NGHIEN C U U DA DANG DI TRUVfiN T A P DOAN LLJA NEP VA LUA N U O N G BAN DIA V I E T N A M BANG CHI T H ! PHAN TU SSR ( M I C R O S A T E L L I T E )
Nguyin Thj Phuong Doai', KhuSt HQu Trung', Nguyin Thiiy Diep', Ha Minh Loan', TrSn Danh Siiu", Dang Trong Lu'ong'
' lien Di truyen Ndng nghiep 'Trung tdm Tdi nguyen Thirc vdt
TOMTAT
Ki lliuat SSR dugc sir dung de phan lich da dang di lru\'cn. mdi quan he di lru\'cn cung nhu phan loai ciic gidng Ilia nep \a kia nuoiig a niirc dudi loeii mc)l each chinh \ac. Su dung 29 cap miii ihu dugc 786 bang DNA lluigc 96 loai allele khac nhau (trung binh 3.310 allele/cap mdi). He sd PIC dao dgng tir 0 den 0.808 (Irung binh 0.474). Cac gidng lua nep \a liia nuong co do ihuan di truyen rat khac nhau, ti le di hgp cua cac mau nghien ciru dao dgng tir 0% den 25% (trung binh la 4.34). He sd luang ddng di truyen giira Ciic gidng dao dgng trong khoang tir 0,06 den 0.84. 27 gidng kia nep \'a lua nuong nghien ciru dugc chia ihanh 2 nhom Ion: Nhdm I gdm cd 6 gidng kia thugc loai phu liidicci co he sd tuong ddng di truyen dao dgng lir 0.32 den 0.63; Nhdm 11 bao giim 21 gidng ihugc loai phu ./iiponica co he sd tuong ddng di liuyen dao dgng iCr 0.20 di'ii 0.84. Cac kcl qua llui dugc con rat huu ich trong \ iec Xiic dinh cac allele hiem. nhan dang nhung nguiin gen co ciic diic linh ndng hgc quy phuc \u phuc \ii cho cdng Uic thu Ihap. bao Idn \;i lai lao gidng.
Tir khda: chi ihi .S'.ST?. he sd PIC. da dgng di Iruycii. liui iicji vd liui nuong ban dia
MO DAU
() Viet Nam, kia nep vii liia nuong ludn dugc xem la loai san pham truyen thdng, nhieu tap tuc \ an hda ciia ngiriri dan viing niii da gan lien vdi \iec canh tac vii su dung kia nep va kia nuong (Luu Nggc Trinh, 2001). Tai nguyen liia nep va kia nuong ban dia cua nude ta rat da dang va phong phii, nhung chua dugc tu lieu hda mdt each cd he thdng, nen viec dat ten cho timg gidng van rat Idn xgn, cd rat nhieu gidng do viec di chuyen giira cac viing da gay ra su nham lan va hieu sai ve xuat xir, ngudn gdc ban dia va mdi quan he di truyen giua cac gidng, nhieu gidng do ggi theo ten dia phuong nen bi triing nhau. Mat khac, mdt sd gidng cd cac dac tinh ndng hgc quy nhu: ham lugng amylose thap, protein cao, pham chat gao ngon, huong thom dac trung; nhieu ngudn gen cd kha nang chdng chiu vdi nhung dieu kien bat thuan cua mdi trudng nhu: chiu ban, chiu lanh...chua dugc nhan biet va sit dung.
