KHOA HOC CONG NGHE
LnXIG DUIXIG CHI TH! MICROSATELITE PHAIXI BIET
QUAIXI DAIXI T d m HUM XAIXIH {Paruiinus itamaru^
a X/ICT IXIAM V A s m LAIXIKA
Nguyfin Nguyen Thanh NhonS D^ng Thiiy Binh^
Trdn Quang Sang^, Nguyfin Minh ChauS Ph^im Thi Hanh^
T6MTAT
Tom hiun xanh (Paruhnus homaruS) phan bd rong rai o Viet Nam, la hai san co gia tn kmh te cao va ddi tuong nuoi a cac tinh mien Trung. Nam (05) chi thi Microsatelite dugc su dung dd khao sat da dang, khac bifit di truydn va cau tnic qudn the torn hiim xanh a Viet Nam va Sri Lanka. Tdng sd 50 alen duoc phat hifin tren ca 2 quan thd Vifit Nam va Sri Lanka (42 6 Viet Nam, 38 6 Sri Lanka), 9/10 vi tri microsatelUte khong tuan theo dinh luat can bang HW. Quan the tom hiim xanh Viet Nam the hien da dang di truyen cao hon quan thd Sn Lanka. O qudn thd torn hiim Viet Nam co sd alen tning binh - va he sd di hop ttf quan sat ldn loot la 8,4+1,778 va 0,540+0,108, trong khi quan th^ torn hiim xanh ciia Sri Lanka co sd alen trung binh la 7,6±1,166 va h6 sd di hpp tu quan sat la 0,367+0,077. Gia Ui khac biet di tinydn Fst la 0,021 (P<0,05) phan tich DAPC cho thay su phan tach biet giua hai quan the, trong khi ki^m tra AMOVA lai kh6ng co su sai khac CO y nghia (P=0,99). Nghifin cuu hifin tai cung cap dSn Ueu ve dac trung di truyfin quan th^ torn hiim xanh 6 Vifit Nam tren co so so sanh voi quin th^ Sri Lanka. Thu thap miu tom hum xanh theo pham vi phan bo va khao sat cac chi thi microsatellite voi so lirong Ion la vii cdn thiet de xay dimg dinh huong bao tdn va quan ly ngudn loi tfim hum xanh b Viet Nam ben vung.
Ttt kbba: Di truyen, microsateUtes, guan the. tom bum xanh.
I.DATVANDE
Tdm hiim la mgt hong nhung hai san cd gia hi thuong mai cao va duge hfiu thu hfin toan the gidi.
Tdm hum phan bd chu ydu dgc theo khu vue An Do Duong va Thai Binh Duong (PhiUips, 2013). Trfin tiie gidi, san lugng eua tdm hum khoang 270.000 t ^ / n a m hong do cd den 9?>% la dang tir khai thac tu nhifin cdn lai la do nuoi hdng. Gidng tom hiun gai (Panuhws spp.) chiem 28% (FAO, 2015). Viet Nam la mdt hong nhimg nha cung cap tdm hum ldn vdi san lugng khai tiiac hang nam khoang 1.500 - 2.000 t&i/nam, tuong duong 3 - 4 nghm ti ddng CTdng cue Thuy san, 2015), chidm 2% tdng san lugng t6m hiun gai hfin tiid gidi. Tdm hum xanh va torn hiun bong la 2 loai dugc nuoi phd bidn d viing Nam Trung bg, Vifit Nam.
Ky thuat MicrosatelHtes hay chudi lap lai don gian (Sunple Sequence Repeats-SSR) la cdng cu hiru
ich, dugc ling dung rfing rai hong danh gm da dang dt truyen, dinh loai tom hum, hfin ket vong ddi va nghifin ciiu cau tnic di truydn quan thd ODao et al, 2015; Delghandi etal., 2017; EUis etal., 2017). Cho den nay, m6t sd ehi tin microsatelUtes dae trung cho tom hiun gai dugc xae dinh va cd thd ling dung hfin nhieu loai va cac khu vuc dia ly (Ben-Horin et al., 2009; Dao etal., 201S; Delghandi etal., 2016; Santos e^^.,2018).
