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Báo cáo " Xác định các chỉ thị AFLP đặc trưng cho bảy dòng cá rô phi nuôi ở Việt Nam "

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Xác định các chỉ thị AFLP đặc trưng cho bảy dòng cá rô phi nuôi ở Việt Nam

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  1. Tgp chi Cdng nghi Sinh hgc 7(4): 455-462, 2009 X A C D I N H C A C CHI THI A F L P D A C T R L T N G C H O BAY D O N G CA R O PHI N U O I d VIET NAM Trin Thj Bich Hong', Nguyin Thi Td Nga', Ngo Thj Kim', Quyen Dinh Thi', Nguyin Hfru Ninh^ Pham Anh Tuin^ 'Vien Cdng nghi sinh hgc ^Viin Nghiin cim nudi trdng Thiiy sdn I TOM TAT Bay ddng ca rd phi nguyen lieu ngoai nhap tir cac nudc khac nhau thupc 2 loai Rd phi xanh (Oreochromis aureus) va Rd phi van (Oreochromis niloticus) dugc sir dung de nghien cim tinh da dang di truyen, tim marker AFLP phan biet cac ddng ca rd phi. DNA cua 7 ddng dupc ghep vdi muc dich giam sd lugng mau ma khdng anh hudng den chat lugng, muc dich nghien cim. Sir dung NTSYSpc cho thay he sd tuong dong giira cac ddng rat cao > 0,85, dac biet la giira cac ddng Rd phi xanh hoac giira cac ddng Rd phi van (> 0,97). Cd 13/16 td hgp mdi deu the hien su khac nhau giiia Rd phi van va Rd phi xanh ve mat di tmyen va 6 td hgp mdi xuat hien bang dac trung cho mot so ddng. Vdi tdng sd 375 bang thu dugc (tmng binh mdi td hgp mdi co 23,4 bang), chi cd 64 kieu bang da hinh trong do 40 bang cd the dung nhan dang Rd phi xanh hoac van va 12 bang dac tnmg cd the dung nhan 7 ddng nhu: AF25/220, AF26/210, AF26/350, AF29/330, 'AF42/180, AF42/170, AF43/170, AF43/225, AF43/250, AF43/160, AF54/125, AF54/180. Day la cac bang dac trung cd thg dung lam nguyen lieu cho viec phat trien cac chi thi phan tir trong chpn gidng d ca rd phi. Tit lihoa: AFLP, bdng dac trung, Oreochromis aureus, Oreochromis niloticus, ro phi MODAU Anh Tuin, 2005). Cac ddng thuan dugc luu giii chu yeu d cac trung tam gidng. Rd phi canh tac la cac Ca rd phi d Viet Nam cd rat nhieu loai nhu Rd ddng lai tao cd dugc tir cac chuang trinh chgn gidng phi van {O. niloticus), Rd phi xanh {O. aureus), hay Norivay nhu NOVIT 1, 2, 3... dugc tao tu ddng rd Rd phi den {O. mossambicus). Do ddng Rd phi den phi bd me GIFT (Genetic Improvement of Farmed (nhap tir Philippines) de qua day, lai cham ldn, kich Tilapia, 1994) (Le Thanh Lim, 2000). Trong cdng thudc ca the bi giam dan, nen ddng rd phi nay khdng tac chgn gidng, chii yeu nghien ciiu tim kiem rd phi dugc phat trien. Trudc day, vdi dieu kien canh tac d sinh trudng tdt frong cac dieu kien Viet Nam theo Viet Nam, viec luu giir gidng thuan khdng bi tap hudng lai tao cac ddng hien cd de tim cdng thirc lai giao giiia cac ddng la van de rat khd khan cho cac