KHOA HOC CONG NGHE
DA DAIVG Dl TRUYEIV CUA CAC GIOIVG LLIA DfA PHUOIXIG {Oryza satii/a L.) DAIXIG CHJ TH! PHAIXI TU S S R
Nguyfen Pham Anh Thi', La H o ^ g Tnic NgSn', Tnrong Thi Bich Van', Nguyto T ^ Pha' T6MTAT
Nghien cihi nay dirgfc Ihuc hien de danh gia su da dang di truydn tren 30 giong liia dja phuong duor cung cap bai Vien Nghien cuu Phat tnen d6ng bang song Ciiu Long bang viec su dung 25 ci\p nioi SSR. Ket qua cho thay trong tong so 25 cap moi SSR, co 22 cap moi co tinh da hinh U^,s%). Cac cap m6i SSR phat hi^n tong sd 62 bang da hinh trong long so 70 bang duoc khuc'ch dai. He sd da dang di Iruygn (PIC) tren cac locus dao dpng trong khoang tir 0.0605 ddn 0.7010. Dira vao phucmg phap UPGMA co th^ phan nhdm 30 gidng Ilia dia phuong thanh 3 nhom chinh. Muc tuong dong di truyen ciia chiing dao dong tir 56% den 94%.
Nhung dir heu trong nghien ciru nay gop phAn cung cap thong tin di truydn lam ca so cho lai tao gidng Irong tuong lai.
Tir khda; Da dang di livyen, lua miia. SSR.
1 . DAT VAN BE
Ltia mua dia phuong co lich sir canh tac lau doi.
dupc trong voi san Iirgng km a cac tinh Ca Mau, Bac Lieu, Tra Vmh va Kien Giang (Tong cue Tlidng ke, 2017). Liia dia phuong co ngudn gen da dang, mang linh img dung cao Irong cong tac lai tao cac gidng liia moi boi nhieu dgc tinh noi bat nhu co kha nang thich nghi tdt trong cac khu vuc thd nhuong bi nhi^m phen, nhi^m man, chdng chiu vcn nhi6u loai sau b^nh, cho pham chal gao ngon, co gia tri kinh te cao (Nguyen VSn Tii va ctv, 2011; Tran Huu Phiic va ctv, 2012). Tuy nhien, cac gidng liia dia phuong thuong CO than cao, dd dd nga, chiu tac dong cua quang ky, vu miia dac trung va phu (huge nhidu vao thoi tiet, do do san lugng va nang suat liia dia phuong tuong ddi tliSp so voi cac gidng liia lai mdi. Trong nhirng nam gan day, duoi tac dgng ciia cudc Cach mang xanh nam ciing vdi nhimg tac dgng tir bien doi khi hau, dien tich canh (ac liia dia phuong ngay cang bi thu hep, thay the bang nhiiiig gidng lua men co thoi gian sinh tnicmg ngan, canh tac dugt: 3 vu liia/nam, mang lai hieu qua kinh te cao (Tran Van Dat, 2010). Dieu nay dan ddn sir thu hep ve mat da dang di truydn, nhidu gidng liia chat lugng dia phuong da khdng con ti'ong san xuit, de doa su giam Slit sd lirgng va thanh phdn loai cua tap doan liia miia dia phuoTig (Khuat Huu Tning va ctv, 2012).
' Vien Nghien cuu va Phat tricn Cdng nghc sinh hpc, Trudng Dai hoc Can Tha
Email: npalhiig'cui cJii.vn
De tiep can va danh gia vdi ngudn vat lieu liia dja phucmg. ky thuat SSR dugc danh gia cd tam quan trgng trong nghien cuu vd di truydn va chgn giong, cho hieu qua vugt trdi so vdi cac ky thuat dua vao hinh thai hgc truydn thdng boi tinh da hinh cao. ban chat dong trgi, co tinh lap lai cao, pham vi bao phu khdp bg gen va cd vi tri dac hieu tren nhidm sSc thd, dd dang phat hien bang ky thuat PCR (Kaha va ctv, 2011; Le Y Phung va ctv, 2018). Hien nay, hon 20.000 chi thi phan tu SSR da dugc tao ra va ung dung trong phan tich kieu gen ciia cac gidng liia (International Rice Genome Sequencing Pi'oject va Sasaki, 2005;
Miah va ctv, 2013). Nhieu nghien cuu su diing ky thuat SSR dd nghien cuu da hinh gen nhu Ngd Thi Hdng Tuoi va ctv (2014) da su dung ky thuat SSR de phan tich da dang di truydn ciia 46 gidng liia cam dia phuong, Pham Quang Nghia va ctv (2015) diing 4 mdi SSR de nghien cuu da hinh tren 6 gidng liia thom, Tran Thi Luong va ctv (2013) sir dung 40 dau SSR de phan tich mdi quan he di truydn ciia 60 gidng Ilia dac san,... cho nhirng ket qua dang tin cay. Vi nhijng ly do tren, nghien cim dugc thirc hien voi muc tieu nham danh gia su da dang di truydn cua cac giong Ilia dia phucmg, tir dd phan nhom duoc ngudn vat Heu ltia dia phuong, lam dan heu cho qua trinh lai gidng trong tuong Iai.
