Tgp chi Cdng ngh$ Smh hgc 10(3): 457-463, 2012
D A N H G I A D A D A N G DI T R U Y E N C A G I O {RACHYCENTRON CANADUM) B A N G C H I T H I P H A N T U M I C R O S A T E L L I T E
Lira T h | Hik Giang, TrSn T h | Thiiy H&, Nguyen Hfru Ninh, Do Xuan Hai Vien Nghien ciru vd nudi trdng thiiy san 1
TOM T A T
Cong tic chpn t^o gidng ca Gio la moi quan tam hang dau Irong cong nghe sinh san va san xual ca Gio thuang pham. Tim hieu da dang di truyen. cau Iriic quan the ciia cac quan dan ca Gio la rat can thiet trong viec nang cao sire sinh trudng va chon tao gidng, Viec su dung chi thi phan lu microsalellite trong nghien ciru cau triic quan the va xac dtnh bien di va sai khac di truyen giiTa cac quan dan ca Gio da dugc nghien cuu va cong bo trudc day. Trong nghien cuu nay, 8 locus microsalellite dugc diing de phiin lich da dang tren 3 quan dan ca Gio Viet Nam, Trung Quoc va Din Loan Tong sd 54 allele da dugc xac dinh Gia tri thong tin da hinh PIC deu trong khoang tir 0,662 ± 0,146 den 0,692 ± 0,087 Mire di hpp tir quan sat va mire dj hpp tiJr ky vpng a cac locus trong cac quan dan co chenh lech, nhimg qua kiem dinh thi cac locus microsalellite khong co sir sai khac so voi quy luat phan ly Hardy-Weinberg Co canh bao thieu hyt dj hop lu trong quan the ca Gio Viet Namthe hien qua he so can huyet (Fis = 0,247). Mire do sai khac di truyen gi&a cac quan dan la Irung binh (trong khoang lir 0,073 den 0,094) va hai quan dan ca Gio Viet Nam, Trung quoc co muc sai khac it hern so vdi quan dan ca Dai Loan,
TH- khoa Cd Gid. di hop tii. da dang di Iruyei crosaiellile. lan so alelte M O D A U
Ca Gid {Rachycentron canaditm). phan bd nhieu d khu vyc Dai Tay Duong, Thai Binh Duong va la mdt yong nhOng loai thuy san nudi bien cd gia yj kinh te cao va dupe nudi phd bien d mdt sd nude chau A. O Viet Nam san xual ca Gid khwig nhimg cung cap cho thi trudng Ucaig nude ma cdn phuc vu cho xuat khau.
Thii ca Gid yang, ihom ngon, dugc coi nhu la dac san, Ca cd gia tri dinh dudng cao ham lugng acid beo khdng no EPA va DHA cao hon so vdi nhieu ddi tugng nudi khac (Su et al., 2000). Viel Nam dugc xem la nude dimg thu 3 tren the gidi ve san xual va nudi ca Gid (Svennevig, 2001). Tuy nhien, ly le ca sdng sau giai doan uong nudi thap, nen lugng ca gidng van khdng du dap img cho nhu cau nudi ca Gio trong nude va van thucmg xuyen phai nhap mdt Iupng Idn ca gidng tu Trung Qudc va Dai Loan.
Ngay nay khi di truyen phan lir phat trien, viec img dung cac chi ihi phan lir vao danh gia vai lieu khdi dau va Up giup chpn gidng ngay cang nhieu.
Mdt sd nghien cim da xay dyng dugc chi thi phan tu phuc vy chgn gidng thuy san nhu. xac dmh ddng, loai va con lai; da dang di yuyen va xac djnh ngudn gdc ciia cac quan dan thuy san; xac dinh huyet thdng va ddng gop vat chat di yuyen; chi thj lien kel vdi vi yi gene quy djnh linh trang sd lugng, chi thj trg giiip chgn loc; lap ban dd di iruyen.
Tren the gidi, nhieu nghien cim su dung chi ihi microsalellite Uong danh gia da dang di truyen, bien di di truyen de phuc vu cho cdng tac chpn tao gidng cung nhu bao tdn ngudn gen quy tren nhieu ddi tuong thuy san. Nghien ciru ciia Pruett va cgng sy (2005) da cdng bd Idng sd 20 vi yi microsalellite da dugc tach ddng tir 24 con ca Gid thu tir ly nhien d Mississippi.