Nghien ciiu nay iing dung cac chl thi MicrosteUites nham khao sat da dang di truydn va dac tnmg quan thd ciia tdm hiim xanh (Panuhrus homarus) doc theo bd bidn mien Trung, Viet Nam va phan dinh vdi qudn tiie d Sri Lanka, lam co sd eho edng tac bao tdn, quan ly ngudn lgi, ciing nhu di huyen chgn gidng dmh hudng phat ti-ien nghe nuoi ben vimg.
2 . DOI TUONQ VA PHUONG PHAP NGHBU CUU 2.1. Ddi tugng, dia didm nghifin cihi va phuong phap thu mau
Tom hiim xanh {P. homarus) tu nhifin dugc thu d Viet Nam va Sri Lanka, mdi khu vuc thu 30 ca the.
Tai Viet Nam, mau dugc tiiu d phudng Due Long, ' Vi?n Nghien cihi Nuoi Uong Thuy san III
^ Vien Cong nghe sinh hoc va M6i tniong, Tnrong Dai hoc Nha Trang
' Vien Nuoi tr6ng Thuy san, Tnrdng Dai hpc Nha Trang Email: [email protected]
85
KHOA HOC CONG NGHl thanh phd Phan Thiet, tinh Binh Thuan; xa Van
Hung, huyfin Van Nmh, tinh Khanh Hoa; xa Cat Khanh, huyfin Phii Cat, tinh Binh Dinh; xa Tmh Ky, huyfin Son Tinh, tinh Quang Ngai; phudng Phudc My, ban dao Son Tra, thanh phd Da Nang dai difin cho khu vuc phan bd d mien Trung, mdi tinh chon 6 ca thd. Do dac thii khoang each dia ly, mlu tom hiun chi duge thu tai mdt dia diem (Negombo) d Sri Lanka. Dia diem thu mau dugc trinh bay d hmh 1.
Mau t6m dugc thu true tiep tir eac ngu dan danh bat tdm hiun. Sau khi dinh danh chinh xae dua hfin dac didm hmh thai (Geoge va Holthius, 1965), mau eg va chan boi duge bao quan hong cdn tuyet ddi, sau dd chuyen vd phdng thi nghifim va su dung eho cac nghifin ciiu tiep theo.
2.2. T ^ h chidt ADN, khudch dai dua hfin chi thi microsatellite
ADN dugc tach chiet tir 25 mg mau mo co ehii ydu vdi mau torn hum tu Sri Lanka) eua timg ca thd tdm hiim bang bfi kit Wizard Genomic DNA Purification (Proraega) theo hudng dan ciia nha san xuat.
Nghifin ciiu hifin tai su dung 5 chi thi phan hi microsatellite dupc lua chgn tir nghifin ciiu eiia (Delghandi et al, 2016). Ddi voi mdi cap mdi, mdi xuoi dugc gan chi thi huynh quang FAM hoac HEX a ddu 5'. Ddng thdi, nghifin eiiu tdi uu td hpp mdi cho
phan ling PCR da moi gdm PCR I (Pho^45 va Pho- G62), PCR II (Pho<;74 va Pho-G91) va PCRIH (Pho- GlOO). Thong tin chi tidt vfi trinh tu doan moi, mau huynh quang, kieu lap cua mdi dugc trinh bay d bang 1.
Phan ung khudch d^ PCR dugc tiiuc hifin vdi tii^
tich 25 pi gdm 12,5 pl GoTaq Master Mm 2X (Promega), 1 pl mdi loai mdi (10 mM), 5 pl m^u DNA va nuoc cho du tiie tich. Phan img dugc ch^y hfin may luan nhifit Icycler (Bio-Rad) theo ehu trinh nhifit d6: bien tinh ban diu tai 94"C trong S phiit; sau dd la 35 ehu ky eua 94"C hong 30 giay, nhifit dp lai (Bang 1) hong 45 giay, 72"'C hong 30 giay; cudi cting la budc kfio dai 72''C hong 5 phiit San pham ciia phan ling PCR dugc difin di hfin gel garose 1,5%, nhudm Ethidium bromide va quan sat kdt qua dudi tia cue tim.