Cac thdng tin ve da dang di truyen la yeu cau quan trgng cho cac chuong trinh bao tdn, chgn tao \'a phat trien ciia cac loai thuc vat. Su' phat trien cua cac chi thj phan tu nhu: RAPD (Random Amplified Polymorphic DNA). RFLP (Restriction Fragment Length Polymorphism), AFLP (Amplytied Fragment Length Polymorphism), SSR (Simple Sequence
Repeats) da ddng gdp mdt pbiTn rat quan trgng trong viec danh gia da dang di truyen ciia cac loai thuc vat ndi chung \'a cay kia ndi rieng. Da cd nhieu cdng trinh cua cac tac gia sir dung cac chi thi phan tu de nghien ciru da dang di truyen cua cac gidng kia dia phuong. Mahmoud va ddng tac gia nghien ciiu bien dgng va quan he di truyen cua 7 gidng kia ciia Ai Cap bang viec ket hgp cua 8 mdi RAPD, 6 mdi SSR va 8 mdi AFLP cho thay mii'c do da hinh tuong iing la 72,2%), 90% va 67,9% va khang dinh rang cac ky thuat neu tren deu cd the ap dung tdt cho nghien ciru da dang di truyen cay liia va SSR la chi thj cho mire da hinh cao nhSt (Mahmoud et al, 2005). Nam 2006, Brondani va ddng tac gia da cdng bd ket qua nghien ciru sir dung chi thj SSR de xac dinh da dang di truyen ciia cac gidng kia truyen thdng cua Braxin (Brondani et al, 2006). Nghien ciru nay da su dung 12 chi thi SSR de md ta dac diem da dang cua 192 gidng kia truyen thdng ciia Brazil. Dua tren cac chi thi SSR nay, tap doan cac gidng liia truyen thdng ciia Braxin dugc phan thanh 39 nhdm. Tdng sd 1 76 allele da dugc xac dinh trong dd 30 allele (tir 23 ddng) la cac allele duy nhat. Sd lugng allele/marker bien dgng tir 6-22 vdi sd lugng allele trung binh la 14,6 allele/locus. Cac tac gia cCing da xac djnh dugc 16 ddng la tap hgp ciia cac ddng thuan hay cac cay di hgp tir. Phan tich bieu do hinh cay da xac dinh dugc 6 nhdm ciia cac ddng gidng nhau vdi cac.ten thudng
ggi khac nhau va chi co mdt nhdm cac ddng gidng nhau cd cung ten ggi, dieu nay cho thay chijihi SSR giup xac dinh mgt each co ban quan he di truyen giiia cac gidng dia phuong.
Chinh vi vay, chung tdi tien hanh de fai; "Nghien cim da dgng di truyen tap dodn lua nip vd lua nuong bdn dia cua Viet Nam bdng chi thi phdn tit SSR (microsatellite^ nham danh gia da dang di truyen ngudn gen liia ban dia quy phuc -vu cho cdng tac bao tdn, khai thac va sir dung cd hieu qua cac ngudn gen lua nep va lua nuong ban dia cua Viet Nam.
V A T
LIEU vA PHUTONG
P H A PNGHIEN CUXJ Vat lieu nghien cihi
Vat lieu nghien ciai la 27 gidng lua nep va lua nuong dugc thu thap d nhieu dia phuong khac nhau, dang dugc luu giir va bao tdn tai ngan hang gen hat ciia Trung tam Tai nguyen Thuc vat (bang 2).
Hai muoi chin cap mdi SSR (bang I) dugc sir dung de phan tich thudc cac locus RM dugc chgn Igc tir 2240 cap mdi, do hang Invitrogen cung cap dua vao cac thdng tin ve trinh tu, kich thudc, sd allele chuan tren mdi locus, vi tri phan bd cua cac locus d tren 12 NST khac nhau da dugc McCouch cdng bd (McCouch era/., 2002).
Phu'OTig phap nghien cihi
Tdch chiet DNA tong so: Mau la cua tirng cay dugc thu thap rieng re va tach chiet DNA tdng sd theo phuang phap CTAB cua Obara va Kako (1998).
Chay PCR: Cac phan irng PCR dugc thirc hien theo chu trinh nhiet; 94°C (5 phut), 35-37 chu ki [94°C (40s); 55°C - eOT (30s), 72°C (3s - I phut)]
va kdt thiic d 72°C (5 phut).
Dien di sdn phdm PCR vd nhuom: San pham PCR dugc bien tinh trudc khi dien di tren gel polyacrylamide 4,5% va dugc phat hien bang phuong phap nhugm bac.
Phdn tich vd xir li so lieu: Ket qua dugc thdng ke dua vao sir xuat hien (1) hay khdng xuat hien (0) cua cac bang DNA (cac allele). Sd lieu dugc xu ly, phan tich bang chuong trinh Excel version 5.0 va He sd tuong ddng di truyen va so do hinh cay dugc phan tich bang phan mdm NTSYSpc 2.0 (Rohlf, 1997).
