IJIMG DLJIMG CdlVG IMGHE C H I T H | P H A I M TIJ TROIVIG CHOrVI TAO GidlMG LUA THUAlM K H A I M G R A Y IMAU CHO
DOIMG BArVlG SdlMG HOIMG
Luu Th| Ngpc Huy^n T6M TAT
Cluin loc (hni vao rar rlii Ihi lini kcM (Marker Assisted ScK'cUon MAS) la mot cniig .u d,: day nhanh hieu qua oiia pluroiig phaji chon giong phan tir. Trong nghicni < ini nay, cac chi tlu SSR lien ket v<ri mot so gicn khang ray nau d;i din.n- su dung trong chon gn'iiK lu-''- ^'••'< dong lua RS, IS1.2, IS2.;', khang ray nau duw sir dung lam dong cho gicn khang. (iiong liia SLIL', IK61 dmrc dung lam gioiig nhan gicn khang. Phan tich di iruyen ciia mot so gicn khang cho ket ciua i ac dong RS. IS1.2 va \S2.:'> mang hai gi<n khang ray. Qua trinh dua hai gicn khang r.iy ii;iu vao c:\c giong liia ini hi SI.IL' va IRiJ-l su dung phuong phap MAS ket hc^} vc'n chon giong tniycn thong d.'i diruc tlurc liirii. C;ic chi Uii I'han tir lidi ket v('ri gien khang ray dirw diing de chon cac dong mang gi(Mi kh.-ing. Cac dong liia kh.ing ray nau DTCl-3, DTE2-3, DTH18, DTR64 da dirrrc chpn loc CO m;ing hai gun khang Bi)h3 v.i H|)hZ, co tinh khang c;io vc'n cac biotype niy nau d DBSH va DBSCL, n:ing suat thuc thu dat duc.rc lu 5,.'>-7,8 t:'in/ha sc s(nii clucrc din v.-'m san xuat.
Tir khoa: Clion Icic dim v.io cic chi thi lien kci (Markrr Assisted ."^election .MAS), khang ray niu. lua. chi tin VI vc linh -S.'^K clu thi phim td-ClVr.
I. DAT VAN DE
Trong nhirng ndm gan day, dich ray nau gay hai nghidm trpng tai nhieu tinh eua ddng bdng Bde bp va ddng bdng song Cuu Long. I h e o sir khao sat muc dp khang ray nau ciia cac gidng liia tidng tai cac tinh thupc ddng bang song Hdng thi phan Ion co mirc nhidm ray nau tir trung binh den mirc cao. Quan the thien dich lai thay ddi dpc tinh theo hudng ngay eang tang dan den tinh trang nhieu rupng lua phun thuoc diet ray 2-3 lan/vu van bj chay ray. Cac phuong phap chpn gidng Iruyen thdng thong thuong de chpn thanh eong mot gidng lua mbi it nhdt phai mat tir 6- 10 nam. Him nura, qua tiinh ebon loc gdp nhieu khd khan, tdn kem. Hien nay, nhd su phdt trien cua eong nghd sinh hoc noi chung va eong nghd fhi thj phan tir (CTPT) noi rieng dd tao ra mpt eong cu hiru ich cho cac nhd chpn gidng. Phuong phap chpn gidng nhd chi thi phdn tii (MAS - Marker Assisted Seletion) su dung cac chi thi phdn tir lidn ket viri cac gien mong mudn vira nang cao hidu qua chpn lpc, vira nit ngdn thdi gian chpn gidng. Muc tidu ciia de tai nay la chpn tao 1-2 ddng liia thuan uu vidt khdng on djnh vdi quan the ray ndu cho ddng bdng song Hdng bdng cdng nghd chi thi phan bi ket hpp vdi chpn gidng truydn thdng.
H. VAT UEU VA PHUONG PHAP NGHIEN CUU
1. Vdt Udu
Dong/giong liia khang niy nau: RS, IS1.2. IS2.;
lam gidng eho gien kh.'ing.
- Gidng liia SL12, IR64 lam gidng nhdn gien.
Cac chi thi phfin tir SSR lien ket gien BphSv BphZ.
Gidng chuan khdng r;i\- ndu Ptb33 va gioni chuan nhiem ray nau TNI tir IRRI.
Ray nau diing cho nghien ciru thu thap tirca khu vuc khae nhau viing ddng bdng song Hdngv:
viing ddng bdng song Ciru Long.