cho ty le gidi tinh due cao (Pham et al, 2002) hay cd nha chgn gidng ca rd phi. Hien tugng tap giao cua tdc do sinh trudng tdt d vimg nudc lg man (Pham gidng gdc vdi cac loai rd phi dai la mdt trong nhiing Anh Tuan et al, 2008), nang cao sire sinh trudng va nguyen nhan lam giam nang suat va chat lugng cua kha nang chiu lanh bang chgn lgc gia dinh (Nguyen mua vu frong nudi trdng thuy san. Kinh nghiem ciia Cdng Dan et al, 2003). Tuy nhien, cac nha chgn cac nha nudi trdng thuy san cho thay neu mudn nudi gidng van khdng ngimg tim kiem cac ddng rd phi cd hieu qua thi phai giii dugc chat lugng ddng thuan, mdi cho hieu qua san xuat cao tu cac ddng thuan sir ddng thdi nghien cim tim bien phap nang cao chat dung phuang phap lai khac loai. lugng di truyen cua chiing va kha nang thich iing vdi dieu kien nudc ta thdng qua qua trinh chgn gidng Tren the gidi, cac nghien cim AFLP tap trung (Dd Diic Hanh, 2006). chil yeu d mdi quan he di truyen, danh gia tinh da hinh quan the (Koeher et al, 1998). Marker phan tit Cac nhdm tac gia nhu Quyen Dinh Thi, Nguyin ra ddi da phan tich true tiep vat lieu di truyen DNA Thi Dieu Thiiy, Nguyen Van Cudng (Nguyin Thu dem lai do chinh xac cao frong viec danh gia va xac Thiiy et al, 2004; Quyin Dinh Thi et al, 2004; dinh cac ddng, loai ca. Mdt sd tac gia nhu Franck, 2005), Nguyen Huu Ninh, Pham Anh Tuin (2003) Rutten, Penman va McAndrew... da sir dung mdi chi sir dung microsatellite, AFLP, RAPD budc microsatellite de nhan dang, xac dinh loai va liet ke diu danh gia da hinh hay mdi quan he di truyin giiia mdt loat cac vi tri du doan lien quan chat che vdi cac cac dan ca nhim tim kiem cac rnarker lien kit gidi allele khac nhau giiip phan biet giira cac loai chu yeu tinh, huylt thdng va pha he (Quyin Dinh Thi, Pham va ddng lai (Franck et al, 1992; Penman, Mcandre'w, 455
  2. Trin Thi Bich Hdng et al 2000; Rutten etal, 2004). Ddi vdi marker AFLP, chi Phu'ffng phap AFLP cd 4 chi thi vl gidi tinh trong dd cd 3 chi thi lien kit Phuang phap AFLP (Vos et al, 1995) cai biln hi NST Y {OniY425, OniY382, OniY227) va mdt lien phuang phap ciia Van Eck va ddng tac gia (1995). Sau kit NST X {OniX420) dugc tim thiy (Ezaz, 2002; khi gin adapter EcoRI (adapter E) va Msel (adapter Ezaz et al, 2004). Nghien cim chgn gidng rd phi M) adapter vao san phim cit han chi DNA. PCR hien nay dang dugc frien khai manh d cac nudc dien chgn lgc sir dung mdi dua tren trinh tu cua ^coRI va hinh nhu Ai Cap, Ecuador, Malaysia, Malawi, Msel va them cac nucleotide chgn lgc tit san pham Norway, Philippines va Viet Nam thdng qua cac du tiln PCR (PCR khdng chgn lgc). Kit qua ciia PCR an NORAD (Norway). Nhiing nghien ciiu ve bang chgn lgc dugc hien thi tren gel polyacrylamid 5%. dac trung hay nhan dang ddng bang AFLP cQng la tiln dl CO sd cho viec tim kiem chi thi phan tir dac trung cho cac QTLs (Quantitative Trait Loci). Phan tich so lieu Su cd mat cua 1 bang dugc ma hda bang sd 1, sir VAT LIEU v A PHUONG PHAP khdng xuat hien dugc ma hda la sd 0. Sd lieu ciia timg cap mdi dugc dua vao excel danh gia va sau dd Mlu xir ly bing NTSYSpc. 