Hinh 1. So dd dia didm thu mlu ( c h ^ trdn mau xanh) ciia tdm hum xanh (Panultus bomarui) tai
Vidt Nam va Sri Lanka Bing 1. Thdng tin vd cac chi thi microsatellite dugc sir dung trong nghifin ciiu To hop
PCR da moi
PCR I
PCRII
PCR III Locus
Pho-G45 Pho-G62 Pho-G74 Pho-G91 Pho-GlOO
Kieu lap
AGAT ATCT AC CCT TCC
Nhiet do lai CC)
56
58
58
Trinh tu moi (5' - 3') FAM F-GGATGGCAGAGGAGGCAAC R- GTTTCTTCGAACCCGGAGACAAATGC HEX F-CGTAGCTAGGACGCCATTTG R- GTTTCTnTGAGCCAACGTCCCAGTC FAM F-ACGTCCACTTACGAGAGCC R-GTrrCTTCCCAGGAATATGCnTGTGCC HEX F-AGAGCAACTCTCGTGAGCC R- GTTTCTTGTCTTGTGACGTCGCTTGG FAM F-CCTCTCGCGGCTACACC R- GTrrCTTATGCGTCATAATCAGGCGG
DO dai (bp)
364-394 174-198 17^204 245-263 228-249 2.3. GiSi ma va xii lyd&h^u trinh tv
Trude khi tifin hanh giai trinh tir, tat ca san pham PCR dugc hnh sach bdi kit Expin™ PCRSV (GeneAll) de dam bao chat lupng cho vific giai trinh tir. Phan tieh doan dugc thue hifin trfin he thdng may
ABI 37S0XL CTbenno Fisher Scientific) tai Cong ty 1'^ Base (Singapore). Phan mdm Peak Scanner v2.0 ( ^ p h e d Biosystems, 2006) dupe sii dung dd ghi nhan kich thudc chinh xac eua cae alen tir file dii l i ^ fasta (.fsa).
NONG NGHIEP VA PHAT TRIEN NONG THON - KY 1 - THANG 3/2020
KHOA HOC CONG NGHfi 2.4. Nghifin ciiu di huydn quin tiid tdm htim
xanh
2.4.1. Khao sit da dang vi sukhac biet di truyen Da dang di truydn cda quan thd torn hiim xanh t^ hai khu vuc dia ly (Vifit Nam va Sri Lanka) dugc khao sat thfing qua cdc gia hi: Sd lupng alen va sd alen trung binh hfin timg locus (Na), sd alen hifiu qua (Ne), di hgp tu quan sat (Ho), di hgp tii mong dgi (He) va ti lfi locus da hinh (%P) su dung phan mem GenAlEx v6.5.1 (Peakall va Smouse, 2006).
Tinh toan cac chi sd khac biet di truydn (Fgr) giira cac qu4n the, va chi sd AMOVA vdi tan sd hoan vi (No. of permutations) la 999 va gia tri tin cay (p- value) thong qua phdn mdm Arlequin v3.5 (Exeoffler va Liseher, 2010) vdi dii Ufiu dau vao duge tao ra tir phdn mem GenAIEx.
2.4.2. )(ay dung cau true quan the tdm hum xanh De xac dinh cau tnic di truydn dac trung ddi vdi quan thd tom hiim xanh, phan tich da bifit thiic cae thanh phan chinh (Discriminant Analysis of Pnncipal Components-DAPC) Qombart et al., 2010) sir dung gdi du hfiu adegenet vl.3.1 (Qombart and Ahmed, 2011) dugc tiiuc hifin bang phan mdm R v3.5.1 (R Core Team, 2017). Tmh nang findeluster dua theo thdng sd BIC (Bayesian Information Criterion) duge
f iL IJ.LI^.^
J L k l .
tidn hanh vdi gia lap sd eau tnic nhdm la K=l-4.
Thdng sd BIC duge bieu difin hfin dd thi nhu mpt ham eua gia hi K, va gia tri BIC tai vi tri giam thap nhat tuong iing vdi sd nhdm (K) tdi uu. Cau tnic di truyen quan the dupc bieu hifin bang tinh nang eompoplot (bieu dd thanh thd hifin xae suat gan nhdm ciia cac ea thd cho cac nhdm) tich hgp hong gdi dii hfiu adegenet