Cac hod chat diing trong nghien cuu dupe mui tir eae hdng (jcnSet, Pharmacia, Sigma. Boehringer Promega. BioRad...
2. Phuong phdp nghidn cuu
Phuong phap danh gid mirc khdng ray tiieo tiei chuan ciia IRRI va Vidn B\'TV vdi thang diem tir U 3...10: Ma dupe gieo theo phuong phdp gieo kho.cJ ddng/gidng lua dupe bd tii ngdu nhidn vdi 3 lan nhii lai. Khi ma dupe 2 la khoang 7- 8 ngay tuoi clii nhidm ray ndu tudi 2 vdi mat dp 5- 6 con/cdy. Sau7- 8 ngdy khi cdy TNI (ddi chung nhidm) chet het,ti^
hanh danh gid dp khang/nhidm ray ndu.
'vien Di truyen Nong nghidp
18
NONG NGHIEP VA PHAT TRIEN NONG THON - KY 1 - THANG 2 / 2 i
Phuong phap lai hdi giao de qui tu gien kh.ing ray tir ddng/gidng cho gien vao gidng lua lai viet bdng phuong phap lai huu tinh. Tien hanh khir due cay me khi lua bdt dau trd bong. Mdi bong chi chpn 15 25 hoa d giua bong. Khir due phai tien hanh khi vach bao phan chua md. Tien hanh thu phan sau khi khir due tir 12h den 2 ngay. Sau 25 - 30 ngay co the thu hat lai.
- Phuong phap chpn tao eae ddng lua kliang ray nau tren co sd sir dung eong nghd ehi thj phan tir.-Sir dung cac ehi thj phan tir SSR lien ket vm gien khang de phat hien cac ea the mang cac gien khang ray nau can thiet tir the he F2 va cac d()ng lai BC.
Chpn tao cac dong lua khang ray nau Ixing phuong phap chpn gidng truyen thdng.
Ul. KET QUA NGHIEN CUU
1. Ket qua thidt lap cdc quan thd chpn gidng liia khdng ray ndu
Cac quan the chpn gidng bdng CTPT da dupe thiet lap thong qua phep lai bun tinh nhu dupe mo ta d phan phuong phap nghien ciru.
Trong nghien cim nay, cae dong/gidng RS, 1S1.2, IS2.3 la ddng cho gien khang him hieu voi ray nau d viing sinh thai nghien ciru nhung chua dap inig dupe ve cac chi tieu nong sinh hpc nhu chat lupng gao, thdi gian sinh trudng, dp cung cay, nang suat bat. Gidng liia SL12 vd IR64 lam vat lieu nhan gien.
Kiem tra sir boat dpng cua gien kliang ray trong cac :-'ddng cho gien khang nhd phdn tich kieu hinh. Bdng :phuong phap lai hiru tinh, cac quan the ebon gidng lir :ac td hpp lai RS, IS1.2, IS2.3/SL12, 1R64 da dupe ::hiet lap.
i 2. Su dung chi thi phdn tir trong chpn gidng liia ihdng ray ndu
Ddng IS1.2, IS2.3 va RS la cdc ddng lua da dupe ,qui tu hai gien khang rdy ndu Bph3 vd BphZ tir gidng lua IR72, Rathu Heenati va GC9. Hai gien nay deu dd lupc xdc dinh la khang vdi qudn thd ray ndu o DBSH ./a DBSCL. Phan tich da hinh cdc chi thi giira cdc
idng cho gien vd ddng nhan gien da xdc dinh dupe
• ;ac chi tiii Udn ket vdi cdc gien khdng hidu qua dupe ':,fU dung ldm chi thi trp giiip trong chpn gidng lua
chdng ray nau cho DBSH. Cdc chi thi dupe lira chpn jdm: RM586, RM588, RM589, RM190 Udn kd't gien 5ph3 trdn NST sd 6; cdc chi thi RM3524, RM1388, yVI3367, RM3735, RM5757, RM6997 lidn ket gien 5phZ ti-dn NST4.
Trong nghien cun n.iy, gien BphZ da dupe danh gia la khang ray nau hieu qua trong nhiing nghien ciru trmVe eua Phong Sinh hpc Phan lir Vien Di truyen Nong nghiep va da dupe lap ban dd tren NST sd 4 ciia liia. Ciicn nay da ducn: qui tu vao (long liia IS 1.2, 1S2.3 va RS tir dong lua GC9 (ci) ngudn gdc tu ddng liia Swanalata ciia IRRI).