189 miu vay ca cua 7 quan dan thudc cac ddng ca nhap tii cac nudc Thai Lan, Tnmg Qudc, Dai KET QUA VA THAO LUAN Loan, Philippines va Israel dugc luu giii tai Vien Nghien cim Nudi trdng Thiiy san I (Dinh Bang, Tit Ba muoi mau DNA cua tirng ddng ca rd phi San, Bic Ninh). Do nam 2008 thdi tiet khac nghiet dugc ghep lai vdi nhau trong ndi bd ddng dd de giam nen ddng ca Rd phi van Thai Lan bi chet chi cdn 9 ca so lugng miu nghien citu. Cac bang DNA dac tnmg thl (Bang 1). cho ddng khdng chi dimg de nhan dang ddng ma cdn cd the ket hgp vdi cac dac tinh kieu hinh uu viet ciia Bang 1. Ten va ki hieu ciia 7 ddng ca rd phi. ddng de tim kiem mdi lien he va phat trien thanh chi thi phan tir. Dong ca Ki hieu So mau Phuang phap ghep mau (bulked segregant 0. aureus Philippines Oa.PL SO- analysis) dugc cac nha nghien ciiu sir dung phd bien O. aureus Trung Qudc Oa.TQ SO frong viec tim kiem marker phan tir. O rd phi, nhieu O. aureus Israel Oa. Is 30 tac gia ciing sir dung phuang phap ghep mau d O. niloticus Thai Lan Oni.TL 9 microsatellite de phat hien mgt sd chi thi lien quan 0. niloticus Israel On/. Is 30 den CO che xac dinh gidi tinh (Lee et al, 2003; Ezaz 0. niloticus Trung Quoc On/.TQ 30 et al, 2004) va nghien ciiu mdi quan he di truyen (Hussain et al, 2005). 0. niloticus Dai Loan On/.DL 30 Sau khi khao sat AFLP vdi 64 td hgp mdi (Bang 3) vdi miu Oa.TQ. 1 va DNA chuan cua hang Phironng phap ghep mau Invifrogen va tham khao cac ket qua ciia Koeher frong viec xac dinh ban do lien ket (Koeher et al, De giam sd lugng mau nghien cim, cac mau DNA 1998) chiing tdi chgn dugc 16 td hgp mdi cho nhieu ciia 1/3 sd lugng ca the frong timg ddng dugc trdn lai san pham nhat. thanh cac mau DNA. Cach ghep nhu sau: (1) DNA cua 7 ddng ca rd phi dugc ghep theo 10 ca thl thanh 1 Phan tich da hinh miu ki kieu la Oni.TLA, Oa.PL. 1, Oa.PL.2, Oa.PL.3, Oa.TQ.l, Oa.TQ.2, Oa.TQ.3, Oa.Is.l, Oa.Is.2, Vdi 16 td hgp mdi da chgn lgc thu dugc 375 Oa.ls.3, Oni.lsA, Oni.ls.l, Om.ls.3, Oni.TQA, kilu bang (Bang 5), trong dd chi cd 64 kieu bang da Oni.TQ.l, Oni.TQ.3, Oni.BL.l, Oni.QL.2, Oni.DL.3. hinh (chilm 16,58%) va tdng sd bang la 7033. T6 (2) DNA ghep dugc do OD va pha loang vdi nong do hgp mdi AF42 cho nhieu nhit 42 kilu bang, AF36 cho it nhit vdi 15 kieu bang. thich hgp cho phan iing AFLP (25 ng). (3) Tiln hanh AFLP fren cac td hgp mdi chgn lgc. Nhu vay, mdi Phan tich NTSYS cho thiy he sd tuang ddng ddng rd phi cho 3 mau DNA ghep, rieng ddng Oni.TL giua cac ddng rat cao 0,852 - 0,981. He sd tiJong chi cd 1 san pham ghep. ddng giiia cac ddng Rd phi xanh la 0,976 - 0,987 va 456