2. VAT UBJ VA PHUDNG PHAP NGHIBU CUU 2.LVatMu
Thi nghiem dugc thuc hien trdn 30 gidng liia dia phuong duoc thu thap va luu trij tai Vien Nghien ciiu Phat tridn ddng bang sdng Cuu Long, Trudng Dai hoc Can Tho (Bang 1).
Da su dung 25 chi lhi phan ui ssK dmh vi tren nhidm sic Ilu- so 1. 3. li. 9.10 va 11 ciia cay lua. Theo cac nghidn cuu tnroc day vi- da dang di truydn cho thaiy phan Idn cac c§p moi trong nghien ohi na>- cho chi SI) da hinh cao tren cay liia trong b nhieu viing khac nhau nhir An Dd, \idi Xam. chau Phi (Davia va
Bang 1. Danh sAch 30 giong liia dia phuong nghidn a m
c!v. 2013: Khuat Hiru Tmng va ctv, 2011', N.T«
Hong Tuoi va ctv 2014: Tran Thi Luong \ a Dao va ctv, 2018). Thdng tin chi tioi ve c.k duoc thu thap tir co sa du heu (tt-\w.graniene.org) (Bang 2).
Till 2013;
",iKMSSR Graniene
STT
1 Lun can do lim can Irang Lim phdu M<ii bill km Mgl bui luu I Mgt bui trang Nang quot bidn Ndp sira Soi lun
STT 17 IS 19 20 21
•ri
• 23 i 24
Tdn gidng Tai nguven Thanh Tn Tai nguven Lun phen ,Tel ran , Thom man
Thom lim man ,Pokkah .IR28 ' Lun cao san do
Ngoc nir 25 ! Trang bo cau
Tra long 26 'Trang trdn
Ba bui ' Nang thom
Nang quot bidn 1 Bir liep
28 iMdtbuLcao 30
29 Liin phdng Lun phen hat nho TT
1 2 3 4 5
()
s
Ten moi RM10(»4 RM243 RM 10745 RM 10748 RM 10793 RM10825 RM10843 RM54in
NST 1 1 1
3Sng 2. CSc c$p mdi SSR dupc su dunpr trong nghii Tnnh tu moi 5' - 3'
|.-. TTIC CCTGGTTTCAAGCTTACG R-AGTACGGTACCITGATGGTAGAAAGG F- GATCTGCAGACTGCAGTTGC R- AGCTGCMCGATGTTGTCC F-TGACGAAITCiACACACCGAGTACG R- ACTIVACCt, 1CGGCAACATGG 1 11-- cATCGci'GAi. c A c c r r n r c
R-CLTGR-AIC TAICICCCTCAAGC 1
1 1 1
;) | K M : ! 4 1 2 • 1
in 11
KM707:i . 1
F-GAClTGCCAACrcrnCAAlTCG R-TCGTCGAGTAGCTTCCC TCTCTACT, F-GGACAG AAGTCCATGATCa ATCC R- GTnCCrrTCCATCClTGlTGr F- CACCTCTTCTGCCTCCTATCATGC R- GTTTCTTCGCG.'WATCGTG TGG F- GCCTCCCATAT.VAACCGGC R-AGCGAAGGG.AACAC(;ACAG F- AAAGCACK ;TTTTCCTCCTCC R- CCCATG rGCAATGTGTCTTr F- TATGG AC 1 GGAGCAAACn C R-GGCACAC.CACCAATGTCTC
RMSaili 1 I"-A(,TAC(;,VnTCTGTCAGCGTTGaTA(;T nciru
Nhiet do bat cap
58 55 58 58 58 56 58 58 55 55 58
Motif lap (AC) 18 (CT)18 (TATG)9 (AG) 14 (ATAG)7 (AAG)10
rrc)io
(TC)19 (a-)i7 (ACAT)l;!~1 aA)2«
1
KHOA HOC CONG NGHE IT'
12 13 14 15 16 17 18 19 20 21 22 23 24 25
Ten nun
RM1287 RM140 RM 10864 RM3252 RM493 RM562 RM283 RM302 RM413 RM122 RM253 RM215 RM333 RM206
NST
1 1 1 1 1 1 1 1 5 5 6 9 10 11
Tnnh tu moi 5' - 3'