S . KET QUA VJ« THAO LUiUV
3.1. Da d^ng di truydn qudn th€ tern hum xanh Su da dang di truyen quan the tdm hiim xanh P.
homarus theo timg locus va giiia cac khu vue nghifin ciiu duge the hifin a bang 2. Nam microsatelUte chgn lgc hong nghifin ciiu deu thd hifin tinh da hinh cao vdi tdng cpng 50 alen trfin hai quan dan duge xae dinh vdi kieh thudc dao dong hi 181 bp den 375 bp (Hinh 2). Cac alen xuat hifin vdi tan sd cao d ea 2 quan the nam rai rae d cac vi tri nhu alen 366, 370 Cocus Pho-G45), alen 181 Oocus Pho-G45), alen 200 aoeus Pho-G74), alen 247 Oocus Pho-G91), alen 234 Ooeus Pho-GlOO). Cac alen dae thii xuat hifin nhidu d quan dan Vifit Nam (12 alen, chii ydu a cac locus Pho-G45, Pho-G74, va Pho-GlOO, hong khi dd Sri Lanka cd 8 alen dac thii, chu ydu d locus Pho-G62, va Pho-GlOO (Hinh 2).
Hinh 2. A Tdn sd alen tgi 5 vi hi microsatellite hfin qudn dan tdm hiun xanh t ^ Vidt Nam va Sri Lanka. Cac alen dgc thu cho qudn tii^ tgi cdc vi tri dugc m d ^ . B: Hinh anh d ^ didn alen hfin mdy ABI da phan tich vdi 2
dai mdu FAM va HEX
87
KHOA HOC CONG NGH|
Da dang di truyen cua quan th^ torn hum xanh a Vi&t Nam cao hon Sri Lanka. Tong so alen tren cac vi tri khao sat lan lugt la 42 (trung binh 8,4+1,778/locus) va 38 (hung binh 7,6+1,166/locus);
sd alen hieu qua trung binh (4,26+0,896 va 3,63+0,667); sd di hop quan sat/di hop tu mong doi (0,540+0,108/0,696*0,09 va 0,453±0,069/0,687±0,053).
Sd luong alen tren mdi locus ghi nhan duoc cd su tuong dong nhat dinh d' ca hai quan thd, trong dd quan the Viet Nam tu 3 - 13 va quan the Sn Lanka la 6 - 12. Sd Iuong alen cao nhat duoc ghi nhan d locus Pho-GlOO (12-13 alen) va thap nhat a locus Pho-G62 ciia quan the Viet Nam (3 alen). Trong tdng sd 5
locus microsatelhte, 3 locus (Pho-G45, Pho-G74 v4 Pho-GlOO) cho thay da dang di truyto cao hon 6 quan th« tdm hiun xanh cua Viet Nam so vdi Sri Lanka. Sd alen lan luot la 9,11 va 13 d Viet Nam, so vdi 6,8va 12 dSri Lanka; sd alen hidu qua- sd di hop tir quan sat/mong doi lan luot la 4,885 - 0,3/0,794 vJ 4,950 0,233/0,798 Oocus Pho-G45); 4,775 0,787/0,791 va 4,918 - 0,267/0,797 Oocus Pho-G74);
6.897 - 0,800/0,855 va 4,327 - 0,600/0,769 Oocus Pho-GlOO). 6 locus Pho-G91, sd alen bang nhau, nhung da d^ing di tmyen cua quan thd Vidt Nam cao hon (Bang 2).
Bang 2. Da dang di liuy&l 6 c ^ quin th^ tfim hum xanh theo titng locus v i giOa c ^ khu vuc Quan the
Vidt Nam 0^-30)
Sri Lanka ff<-30)
Ca hai quan dan 0^-60)
Locus Pho-G45 Pho-G62 Pho-G74 Pho-G91 Pho-GlOO
Tdng (TB+SE) Pho-G45 Pho-G62 Pho-G74 Pho-G91 Pho-GlOO
Tdng (TB+SE)
Tdng (TB+SE)
Na 9 3 11 6 13 42 (8,4+1,778)
6 6 8 6 12 38 (7,6+1,166)
50 (8,0±1,011)
Ne 4,865 1,542 4,775 3,220 6,897 4,260+0,896
4,959 2,187 4,918 1,925 4,327 3,66+0,667 3,96+0,536
Ho 0,300 0,533 0,767 0,300 0,800 0,540+0,108
0,233 0,600 0,267 0,233 0,600 0,367+0,077 0,453+0,069
He 0,794 0,352 0,791 0689 0,855 0,696+0,09
0798 0,543 0,797 0,481 0,769 0,677±0,069 0,687+0,053
%P
100
100 HVVE test
...
ns
***
•"
..
...
...
...
... ...