  3. Tgp chi Cdng nghe Sinh hgc 7(4): 455-462, 2009 giiia cac ddng Rd phi vin dao ddng 0,971 - 0,981 ciia hai nhdm Rd phi van va xanh (Hinh 1), khoang (Bang 4). He sd tuong ddng giiia cac ddng Rd phi each di truyin giiia cac ddng rat thap, cao nhat la xanh vdi cac ddng Rd phi vin thip ban 0,852 - giiia cac ddng xanh vdi cac ddng van thap nhat la 0,873. Dieu nay phu hgp vdi ngudn gdc di truyen vdi giiia cac ddng xanh vdi nhau (Oa.TQ vdi Oa.Is cac ddng. (0,013) hoac giiia cac ddng vang vdi nhau. Tai gia tri 0,85, cay mdi phan nhanh va chia thanh hai nhdm Rd Bardakci va Skibinski (1994), Dinesh va ddng phi xanh (Oa.PL, Oa.TQ, Oa.Is) va Rd phi van tac gia (1996), Sherif va ddng tac gia (2009) sir dung (0«z.TL, Oni.ls, Oni.TQ, On/.TQ). Day la hai loai cd RAPD udc lugng do tuang ddng di truyen giiia O. the phan biet bang hinh thai. He sd tuang ddng giiia niloticus va O. aureus la 0,698, 0,46 va 0,612. cac ddng trong nhdm Rd phi van thap hon he sd Nghien ciiu ciia Sodsuk and McAndrew (1991) fren tuang ddng giiia cac nhdm Rd phi xanh, cho thay Rd chi thi isozyme cho do tuong ddng la 0,744. Do phi van da hinh han Rd phi xanh. Ket qua cQng lap tucmg ddng cao trong ky thuat AFLP giup ngudi lai nghien ciiu cua Agnese et al.(1997) va phii hgp nghien cuu de dang nhan ra cac bang dac trung. vdi ket qua tir phan tich mtRFLP va microsatellite Tren cay di truyen cd the thay rd su phan nhanh ciia nhdm tac gia. Bang 2. Trinh tu cac adapter va mdi du-gc sir dung trong phan tich AFLP. Enzyme han che EcoRI [5' - 3T Mse\ [5' - 3'] Adapter CTCGTAGACTGCGTACC GACGATGAGTCCTGAG AATTGGTACGCAGTC TACTCAGGACTCAT Mdi preselective GACTGCGTACCAATTCA GATGAGTCCTGAGTAAC Moi selective GACTGCGTACCAATTCAN N GATGAGTCCTGAGTAACN N Bang 3. Ky hieu ciia cac cap moi chpn lpc IVIoi chpn loc M-CAA M-CAC M-CAG M-bfiT M-CTA M-CTC M-CTG M-CTT E-AAC 1 2 3 4 5 6 7 8 E-AAG 9 10 11 12 13 14 15 16 E-ACA 17 18 19 20 21 22 23 24 E-ACC 25 26 27 28 29 30 31 32 E-ACG 33 34 35 36 37 38 39 40 E-ACT 41 42 43 44 45 46 47 48 E-AGC 49 50 51 52 53 54 55 56 E-AGG 57 58 59 60 61 62 63 64 Ghi chu: * AF la ki hieu chung che tit ca cac to hgp mdi, in dam la cac to hgp mdi dugc sir dung trong nghien ciru. Bang 4. He so tuong dong glQ-a cac ddng ca rd phi. Dong Oa.PL Oa.TQ Oa. Is Oni.TL On/.ls On/.TQ On/.BL Oa.PL 1 Oa.TQ 0,976 1 Oa. Is 0,984 0,987 1 Oni.TL 0,868 0,865 0,873 1 On/.ls 0,857 0,860 0,862 0,974 1 On/.TQ 0,860 0,852 0,855 0,971 0,981 1 On/.DL 0,862 0,855 0,857 0,974 0,974 0,981 457