R-GGATGAAAGTIGATGGATGATCTACTTGTT F- GTGAAGAAAGCATGGTAAATG R- CTCAGCTTGCTTGTGGTTAG F- TGCCTCTTCCCTGGCTCCCCTG R- GGCATGCCGAATGAAATGCATG F- GAGGTGAGTGAGACTTGACAGTGC R- GCTCATCATCCAACCACAGTCC F- GGTAACTTTGTTCCCATGCC R- GGTCAATCATGCATGCAAGC F- TAGCTCC AACAGGATCGACC R- GTACGTAAACGCGGAAGGTG F- CACAACCCACAAACAGCAAG R- CTTCCCCCAAAGTTTTAGCC F- GTCTACATGTACCCITGTTGGG R- CGGCATGAGAGTCTGTGATG F- TCATGTCATCTACCATCACAC R- ATGGAGAAGATGGAATACTTGC F- GGCGATTCTTGGATGAAGAG R- TCCCCACCAATCrrGTCTTC F- GAGTCGATGTAATGTCATCAGTGC R- GAAGGAGGTATCGCTITGTTGGAC F- TCCTTCAAGAGTGCAAAACC R^ GCATTGTCATGTCGAAGCC F- CAAAATGGAGCAGCAAGAGC R- TGAGCACCTCCTTCTCTGTAG F- GTACGACTACGAGTGTCACCAA R- GTCTTCGCGATCACTCGC F- CCCATGCGTrTAACTATTCT R- CGTTCCATCGATCCGTATGG
Nhiet dp bat cap
55 58 58 56 56 60 56 55 58 56 55 55 55 58
Motif lap (AC) 16 (AG) 17 (CT)12 (GT)27 (CD 13 (CTD9 (AAG)13 (GA)18 (GT)30 (AXIS (AG) 11 (GA)11 (GA)25 (CD 16 (TAD 19 (CTD19 (CD21 2.2. Phuong phap nghifin ciiu
2.21. Ly ti-ich DNA
Hat gidng liia dugc ii trong tii u (37"C) den khi giai doan hat nay mam. Hat liia nay mam dai tir 1,5 - 2,0 cm se dugc trong vao chau dat, bd sung nuoc tuoi moi ngay den giai doan la non 10 - 12 ngay tudi thi tien hanh Iy trich DNA. La non dat tieu chuan dugc lau sach bang cdn 70" va thuc hien ly hich DNA theo quy dinh CTAB dugc md ta boi Roger and Bendich (1988) cd hieu chinh cho phii hgp voi didu kien phong thi nghiem. DNA sau khi dugc ly trich se dugv kiem tra chat lirgng bang gel agarose 1%. DNA dii tieu chuan se duoc su dung trong cac thi nghiem tidp theo.
2.2.2 Phin ting PCR
Phan ling PCR dugc thuc hien voi tong the tich 15 pL hdn hgp bao gdm 1 pL DNA mau; 9,65 pL
BiH.O; 3 pL Buffer 5X; 0,6 pL mdi xudi (10 pM) va 0,6 pL mdi ngugc (10 pM); 0,15pLTaq Polymerase (5U/pL). Phan ung dugc thuc hidn theo chucmg trinh nhiet nhu sau: 94"C trong 2 phtit, 30 chu ki lap Iai vdi 3 buoc chinh: Bidn tinh DNA khudn b 94"C trong 30 giay, gan mdi d 50 - 65''C trong 30 giay tiiy thugc vao timg cap mdi SSR, va keo dai chudi O' 72"C trong 45 giay. San pham PCR dugc giu trong 5 phtit a 72"C va sau do dugc trir lanh 6- 4"C.
San pham PCR duoc kiem tra tren gel agarose 2% a hieu dien the 140V bang ki thuat dien di vdi dung dich dem TBE IX, thdi gian dien di phij thugc vao kich \h\xaQ cua san pham PCR, dao dgng tir 40 phtit ddn 1 gio. Thang chuan Ladder 100 bp cua Cdng t\- Thermo Fisher Scientific dugc su dung de uoc lirgng kich thuoc cac doan san pham PCR. Ket
Da SLT dung 25 chi thi phan tu SSR dinh vi tren nhidm sac the sd 1. 5.6. 9.10 \-a U cua cay lua. Theo cac nghidn cihi truoc day ve da dang di truyen cho thay phan Ion cac cap moi tmng nghien cuu nay cho chi Sll da hinh cao tren cay lua in'ini; a nhieu \img khac nhau nhu: An Dd. \iv\ Nam. chau Phi (DavIa va
Bang 1. Danh sach 30 girfng liia dia phuong nghidn cuu
ctv. 2013; Khuat Him Trung va ctv, 2012:
Hdng Tuoi va ctv 2011; Ti-an Tlii Lutmg va Dao \"a civ. 2iU8) Thdng tin chi Iiet ve c u duoc thu thap tir co so dir lieu (w^v-^v.gramene.org) (Bang 2).