He sd PIC (Polymorphic Information Content) dugc coi la thudc do tinh da hinh cua cac allele d timg locus SSR. He sd PIC dugc tinh theo cdng thiic
cua Nei (Nei, 1973) nhu sau:
PIC =1 - SPj- (trong dd Pj la tan sd xu4t hien cua allele thii i). Ty le di hgp (H) cua mdi mau dugc tinh theo cdng thirc;
/ / % = •
X K-Y
Trong do: X: la tdng sd mdi cd xuat hien 2 alIele/1 locus SSR. K: la tdng sd mdi dugc su dung trong nghien ciiu. Y; la tdng sd mdi SSR cd xuat hien bang DNA.
Ty le khuyet sd lieu (M) dugc tinh bang cdng thirc:
M% = — K
Trong do: Z la tdng sd mdi khdng xuat hieo bang DNA. K la tdng sd mdi dugc sir dung trong nghien ciru.
KET QUA VA THAO
L U A NHe s6 PIC, s6 allele va tdng so bang DNA the hien tren tirng cap moi
Phan tich 29 cap mdi SSR tren tap doan 27 gidng lua nep va lua nuong thu dugc tdng sd 786 bang DNA thudc 96 loai allele khac nhau. Duy nhat cap mdi RM9 cho locus don hinh (chi thu dugc I loai allele), 28 cap mdi cdn lai cho cac locus da hinh.
Trong sd 28 cap mdi cho cac locus da hinh cd: 9 cap mdi thu dugc 2 allele, 9 cap mdi thu dugc 3 allele, 7 cap mdi thu dugc 4 allele va 2 cap mdi thu dugc 7 allele va I cap mdi thu dugc 8 allele (Bang I).
He sd PIC (Polymorphic Information Content)
dugc coi la thudc do tinh da hinh cua cac allele d
timg locus SSR. Sd lieu d bang I cho thay; tap doan
lua nep va lua nuong ban dia cua Viet Nam rat da
dang ve thanh phan cac allele d nhirng locus gen
nghien ciiu. He sd PIC cua 29 cap mdi thay ddi tir
0,0 (d cap mdi chi xuat hien bang don hinh - RM9)
den 0,808 (d cap mdi xuat hien 8 loai allele -
RM24I). He sd PIC trung binh cua 29 cap mdi
nghien cira la 0,474. Ket qua nay tuong tir vdi mgt
sd ket qua nghien ciru cua mdt sd tac gia tren the
giai, chang han nhu: Raj va ddng tac gia khi sir dung
12 cap mdi SSR de danh gia da dang di truyen cac
gidng lua cua An Do da chi ra he sd PIC dao ddng tir
0 den 0,830 (Raj et al, 2006); dd danh gia da dang di
truyen cua 179 gidng lua d 19 dja phuong ciia Bd
Dao Nha, Jayamani va ddng tac gia da sir dung cac
chi thj SSR va chi ra he sd PIC dao ddng tir 0,179
ddn 0,894 (Jayamani et al, 2007).
Bang 1. S6 allele the hien va he so PIC cua 29 cap mdi SSR.
TT
1 2 3 4 5 6 7 8 g 10 11 12 13 14 15
Ten cap moi RM 104 R M 9 RM259 RM318 RM 135 RM 143 RM156 RM241 RM289 RM153 R M 1 3 RM 190 RM314 RM340 RM345 Trung binh
VI tri tren NST 1 1 1 2 3 3 3 4 5 5 5 6 6 6 6
8 6 allele the hien 2 1 7 4 4 2 3 8 2 4 4 3 3 2 3
H e s d PIC 0.311 0.000 0.738 0.630 0.612 0.403 0.431 0.808 0.346 0.558 0.422 0.479 0.416 0.482 0.573
TT
16 17 18 19 20 21 22 23 24 25 26 27 28 29
Ten cap mdi RM 125 RM 172 R M 3 4 6 RM 152 R M 3 3 7 R M 2 4 5 R M 2 9 6 RM171 R M 2 3 9 R M 3 3 2 R M 2 2 9 R M 2 2 4 R M 3 0 9 R M 2 7 7 Tong
Vi tri tren NST 7 7 7 8 8 9 9 10 10 11 11 11 12 12
S6 allele thi hien 3 3 . 2
4 .2
«
3 3 2 4 7 4 2 2 96 3.310
He sd PIC 0.449 0.491 0.324 0.358 0.142 0.582 0.618 0.582 0.493 0.474 0.686 0.565 0.346 0.417 13.735 0.474 Ghi chu: PIC: Polymorphic Information Content, NST: Nhiem sac the.