Renshaw va cdng sy (2005) sir dung microsatellite nghien cim da dang di yuyen tren 3 loai ca Trdng dd, ca Chi vang, va ca Gid da xac dinh dupe cac chung lai can huyet, xay dyng pha he va lap duoc ban dd locus cac linh trang sd Iupng Salan va cdng sy (2009) da img dung microsalellite nghien cim da hinh quan dan ca Gid d Iran. Qua phan lich di yuyen, sy khac nhau trong va giua cac qiuin dan hoang da ciia ca Old o hat viing bien Iran qua 10 locus da xac dinh dugc il nhal cd 3 quan dan khac nhau.
Trong nghien cuu nay, chi thi phan lu microsalellite duoc sir dung de danh gia muc dp da dang dl truyen va eau Iriic quan the cua ca Gid d vung bien Viet Nam, Trung Qudc va Dai Loan.
V A T L I E U VA PHUONG PHAP NGHIEN CUU Thu m i u thi nghiem
Mau vay ca Old ciia ba quan dan Viel Nam, Trung Qudc va Rai Loan dugc thu d Cat Ba - Hai 457
L u u T h i H a G i a n g et ai
P h d n g . T h u ng?ui n h i c n 3 0 m a u ca l i r m o i qui\n d a n , E l h a n o l 1 0 0 % , l y l a m g i i r l a i k e l l i i a , _ D N A ket tiia cat v a y va biio q u i i n I i o n g e l h a n o l 1(10"" o i i h i p l d p d u p e r u a hai lan b a n g E l h a n o l 7 0 % M a u D N A dupe 4 " C c h o dc-n k h i sir d p n g boa lan b.ing d u n g d j c h d p m 1 x T E va bao q u a n d 4 " C . C i i c c a p m o i e h o P C K
N g h i e n c i r u d u g c I i c n h a n h I r c n 8 l o c u s m i c r o s a l e l l i t e \ d i t r i n h l y ' " d i , I r i n h l u l i i p \ a n h i p l d d g i i n m d l dtroc t r i n h b;'i> o b.iiig I.
I ; i e h c h i i - t D f N A l d n g s d
I'lirdc hOl m a n vfiy d i i g t n g a m IS g i d I r o n g l , 7 n i l d u n g d i c h d e m l \ I I ( 1 0 m M r i i s - b n s e , 0,1 i n M F D l A , p l U , 0 ) M a i l d u g c u o (.0"C t r o n g 7 2 0 fil C e l l l \ M s S n l u l i o n ( 1 0 i n M I n s . 100 i n M I ' D T A , 2 ' l „ _ s i ) S p i I 8.0), 5 p i PioieiiKi'^e-K ( 2 0 m g / i n l ) dc-n k h i m a i l lan hoi'iii l o i i i i I r o n g d u n g d i c h , I h e m v a o d u g c h u a m i n i 2 4 i i ( i l d u n g i h c l i P i o l e i n P i c c i p i l a l c S o l i i l i o n (7,5 M \ m m i > r n i i i n A L C I . I I C ) . ihc l i i n i d i - i i m a i l \ , i dc o 4 " C I r o n g 2(1 phi'it. M . u i d u o c l y l.lin \ i r i tdc d p 12(10 \ d n g p h i i t va l;iy p h a n d u n g d i c h p l i i a tren D I I I I L ; d i c h LI> chira D N A d u o c kel tiia b a n g
I ' h i i n m i ) ; P C R
Phiin i m g P C R v d i i d n g I h e l i c h l O y l bao g d m 4 , 1 5 f i l N u c l e a s e free w a t e r , 2 p l I X B u l l e r colorless g o I'iK] l l e x i ; 0 . 8 p l d N T P s 2 0 0 y n i ; 0 , 5 p l m d i loai inoi (0.5(1111); I p ' ^^^'^1 l m M ; 0 , 0 5 M l ( i o taq l l e x i b,25U va I f i l D N A l o n g sd. C l u i t r i n h nhipt ciia p h a n ung b i l d i i u d 94"C' I r o n g 3 p h t i l , t i e p I h e o la 3 0 c h u k y gom 9 4 " C i r o n g 3 0 g i i i y , nhiet d o g a n m d i ( r i e n g cho lung m o i ) I r o n g 45 gii'iy. g i a i d o a n l o n g h g p 7 2 ' C Irong v d i i g 6 0 g i a y , va c u d i c i i n g lii g i a i d o a n k e o dai chuoi U o n g v i m g 10 p h i i l o 72 " C . K e l t h i i c phan i m g , san p l i i i n i P C R d t r o c g i i r i r 4 " ( d e n k h i sir d u n g .
