J Sci &Devel Vol 11 No.2:184-193 TepchlKhoahpcvfiPhfittri6n2013.T$p 11,s62:184-193 www.hua.edu.vn
CHQN LQC DONG NGO Cd KH& N A N G CHjU HAN DlTA TRiN KIEU H I N H VA MARKER P H A N TUT
Phan Dire Thjnh, Hoing Thi Thiy, Phpm Quang Tufin, VQ Thj Bich Hanh, Nguyen Thj Hfin, VQ Vfin Liet*
Vi$n Nghidn cttu vd Phdt trien cdy trSng, Trttimg Dpi hpc Ndng nghidp Hi Npi E-mail*: [email protected]
Ngfiy gfli bdi: 19,02.2013 Ngdy chAp nh§n: 22.04.2013
TOM TAT
28 dflng ty phdi cua ngd (Zea mays I.) cfl ngudn gdc khfic nhau dug-c dfinh gifi dh chgn dflng bd mg cfl kha nfing chju hgn phyc vy cho tgo gidng ngfl uu thd lai. Nhung ddng nfiy dug-c phfit Iri^n tu' gidng ngfl dja phuo-ng vS tir cfic dflng ngudn gen ngfl M? vfi Trung Qudc, Thf nghi$m dfinh gifi khfi nfing chju hgn trdng trong chfiu plaslic, gid fli§
Ifi cfit sgch gfiy hgn nhfin tgo vfi thao dfli cfic linh trgng nhu didn tich !fi/cfiy, t h i tfch rS, chi4u dfii r l , chilu cao cSy, khd \\j<yng r l khfl, khdi lug'ng thfin khfl, Si> dyng marker phfin tu SSR d l dfl tim cfic gen vfi QTL (locus tlnh trgng so lugng) dilu khien mgt s6 tinh trgng iifen quan d i n khfi nfing chju hgn. Ket qua cho thay trong dilu kign gfiy hgn nhan tgo, mflt s6 tinh Irgng cua re cfl tuong quan chfit v6i nfing sudt. Ket qua dfi chi ra rfing cd the si> dyng cfic llnh trgng nfiy d l xfic djnh kha nfing chju hgn a ngd. Marker SSR vdi 3 mdi dfic hidu Ifi umc1862 lidn k i t vfl-i gen QTL nfing suiit dudi dieu kign hgn, umc2359 lidn k i t vfl'i chi s6 chju b^t thufin vfi nc 133 lidn k i t chi s6 chju hgn dS nhfin biet 26 ddng cd gen QTL dieu khien nfing su4t ngfl du6i dilu kidn bit thufin nufl'c (YS) vfi QTL chi s6 chdng chju bSt thugn (TOL), 14 dflng mang QTL chdng chju v6i dieu ki§n bfit thufin (TOL). Dya trdn k i t qua dfinh gifi kilu hinh vfi marker phfin tLP da chgn dug-c 5 ddng ifi TP17, TP12, TP2, TP5 vfi TP24 cfl t h i s u dyng cho chgn tgo gidng ngo lai chju hgn,
Tif khfla: Ddng thu^n, ch|u hgn, tinh trgng re. marker phfin tu.
Selection of Inbred Maize Lines for Drought Tolerance Using Phenotyic Evaluation and Genetic Markers
ABSTRACT
Twenty eight maize fZea mays L ) inbred lines of different origin were evaluated to select the drought tolerant lines as parents for hybrid maize breeding. These inbred lines was developed from ihe local open-pollinated cultivars and exotic gerplasm from China and Amenca, To preliminarily identify drought tolerant inbred lines, the lines were grown in plastic pots containing stenlized fine sand and the following traits were observed, leaf area per plant, root volume, root length, plant height, freshand dry root weight, and dry stem weighL The genes or QTLs associated with drought tolerance were analyzed using three specific SSR markers Data analysis showed that nDot traits are conelaled with yield under drought condition. And cound be considered as primary indicators to detemnine the drought-tolerance of maize, umc1d62 marker was found to be associated with grain yield under stress condition (Ys), nc133 marker associated with the stress tolerance and umc 2359 associated with mean productivity, 28 inbred lines investigated have QTL controlling yield under strees and tolerance. Of these 14 lines that have QTL associated with strees tolerance index. Combining phenotypic evaluation and genetic markers. 5 tines, i e TP17, TP12, TP2, TP5 and TP24 are suggested for combining ability testing for future development of hybnd maize varieties wiOi tolerance.