NL'O Til!
.,v.2m3:
noi SSK .tramcne
Ten gidng STT Ten gidng
1 Lun can do lam can trang I^n pheu
16 Tai nguven Thanh Tn Tai nguyen IS Lun phdn , Miii^uijiin
_\h»t biij^lim C,i M:iu M'll bgi trang
19 Tet ran 20 Tlimii man
Th<mi lun man
Nang quot bien Pokkali
8 '1 10 11 12 13 II 15
Xep sua Soi km N.yoe nu
Tra lon^
\ia bui Nan.i,' qu Ho lic2^
Cao san
Lun cao san do ranK bo can I'ranjj tron 'Win.mlioin Mulbiiicao
"l.mplioiv;
Lull pi ihal nho Bang 2. C^c cgp moi SSR dupc sir dung trong nghl&n cuu
RM 10793
Tnnh tu nioi f - r F- ITrCCCI{,GL!'ICA-\(,CI'L\C(
ItA(iLACC,(,IACCI"l(,AI<,(,IA(,A'U(,(
F-GA'I'C IGCAGACJCiCAGLLC ,C R-AGCr(,CA\C(,\l(,LI(.ICi F-TGACGAVIT(,ACACAL CC;A(;TACG R- AcnTCACCC, I C(,Gl. A \ t ' VI(,(
I--CATC(i(.n;Ai.CAtC'LK I \ i R- CCTGTCA IC IAI CI Ci. i I CAA( ;c F- (.AC
R-'LC(i
TGCCA\CIVcTU V-Vnct, 'CC,A( .TAGCn Ll- C rcTC lACC F-CitiAC \CAAGTCCATGATCCT\IX I R- GTTTCCTTTCCATCCTTGTI (,C F- CACC I c n CTGCCTCCTATCAI ( , t R^CriTLCITCl.CGAMTCGTX, r(,(_, Y. GCCIVCC VL\1 AV\CC( ,(,C K- \(;CC,A\C,C,C,A\CAC(,\LA(, F-AV\(;C XGGTITTCCTCCTCC K-CCC\l(ri(.CA\LGI(,LL I'IL F- I'ALGGAC rCiGAGCAA-Wa c K-(;t,C\C\(,CAH A M G k k 1^- u . j \ c i AI'nc'K.ri. V,L(,£iGCTI'A(;T
G
Nhiet dp baicap
58 55 58
"Is
58 56 58 58 55
Motif lap (AC) 18 (C018 ( T A T ( ; ) 9
(Afirfl (ATAG)7 (AAG)IO (TC)IO aC)19 ICDLT
(ACAD 13 _nA)28"
KHOA HOC CONG NGHE TT
12 13 14 15 16 17 18 19 20 21 22 23 24 25
Ten moi
RM1287 RM140 RM10864 RM3252 RM493 RM562 RM283 RM302 RM413 RM122 RM253 RM215 RM333 RM206
NST
1 1 1 1 1 1 1 1 5 5 6 9 10 11
Trinh hr moi 5' - 3'
R-GGATGAAAGTTGATGGATGATCTACTTGTT F- GTGAAGAAAGCATGGTAAATG R- CTCAGCTTGCTTGTGGTTAG F- TGCCTCTTCCCTGGCTCCCCTG R- GGCATGCCGAATGAAATGCATG F- GAGGTGAtJTGAGACTTGACAGTGC R- GCTCATCATCCAACCACAGTCC F- GGTAACTTTGTTCCCATGCC R- GGTCAATCATGCATGCAAGC F- TAGCTCCAACAGGATCGACC R- GTACGTAAACGCGGAAGGTG F- CACAACCCACAAACAGCAAG R- CTTCCCCCAAAGTITTAGCC F- GTCTACATGTACCCTTGTTGGG R- CGGCATGAGAGTCTGTGATG F- TCATGTCATCTACCATCACAC R- ATGGAGAAGATGGAATACTTGC F- GGCGATTCTTGGATGAAGAG R- TCCCCACCAATCTTGTCTTC F- GAGTCGATGTAATGTCATCAGTGC R- GAAGGAGGTATCGCTITGTTGGAC F- TCCTTCAAGAGTGCAAAACC R- GCA'ITGTCATGTCGAAGCC F- CAAAATGGAGCAGCAAGAGC R- TGAGCACCTCCTTCTCrGTAG F- GTACGA(rrACGAGTGTCACCAA R- GTCTTCGCGATCACTCGC F- CCCATGCGTTrAACTATTCT R- CGTTCCATCGATCCGTATGG
Nhiet do bat cap
55 58 58 56 56 60 56 55 58 56 55 55 55 58
Moliflap (AC) 16 (AG) 17 (CD 12 (GD27 (CD 13 (CTD9 (AAG)13 (GA)18 (GD30 (ADS (AG) 11 (GA)11 (GA)25 (CD 16 (TAD 19 (CTD19 (CD21 2.2. Phuong phap nghi&n cuu