San p h a m P C R d u g c c h ; i \ d i c n d i Iren gel P o l y a c r y l a m i d c (>"o \ a n h u d i i i b o i d u n g dich I v l h i d i t i i n b r o m i d e 0 5 % I r o n g 15 pht'it \;'i soi kiem tra I r c n d e n U V
g 1 Tnnh l u lap ua nhiet do gan n a 8 chi thi microsalellite
Microsatellite T r i n h t u ' m o i ( 5 ' - 3 ' ) So hieu
g e n b a n k
CATATCAAGTCAATATCACAGACC CCACGGAATAGCAGACTTTCTC
(CA)3GA (CA)5A(CA),6
N h i f t do gan r"^'(°C) Rca IB-F06
Rca I-A04
R e a l - A l l
Rca1-E09
Rca1-E06
Rca1-C04
Rca1-B12
CAAGCAAATGCGTGGCCGA CGTTAGCAACCACACGAGCTTG CACGCACATGCACTACTTTAACC GCTGTTGATGTGGCGAAGCAAC CTACAGTGGTGTTCCCTGTTAG CAGTACATAGAGAAACAGGAGG CATGTTATTCTCCAACTCATGG GTGTATCCGCATACTTTCAG GGCACCAATCACTCACTACTG TGTTGAGGTCTATCAGTGCC GACATCAAGTGGCACTTTG CACTAAACTTGTTCCTCCTG GCTTCAGGCAAGTGAGACC GGGAGGTAATTATGTCCTGT
(CTAT),5
CA)9(CACT).
(GT)2.
(GATA)j,
(CA)33
(GT),r
(AC)9
AY721668
AY721672
AY721673
AY721671
AY721679
AY721675
AY721674 55
60
55
48
46
48
55
P h a n l i c h so l i e u i h e o k i c h t h u d c c u a cac d o a n d u g c n h a n len ciia mpl l o c u s m i c r o s a l e l l i t e t r e n ca 3 q u a n d a n ca, cd bao B a n gel d u o c c h u p a n h v a g h i ket q u a . C a c v a c h n h i c u d o a n ed k i c h t h u d c k h a c n h a u t h i c d bay nhieu b i i n g tren l u n g ban g e l d u g c d d i c h i e u v d i t h a n g a l l e l e . C a c k i e u g e n m i c r o s a t e l l i t e d u g c ma hoa c h u a n va d d i c h i e u v o i n h a u , C a c a l e l l e I r o n g locus i h t i n h diir l i e u d ^ u v a o c h o p h a n m e m G e n e p o p 4,0 m i c r o s a t e l l i t e tai d u g c m a hda t h e o k y h i e u b a n g c h u ( R o u s s e t , 2 0 0 8 ) , &h t i n h l o a n l ^ n s d a l l e l e , mire di cai i n h o a ( A , B, C , . , ) , C a c a l l e l e d u p e xac d i n h d y a h g p l u q u a n sat ( O b s e r v e d H e t e r o z y g o s i t y = HQ),
Tap chi Cdng nghe Sinh hoc 10(3): 457-463, 2012 mire dl hgp tir ky vpng (Expected Heterozygosity He), va kiem Ira do lech tir trgng thai can bang Hardy-Weinberg. H? sd sai khac di Iruyen (Est), hp sd can huy^l (Fis) va lugng thdng tin da hinh (PIC) dugc tinh loan bang got phan m^m Arlequin 3,0 (Excoffier c? n/, 2005),
KET QUA VA THAO L U A N
T i n h d a d a n g d i t r u y e n t r o n g c^e q u a n the ca (^iii Sau khi xu ly sd lieu bang phan mem Arlequin va Geiicpop, chimg lot da long hpp cac giti tri \ e l.in sd allele, sd allele tren locus (Na), di hop lu phat hien (Ho), di hgp tu mong doi (He), chi sd can luijei FIS, he sd tuong quan di truyen Isi. lugng thdng tin da hinh (PIC) Ciic chi sd n.i\ duoc phan tich \;i th;io tuan ophan Hep iheo.