Ghi chu: TB+SE: gia tii tiimg binh ± sal sd chuan; ns: Sal khae khdng cd y nghia so vdi dinh luit Hardy- Weinberg; (*): Sai khic cdy nghia so vdi dinh luat Hardy-Wemberg (* P<0,05; ** P<0,01; ***P<0,001);
N: sd ci thd; Na: Sd luang alen/locus; Ne: sd alen hieu qui; Ho: di hop tir quan sat; He: di hgp tir mong d(ri;P: vi tri da hinh; HWE test kiem tra qui luit Hardy Weinberg.
Cac kiem dinh di truyen qudn the tom hiun xanh vdi timg vi tri microsatelhte chi ra do lech dang ke (9/10 vi tri microsatellite hfin hai quan dan nghifin eiiu la sai khae cd y nghia thdng kfi vdi gia tn P<0,01) so vdi can bang Hardy Weinberg, ngoai tni locus Pho-G62 d qudn the Viet Nam la tuan theo dmh luat can bang HW (Bang 2).
3.2. Sv khde bi^t di truydn giiia cdc qudn th^ tdm hum xanh
Ket qua phan tieh cho thay su sai khae di truydn (Fst) ed y nghia tiiong kfi giiia hai quan thd tdm hiim xanh Viet Nam va Sri Lanka (Fst=0,021; P=0,0018
<0,05). Kdt qua phan tieh AMOVA (Bang 3) eho tiidy da dang di truyen d miic dp phan tii chu yeu la giiia cac ea thd ti-ong quan the (53,87%) va giira eac ea tii^
(45.73%) va cd y nghia thdng kfi (F|s=0,54. P<0,001;
FI 1=0,54, P<0.001). Trong khi miic dg da dang cua 2 qudn the tom hiim xanh khi so sanh voi nhau Id rat thap (0,4%) va khong cd y nghia thdng kfi (P>0,05).
KHOA HOC C 6 N G N G H |
Bing 3. K^t qui phin tich phuong sai Ngudn bien ddng
Giiia cac quan thd Giiia cac ca thd trong quan thd Giiia cac ca thd
Do hi do
1 58 60
Tdng binh phuong
4,392 220,6 68
phin tir (AMOVA) oia cic quit Thanh phan
bien ddng 0,009 1,335 1,133
Phan tram bidn dong
0,4 53,87 45,73
tfa6 tdm h(mi xanh Fixation
mdex Fs,-0,004
F,s=0,54 Fn-0,54
G i a t n P 0,99
<0,001
<0,001 3.3. Cdu tnic qudn thd tdm hiim xanh
Phan tieh DAPC ve cdu tnic di truyen quan the tom hum xanh dugc trinh bay a hmh 3. Thong sd BIC cho thay sd nhdm quan the tdi uu la 2 (Hinh 3A). Bidu dd tiianh thd hidn xac xuat gan nhdm cung thd bifin 2 nhdm quan thd rifing bifit, dieu nay eho thay ed su phan tach ro ret gitia quan dan tfim hiun xanh cua Viet Nam va Sri Lanka (Hinh 3B).
Hinh 3. Phan tich DAPC cdu hue di truydn qudn thd tdm hiim xanh t?u Vi^t Nam vd Sri Lanka.
A. Gia tri BIC (Bayesian Information Criteria) thd bien sdnhdm tdi uu; B. Sadd thanh thehiinxac xuat gin nhdm eua cic ci thd.
Tdm hiim xanh Panuh'ms homanis phan bd rgng d An Dd Duong - Tdy Thai Binh Duong, tir Ddng Nam chau Phi ddn bidn A Rap va phia Dong ddn Nhat Ban, Indonesia va Bdc Aushaha (Phillips, 201S). Nghifin ciiu vd lich su hen hda va dia sinh hda ghi nhan mgt sd phdn loai eiia Panuhrus homanis (Al-Breiki et al., 2018; Farhadi et al, 2017; Reddy, 2013), Cac phan tieh sii dung chi thi phan tii cua ti thd, gen nhan va chi tiii microsatehte cho thay tdn tai it nhat ba hoac bdn phan loai cd pham vi phan bd dia ly khac nhau, vi du nhu Panuhrus homams mbellus phan bd d Tay Nam An Dp Duong; Panulirus homarus "brown" ehi ed d quan dao Marquesas, tinng tam Thai Binh Duong; va P. h. homarus cd vimg phan bd rgng rai d cac khu vuc edn \ax; phan loai P. h. megasculptus a Tay Bdc An Dp Duong (chu ydu la bien A Rap) vdn chua dugc kidm chrnig (Al-
Breiki etal., 2018; Farhadi etal., 2017; Lavery etal., 2014; Singh et al., 2018). Viet Nam tiiuge khu vuc nhiet ddi eua An Dp Duong - Tay Thai Bmh Duong, chu yen tdn tai phan loai P. h. homarus.