2.2.1. Ly tiich DNA
Hat gidng Iiia dugc li h'ong tii ii (37"C) den khi giai doan hat nay mam. Hat liia nay mdm dai tir 1,5 - 2,0 cm se duoc U'dng vao chau dat, bo sung nude tudi mdi ngay ddn giai doan la non 10 - 12 ngay Uioi thi tidn hanh ly trich DNA. La non dat tieu chuan duoc lau sach bang cdn 70" va thuc hien ly tiich DNA theo quy trinh CTAB dugc md ta bdi Roger and Bendich (1988) cd hidu chinh cho phii hop vdi dieu kien phdng lhi nghiem. DNA sau khi dugc ly trich se duoc kiem tra chat lugng bang gel agarose 1%. DNA dii tieu chuan se dugc sir dung trong cac thi nghiem tiep theo.
2.2.2 Phin ting PCR
Phan ling PCR duoc thuc hien voi tong the tich 15 pL hon hgp bao gom 1 pL DNA mau; 9,65 pL
BiH>0; 3 pL Buffer 5X; 0,6 pL moi xudi (10 pM) va 0,6 pL mdi ngugc (10 pM); 0,15pL Taq Polymerase (5U/pL). Phan ting duoc thuc hien theo chuong trinh nhiet nhu sau: 94"C trong 2 phiit, 30 chu ki lap lai vdi 3 bude chinh: Bien tinh DNA khudn b 94"C trong 30 giay, gdn mdi a 50 - 65''C trong 30 giay tiiy thugc vao timg cap moi SSR, va keo dai chudi (> 72"C trong 45 giay. San pham PCR dugc giir trong 5 phiit b 72"C va sau do duoc trir lanh a 4"C.
San pham PCR dugc kiem tra tren gel agarose 2% a hieu dien the MOV bang kl thuat dien di vcri dung dich dem TBE IX, thdi gian dien di phu thudc vao kich thudc ciia san pham PCR, dao ddng lu 40 phtit ddn 1 gio-. Thang chuan Ladder 100 bp ciia Cdng ty Thermo Fisher Scientific duoc sir dung de uoc luong kich thud'c cac doan san phdm PCR, Ket
qua dien di dugc ghi nhan bang may chup hmh gel Biorad UV 2000 (IS.A).
2.2.3. Phan Uch so lieu
Dua vao hmh anh didn di san pham PCR. su xuat hien ciia cac bang di?n di dupc udc lugng kich thirdc dua vao thang chuan. Cac bing dugc ma hoa duoi dang nhi phan, 1 (ddi vm truong hgp xual hien bang) va 0 (ddi vcri trucmg hop khdng xuat hien bang). Sd heu cac bang ghi nhan dugc luu Iriir irong phan mdm Excel. Hg sd luong ddng di truyen Jaccard, phan nhom di truyen va so dd hinh nhanh so sanh mdi quan hd di truyen giiia 30 gidng lua duoc xac dinh dua theo phuong phap I'PCiNLA (Sneath va Sokal, 1973) Irong phan mem Uidng ke NTSVSpc 2.1
Ham lirgng thdng lin da hinh (PIC- Pol>Tnorphic Information Content) dugc tinh toan theo phuong phap ciia Weir (1996).
PlC(i) =1-IP„^
Trong dd; P^ la tan xuat alen Ihu j vdi locus SSR thu i. Pham vi gia iri PIC lu 0 (khdng da hinh) loi 1 (da hinh hoan loan).