  4. Trin Thi Bich Hdng et al Nhan dang cdc ddng cd ro phi bang AFLP dung (Koeher ef a/., 1998). Bang dac trung dugc xac dinh la cac bang chi xuit hien d ddng nay ma khdng xuat hien d mdt sd Nhdn dgng cdc ddng Rd phi xanh O. aureus ddng khac hoac bang khdng xuat hien d ddng phan Bang AF43/17/170bp (E-ACT/M-CAG) chi xuit biet iihung xuat hien d cac ddng khac. hien d ddng Oa.PL, dac trung cho ddng rd phi Oa.PL (Hinh 3). Bang AF29/16/330bp (E-ACC/M-CTA) Nhdn dgng cdc ddng rd phi O. niloticus vd O. aureus xuat hien d ddng Rd phi xanh Oa.PL va cac ddng Rd phi van khac. Trong 16 td hgp moi cd 14 td hgp mdi cho phep Bang AF26/17/350bp (E-ACC/M-CAC) xuit phan biet hai ddng Rd phi xanh va van vdi tdng sd hien d ddng Oa.TQ va cac ddng Rd phi van. Bang 40 bang chi cd fren ddng xanh hoac van nhu nhu AF43/16/160 (E-ACT/M-CAG) chi xuit hien d ddng bang AF3/19,26, bang AF42/21, 27, bang AF43/24, Oa.TQ, dac trung cho ddng Oa.TQ. bang AF54/12,13. Sir dung cac bang dac tmng cho loai Rd phi xanh hoac van danh gia su di truyen ket Bang AF43/28/250bp (E-ACT/M-CAG), xuit hgp vdi cac dac tinh ciia mdi loai de nghien cim img hien d tat ca cac ddng rd phi tni ddng Oa.PL. Cd the dung frong cac phep lai xa giua hai loai rd phi, cac dimg bang nay de nhan dang ddng Oa.PL vdi cac phep lai nay dang dugc cac nha chgn gidng rd phi ap ddng Rd phi xanh khac. - Oni.TL i—On/.ls On/.TQ On/.DL Oa.PL Oa.TQ Oa. Is Hinh 1. Cay di truyin cua 7 ddng ca rd phi. Bang 5. S6 lugng bang cu the cua timg to hgp mdi. To t\cf
  5. Tgp chi Cdng nghi Sinh hgc 7(4): 455-462, 2009 Nhdn dgng cdc ddng Rd phi xanh O. niloticus nao nhung cd thl kit hgp vdi cac td hgp mdi khac dl 0«/.DL; bang AF26/12/210bp (E-ACC/M-CAC) Nhdm Rd phi van cd tinh da hinh cao ban, cd nhan bilt cac ddng nhu bang AF54/12/180bp nhieu bang dac trung ban nhu: bang AF42/17/180bp (E-AGC/M-CTC) chi xuit hien d ddng vin 0«/.TQ (E-ACT/M-CAC) xuat hien d cac ddng vin trir ddng va xuit hien d 0«/.TL va cac ddng Rd phi xanh, 0«z.ls (Hinh 3.17), dac tiimg cho ddng Oni.ls; bang khdng xuat hien d 0«/.Is va Oni.TQ. AF25/19/220 bp (E-ACC/M-CAA) chi xuit hien d ddng Ow/.TQ. Bang AF54/6/125 (E-AGC/M-CTC), Trong mdt nghien ciiu cua Koeher va ddng tac bang AF42/18/170 (E-ACT/M-CAC), bang AF43/25/225bp (E-ACT/M-CAG) chi xuit hien d gia (1998), mdt sd bang dac trung chimg tdi phat ddng 0«/.TQ. hien da dugc dinh vi fren cac nhdm lien ket nhu: AF43/225 (thudc nhdm lien kit sd 16), AF43/250 Mdt sd bang tuy khdng dac trung cho mdt ddng (thudc nhdm lien ket sd 7). M On/.TL Oa.PL Oa.TQ Oa.ls On/.ls Oni.TQ Oni.BL 200 bp — 150 bp—^, Hinh 2. Bang dac trung AF54/12 va AF54/13 cho dong 0. niloticus va O.aureus. M Oni.TL Oa.PL Oa.TQ Oa.ls On/Is Oni.TQ On/.DL 200bp--.i^iiSS3^ 200 bp — ^ n ^ ••'• Hinh 3. Bang dactru-ng ciia to hgp mdi AF43. Bang AF43/17 ciia ddng Oa.PL, bang AF43/16 ciia ddng Oa.TQ. Oni.TL Oa.PL Oa.TQ Oa.ls Oni.\s Oni.TQ On/.DL M 200 bp 150 bp Hinh 4. Bang dac tru-ng cue to hgp mdi AF42. Bang AF42/17/180bp cua ddng On/Is va bang AF42/18/170 bp ciia ddng On/.DL. M. Marker. 459