Nghifin ciiu nay khao sat da dang di truyen va cau tnic ciia tfim hum xanh d Viet Nam (thugc Tay Thai Bmh Duong) va so sdnh vdi quan the tdm hiun ff Sri Lanka (thugc An Do Duong) vdi 5 hong sd 15 ehi thi Microsatehte cong bd bdi Delghandi va es (2016) dugc lua chgn. Qua khao sat 96 ea the torn hiun xanh tir Vifit Nam, (Delghandi et al., 2016) eho thay sd alen dao dgng hi 3-10, vdi di hgp hi quan sat/mong dpi la 0,021-0,818 va 0,021-0,858 hfin 15 chi thi. Kdt qua nay cd su tuong ddng vdi cac thong sd ve da dang di truydn tfim hum xanh Vifit Nam (sd alen 3-12, di hgp tii quan sat/mong dgi 0,540±0,108/0,696±0,09, n=30). So sanh vdi tom hiim xanh d Indonesia (n=7), tom hiim Vidt Nam cung the hifin su da dang di truyfin cao hon, tuong tu nhu ket qua hien tai vdi tdm hiim xanh d Sri Lanka (n=SO) (Bang 2). Dieu nay cd the giai thich do sd lugng thu mdu nhd (hong trudng hgp d Indonesia), hoae gidi han dia diem thu mau (1 dia diem ea d Indonesia va Sri Lanka), hong khi dd tdm hiim xanh d Viet Nam dugc thu hi cac dia diem khac nhau dpc theo bd bien mien Trung, Viet Nam. Theo nghifin eiiu cua Delghandi va cs (2016), 10 hong tdng sd 15 locus cho thay su sai khae cd y nghia vdi djnh luat HW va de xuat nguyfin nhan co the do sd lugng ca the nhd, su hifin dien ciia null alen hoac troi dat di truydn. Kdt qua hifin tai ciing ghi nhan 9/10 vi ti-i microsatellite cd su sai khae, va chi cd mgt vi tri (Pho-G62) d qudn tiie Viet Nam la tuan theo dinh luat HW (Delghandi e? a/., 2016).
Gia tri khae bifit di truyfin Fst cho thay su phan tach ed y nghia thdng kfi giira 2 quan tiie d Viet Nam va Sri Lanka. Tuy nhifin, phan tieh AMOVA khdng the hien su sai khac cd y nghia giua hai quan the (0,4%, P>0,05). Su kfit ndi di tiiiydn d ca pham vi phan bd hep va d qui mo ldn dugc quan sat a tom hiun
KHOA HOC CONG N G H l bong (Dao etal., 2015), tom hiim xanh (Al-Breiki et
al., 2018; Galmdo et al, 2006) vd tom hiun chau Au (Ellis et al., 2017). Rat nhieu yen td nhu ddng ehay dai duong, giai doan au trimg hoi ndi, su luu giti au trimg tai noi smh ra (\awsX retention) ed the ddn ddn sir hfin ket rgng rai ciia ede qudn the tom hiim a cac khu vuc dia ly cdch xa nhau. Trong nghifin ciiu hifin tfii, tom hiim xanh d Vifit Nam va Sri Lanka thugc 2 dai duong vdi eac dac tinh khac bifit, the hifin su phan tdch ro rfit dua vao phan tieh DAPC (hinh 3).
Su phan tach quan the smh vat giua hai dai duong dugc ghi nhan hen nhidu sinh vat bien (Postahe et al., 2017; Salas etal., 2019), tuy nhifin, ehua ghi nhan su phan tach ciia cae loai tom hum. Nam 2013, Reddy (2013) phat hifin cac qudn the phu (subpopulation) cua phan loai P. h. mbellus phan bo a bd bien chau Phi, lifin quan mat thiet den ddng chay be mat.