S. KFT QUA Vd THAO LUAN
3.1. Su da hinh cac cap m6i SSR ddi vol 30 gidng hia dia phuong . . •
Trong 25 cap mdi SSR dugc su dung cho phan ung K R tren 30 gidng lua dia phuong. cd 3 cap mdi RM1074S. RM283, RM243 khdng the hien tinh da hmh (Hinh 3) va 22 cap mdi (88%) cho da hinh. Cac cap mdi khudch dai sd lugng alen kha ldn vdi 67 alen.
trong do cd 62 alen da hinh. dao dgng hi 2 den 5 alen tren mdt locus. Sd alen tnmg binh h'en mgt locus la 3.05 alen, ket qua nay cao hon nghien cuu ciia Singh va ct\- (2015), tuy nhien tuong ddi tliap hon so vdi nghien ciiu ciia Vu Thi Bich Huydn va ctv (2013), Ashraf va ctv (2016) va Dao va ctv (2018). Kich thudc san pham PCR dao ddng tir 70 bp (RM413) den 600 bp {RM;'.:-;:!) cap mdi RM3252 cho sd alen nhieu nhat voi 5 alen, cd 8 cap mdi cho sd alen it nhat vdi 2 alen (RM10825, RM10843, RM8046, RM140, RM215.
RM5461. RM1287, RM10684). Da sd cac cap mdi cdn lai khuech dai lir 3 den 4 alen (Bang 3, hinh 1 va 2).
STF 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
9 9
1
Ten moi RM3252 RM10793 RM3412 RM10694 RM493 RM10745 RM562 RM7075 RM302 RM10864 RM140 RM10825 RM 10843 RM8(H6 RM5461 RM12S7 RM413 RM122 RM253 rRM215 RM333 RM206
To rnin,£
N'ST
1 _|
1 1 1 1 1 1 1 1
So alen
D
4 4 4 3 3 4 3 3 1
1 1 1 1 1
- -
So alen da hinh 5
1 4 4 3 3 4 3 2 2 2 -' 1 1
> 1 2
2 2 2 1 ' 2 1 5
5 6 9 10 11 ng
binh
4 3 3
4 3 3 2 1 1 4 4 67 3.05
4 4 ' 62
2.82
Chi so PIC 0,7010 0,6682 0,6425 0,6374 0,5870 0,5581 0,5492 0,4890 0,4607 0,3581 0,3557 0,2772 0,2149 0,2044 0.1595 0,0605 0,6519 0,4377 0,5427 0,0605 0,5,562 0,6830 0,4479
Kich thiroc san pham PCR (bp) 150-500 125 - 250 175-275 200-275 225-350 150-200 100 - 300 150-200 200-350 200-350 100-150 100 -150 150-200 150-200 1.50-200 150-200 70 150 200-275 150 - 200 ___ 150- 200 150-600 i 400 - 500
T
KHOA HOC CONG NGHE
He sd da dang di truydn PIC cua 22 cap mdi dao ddng tu ddng 0,0605 (R.M215. RM1287) ddn 0,7010 (RM3252); trung binh dat 0,4479 (Bang 3).
Ket qua cho thay cd 7 cap moi SSR cho he sd da dang di miydn Ion hon 0,5 (RM3252, RM10793, RM3412.
RM10694. RM493, RM10745). He so PIC lon hon hoac bang 0.5 se cho su khac biet cao vd ty le da hinh cua cap moi SSR dugc sii dung. He sd da dang dl truyen trong nghien cuu nay thap hon so vdi mdt sd nghidn cuu trudc day nhu nghien cuu ciia Davla
va ctv (2013) sir dung 39 cap mdi SSR cho chi sd PIC la 0,670; hay nghien cuu oJa Tran Thi Luang va ctv (2013) sii dung 40 cap moi SSR cho chi sd PIC trung binh la 0,60. Tuy nhien. ket qua nghien cuu kha tuong ddng vcn nghien ciiu ciia Vu Thi Bich Huyen va ctv (2013), Ngo Thi Hdng Tuoi va ctv (2014).
Singh va ctv (2015) vdi cac he sd PIC Idn lugt la 0,427; 0,46 va 0,47 va cao hon nghien ciiu cua Dao va ctv (2018) sir dung 59 cap moi cho hd sd PIC 0.394.
Hinh 2. San pham PCR vdi cap m6i RM3412
200 b f i -
Hinh 3. San phdm PCR vdi cgp m6i RM283
(Ghi chii: M- tiiang chuan Ladder 100 bp, sd thd ttr dien di tiieo danh sich gidng lua duac tiinh biy tiong bingl).
3.2. Quan hd di truydn giiia cac gidng liia dia phirong nghi&n cihi
Dira tren su da hinh ciia cac cap moi SSR, mdi quan he di truydn giira 30 gidng ltia dia phuong da dugc thidt lap theo phuong phap UPGMA dugc trinh bay trong hinh 4.
Ket qua nghien cuu cho thay he sd tuong ddng di truyen giira 30 gidng liia dia phuong dao dgng tir 0,56 den 0,94. Tir so dd hinh nhanh the hien quan he di truyen giiia cac gidng ltia, d' miic tuong dong di truydn 60,9%, tap doan 30 gidng ltia dia phuong co the phan thanh 3 nhdm chinh rd ret: Nhdm I gdm 2 gidng Ilia, nhdm II gdm 11 gidng liia va nhdm III gdm 17 gidng liia cdn lai.