Keywords: Drought tolerance, maize inbred lines, molecular markers, root traits.
l . D A T V A N D E . . , - W T . * ^ nnnm T„V difdc nhu cfiu ciia con ngUdi (FAO, 2009). IW San lUdng cay lUdng thdc toan ciu den nfim nhien, san xufi't liidng thiJc ndi chung cung nhif 2050 c i n viidt qua 400 t r i l u tfi'n mdi dap iing san xufi't ngd nfli rieng dang phai dli mat vdi 184
Phan Dlic Thjnh, Hofing Thj Thuy, Phgm Quang Tufin, VO Thj B(ch Hgnh, NguySn Thj Hfin, Vu VSn Liet
thach thtic ldn nhS^t Ifi dilu kien btft thu^n sinh hgc (sau binh hai) vfi bfi't thufin phi sinh hgc (han, d^t nghdo dinh dudng, dfi't chua vfi ngfip nddc). Nhiing thfich thilc nfiy d$c biet tac ddng mgnh dH vdi ndng dan san xufi't nhd, nghio tfii nguyen vfi dau tii thfi'p (Weiwei- Wen vfi cs., 2011). Cfic nhfi nghi&n ciiu tai CIMMYT cho r^ng each til'p cfin duy nhfi't di cai til'n kha nfing chiu han b ngd Ifi danh gifi mdt s l lUdng Idn quin the de nhfin biet va s\i dung nguon gen chiu ban cho chuong trinh cai tie'n gid'ng (Banziger va cs., 2000).
Theo Yunbi Xu (2010), ban d l di truyln phan tii cua cay ngo da dUgc xay diing bing cfic marker SSR, RFLP va d l nhfin bil't cfic locus tinh trang so' Iiidng (QTL) lien quan dl'n kha nfing chiu ban). Trong nhiing nfim gdn day, cac chi thi phan tii da ditdc sii dung de danh gia, nhfin bil't tinh trang phuc vu chgn tao gid'ng ngd chiu han va kJ thufit nfiy phat trien manh me (Nathinee Ruta, 2008; Rahman, 2011;
Zahrar va Jahad, 2011; Weiwei Wen, 2011).
Nhflng vung trong ngd chinh b miln Bfic niidc ta chii yeu vung niii, hai ven song canh tac nhd niidc trdi, Tinh trang thieu niidc se ngfiy cfing nghiem trgng hdn do bien doi khi hfiu, Vi vfiy, chgn tao giong ngd liu the lai cd kha nang chiu han Ifi mgt giai phap d l san xufi't ngd on dinh vfi bin vflng.
2. PHUONG PHAP N G H I E N CUtJ Vat lifiu nghien cflu gom 28 ddng tii phoi tfl ddi S3 din Si2, kj^ hieu tii TPl den TP2S, gom 17 dong tfl phdi phfit trien tfl giong ngo dia phfldng (TPl, TP2, TP3, TP4, TP5, TP12, TP13, TP14, TP15, TP17, TP18, TP19, TP20, TP22, TP23, TP24 vfi TP25); 4 ddng tii nguon gen ngfl cua My (TP6,TP16, TP27 va TP28) va 7 ddng tfl ngudn gen ngd cua Trung Qu6i: (TP7, TP8, TP9, TPIO, TPll, TP21 va TP26). D61 chflng la LCH9.
Nghiin cflu dfldc til'n hfinh tai Viin NghiSn cflu vfi Phat triln cay trong, trfldng Dai hgc Nong nghiep Ha Ngi vu xuan vfi thu dong 2012.
Cac dgc dilm nong sinh hgc, nfing sufi't vfi yiu to' tao thfinh nfing sufi't cua cac ddng dfldc dfinh gia bang phflOng phap thi nghiem ddng
rugng, hai Idn Ifip lai, di^n tich moi d thi nghiam Ifi 14m^.
