PD [14 ],,.,,,. Gst Shannon AMOVA 3. Nei [21 ] Gst 1,. Gst ,. Wright [ 22 ] Fst, Fst > 0. 15,. Fst ( ) ( p < 0. 05,),. A FL P [23 ]. A FL P, Xu [24 ] 4 ( % %) [25 ] ( %) [26 ] ( %) ( %) [27 ] ( Ci rrhi nus molitorell a) (41. 5 %) [28 ] (V eras permoseri) ( %) ( %). Liu [29 ] ( Paralicht hys oli v aceus ) ( % %) [ 30 ] ( Pa g rus m a2 j or) (58. 4 % 64 %) Chen [ 31 ] ( Cy noglossus semil aevis) (40 % 77 %).,,.,, % 48 %, [ 32 ],,.

4 6 :AFL P 893,,,. : [1 ],. [ M ]. :,1983. [2 ],,. [J ].,2007,3 (4) : [3 ]. [J ]., 2008,23 (1) : [4 ],,. [J ].,2006,6 (2) : [5 ] Broughton R E, Gold J R. Microsatellite development and survey of variation in northern bluefin tuna ( Thunnus thy nnus) [J ]. Mol Mar Biol Biotechnol, 1997, 6 : [6 ] Takeyama H,Chow S, Tsuzuki H,et al. Mitochondrial DNA sequence variation within and between tuna Thun2 nus species and it s application to species identification[j ]. Journal of Fish Biology,2001,58 : [7 ] Chow S, Inoue S. Intra2and interspecies restriction frag2 ment length polymorphism in mitochondrial genes of Thunnus tuna species [ J ]. Bulletin of the National Re2 search Institute of Far Seas Fisheries, 1993, 30 : [8 ] Terol J,Mascarell R,Fernandez2Pedrosa V,et al. Statisti2 cal validation of the identification of tuna species : boot2 strap analysis of mitochondrial DNA sequences [ J ]. Agr Food Chem,2002,50 : [9 ] Ward R D. Population genetics of tunas [J ]. Journal of Fish Biology,2006,47 : [10 ] Scoles D R, Graves J E. Genetic analysis of the popula2 tion structure of yellowfin tuna ( Thunnus albacares ), from the Pacific Ocean[J ]. Fish Bull U S,1993,63 : [11 ],,. [J ].,2007, 31 (12) : [12 ],,. [J ].,2003,20 (3) : [13 ],. [J ].,2005 (3) : [14 ],,,. [J ]. :,2008,47 (4) : [15 ] J. [ M ]. :,2002. [16 ] Vos P, Hogers R,Bleeker M,et al. A FL P : a new tech2 nique for DNA fingerprinting [ J ]. Nucleic Acids Res, 1995,23 (21) : [17 ],,,. [J ].,2007,31 (2) : [18 ] Salhi Hannachi A, Chat ti K, Saddoud O,et al. Genetic diversity of different Tunisian fig ( Ficuscarica L. ) col2 lections revealed by RA PD finger print s [J ]. Hereditas, 2006,143 : [19 ] Liu Y G,Chen S L,Li B F,et al. Analysis of genetic var2 iation in selected stocks of hatchery flounder, Paralich2 thys olivaceus, using A FL P markers [ J ]. Biochemical Systematics and Ecology,2005,33 : [20 ] Han K, Ely B. Use of AFL P analyses to assess genetic variation in morone and thunnus species [J ]. Marine Bio2 technology,2002,4 : [21 ] Masatoshi Nei,Aravinda Chakravarti. Drift variances of FST and GST statistics obtained f rom a finite number of i2 solated populations [J ]. Theoretical Population Biology, 1977,11 (3) : [22 ] Wright S. The genetical structure of populations[j ]. An2 nals of Eugenics,1951,15 : [23 ],,,. A FL P [J ].,2002,9 (3) : [24 ] Xu Xiaofei,Zhang Quanqi,Wang Zhigang,et al. Assess2 ing genetic diversity of wild populations of J apanese flounder using A FL P markers [ J ]. Acta Oceanological Sinica,2006,25 (3) : [25 ],,,. [J ].,2004,34 (5) : [26 ],,,. AFL P [J ].,2006,30 (5) : [27 ],,,. A FL P [J ].,2007,3 (3) : [28 ],,,. A FL P [J ].,2008,32 (3) : [29 ] Liu Y G,Chen S L,Li B F,et al. Analysis of genetic var2 iation in selected stocks of hatchery flounder, Paralich2 thys olivaceus, using AFL P markers [J ]. Bioch System Ecol,2005,33 (10) :

5 894 ( ) 2009 [30 ],,,. A FL P [J ]., 2001,25 (4) : [31 ] Chen S L,Li J,Deng S P,et al. Isolation of female2spe2 cific A FL P markers and molecular identification of ge2 netic sex in half2smooth tongue sole ( Cy noglossus sem2 ilaevis) [J ]. Mar Biotechnol,2007,9 (2) : [32 ],. (3) [J ].,2002,17 (11) : Genetic Diversity of Thunnus t hynnus and Thunnus albaca res Stocks by AFLP WAN G Ding,SU Yong2quan,MAO Yong,XU Bin,WAN G J un 3 (College of Oceanography and Environmental Science,Xiamen University,Xiamen ,China) Abstract : T hunnus is a important commercially marine fish species,which is widely distributed. Thunnus has been intensively studied in China,including it s basic biological characteristics in fishery and artificial reproduction. However, very few studies on Thunnus were focused on genetic diversity,which has become an obstacle to f urther researches on the protection,management,rational exploi2 tation and sustainable utilization of Thunnus resources. The genetic diversity of wild Pacific blue2fin tuna ( Thunnus thy nnus) and Pa2 cific yellow2fin tuna ( T hunnus albacare) were investigated by using amplified f ragment length polymorphic (AFL P) in the present pa2 per. A FL P fingerprinting was regarded as one of the most usef ul techniques in nowadays for DNA fingerprint analysis. The result s showed that a total of 675 DNA amplification bands ranging from bp were produced using 9 pairs of primer combinations, and polymorphic loci were detected per primer combination. The percentage of polymorphic loci and Shannon s information index of the blue2fin and yellow2fin tuna were %, and %, , respectively. Both populations were at the same high level of genetic diversity. AMOVA analysis ( %) indicated that a certain extent of differentiation in two species. There were % of variance was among the populations and % of variance was within a population. Compared with other fi2 shes,both tuna species possessed rich population genetic diversities. The information of genetic diversities in different populations will give us theoretical guidance in breeding and genetic improvement. Key words : Thunnus thy nnus ; T hunnus albacares ;A FL P ;genetic variation ;genetic resources