Nhum I: Gdm 2 gidng Soi lun va Lun phen voi muc Itrong ddng di truyen ihap (67.9%). Nhom I co so lugng gidng it nhal U'ong ba phan nhom va cung CO muc tuong ddng kha thap so voi hai nhdm con Iai (60.9%). Ket qua nay cho thay hai gidng lua Soi Lun va Lun phen co mdi quan he di truyen kha xa cac ginng con Iai, co the sir dung lam ngudn nguyen lieu tao chgn gidng lua moi khi ket hgp voi cac gidng lua nhdm II. III. Nhom II: Q muc tuong ddng di liuycn 64.3%. nhom II dugc phan thanh hai nhom phu: Nhom phu IIA va nhom phu II.B. Nhom phu
IIA a mrrc luong ddng 65,1% bao gdn. .i-'i^"^
iNep siia. Tra long, .Nang quot bidn i. Ba b^'- liep. Lun cao san dd) vdi hai gidng c^' m"^" ^'^'^^^
(lunjzcaonhatlaBabuivaBoUep 185., > CAO^^^^^^
Cao san Uid hien muc tuong ddng thap aliii' ('t,A%) so voi 6 gidng cdn Iai trong nhdm. Nliom plw ILB gdm 4 gidng (Ngoc nir, Liin phdng, Trang trdn. Nang thom) cd muc tirong ddng 70,7%; trong ^o. Trang iron va Nang thom ia hai gidng cd muc tuong dong cao nhat (79,5%).
(Ghi chii: Sc i tiiu tu trong sa do quan be di truydn tiieo danh sich gidng liia duoc trinh bay tiong bang 1).
Hinh 4. So d6 hinh nhinh th^ hi$n mrfi quan h^ di tniyte ciia 30 gidng Iiia dia phuong dtra trfin s6 U^u phan tich DNA vdi 22 chi thi phSn tu SSR
Nhom III; O muc tuong dong 66,9%. cd so luong gidng nhidu nhal vdi 17 gidng liia. duoc phan thanh 3 nhdm phu. Nhdm phu lUA gdm 7 gidng lua mua (Lim can dd. Lim can trang. Mot bui lim, Thom liin miia. IR28. Pokkah, Tet ran), cd mirc Urong ddng dao
bui uang CO miic do tuong ddng cao nhat (94,0%), tiep den la cap gidng Lun can do va Liin can trang (92.1%). Ddng thdi, hai cap gidng nay cung cd miic tuong ddng cao nhat trong 30 gidng Ida nghien ciiu.
Nhu vay, so dd quan he di truydn cua 30 gidng ddng tir 70.8% ddn 92.1% Nlium phu III.B gom 9 '"^ nghien ciiu cd mirc dd tuong ddng kha cao.
Gidng Ilia Mdt bui lim va Mdt bui trdng Ca Mau co muc tuong ddng cao nhat (96%), chung to hai giong Ilia nay cd ngudn gdc gan nhau hon cac nhdm khac.
Trong khi dd, hai gidng liia Lun can dd va Liin phen cd muc tuong ddng thap nhat (56%).
4. KET LUAN
Trong sd 25 chi thi SSR sii dung irong nghien cuu da dang di truyen ciia 30 gidng lua mua thi ro 2?
chi thi cho cac bang ADN da hinh, Ihu duac: 67 alen khac nhau, sd alen dao ddng lu 2 ddn fi .(Icn/ln.-, gidng (Lim pheu. Mdi bui liin Ca Mau. Mot bui
trang, Tlioni man. Tai nguyen Thanh Tn. Nang quot bien, Trang bd cau. -Mot bui cao 1. Lim phdn hat nhd) CO he so tuong ddng dao ddng hi 0.(38 den 0.94.
Nliom phu III.C chi gdm 1 gidng ltia Tai nguyen diap nhat {66.9%) so voi 16 gidng Iiia cdn lai va nam neng re mgt nhanh. He so luong ddng ciia nhom III dao dgng hi 0,67 den 0.94. chiing to cac gidng Iiia trong nay cd quan h? di truydn tuong ddi gan giii. Trong nhdm III. cap gidng liia Mdt bui lim Ca Mau va M<ii
N O N G NGHIEP VA PHAT TRIEN N O N G THON - KY 2 - THANG IO/20
KHOA HQC CONG NGHl sd alen trung binh dat 3,05 alen/locus. He sd PIC oia
22 cap moi nam trong khoang 0.0605 den 0,7010, trung binh dat 0,4479. Tap doan 30 giong liia dia phuong cd miic tuong ddng di truydn tuong ddi cao (tren 56%). He sd tuong ddng di truydn dao dgng trong khoang tir 0.56 den 0,94. O miic tuong ddng di truydn 63.3% cac gidng Iiia nghien ciiu dugc chia thanh 3 nhdm chinh. Nhdm I gdm 2 gidng liia, nhdm II gdm 11 gidng liia, nhdm III gdm 17 gidng liia.