Da hinh alleles o' mdi locus microsatellite Innig eae quan dan cii Gid
S u d a hinh cua ciie locus duoc the hien tren bang sd lieu cung \oi muc dp sai khac cac tan sd allele cho ihay sy khac nhau ve di Iruyen cua cac quan dan ca Gid. Tam locus microsatellite da lao ra long sd 54 allele, trong dd quan sal duoc d locus Rca IB-E09 va Real - C04 la 9 allele. Real - E06 la 10 allele, quan sat dugc 6 allele d locus RcalB-F06 va R e a l - A l l , locus Real- BI2, Rcal-A04 la 5 allele, d locus RcalB - E08A chi quan sal duoc 4 allele. Nhu vay, cac locus RcalB - E09. Real- E06, Real - c 6 4 , RcalB-F06 va locus R e a l - A l l cd dd da hinh cao hon locus Rca I-A04, RcalB - EOS A va Real - B12.
Nghien ciru ciia Pruett (2004), ciing chi ra cac locus Rca I-A04, Rca IB - EOS A va Rca 1 - B12 cd mire da hinh Ihap hon cac locus cdn Igi.
d locus RcalB-F06 cd allele RcalB-F06" voi lan sd xuat hipn cao nhal 0,321 o quan dan ca Vi^l Nam va 0,429 d quan dan ca Diii Loan, Tan sd allele thap nluillti RcalB-FOd", chi Ihiiy xual hien o quiin dan ca Viel Nam vdi liin sd 0,071. Xuiit hien vdi lan sd cao 0,214, nhung allele RcalB-i-06'* lai chi Ihiiy CO o quan dan cii Viel Nam iiiii khiing cd o hai qtiiin dan ca cdn lai.
T,^n sd allele cao nhiil o locus Rcal-Al 1 In o Rca 1 -A 1 I" irong khoang 0,100 a qu;iii dim Dm Loan Vii 0,467 o quiiii dim Viel Nam, Tiin sd allele Ihap nliiit la Rcal-Al l' trong khoang 0,000 d quan dan Dai Loan \.'i 0,033 d quan dim Viel Nam, Allele Rcal-Al f^chi Ihiiy xuiil hicn o quan dan Dai Loan vdi tan sd luinig ddi cao 0,476 mil khdng thay cd d quan dan Trung Quoc va Viel Nam,
0 locus Real-A04 lan sd ciia cac allele Rcal- A04 ' va Rca I-A04 deu rat cao, len tdi 0,633 a qLi;lii dan Dai Loan. Nhung d eae allele Rcal-A04'^ va Rcal-A04" thi t,in sd allele lai lh5p hon, chi la 0,000 den 0,400 Quiiii dan ca Trung Qudc khdng thay xuiit hien hai allele n<iy
Tai locus Real - E09, allele Real - E 0 9 ' ' c d t a n sd cao nhat la 0,536 tai quan dan Viet Nam. Quan dan Viel Nam khdng thay cd sy xual hien cita allele RcalB - E09'^ va RcalB - E09'. Allele RcalB - E09' chi thay xuat hien Iren qiuin dim Trung Qudc vdi lan sd 0,017.
300bp
G i 4 n g 1 ^ 1 5 M l u c ^ g i b Viet Nam, M ladder lOObp
Hinh 1. San p h i m PCR m6i R c a l - A l 1 qu^n dan ca Gio ViSt Nam tr§n gel Acrylamide 6%
Locus Real - E06, tan sd allele Real - E06 tat quan dan Trung Qudc den 0,224 Uen quan dan kha cao d ca 3 quin dan n i m trong khoang 0,117 Viet Nam. Real - E06'^ va Real - E06' deu khdng
Luu Thj H^ Giang elal
ih^y xukx hipn l i o n g qii;\n diin Vifl Nam. Trong Tuy nhifin tren qu3n dan Vipl Nam Igi thay tan s6 khi dd allele Real liOd" liii xu:)l hipn o qiit^n dim allele Real - EOd*^ dai 0,138 Irong khi nd chi dai Trung Quoc \ o i xCm sd 0,100; cdn allele Real 0,054 Iren quan dan Dai Loan va 0,000 tren qu5n E06' cd lAn sn lii 0,161 lai quiin diin Diii Loan dim T r u n g Q u d c .