  6. T r i n Thi Bich H d n g et al Bang 6. Mpt sd bang dac trung cho ddng. ThCp tiF To hap moi Kich thiFO'c bang (bp) Dac trirng cho dong Ki hieu 1 E-ACG/M-CTA -330 Oa.PL AF29/330 2 E-ACT/M-CAG -160 Oa.TQ AF43/160 3 E-ACT/M-CAG -170 Oa.PL AF43/170 4 E-ACT/M-CAG -250 Oa. PL^ AF43/250 5 E-AGC/M-CTC -180 On/.TQ, On/DL AF54/180 6 E-AGC/M-CTC -125 On/.DL AF54/125 7 E-ACC/M-CAC -350 Oa.TQ AF26/350 8 E-ACC/M-CAC -210 On/.TL AF26/210 9 E-ACC/M-C/\A -220 On/.TQ AF25/220 10 E-ACT/M-CAC -170 On/.DL AF42/170 11 E-ACT/M-CAC -180 On/Is' AF42/180 12 E-ACT/M-CAG -225 On/.DL AF43/225 Ghi chu: 'Khdng xuat hien d ddng, nhu-ng dac trung cho ddng, AF54180 la bang cd kich thudc khoang 180 bp cua to hop mdi AF54. KET LUAN Tilapia (Oreochromis niloticus L.): A molecular genetics approach. PhD. Institue of Aquaculture, University of Phan tich A F L P bang 16 td hgp mdi tren 19 mau Stirling, Stirling, Scotland. D N A ghep cua 7 ddng ca rd phi cho thay 13 frong sd 16 Ezaz MT, Harvey SC, Boonphakdee C, Teale AJ, mdi cd bang dac trung cho ddng Rd phi xanh hoac van, McAndrew BJ, Penman DJ (2004) Isolation and physical 6 mdi cho 12 bang dac trung nhan dang dugc 7/7 ddng. mapping of sex-linked AFLP markers in nile tilapia (Oreochromis niloticus L.). Mar Biotechnol 6(5): 435-245. T r o n g tdng sd 64 bang da hinh A F L P , cd 12 Franck JPC, Wright JM, McAndrew BJ (1992) Genetic b a n g cd kha n a n g diing nhan dang ddng; trong dd variability in a family of satellite DNAs from tilapia. d d n g Ort/.TL cd 1 b a n g (AF26/210); O a . P L cd 3 Ge«ome 35: 719-725. b a n g A F 2 9 / 3 3 0 , A F 4 3 / 1 7 0 , A F 4 / 3 2 5 0 ; ddng O a . T Q cd 2 b a n g A F 2 6 / 3 5 0 , A F 4 3 / 1 6 0 ; O/n'.TQ cd 1 bang Hussain A Elghobashy, Ashraf A Ramadan, Ibrahem H A F / 2 5 2 2 0 ; 0 « / . D L cd 3 b a n g A F 5 4 / 1 2 5 , A F 4 2 1 7 0 , Ibrahem, Mohamed A Rashed (2005) Phylogenic relationship for some Tilapia species using elactrophoresis. A F 4 3 / 2 2 5 . Cac b a n g dac t n m g nhan dang ddng la Egypt J Exp BiolX: 57-63. n g u y e n lieu cho qua trinh tim k i e m marker p h a n tii sau nay Koeher TD, Lee WJ, Sobolewska H, Penman D, McAndrew B (1998) A genetic linkage map of a cichlid Ldi c a m o n : Cdng trinh cd su hd trg ciia di tdi fish, the tilapia (Oreochromis niloticus). Genetics 148(3): 1225-1232. "Nghien cieu phdt trien cdc marker phdn tir phuc vu Le Thanh Liru (2000) Nhirng tiln bp khoa hpc cong nghe chon gidng vd bdo tdn quy gen a cd tra vd cd rd thuy san ciia vien