13
^'i-s f
Spf
A
f '1
% ^ 5JOOljp^PC
1
"3
; B
r. J
5^
1
•I
i5 V
1
'R3
=^' 5 -.-cr
2
- J2-
1
•if -a
1
5
litbp • »
RM 332 (162'IS3)
Hinh 1. Anh dien di san pham PCR ciia cac giong liia nghien ciru vdi cap moi RM332 (M: Marker 0X 174/Hinfl).
Ty le khuyet so lieu (M) va ty le dj hop tii' (H) ciia cac giong liia nep va lua nuong nghien cuu
Sd lieu d bang 2 cho thay: Ty le khuyet sd lieu (M) cua 3 gidng Ka tieu, Khau giang cam va Khau lao cao nhat (khuyet sd lieu 4 trong tdng sd 29 cap mdi nghien cim = 13,79%o). Gidng lua Khau didn lu khuyet sd lieu d 3 cap mdi tuang iing vdi ty le 10,34%. Gidng Ndp bd hdng Hai Duong va Kh§u nua nuong cd ty le khuyet sd lieu la 6,9%. 12 gidng cd ty le khuyet sd lieu la 3,45%) va 9 gidng cdn lai
khdng bi khuyet sd lieu. Ti le khuyet sd lieu trung binh cua 27 gidng la 3,96; khdng cd gidng nao cd ti le khuyet sd lieu Idn hon 15%). Nhu vay, ca 27 gidng nghien ciiu deu cd y nghTa phan tich thdng ke.
Ty le di hgp tii (H) cao nhat d gidng liia Plau tau
dang dang 2 la 25%o, tiep den la gidng Nep cai dd
(21,43%), Tan lanh (13,79%), Nep cai nuang
(10,34%)). Hai gidng Khau khinh va Khau dang danh
cd ty le di hgp la 6,9%). 9 gidng cd ty le di hgp trong
khoang tir 3,45%o den 4,0%). 12 gidng cdn lai cd ti le
di hgp tir la 0%o (ddng hgp d ca 29 locus nghien ciru).
Ti le di hgp tu trung binh cua ca tap doan la 4,34.
Ket qua nay cQng phu hgp vdi nhiing nghien ciiu cua Olufowote va ddng tac gia. Cac tac gia nay da sir dung chi thi phan tir SSR va RFLP dd nghien ciru da dang di truyen cua cac gidng lua dia phuong va cac gidng lua cai tidn va da ket luan rang cac gidng lua dia phuang thudng cd miic do di hgp tir cao hon cac
gidng lua cai tidn (Olufowote et al, 1997). Nghien cim ciia Alvarez va dong tac gia da sir dung chi thi SSR de nghien cim 39 gidng lua dia phuang va 11 gidng liia dugc trdng phd bien d CuBa. Cac tac gia da dua ra ket luan riing cac gidng lua dia phuong cd ty le di hgp tir cao han cac gidng lua dang dugc trdng phd bien. (Alvarez et al, 2007).
Bang 2. Ty le khuylt so lieu (M) va ty le di hgp tir (H) ciia cac gidng lua ndp va lua nuong nghien ciru.