!. Tin sd alleles Irfin cdc locus
Rca1B- F06
R c a l - A11
R c a l - A04
R c a l - E06
A B C D E F A B C D E F A B C D E A B C D E F G H 1 K
Vl^t Nam 0,214 0,321 0,036 0,071 0,375 0,000 0,000 0,333 0,033 0,467 0,033 0,133 0,000 0,400 0,167 0,267 0,167 0,035 0,000 0,069 0,034 0,224 0,000 0,155 0,155 0,189 0,138
T r u n g Qudc 0,000 0,400 0,167 0,000 0,267 0,167 0,000 0,393 0,000 0,214 0,107 0,286 0,000 0,000 0,214 0,571 0,214 0,100 0,100 0,250 0,167 0,117 0,050 0,000 0,133 0,083 0,000
Ddi Loan 0,000 0,429 0,071 0,000 0,393 0,107 0.467 0,167 0,267 0,100 0,000 0,000 0,067 0,000 0,267 0,033 0,633 0,018 0,018 0,179 0,178 0,161 0,161 0,036 0,179 0,018 0,054
T r u n g b i n h t S D 0,071 ±0,124 0,38340,056 0,091 ±0,068 0,024±0,041 0,345±0,068 0,091±0,085 0,156±0,27 0,298±0,117 0,100±0,146 0,260±0,188 0,047±0,055 0,140±0,143 0,022+0,039 0,133±0,231 0,216±0,05 0,290±0,27 0,338±0,257 0,051±0,051 0,039±0,039 0,166±0,166 0,126±0,126 0.167*0,167 0,07±0,07 0,064±0,064 0,156±0,156 0,097±0,097 0,064±0,064
R c a l - C04
R c a l B - E08A
R c a l - B12
R e a l - E09
A B C D E F G H 1 A B C D A B C D E A B C D E F G H 1
Vl^t Nam 0,071 0,196 0,089 0,036 0,125 0,268 0,214 0,000 0,000 0,258 0,396 0,052 0,293 0,083 0,217 0,000 0,233 0,467 0,000 0,178 0,250 0,304 0,071 0,125 0,536 0.018 0,000
T r u n g Q u 6 c 0,133 0,100 0,100 0,017 0,017 0.050 0.083 0,367 0.133 0,300 0,517 0,000 0,183 0,035 0,172 0.172 0.603 0.017 0,069 0,172 0,172 0,086 0,189 0.138 0,138 0,017 0,017
Dai Loan 0,000 0,017 0.103 0,069 0,052 0,069 0,052 0,189 0,448 0.300 0,367 0,300 0,033 0,000 0.155 0,138 0,155 0,552 0,067 0,100 0,167 0,150 0,100 0,367 0.017 0,033 0.000
T r u n g b I n h i S D 0.068±0.067
0,104 ±0,09 0,097±0.007 0.041 ±0,026 0,065±0,055 0,129±0,12l 0,116±0,086 0,185±0,184 0.194±0.23 0,286±0,024 0,427±0,08 0,117±0,16 0.170±131 0,03910,042 0.181 ±0.032 O,1O3±0,091 0.330±0,239 0.345±0,288 0.04510.039 0,150±0.O43 0,19610,047 0,180±0,112 0.120±0,061 0,210±0,136 0,230±0,272 0,023±0.009 0,006±0,01
Allele Real C04' cd tan sd allele cao nhat Iai locus Real - C04 la 0.44S IT quan dan Dai Loan, 0,133 tren quan dan Trung Qudc. Tuy nhien, allele nay lai khdng ihay xual hien d quan dan Viet Nam.
Ngoai ra, tren quan dim Vicl Nam cdn khdng thay sy cd mat cua allele Real - C04", trong khi dd allele nay lai xual hien tren hai quan dan kia vdi tin so kha
ciio \ o i 0,189 (Dai Loan) va 0,367 (Trung Quoc).