nghien cihi NTTS I giai doan 2000 - phi", thugc chuang trinh phdt trien vd irng dung 2006 cd the ap dung tai dong hhng BJc Bp. Hpi nghi cong nghe sinh hoc trong linh vuc thuy sdn din ndm KHCN 2006-07 cdc tinh mien Bdc. http://www.vnast.eov. 2020, Bg Ndng nghiep vd Phdt trien ndng thdn ndm vn/default.asvx? ChannelID=274 2008 - 2010. Lee BY, Penman DJ, Koeher TD (2003) Identification of a sex-determining region in Nile tilapia (Oreochromis T A I L I E U THAM KHAO niloticus) using bulked segregant analysis Anim Genet 34(5): 379-383. Do Due Hanh (2006) Cdng trinh KHCN cua nganh thuy Nguyen Cdng Dan, Phan Minh Quy, Trin Dinh Luan san tham gia Giai thuang Sang tao KHCN Viet Nam nam (2003) Chpn gidng ca ro phi (0,-eochomis niloticus) ddng 2005. Tgp chi Thuy sdn 6(3): 13-16. GIFT nham nang cao sue sinh tmong va kha nang chiu Ezaz MT (2002) Analysis of sex determination in Nile lanh. Tom tat Bdo cdo Hpi nghi Khoa hoc Todn qudc vi 460
  7. Tgp chi Cdng nghe Sinh hgc 7(4): 455-462, 2009 Nuoi trong Thuy sdn 24 - 25/11/2003: 39. Quyin Dinh Thi, Dao Thi Tuylt, Pham Anh Tuin (2004) Danh gia tinh da hinh cac quin dan ca tra nudi (Pangasius Nguyin Thi Td Nga, Ngd Thi Kim, T r k Thi Bich Hdng, hypophthalmus) a Viet Nam bang 4 mdi ngiu nhien chpn QuySn Dinh Thi, NguySn Huu Ninh, Pham Anh Tudn trudc. Bdo cdo Khoa hpc, Hpi nghi Khoa hpc Todn quoc (2009) Biet hda cac ddng ca rd phi bing cac chi thi phan tii 2004. NhOng van ii NCCB trong KHSS dinh huang Ndng ve tinh. Tap chi Nong nghiep vd Phdt trien nong thon Ldm nghiep mien niii: 885-888. 134(5): 64-68. Nguyen Thu Thiiy, NguySn Thi Dieu Thiiy, Nguyen Van Quyin Dinh Thi, Dao Thi Tuylt, Pham Anh Tuin (2005) Cudng (2004) So sanh da hinh AFLP giiia hai nhdm ca tra Sir diing chi thi phan tu RAPD va mtDNA -RFLP de phan (Pangasius hypophthalmus) cd trpng lupng phan biet. Tgp tich da hinh cac quan dan ca tra (Pangasius^ chiDi truyen vd Ung dung 3(4): 17-20. hypophthalmus) nudi d Viet Nam. Hpi thdo todn qudc ve nghien cim vd img dung khoa hpc cong nghe trong nuoi Penma.i DJ, McAndrew BJ (2000) Genetics for the trong thuy sdn: 919-939. management and improvement of cultured tilapias. Tilapias: biology and exploitation 227-266. Quyin Dinh Thi, Pham Anh Tuin (2005) Su dung cac ky thuat sinh hpc phan tii ti-png chpn gidng ca rd phi. Hpi thdo Pham Anh Tuan, Le Quang Hung, Nguyen Thi Tan. (2008) Danh gia lua chpn vat lieu chpn gidng nang cao tdc todn quoc ve nghien ciru vd img dung khoa hpc cong nghe dp sinh tnrdng ca rd phi nudi vung nudc lg man. Tgp chi trong nuoi trong thuy sdn: 841-865. KhOa hpc vd Phdt triin 6(2): 161-165. Rutten MJ, Komen H, Deerenberg RM, Siwek M, Pham Anh Tuan, Nguyen Hiiu Ninh (2003). Nghien ciiu Bovenhuis H (2004) Genetic characterization