,SL12IS2,3 1 2 T) f; 7 8 !) 10 11
• 4 / ; '•t??.
''sA>'
Hinh 1. Minh hpa tiii nghidm sir dung CITT RM5757 lidn kdt gien BphZ chpn lpc cdc cd thd mang gien
khdng cua ddng DTli2-3. Cac cd the tir 1-11 mang bdng ADN cua chi thi Udn ket gien khang Tren hinh 1 minh hoa thi nghiem sir dung ehi thi phan tir trong phan tich x.ic dinh ea the mang bang chi thi lien ket gien khimg BphZ d td hcy[-) SL12/1S1,2 la cac ca the ciia dong trien vpng DTE2-3 mang gien khang ray nau BphZ tir dong 1S1.2.
Hinh 2 minh hpa thi nghiem phan tich sir co mat ciia gien Bph3 trong cac ddng DTR64 (tir td hcyp lai giira 1R(34 voi 1S2.3) sir dung chi thi RM190 lien ket gien Bph3. Dua vao ket qua phan tieh tren gel polyacrylamit, da nhan tii.ay co 7 ca the mang gien khang ray nau Bph3.
1S2.3 1R64 1 2 3 4 5 6 7
i-i^ yik
^'4 -
Hinh 2. Muih hpa thi nghidm sir dung chi tin RM190 Udn kdt Bph3 trong chpn lpc cdc cd thd mang gien khdng ciia ddng DTR64. Cdc cd thd tir 1-7 mang bang
ADN ciia chi thi Udn kdt gien khdng
Hinh 3 minh hpa thi nghidm phan tich sir cd mat cua gien Bph3 trong cdc ddng DTH18 (tir td hpp lai giira H2 vdi IS1.2) sir dung chi thi RM190 lidn ket gien Bph3.
2/J6NG
NGHIEP VA PHAT TRIEN NONG THON - KY 1 - THANG 2/2012
19H2 IS1.2 1
<Si!^>l0f)t-''<-'f'^"--^''
6 7
Hinh 3. Minh hpa thi nghidm sii dung chi thi RM190 Udn kdt Bph3 tiong chpn lpc cdc cd tiid mang gicn
khdng cua dong DlTl 18. Cdc cd tiid tir 1-7 mang bdng ADN ciia chi thi lidn kdt gien khdng
Dim vao ket qua i)han tich tren gel
khiuig ray nau Bph3 tiong thd he con lai cua cac ddng d i i w chpn.
3. Chpn gidng truydn thdng vd khao sdt ddc tinh khdng rdy ndu ciia cdc ddng tridn vpng
1 )e chpn lpe ducyc- nhimg di)ng lua trien vpng cho ndng suat c i o , ehat lirc.mg tdt, da tien hanh chon gidng bang phuimg ph.i]) truyen thdng ket hpp chi thj phan tir. Ket qua da chpn ducyc 6 ddng lua khang ray nau trien vpng eho ddng bdng song Hdng. Danh sach cac ddng trien vpng viri cac ehi tieu nong sinh hpc chinh va d.ie tinh khang ray nau trong vu xuan ndm 2011 d d d n g bdng song Hdng dufn: trinh bay a bang 1.
polyaciylamit, da thu duix" nhimg ca the ni.'ing gien
Bang 1. M6t so chi tidu ndng sinh hpc vd didm khdng rdy ndu ciia cdc dong tridn vpng TT
1 2 3 4 5 6 7 8
Tdn dong DTH18 DTRt^
DTC 1-3 DTE2-3 1R(54 H2 SL12 LSI.2
TGST vu Miia 2010 (ngdy)
100 -105 105 -108 108-112 105 -108 105 -108 105- 110 125-128 105-110
TGS^r vu xudn 2011 (ngdy)
140-145 140-145 140-145 140-145 135-140 135-140 150-155 135-140
Cao cdy (cm) 118-120 118-120 115-117 111 113 105-110 125-128 115-120 135-140
Ddcimg cdy (didm)
.3
Didm khdng rdy
2
3 2
1 3
:\ 2 1 5 ' 7 !
1 1 4
1 7 9 2 Trdn bang 2 la cac chi tidu cau thanh ndng suat trong vu xudn ndm 2011 cua cac ddng trien vpng dddng bdng song Hdng.