TT
1 2 3 4 5 6 7 8 9 10 11 12 13 14
Ky hieu Nl N2 N3 N4 N5 N6 N7 N8 N9 N10 N i l N12 N13 N14
Ten giong
Ka tieu Khau khinh Khau giang cam Khau tan pdn Nip ICin
Plau tau dang dang 2 Tam ngi
Khau meo
Nip bo hdng Hai Duong Ua tdn
Nip rau Khau nua nuong Nip nuong cam Tan lanh
M (%) 13,79 0,00 13,79 3,45 3,45 3,45 3,45 0,00 6,90 3,45 3,45 6,90 3,45 0,00
H (%) 0,00 6,90 4,00 3,57 3,57 25,00 3,57 3,57 0,00 0,00 3,57 0,00 0,00 13,79
TT
15 16 17 18 19 20 21 22 23 24 25 26 27
Ky hieu N15 N16 N17 N18 N19 N20 N21 N22 N23 N24 N25 N26 N27 Trung
Ten giong
Nip cai nuang Khau mac bugc Te nuong Ld dip cam Khau cam pi Khau dang danh Nip cai dd Khau lao Blao dang Khau diln lu Nip cam Ne nuong Nip cam den binh
M (%)
0,00 0,00 0,00 3,45 3,45 0,00 3,45 13,79 3,45 10,34 0,00 3,45 0,00 3,96
H (%) 10,34 0,00 0,00 0,00 0,00 6,90 21,43 0,00 3,57 3,85 3,45 0,00 0,00 4,34
Ket qua phan tich moi quan he di truyen ciia cac giong Ilia nep va liia nu'ong nghien ciru
Cac ket qua thdng ke va sd lieu d hinh 2 cho thay; he sd tuong ddng di truyen cua 27 gidng lua nep va lua nuong nghien cuu dao ddng trong khoang tir 0,06 den 0,84. C) mire tuong ddng di truyen 11%, 27 gidng lua nep va lua nuong nghien ciru dugc chia thanh 2 nhdm nhu sau:
Nhdm I gdm cd 6 gidng; Nep bd hdng Hai Duong (N9) va Ndp CSm (N25), Ngp liin (N5), Khiu lao (N22), Kh4u meo (N8) va Tan lanh (N14). He so tuong ddng di truyen giii'a cac gidng trong nhdm I dao dgng tir 0,32 (giira hai gidng Nep liin va Nep Cam) den 0,63 (giiia hai gidng Nep liin va Khau lao).
Nhdm II gdm 21 gidng cdn lai. Trong dd, hai gidng Ndp cai dd (N21) va Ka tieu (Nl) cd he sd tuong ddng di truyen thap nhat la 0,20 va hai gidng
Khau tan pdn (N4) va Tam ngi (N7) cd he sd tuong ddng di truyen cao nhat la 0,84. Nhdm II dugc chia thanh 3 nhdm phu sau:
- Nhdm 2.1 chi cd I gidng la Nep cai dd (N2I).
He sd tuong ddng di truyen cua gidng Nep cai dd vdi cac gidng liia cdn lai trong nhdm II kha thap dao ddng tir 0,20 (giira gidng Nep cai dd vdi gidng Nep Ka tieu) den 0,38 (giira gidng Nep cai do vdi gidng Plau tau dang dang 2).
- Nhdm 2.2 gdm 4 gidng: Blao dang (N23) va Khau dien lu (N24), Ne nuong (N26) va Nep cam den (N27) vdi he sd tuong ddng di truyen giiia cac gidng trong nhdm phu nay dao ddng trong khoang 0,36-0,65.
- Nhdm 2.3 gdm 16 gidng: Ka tieu (Nl), Khau
khinh (N2), Khau giang cam (N3), KhSu tan pdn
(N4), Piau tau ding dang 2 (N6), Tam ngi (N7), Lia
ton (NIO), Nep rau ( N i l ) , KhSu nua nuong (N12), Nep nuong cim (N13), Nep cai nuong (NI5), Khau mac budc (NI6), Te nuong (NI7), Ld ddp cSm N18), Khau cam pi (NI9), Khiu dang danh (N20). He so tuong ddng di truyen giira cac giong lua trong nhdm nay dao ddng tir 0,36 (giiia gidng Khau giang cam va Khau cam pi) den 0,84 (giira gidng Khau tan pdn va Tam ngi).
Ket qua phan nhdm bang chi thi phan tit SSR hoan toan triing khdp vdi ket qua phan loai cac gidng lua nep va lua nuong dua tren doan trinh tu cua DNA luc lap va sir dung phuang phap phenol (Ha Minh Loan et al, 2010). Ket qua phan loai d mire dudi loai tap doan 27 gidng lua nep va lua nuong nghien ciiu va so sanh vdi hai gidng ddi chimg Nipponbare (liia Japonica) va Kasalath (lua Indica) dua tren sir khac biet DNA luc lap ciia hai loai phu Indica va Japonica bang cap mdi (5'-
gtccactcagccatctatc-3' va 5'-ggccatcattttcttctttag-3') dugc thidt ke de khuech dai doan DNA luc lap nam tren vung ORE 100 ciia doan Pst-\2 cho thay: trong sd 27 gidng lua nep va lua nuong nghien ciru cd 6 gidng thudc loai phu Indica va 21 gidng thudc loai phu Japonica. Cac gidng lua trong phan nhdm I tuong duong vdi 6 gidng thudc loai phu Indica vk cac gidng thugc phan nhdm II tuong duong vdi 21 gidng thudc loai phu Japonica.