Kha nfing chiu han ciia cac ddng dfldc danh gifi theo Camacho vfi cs. (1994), Ngo dfldc trong trong ch§u plastic dfldng kinh 20cm, cao 35 cm, cd t b i rut nfldc chii dgng). Moi ddng trong trong 3 ch4u, mSi ch&u trdng 3 cfiy, Gia the trdng Ifi cat sach... Sau gieo 4 tudn, thu hoach biing each nhd ca cfiy vfi theo doi cac chi tilu liln quan den kha nfing chiu han gdm: Dien tich la /cay; the tich r l (RV) dfldc xac dinh bfing each cho re vao ong dong, dd nfldc ngfip ghi the tich ta cd gia tri the tich td'ng (V(tdng)) sau dd vdt re ra ghi the tich Ifi the tich nfldc V(nfldc), cdng thflc tinh the tich r l RV = V(tong) - V (nfldc); chilu dai re dai nha't (- (LRL), chilu cao cay (PH) dfldc do bfing ddn vi cm; khdi litdng re tfldi (RFW); khoi Ifldng re kho (RDW); khoi Ifldng than khd (SDW) dfldc can bfing cfin dien tii va do chinh xac 0,01g; ty 16 RDW/SDW, Kieu gen nao cd cac chi tieu tren cao hdn thi dfldc danh gia cd khfi nfing chiu han tot hdn.
Cac ddng cd mang QTL chiu han ciia 3 tinh trang la: (i) nang sufi't diidi dieu kiln han,; (ii) chi so'chiu ban va (iii) kha nang chiu han dfldc nhfin hilt thdng qua cac cap moi dfic hieu cd nguon goc tfl Dflc do khoa Cdng nghe sinh hgc mua tfl nfim 2011 vfi dfldc tham khao tfl nghien cflu cua Mohammadreza Shiri (2011). Ten va trinh tfl moi trinh bay d bang 1.
DNA dfldc tach chiet tfl 3 la non cua cay d giai doan 4 din 5 la that bfing phfldng phap CTAB (cetyl trimethylammonium bromide theo Saghai-Maroof vfi cs. (1984). So Ifldng va cha't Ifldng DNA danh gia tren may quang phd UV- spectrophotometer. Sii sung lOul hon hdp DNA cua cac cfi the, lOpl tong so the tich DNA phan flng PCR cho vao dem phan flng IX PCR chfla 2,0mM MgCIa; 0,2 mM dNTP mix; 0,4 ^M mdi bd primer 0,5 U Taq DNA polymerase (SIGMA) va dieu chinh lOul bfing nfldc cfi't 2 Ian (ddH^O).
Khuyech dai trong may PCR vdi chu k^ nhilt nhfl sau chu k^ 1: lfim bie'n tinh tai 94''C trong 5 phiit, tiep theo 35 chu k>' tai nhiet do 94''C trong 30 giay; nhiet do 55°C trong 1 phut va 72°C trong 1 phut va 30 giay, cuoi cung la chu k)> md rdng tai 72"C trong 10 phiit. San ph^m PCR dfla
Chgn Igc dflng ngfl cfl khfi nfing chju hgn dya tr^n kilu hinh vfi marker phdn ti>
B&ng 1. Tgn vfi trinh tii mdi
Trlnh ty mil Dfl tim gen ho^c QTL
umcl 862 forward: ATGGGCACATGAAAAAGAGACATT Nang suit dir(i'i diiu ki$n hpn umcl 862 reverse- CCCATGAGAAGAGTGAAGACAACA Nfing su^t dviei diiu kl$n han
net 33 forward. AATCAAACACACACCTTGCG Chi sd chju h^n
GCAAGGGAATAAGGTGACGA Chf sd chju h^n
umc2359 reverse:
umc2359 forward-
GCCTGACATGAATGTTACATGAGC CTGGATCAGATGAAAAAGAAGGGA
Khi nfing sinh sfin tmng brnh duM diiu klin hpn, chf s6 chju bit thu$n Khfi ndng sinh sfin trung binh dirdl diiu kl^n hpn, chi s i chju bit thudn len gel agarose 2,0% trong d^m IX THE (89 mM