Allele Real - 0 0 4 " cd t i n sd la 0,000 tren qu^n dan Dai Loan, 0,071 lai quSn dan Viet Nam va 0,133 6 quIn dan Trung Qudc, Tai locus nay thi quan dan Trung Qudc la cd su xuat hi?n ciia nhieu allele nhal hiiy ndi each khac la tren quiin dan Trung qudc Real - C04 cd sy da hinh n h a t
Tgp chi Cdng nghe Sinh hgc 10(3): 457-463, 2012 Ddi vdi locus Real - BI2 thi quiin dan Trung Qudc xual hien 5 allele, nhieu hon so vdi cac quan dim khac. Real - B12" la allele cd tin sd xuil hien cao nhat (0,603) va dugc the hien Ihdng qua quiin dim Trung Qudc. Quan dan Viet Nam va Dai Loan d§u cho thay sy cd mat ciia allele Real - BI2' la cao nhat vdi tan sd luong irng \h\ lirgt la 0,467 va 0,552 Tai quan dan Viet Nam khdng Ihay sy xult hi^n cua allele Real - B12^ Cdn tren quiin dan Din Loan lai khdng thay sy xuiil hien cua allele Real - B I 2 \
Locus RcalB - EOS'^ la locus kem da hinh nhiil trong cac locus nghien ci'ni, chi Ihiiy Miiil hien co 4 allele iren quan dim Viet Nam vii Dili Loan, cdn qufin dan Trung Quoc lai chi cd 3 allele, khdng ihay xiiiil hien allele RcalB - EOSA^ Allele cd mat nhi^ti nln^t lii RcalB - E O S A " \ O I tSn sd 0.^17 Iren qiuin diin Trung Qucic, 0.396 a quan dan Viet Nam vii 0.367 Irong quan diin Dili Loan,
Cd the nhgn ihiiy d ca 8 locus cua ba quiin dtin ci Gid nghien ciru deu ton lai ctic allele cd lan sd xuat hien rat Ihap, vdi gia trj nhd hon 0.1. Cac allele nay deu cd the do dang bi mat di neu khdng lien hanh lai ba quan dan ca Gid nay vdi nhimg quan dim khac cd bien di di truyen cao hon hoac lai cheo giua cac quan dan nham lam giam nguy co bi mat ciic allele nay, lu dd ciing giiip han che sy thieu hut allele o ba quan dan ca Gid nghien cuu.
He so sai khac di truyen
Su dung gia Iri Fsl de xac dinh mdi quan he giira cac quan the nghien cim. He sd nay cd the danh giii mdi lien he khdng ngau nhien giua cac allele trong quan the Fsl luong img vdi tan sd allele khac nhau giua cac quan the. Neu Fsl< 0.05 thi sai khac di truyen nhd. Neu 0.05<Fsl<0.15 ihi sai khac di truyen la trung binh. Neu Fsl>0,15 thi sai khac di truyen lii ldn(Nie, 1978)
Gia In he sd sai khac di truyen ihu dugc lu 3 quan dan nghien cuu (Fst) cho thay rang su sai khac dl Iruyen giua cac quan dan d mire Irung binh (Irong khoang tu 0,073 den 0,094), va hien luong ddng chay gen giua cac quan the d mire ihap.
Bang 3. He s6 sai kh^c di tmyen (Fsl) cua ba quIn dan ca Gio
Viet Nam Trung Qudc Dai Loan
Viet Nam 0,000
Trung Quoc 0,087 0,000
DSiLoan 0,073 0,094 0,000
K i ^ m t r a cSn b 3 n g H a r d y - W e i n b e r g Trung binh lugng ihdng tin da hinh PIC Irong loan bp ba quan dan deu >0,5 (>0,662) khang dmh tinh da hinh giOa cac quan dan vdi nhau rat cao Tan sd di hop lir mong doi ddt vdi irung binh cac locus phan bd Irong khoting giii tri ihiip nhal la 0,633±0.32 d quiin diin Diii Loan cho den cao nhiil la 0,709±0,19 d quan dim Vipl Nam. Trung binh loan bd muc dj hgp lu quan s;il vii mire di hgp lit k> \ img khdng qua chenh lech nhau, Mi'rc di hgp tu quan sal nhd hmi mire di hgp lir du kien, chi cd o locus RcalB-F06, locus RcalB-EOKA o c a hai qiitin dim vii locus Rcal- C04 o- quiin diin Diii Loan cho nu'rc dj hgp lu quan Siil Idn hon mire dj hgp lu ky vgng, luy nhien su .sai khiic nii\ khdng cd y nghTa Kiem djnh cho ihiiy o Ciic locus microsalellite cua cac quiin dan ca Gid khdng cd sy sai khiic so vdi quy luat phan ly Hardy- Weinberg vdi mire y nghTa P<0 01,
He sd ciin hiiycl cua quan diin Viet Nam cao nhal 0,247±0,2 the hien sy thieu hut dj hgp lu cao dac biel la d hai locus Rcal-E09, Rca I-B12 (Fis=
0,51 va 0,663) He sd can huyet ihay ddi khac nhau giua cac locus Irong quan dan Dai Loan, nhung cd trung binh Fis=0,075 ± 0,43 nen sy thieu hut di hgp lir la khdng dang lo ngai, Trong khi dd he sd can huyet tai quan dan Trung Qudc la nhd -0,017 ± 0,24, the hien sy vugt Irdi cita ca the mang allele di hgp tir so vdi o hat quan dan Viet Nam va Trung Qudc Hien tugng giao phdi can huyet xa> ra vdi ty le kha cao trong quan dan Viet Nam va Dai Loan dan den giam linh da dang ciia cac quan the. Viec canh bao ve mire do can huyel la dieu can thiet trong chuong trinh chpn gidng bdi vi neu quiin the muc dp can huyel cao se dan den tinh trang thieu hut di hgp lu, khdng dam biio lieu chuan chpn linn con gidng de lien hanh lai lao Iren mdl sd tinh trang dang va se lien hanh chon Igc.