Bang 2. Mpt sd chi tidu cdu STF
1 2 3 4 5 6 7 8
Tdn ddng DTH 18 DTR64 DTC 1-3 DTE2-3 IR64 H2 SL12 IS1.2
Sd bong/
khdm 5,32±1,14
5,7±0,59 4±0,57 6,7±0.59 5,75±1,1 5.33+0.76 5.67±0,56 6.25±1,1
thdnh ndng suat vd ndng sudt a i a cdc c Sd
khdm/
m2 50 50 50 50 50 50 50 50
Sd hat/ bdng 160,2±22,24
160,6±9,79 165,83±13,29 190,67±17.13 102,5±4,95
146±5,92 181.33±7,8
98,5±4,95
Ti Id hat chdc
(%)
85,86 92,90 91,06 82,80 96.47 91.66 88,22 86,47
dng tridr PIOOO
h?it (g) 24.56 22.81 24,24 24.09 22,31 24.57 24,56 23,31
1 vpng vu xudn 2011 NSLT
(ta/ha) 89,85 9(S.94 73.15 127,4 63.42 87.62 111.3 62,04
NSTT (ta/ha) 68,0 66,70
58,0 62,0 49,80 67.20 74.30 46.70 Ket qui khao sit die tinh khang ray cua cac
ddng tiien vong a mdt so viing sinh thii
Trong vu he tiiu ndm 2010 vd vu xuan ndm 2011, 4 ddng trien vpng mang gien khdng ray dd dupe ddnh gid muc khdng ray ndu tai Vidn Bao vd Thuc vdt vdi cdc ngudn ray ndu tiiu thdp tai Hd Npi. Nam
Dinh, Nghd An, Long An va Can Tha. Ket qua cho thay ca 6 ddng tridn vpng ddu tiid hien tinh khang cao ddi vdi cac ngudn ray ndu trdn. Kdt qua danh gia tinh khdng ray ndu cua cac ddng/ gidng liia tiong nha ludi duoc thd hidn or bang 3.
20 NONG NGHIEP VA PHAT TRIEN NONG THON - KY 1 - THANG 2 / 2 0 |
Cdc ddng liia mang gien khang rdy nau chpn tao tai Vidn Di truyen Nong nghidp da dupe dua vao viing Ddng Thap Mudi thupc tinh Long An. Day Id viing cd dich ray ndu hai lua (cdc ndm 2007, 2008.
2009, 2010). Quan the ray ndu b day mang doc tinh cao. Voi mat dp ray Id 3500 - 4000 con/ khdm d vu he thu 2007 trong khi cac gidng nhidm bj chay thi cac ddng Ilia cua Vidn Di truyen Nong nghidp mang 2 gien khang vdn cho thu hoach. Ket qua danh gid phan img khang ray ndu d trdn cho thay trong 6 ddng thir nghidm, cd 4 ddng khdng bdng ddi chimg khdng Ptb33 mang 2 gien khang cua IRRI, ddng DTE2-3 cd diem khang tuong duong vdi ddi chimg khdng. Chi b ddng DTC 1-3, cd diem khang b mirc khdng vira. Nhu vay, gien khdng ray trdn cac ndn gien khdc nhau cung the hien tinh khdng b mirc dd khdc nhau.
Bang 3. Kdt qua ddnh gid phan ung khdng rdy ndu ciia mpt sd ddng liia tridn vpng ndm 2010 STT
1 2 3 4 5 6 7 8 9
Tdn ddng DTE 2-3 DTC 1-3 DTH 18 DTR64 IR64 IS1.2 SL12 Ptb33 TNI
—..
Did Hd Npi 2 2 2 2 4 2 7 2 9
m khdng tai cdc ngudn ray Nam
Dinh 3 2 2 2 4 2 8 2 9
t , WK^a-V*"*
Nghd An
2 2 2 2 5 2 7 2 9
_-,.
Ddng Thdp
2 2 2 2 5 2 8 2 9
Can Tha 4 4 4 4 7 4 7 4 9
, ^
Hinh 4. Ddnh gid phan irng khdng ciia mdt sd ddng F2 mang gien khdng tai vung dich rdy ndu - Long An
2007
Hinh 5. Ddng liia khdng ray DTR64 h-dn rudng tiii nghidm tai Thai Binh vu he thu nam 2010
IV. KET LUAN
Sir dung chi thi phan tir tiong chpn gidng khang ray nau da cho thay hieu qua cao khi dua vao va kiem tia sir cd mdt cua 2 gien kliang ray nau ciing tren mot nen gien uu tu. Nhd chpn gidng truyen thdng ket hpp chi thi phan tir, da chpn ducyc 4 ddng trien vpng DTR64, DTE2-3 va DTH 18 va DTC 1-3 m.'ing 2 gien khang ray nau Bi)h3 va BphZ cd dac tinh khang ray cao ddng thdi mang cac dac diem nong sinh hoe tdt, dap ling muc tieu chpn gidng liia khang ray nau cho ddng bdng song Hdng.