Nguyen Dire Thanh va ddng tac gia (1999) da sir dung mdt sd mdi RAPD, 20 cap mdi SSR va 15 cap mdi AFLP de nghien ciiu mdt sd gidng lua nep va lua nuong dia phuong va mdt sd ddng liia tai sinh tir md khang dich nam dao dn da ket luan rang: ky thuat SSR cho mire do phan biet giiia cac gidng lua cao hon AFLP va cac gidng lua nep va lua nuong nghien cim rat da dang ve phan tir, cd the lam nguyen lieu tdt cho cac nghien cim di truyen va chgn gidng.
H
1
1
1 1
T — I — r
0,44 0.51
Coefficient
— I — T 0.57
T—I—T 0.77
N l •
N2 N6 NIO Nil N12 N13 N15 N16 N18 NI9 N17 -N3 jN4
"IN7 -N20 -N23 -N24 -N26 -N27 - N 2 L -N5 -N22 -N8 -N14 -N9 -N25-
n
— I 0.84
Hinh 2. So d l hinh cay ve mire tuong dong di truyin giua cac gidng liia nip va liia nuang nghien ciru. Ten cac gidng lua dugc ky hieu tir N1-N27 (chi tilt xem bang 2).
KET L U A N
Tap doan liia nep va lua nuong ban dia nghien ciru r4t da dang. Phan tich 29 cap mdi SSR tren 27
gidng Ilia nep va lua nuang thu dugc tdng sd 786 allele, thudc 96 loai allele khac nhau, trung binh 3,310 allele/cap mdi. He sd PIC dao ddng tir 0 den 0,808, trung binh 0,474. Cac gidng lua nep va liia
nuong cd do thuan di truyen rat khac nhau ti le di hgp cua cac mau nghien ciru dao dgng tir 0% den 25%) (trung binh la 4,34). He sd tuong ddng di truyen giiia cac gidng dao dgng trong khoang tir 0,06 ddn 0,84.
Tap doan 27 gidng lira nep va lua nuong nghien cim dugc chia thanh 2 nhdm Idn: Nhdm I gdm cd 6 gidng lua thugc loai phu Indica (Nep bd hdng Hai Duong, Nep Cam, Nep lim, Khau lao, Khau meo va Tan lanh) cd he sd tuong ddng di truyen dao ddng tir 0,32 den 0,63; Nhdm II bao gdm 21 gidng thudc loai phu Japonica cd he sd tuang ddng di truyen dao ddng tir 0,20 den 0,84. Nhu vay, sir dung cac chi thi SSR khdng nhiing cd the danh gia dugc mire do da dang, xac dinh mdi quan he di truyen giii'a cac gidng lua nep va lua nuong ban dia ma cdn cd the phan loai d mire dudi loai mdt each chinh xac phuc vu cho cdng tac chgn va lai tao gidng.
Loi cam gn: Cong trinh duac thuc hien bdng nguon kinh phi ciia de tdi KC.04.16/06-10 "Nghien ciru ddnh gid tu lieu hoa nguon gen cay trong bdn dia a mice do phdn tir de bdo ton vd sir dung chiing mot each CO hieu qud ".
T A I LIEU THAM K H A O
Alvarez A, Fuentes JL, Puldon V, Gomez PJ, Mora L, Duque IVIC, Gallego G, Tohme JM (2007) Genetic diversity analysis of Cuban traditional rice (Oiyza sativa L.) varieties based on microsatellite markers. Genet Moi Biol30(4): 1109-1117.
Brondani C, Borba TCO; Rangel PHN, Brondani RPC (2006) Determination of genetic variability of tranditional varieties of Brazilian rice using microsatellite markers.
Genet Moi Biol 29(4): 676-684.
Mahmoud M Saker, Sa-wsan S Youssef, Naglaa A.
Abdallah, Hany S Ashandy, Ahmed M El Sharkawy (2005) Genetic analysis of.some Egyptian rice genotypes using RAPD, SSR and AFLP. Afr J Biotechnol 4(9): 882-890.