K E T L U A N
Phan lich da dang di truyen cua 3 quan dan ca Gid Viet Nam, Trung Qudc va Dai Loan su dung 8 locus microsalellite cho thiiy quan dan ca Gid deu cd lugng Ihdng tin da hinh cao (>0,662). Gia tri allele Irung binh d ca 3 quan dan nghien ciru la 5,5 allele/l locus ihap hon so vdi quan dan ca Gid trong nghien cim Ciia Pruett va cdng sy (2005) (9,5 allele/I locus) Nhu vay, ba quan dan ca Gid nghien ciru trong de tai cd sy thi&u hut allele, dong Ihdi cung cd sy thieu hut dl hgp tu mong dgi cd y nghTa (P = 0,0001). Ben canh dd, chiing tdi nhan ihay ba quan dim ca Gid nay 461
L u u T h i H a G i a n g CM/,
c d s u k h i i c b i e l d i t r u y e n c h i d m u c I r u n g b i n h , V i p c t r u y e n v a h a n c h e l a c d g n g l i e u e y e c i i a can huyft l a i c h e o g i u a cac quiTii d i m ca G i d c i i c h x a n h a u vc n h u n g van g i u d u g c cac d a c l i n h t d i c u a c;ic d d n g ca d i a l \ . c d I h d la p h u o n g p h a p l o t d e t a n g da d a n g d i n h g p n g i .
B a n g 4 K i t q u i ki6m djnh lung locus microsatellite Iheo dinh l u ^ l Hardy-Weinberg
STT
1
2
3
Q u I n d a n
Vi^l Nam N' Ho' H e ' Fis' PIC' P^
Trung Qu6c N' Ho' He' F I S ' PIG' P^
Dai Loan N' Ho' He' F I S ' PIC' p6
L o c u s R c a l -
A04 R c a 1 - A11
R c a l B -F06
R c a i - E09
R e a l - EOS
R e a l - C04
R c a l - B12
14 0,72 0,86 0,16 0,696
<0,01 15 0,67 1 0,32 0,578
<0,01 15 0,738 1 0,26 0 657
<0,01 30 0,393 0,804 0,516 0 75
<0,01 30 0,621 0,854 0,277 0.819
<0,01 30 0,786 0,829 0,053 0,788
<0,01 30 0.233 0,685 0,663 0.626
<0.01 R e a l B E08A
30 0.552 0,699 0,214 0,620
<0,01 T r u n g binh
24.25 0,709±0,19 0.95310,12 0,24710,2 0,692±0,087
<0,01
15 0,74 0.87 0,14 0,67
<0,01 14 0,73 0,93 0,21 0,661
<0,01 14 0,60 0,43 - 0 , 4 0,464
<0,01 30 0,793 0,869 0,089 0,838
<0,01 30 0,667 0,863 0,231 0,831
<0,01 30 0,767 0.814 0,059 0.779
<0,01 30 0,345 0,585 0.415 0.554
<0,01 30 0,833 0,620
•0,353 0,501
<0.01 24,12 0,69010,16 0,743±0,1B -0,017±0,24 0,662±0,146 eO.OI
14 0,67 1 0,33 0,577
<0,01 15 0.69 0,78 0,11 0,627
<0,01 15 0,54 0,5 -0,08 0,433
<0,01 30 0,067 0,803 0,918 0,765
<0,01 30 0,857 0,863 0,007 0,829
<0,01 30 0,793 0,750 -0.058 0,71
<0,01 30 0,103 0,639 0,841 0,641
<0,01 30 0,833 0,696 -0.201 0,571
<0,01 24,25 0,63310.32 0,76010,15 0,075±0,43 0.644±0,124
<0,01 'n S6 mau nghien c u u , ^Ho IUILTC di hop t u quan sat (Obser>
Heterozygosity), " F I S He S 6 c$n huy^l, 'PIC L u g n g thong tin
L d ' i c a m o*n (.'dug Iritili nay dtroi Itodn thanh voi kinh phi < iia de lui "Ung dung chi thi phan ttr PN i trong danh gid da dang Jt truyen mdl .\d quan ddn t a Gid (Rachycentron ianadum)" thudc chtrang ninh de an Phat trien vd irng dung i dug nghe sinh Itoc Irong linh virc Ihin san. Bd Ndng nghiep vd Phat trien Ndng thdn Dirge time hien Igi phdng lIu nghiem Di truyen vd Chgn gidng. Vien Nghien cim nudi trdng thiiy san I.