Tris, 89 mM Boric acid vfi 2,5 mM EDTA pH 8,0), chfla 0,15 pg/pl thidium Bromide. Quan sfit vfi chup anh dfldi din ci^c tim,
Phan tich phuong sai, phan tich tfldng quan, b^ng chfldng trinh phdn mim IRRISTAT 5.0 va chi so" chgn lgc b^ng chfldng trinh thong k l sinh hgc cua Nguyen Dinh Hien (1995)
3. KET QUA VA THAO LUAN
3.1 Dac d i l m nong sinh hoc cua cac dong nghien ciJfu t r o n g t h i n g h i e m dong r u o n g
Dac diem ndng sinh hgc ciia 28 ddng ngo nghien cflu trong vu thu dong nam 2012 tren dfi't phu sa song Hong khong dfldc bdi hang nfim vfi canh tac nhd nfldc trdi bieu hiln ve thdi gian tfl gieo den chin sinh If ciia cac ddng bien dgng tii 96 den 115 ngay thugc nhdm trung ngay (Bang 2). Trong do, 7 ddng co thdi gian sinh trudng dfii hdn hoac ngang bfing doi chflng, dd la cac dong TP2, TP7, TP14, TP15, TP17, TP25 vfi TP26; cac ddng cdn lai thdi gian sinh trfldng deu ngfin hdn doi chflng LCH9 (108 ngfiy).
Chenh llch tro cd - phun rau la mgt tinh trang liln quan den kha nfing chiu han, ddng giong cd chinh llch tro cd - phun rau ngfin cd kha nang chiu ban td't hdn (Pervez, 2002;
Gemenet, 2010). Tfi't ca 28 ddng tii phm nghien cflu dlu cd thdi gian chenh llch trd cd - phun rau tu 0 dl'n 3 ngay phfl hdp vdi dac diem ddng cd kha nfing chiu ban.
Chilu cao cfiy cua 28 ddng kha biln dgng, nhdm cac ddng cfl chilu cao cfiy cao hon doi chflng gdm TP4, TP7, TP8, TP9, TPlO, TP12, TP13 vfi TP24. Cac ddng thSp cay gom TP19, TP20, TP21, TP22 vfi TP 28, chilu cao dfldi 170cm phii hdp cho phat tri^n giong ngo lai tham canh. Trong dd ddng TPl 9, TP20 vfi TP22 dfldc phfit trien tfl giong ngd dia phfldng Chfim deng vfi Khdu li se ciia ngfldi Mdng, Hfi Giang, Ddng TP21 vfi TP28 phat trien tfl nguon gen nhfip ndi ciia Trung Quoc vfi Mj.
Nfing sufi't va yl'u to tao thfinh nang suat cua cfic ddng ngd tfl phoi trong dieu kien canh tac nhd nfldc trdi vu thu dong 2012 dlu thfiip hdn hofic ngang bfing doi chflng, sai khac gifla cfic ddng khdng b mflc cfl y nghia. So bfip/cay cua cac ddng trong dieu kiln dong rugng bien dgng tfl 0,85 - 1,1 bfip, ddng cd ty 1$ bfip cao la TP2, PT16 vfi Vr 17 (1,1 bdp/cfiy). S l hfing hat trung binh ciia cac ddng ngo thi nghiam nam trong khoang 12 dl'n 16 hfing, so h^t tren hang 28 - 30 h^t, khdi Ifldng 1000 hat bien dgng trong khoang 200 den 250g, vfi ca 3 yiu to t?o thfinh nfing sufi't nfiy khdng sai khac gifla cac ddng d mflc cd J nghia. Nfing sufi't cfi thi chi c6 3 ddng ngang bing d6i chflng Ifi TP15, TP16 va TP17, cdn lai cac dong dlu cfl nang sud't ca the tha'p hdn dll chflng (Bang 3). Tuy nhidn, so sanh vdi doi chiing chi d l tham khao, vi muc dich ddi chiing trong nghien cflu Ifi danh gia kha nfing chiu han cila cac dong so vdi giong lai chiu han (LCH9).