Permana va cs (2019) sii dung chi thi COI ciia DNA ti the ghi nhan su phan tach cua quan the tOm hum xanh d Indonesia vdi eae quan the d An Do Duong (Sri Lanka, An Dp, Omans, han) va bidn Dong (Viet Nam, Dai Loan) (Permana et al., 2019). Ddi vdi tdm hum chau Au {Homarus gammarus.), su phan tach d qui md ldn eiing dupe ghi nhan giiia quan thd oThuy Dien va d Dai Tay Duong (EUis etal., 2017). Nghifin ciiu dgc theo khu vue phan bd cua P. homarus, Farhadi va cs (2017) nhan thay su phan tach cua cae qudn thd ngoai bifin (phdn loai P. h. mbeUus a Nam Phi vd Madagascar, P. h "brown" d dao Marquesas, hong khi eae quan the d An Do Duong-Thai Binh Duong (phan loai P. h. homams) the hifin su ket ndi rdng rai (mac dii cd su phan hda theo khu vuc dia ly) (Farhadi et al., 2017). Nghifin ciiu nay cho thay su phan tach cua qudn thd tfim himi xanh d Vifit Nam va Sri Lanka (gia tn Fst, phan tich DAPC), tuy nhifin, kidm h a AMOVA lai khfing the hien miic sai khac cd y nghia giiia 2 qudn the.
Mac dil cdn han che ve sd lugng va khu vuc thu mdu trong pham vi phan bd rpng eua torn hiim xanh nhung nghifin ciiu nay gop phan danh gid da dang di truyen va phan dmh qudn the tom hiim xanh Viet Nam, lam co sd cho cong tde bao tdn, quan ly ngudn lpi, phat trien nuoi hdng thuy sdn va di truyen chgn gidng.
4.K£rLUiUUVAKi\INGHI
Nam ehi thi Microsatehte hihi hieu hong khdo sdt da dang, khde bifit di truyen va cau tnic qudn the tom hiim xanh d Viet Nam va Sri Lanka. Tdng sd 50
alen duge phat hifin (42 d Viet Nam. 38 d Sri Lanka), 9/10 vi tri microsatelhte khong hian tiieo dinh ludt can bang HW.
- Quan tild tdm hiun xanh Viet Nam tiie hidn da dang di hniyen cao hon quan tiie Sn Lanka (so alen locus - di hpp tii quan sat/mong dgi ldn lugt la 8,4+1,778 - 0,540±0,108/0,69fe0,09 va 7,6+1,166 - 0,S67+0,077/0,677±0,069). Phan tich DAPC tiiong qua so dd thanh (mau dd va mau xanh) cho tiiay su phan tdch ro rfit giua hai quan the.
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MICROSATELITE MARKERS FOR SEPARATION VIETNAM AND SRI LANKA SPINY LOSTER (Panilmus homam^ POPULATIONS
Nguyen Nguyen Thanh Nhon, Dang Thuy Bmh, Tran Quang Sang, Nguyen Mmh Chau, Pham Thi Hanh
Summary
Spiny lobster {Panilinus homarus) widely distributed in Vietaam, is a high-value seafood and major cultured species in the central provinces. 5 Microsatelite markers are used to investigate diversity, genetic differentiation, and population structure of spiny lobster ui Vietnam and Sn Lanka. Total of 50 alleles were detected (42 in Vietnam, 38 in Sn Lanka), 9/10 microsatellite loci deviated from Hardy-Weinberg equilibnum. P. homarus population in Vietnam exhibited higher genetic diversity than Sri Lankan population (number of allele/oocus; observed/expected heterozygous are 8-4+1.778;
0.540+0.108/0.696±0.09 and 7.6+1.166; 0.367+0.077/0.677+0.069). The Fst value (0.021; P<0.05), DAPC analysis showed the disconnectivity between the two populations, while AMOVA test showed no significant difference (P=0.99). The present study provides data on the genetic characteristics of P. homarus population in Vietaam based on comparison with Sri Lanka's population. Collecting P. homarus samples throughout its distnbution range, and examine additional microsatelhte markers is essential to formulating strategies for conservation and sustainable management of fisheries resources.
Keywords: GeneUcs, microsatehtes, population, spmy lobster.
Ngu6i p h ^ bi^n: TS. T h ^ Thanh Binh Ng^y nh4n bM: 6/12/2019 Ngiy thdng qua phan bi§n: 6/01/2020 NgAyduy^tdang: 13/01/2020