T A I L I E U T H A M K H A O
Briggs JC (1960) Fishes o f w o r l d - w i d e (circumlropicaH
/ed Heterozygosity),') H^ Muc di hgp t u ky vpng (Expected da hinh.^'p Sai khac y nghTa so voi Hardy-Weintwrg
distribution, (.••bu 171-181
C h i r s t i n LPruet, f-ric S, M a r k A R , John CP, Caird ER.
John R U (200.>) Microsatellite D N A makers Tor population genetic studies and parentage assignment in cobia, RcK hvcentron canadum Mol Ecol Notes 5: 84-86.
l-erguson M . (1995) The role o f molecular genetic markers in the managent o f cultured llshes. Rev Fish Biol Fish 4(3) 351-373
Ibrahim O, Y i l m a z C (2003) Fish Population Genetics and M o l e c u l a r Markers, I I - Molecular Markers and Their Applications i n Fisheries and Aquacullure. Turk J Fish Aqiial Sci 3. $\-79.
Su MS, Chien YH, Liao IC (2000) Polennal of marine Tap chi Cdng nghf Sinh hgc 10(3): 457-463, 2012
Nei M (!978) Estimation of average heterozygosity Shaffer RV. Nakamura EL (1989) Synopsis of Biological genetic distance from small number of individuals. Data on the Cobia, Rachycentron canadum, (Pisces, Genetics 89:283. Rachycentndae). National Marine Fisheries Serv , Panama Svennevig N (2001) Fanning of cobia or black kingfish ' •"
(Rachycenn-on canadum). Head. International Projects Department SINTEF Fisheries & Aquacullure, N-7463,
Trondheim Norway '-'•'='•' i^quaeukure in Taiwan Cobia culture Iniernalional Svinposiiim on Cage Aquacullure in Asia. Tungkang, Liu ZJ, Nichols AP, Li P, Dunham RA, 1998. biheritance Pintimg (TaiuvnI. 2-6 Nov 1999
and useftilness of AFLP markers in channel catfish
(lctaluriispunctatus),b\ueca.lfish{lcfaluriislurcatiis). and \ci^poor E (1998) Molecular markers and genelic Iheir Fl, F2. and backcross hybrids. Mol Gen Genet managetiiem of tanned fish Biolog}' of Farmed Fish 258(3)- 260-268 Shaffield Acadamic Press, England 355-377.
S T U D Y I N G G E N E T I C D I V E R S I T Y O F C O B I A (RACHYCENTRON CANADUM) U S I N G M I C R O S A T E L L I T E M A R K E R S
Luu Thi Ha Clang*. Tran Thi Thuy Ha. Nguyen Huu Ninh. Do Xuan Hai Research Institute for Aquacullure No 1
SLIMMARY
Selecting seed of cobia is the first concern in reproductive technology and producing commercial cobia.
Studying genetic diversity and structure of cobia population is important to improve growth rate and selecting generation cobia (Rachycenti on canadum) Using microsatellite marker in studying population structure, defining definning vanauon and genetic differences among populations of cobia were carried out, were studied and published. In this sludy, 8 microsalellite loci were used to analyse genetic diversity of three cobia populations of Vietnam, China and Taiwan. 54 alleles were identified Tlie polymorphic information content (PIC) value of each locus ranged fi-om 0,662±0.146 to 0,692±0,087 Observed and expected heterozygosities (y) at loci in these populations were differems. and all three population Hardy-Weinberg ratios. The level of genetic diversity in populations is not high through inbreeding coefficient (Fis = 0,247 and 0,075), that suggests had inbreeding mating in these populations. There were warning heterozygote deficiency in populations of cobia Vietnam expressed Fis - 0.247 The moderate geneiic diversity among cobia populafions was franged from 0 07266 to 0.09368) and two populations of Vietnam, China were less different.s compared with Taiwan cobia population.
Keywords- allele frequence cobia. genetic divers iri: heleroz\gosit\'. microsatellite
' Author for correspondence: Tel: +84-241-3842383: E-mail: hagians&rial.org