LOI CAM ON
Nhom tac gia xin chan thinh gui ldi cam on ddn Vien Nghien cdu va Phit trien Cong nghe Sinh hoc, Tnrdng Dai hoc Can Tha da tao dieu kien ve phong thi nghiem vi tiang thiet bi de nhom co the hoan thinh nghidn ctiu nay.
TAI UEU THAM KHAO
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Pairay G. A., Khan S. and Ganai N. A., 2016. SSR based genetic diversity of pigmented and aromatic rice (Oiyza sativa h.) genotypes of the western Himalayan region of India. Physiology and Molecular Biology of Plants: An International Journal of Functional Plant Biology. 22: 547-555.
2. Dao, S., Timbine, H., Goita, 0 . and Traore. D., 2018. Genetic Variability Assessment in Irrigated Rice (Oiyza sativa and Oryza glaberrima) by PCR- SSR in Mall International Journal of Genetics and Genomics. 6: 22.
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http://www. gi'am ene.org/markers/micro sat/, tiiiy cap ngay 15/4/2019.
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177: 309-334.
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dia cua Viet .\aiii bang cin lhi phan luSSR li ve tmh (MicrosateUite). Tap chi Ndng nghiep va PTNT. 17:
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10. Nguydn Van Tii, Tnrong Trgng Ngdn. Trdn Nhan Dung va Nguydn Vu Linh, 2011. Thanh loc cic gidng Ilia mang gen khing riy nau bing dau phan tu DNA Tap chi Khoa hgc - Trudng Dai hgc Can Tho.
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Extraction of DNA from plant tissues. In: Gelvin, S.B., R A Schllperoort and D. P. S. Verma (Eds.).
Plant Molecular Biology Manual, Springer Netherlands, Dordrecht, pp. 73-83.
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16. Tran Thi Lmmg. Lmi Mmh Cue va Ngii\in Thanh. 2013. Banli gia <la dang di tiruu-n n"'' ^"
Due Thanh. 2013. nan ticb guan he dl tinyen mot so gionff lua bang kf- thuat .\-iA'phuc vu cb' • < * " ' ' " ' giong lua d$c s.ni. cbat luong. ti-ong pho bien & I let lai tao giong chiu ban. Tap chi Sinh hgc. 3 ' S""-"
V.im Ad/,.j.r(./,/rt;pAa/jfl>.\«TapchiSinhhoc. 35: j j Wj.,,. g g jggg Genetic Data-\nalsf'''"- 34S-35<i. Meihods tor Discrete Population Gem-tic Dl'"-
17. \ u Thi Bich Huyen, Le Thi Bich Thuy, Sinauer.^sociates.
N.Kuyen Anh Dung. Hoang Ba Titn ra NguySn Due
GENETIC DIVERSnY IK SEASONAL RICE (OrKzasatfraL.) USING SIMPLE SEQUKNLi;
REPEATS (SSR) MARKERS
Nguyen Pham Anh Thi, La Hoang True Ngan, Truong TTii Bich Van, Nguyen Thi Pha Summaiy
Tliis rest-arcli was earned out to evaluate the geneUc diversity among Uie 30 seasonal rice genotypes from Cuu Lung delta Rice Research using 25 simple sequence repeat (SSR) markers. The results revealed that out of 25 SSR pnmers. 22 pnmers showed dlsUnct polymorphism (88%), indicahng the robust nature of iiiien)s;itellltes in revealing polymorphism. SSk markers produced 62 polymoi-phic bands out of 70 amplifieaUon products. The polymorphism infonnation content (PIC) value for the SSR loci ranged from 0.0(505 10 0.7010. The dendrxigr^m based on UPC'iM.\ classifed the 30 nce accessions into 3 main clusters.
Similarity coefficients ranged trom .56% to 94%. The data obtained m this study provides genetic information for bacl<gronnd seleclions dunng bacltcross breeding programs.
Keywords: CeiieUc iliver^in. se.t«in.-ilnee. SSR NguM phin bi^n: PGS.TS. U Tufa Nghia Ngiy n h ^ b M : 9/8/2019
Ngiy thang qua phin bi$n: 10/9/2019 Ngiy duy^t ding: 17/9/2019