of four bien di RFLPs mpt sd quan dan ca tra (Pangasius strains of Nile tilapia (Oreochromis niloticus L.) using hypophthalmus). Hpi nghi Cong nghe Sinh hpc Todn quoc. microsatellite markers. Anim Genet 35(2): 93-97. Nha xuat ban Khoa hpc va Ky thuat:724-726. Vos Pieter, Rene Hogers, Marjo Bleeker, Martin Reijans, Pham AT, Tran MT, Mair GC. (2002). Sex determination Theo van de Lee, Miranda Homes, Adrie Friters, Jerina and the feasibility of YY male production in the Pot, Johan Paleman, Martin Kuiper, Zabeau Marc (1995). Vietnamese strain of Oreochromis niloticus (L.). AFLP: a new technique for DNA fingerprinting. Nucl Aquaculture 173: 257-269. Acids Res 23(21): 4407-4414. IDENTIFICATION OF SPECIFIC AFLP MARKERS FOR SEVEN TILAPIA BREEDS CULTURED IN VIETNAM Tran Thi Bich H o n g \ N g u y e n Thi To N g a ' , N g o Thi K i m \ Quyen Dinh Thi'' *, N g u y e n H u u N i n h ^ Pham A n h Tuan^ Institute of Biotechnology Research Institute of Aquaculture Noi SUMMARY Seven Tilapia breeds originated from different countries belonging to two species of Oreochromis aureus and Oreochromis niloticus were used to study the genetic diversity and find out the differences expressing in specific bands. AFLP technique and polyacrylamide 5% for screening were used. DNA of every breed was bulked so that 10 inviduals made 1 sample. Bulked segregant analysis is a rapid mapping strategy suitable for monogenic qualitative traits. When DNA often inviduals are pooled, all alleles must be present. Sixteen primer combinations were tested and bulked samples were established from each population and produced distinctive bands. Moreover, the complexity of DNA fingerprinting varied from one species to another, suggesting that the bulked sample can be particularly useful in breeding programs and hybridization. The results generated from the AFLP analysis were used for drawing the genetic relationships among the examined populations.The presence of band scored 1, the absence was 0. NTSYSpc revealed similarity level between 7 breeds was very high (> 85%), especially among O. aureus or O. niloticus (> 97%). In total 375 bands observed from 16 primer combinations, there were 64 polymorphic bands but only 12 specific bands for 7 breeds such as: AF25/220, * Author for correspondence: Tel: 84-4-37568260; Fax: 84-4-38363144; E-mail: auvendt(d),ibt.ac.vn 461
  8. Trin Thi Bich Hdng e/a/. AF26/210, AF26/350, AF29/330, AF42/180, AF42/170, AF43/170, AF43/225, AF43/250, AF43/160, AF54125, and AF54180. These specific bands are source of markers development for breeding selection. Keywords: AFLP, Oreochromis aureus, Oreochromis niloticus, special band, tilapia 462
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