McCouch SR, Leonid T, Xu Y, Lobos KB, Clare K, Walton M, Fu B, Maghirang R, Li Z, Xing Y. Zhang Q, Kono I, Yano M, Fjellstrom R, DeClerck G, Schneider D, Cartinhour S, Ware D, Stein L (2002) Development and Mapping of 2240 New SSR for Rice (Oiyza sativa L.).
DNA Res 9: 199-207.
Nei M (1973) Analysis of gene diversity in subdivided populations. Proc Nah Acad Sci USA 70: 3321-3323.
Jayamani P, Negrao S, Martins M, Macas B, Oliveira MM (2007) Genetic Relatedness of Portuguese Rice Accessions from Diverse Origins as Assessed by Microsatellite Markers. Crop Sci 47: 879-884.
Ha Minh Loan, Tran Danh Siru, Nguyen Thi Lan Hoa, Nguyen Song Ha, Nguyen Thi Phuong Doai, Khuat Him Trung (2010) Phan loai cac gidng lua nep va lua nuong dia phuong bang DNA luc lap va dung dich Phenol, Tgp chi Khoa hpc Cong nghe Ndng nghiep Viet Nam 1(14): 1-5.
Obara OP, Kako S (1998) Genetic diversity and identification of cymbidium cultivars as measured by random amplified polymorphic DNA (RAPD). Euphytica 99:95-1001.
Olufowote JO, Xu Y, Chen X, Park WD, Beachell HM, Dilday RH, Goto M, McCouch SR, (1997) Comparative evaluation of within-cultivar variation of rice (Oryza sativa L.) using microsatellite and RELP markers. Genome 40(3):
370-378.
Nguyin Due Thanh, Phan Thi Bay, Le Hdng Diep (1999) Phat trien va irng dung cac chi thi phan tii' trong nghien cim da dang lua, Bdo cdo khoa hpc. Hoi nghi Cdng nghe sinh hpc toan qudc 1999: 1205-1215.
Luu Nggc Trinh (2001) Tinh hinh va nhiem vu ciia cong tac bao tdn tai nguyen di truyen thuc vat d Viet Nam, Bdo cdo tdng kit du dn: Xdy dung ngdn hang gen cdy trdng Qudc gia.
Raj KJ, Lambodar B (2006) Identification and differentiation of indigenous non-Basmati aromatic rice genotypes of India using microsatellite. Afr J Biptechnol 6 (4): 348-354.
Rohlf FJ (1997) NTSYS-PC: numerical taxonomy and multivariate analysis system. Exeter software. New York.
E V A L U A T I O N O F G E N E T I C D I V E R S I T Y O F G L U T I N O U S R I C E A N D L O C A L U P L A N D R I C E IN V I E T N A M USING M I C R O S A T E L L I T E M A R K E R S
Nguyen Thi Phuong Doai', Khuat Huu Trung'' *, Nguyen Thuy Diep', Ha Minh Loan^ Tran Danh Suu^ Dang Trong Luong'
'Agricultural Genetic Institute 'Plant Resources Center
SUMMARY
In order to provide materials for breeding, the genetic diversity, relationship as well as subspecies taxonomy of glutinous rice and local upland rice was evaluated using SSR techniques. Using 29 SSR primers 786 DNA bands were obtained belonging to 96 different alleles (with an average of 3.310 alleles per SSR loci).
PIC value changed from 0.00 to 0.808 with a mean of 0.474. Upland rice varieties have different homozygosity in 29 SSR loci, the ratio of heterozygosity changed from 0.00 to 25% (with a mean of 4.34). Genetic similarity coefficients of 27 varieties were ranged from 0.06 to 0.84. Twenty seven upland rice varieties were divided into two groups: group I consists of 6 varieties belong to Indica subspecies with genetic similarity coefficients from 0.32 to 0.63. Group II consists of 21 varieties belong to Japonica subspecies with genetic similarity coefficients from 0.20 to 0.84. The obtained results could be useful for germplasm fingerprinting, preeminent selection and proper utilization in breeding program.
Keywords: SSR marker, PIC value, genetic diversity, glutinous rice and local upland rice
' Author for correspondence: Tel: 84-4-37540764; E-mail: khuathuutruns(a).vahoo.com