Da dua khao nghiem qudc gia (VCU) 3 dong, gdm: DTE2-3, DTCl-3 va DTH18. Cac dong nay deu cd thdi gian sinh trudng ngdn, tir 103-110 ngay tai DBSH vu miia nam 2010. Ndng suat TB dat tir 5,5 - 7,8 tan/ha; chat lupng hat trong vd thon dai; diem khdng ray ddi vdi ray ndu thu thap tai 5 viing sinh thai da khao sat dat tir diem 2- 4 theo thang diem cua Vidn Bao vd Thuc vat. Cac ddng lua nay dupe Trung tam Kliao Kiem nghiem gidng, San pham cay trdng va Phdn bdn danh gia la gidng khang ray nau trien vpng.
TAI UEU THAM KHAO
1. Biii Chi Bim va Nguydn Thi Lang (1999/ Ung dung ADN marker trong danh gia quy gien cay liia.
Bio cio khoa hoc- Hdi nghi Cdng nghe Sinh hoc toin qudc, Ha Npi 1999. Nxb. Khoa hpc Ky tiiuat, tr.
1216-1225.
2. Pham Thi Miii (2001). Nghien cuu chpn tao mpt sd gidng liia khdng ray ndu cho viing ddng bdng song Cuu Long. Ludn van Tien si Nong nghidp. Vidn Khoa hpc Ky thudt Ndng nghidp Viet Nam. 176 trang.
3. Nguydn Cong Thudt. Hd Vdn Chid'n vd Nguydn Thi Hudng (1993). Theo doi sir thay ddi biotyp ray ndu d ddng bdng song Hdng vd ddng bdng song Cim Long va tuyen chpn gidng liia khdng biotyp ray ndu mdi. Hpi nghi khoa hpc BVTV, 24- 25/3, Hd Npi. Nxb. Nong nghidp, ti. 19-20.
4. Nguydn Thi Lang, Biii Chi Biru. Ung dung marker phdn tii danh ddu gien miii thom trdn liia. Di truyen hoc va Ung dimg (2) ndm 2004.
5. Jarin J., Phengrat K., Teangdeerith S., Vanavichit A , Toojinda T., 2007a. Mapping of a broadspectrium brov^n planthopper resistance gene Bph3 on rice chromosome 6. Moi. Breed. 19: 35-44.
NONG NGHIEP VA PHAT TRIEN NONG THON - KY 1 - THANG 2/2012
^ 21APPLICATION OF MAS IN BROWN PLANTHOPPER RESISTANT RICE BREEDING FOR I ^ D RIVER DELTA
Summary
Marker assisted selection (MAS) is a tool for enhancing the efficiency in molecular breeding. In this research, the SSR markers bnked to brown i)I.-intho|)i)er resistance genes were yscd in rice breeding. RS, 1S1.2, IS 2,3 rice lines which is resistant to brown i)lanthop|)er were used as donnor. Rccepiens were elite rice lines SL12, IRf>4. Genetics .analysis of resistance gens showed that there were two genes in RS. 1S1.2 and 1S2.3 cultivars against brown planthopper. Inlrogresion two those genes into the elite rice lines SL12 and 1RG4 using MAS integrating with tradidilional breeding were performed. Molerculars markers linked to those genes were used to select plants possessing HPH resistance genes. Tlie rice lines DTCl-3, DTE2-3, DTH18, DTR64, were selected which possesing two genes Bph3 and Bph7. resist.int to brown planthopper biotypes in both Cuu Long and Red river delta rice fields. Screening for BPH H'sistanee in green house were shown high resist.ince level. Re.il revenue yield of the promissing rice lines were from 5..5-7.8 ton/ha.
Keywords: Bremn planthopper, marker as.^istcd selection (MAS), rice breeding, SSR.
Ngudi phan bidn: GS.TSKH. Trdn Duy Quy