186
Phan Dtfc Thjnh, Hofing Thj ThCiy, Phgm Quang Tufin, VQ Thj B(ch Hgnh, NguySn Thj Hfin, VO Vfin Lilt
B a n g 2. Mot so d|ic diem n o n g sinh h9c cua cdc dong nghien cvTu t r o n g vu t h u dong n a m 2012 tai Gia Lam, Ha N6i
STT Ky hi$u dflng CC (cm) CDB (cm) TGST (ngfiy) G-TrC (ngSy) G - PR (ngfiy) ASI (ngfiy) 1
2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 Chii thfch.
TPl TP2 TP3 TP4 TP5 TP6 TP7 TP8 TP9 TPIO TP11 TP12 TP13 TPl 4 TPl 5 TPl 5 TPl 7 TP18 TP19 TP20 TP21 TP22 TP23 TP24 TP25 TP26 TP27 TP28 LCH9 (die) CC. chieu cai G-PR : gieo din phun ra
190,1 190.4 194,0 204,1 195,1 171,7 208,0 201,4 201,2 205,1 191,3 254,9 248,1 198,7 176,3 193,3 197,2 173,9 165,9 162,9 165,6 168,4 191,6 216,7 188,5 181,6 192,5 165,3 194,3 ) cay; CDB- chieu u;ASl. thdi gian
71,4 66,9 90,6 82,7 84,1 67,5 90,0 57,0 53,1 57,1 58,7 125,4 120,3 66,4 65,5 81,6 77,4 69,1 66,8 61,0 68,0 61,1 68,0 74,3 76,2 62,9 68,0 67,0 109,7 'caodont chenh lei
dong bip; TGST: thdi gian sinh trUdng; G-TrC: thdi gian (LT gieo din ti '-phun rau.
3.2. Danh gia k h a n a n g chiu h a n ciia cac ddng b&ng thi nghiem gay h a n n h a n tao
Trong dilu kien gay han trong chfiu plastic, cac ddng TPIO, T P l l , TP12, TP13, TP24, TP 25 vfi TP 26 deu sinh trfldng td't tfldng dfldng vdi ddi chflng LCH9. Cac ddng cfl so la cao hdn ddi chflng la TP12, TP13, So ddng thfi'p hdn ddi chflng la TPl, TP4, TP6, TP7, TP17, TP22 (Bang 4). Cac ddng TP5, TP7, TP12, TP13, TP21 vfi TP25 cd gia tri diln tich la vfldt hdn so vdi doi chflng LCH9, dac hilt ddng TP5 (235,79 cmVcay), TP12 (244,47 cm7cay), TP 13 (238,67
cmVcfiy). Cac dong cdn lai dien tich la thfi'p hdn doi chflng LCH9 (Bang 4).
Trong dilu kien ban, qua tnnh sinh trudng re cay giam, so lUdng re va the tich cua bd re giam dang k l so vdi cay trdng trong dieu kien binh thudng. Trong thi nghiem, nhilu ddng co t h i tich re vfldt hdn so vdi doi chflng LCH9 (2,40ml), cac dflng TP 2, T P l l , TP12, TP13, TP17,TP 22, TP24 va TP 26 deu cd the tich r l ldn hdn 3ml/cay (Bang 5). Pierangelo Landi & cs.
(2010) cho rang nhflngddng cfl tinh trang nay cao hdn, kha nang chiu ban td't hdn. Nhfl vay, cac
Chgn lgc dflng ngfl cfl kha nfing chju hgn dya trfen kilu hinh vfi mariner phSn tO-
ddng cdn lai deu cd t h i tich re thfi'p hdn doi chiing LCH9, cfl t h i kha nfing chiu han kdm bdn.
Chieu dfii re lien quan den kha nfing hut nfldc ciia cfiy; bd r l cfing dfii thi kha nfing dfim xuyin vfi kha nfing hut nfldc cfl hieu qua hdn.
Chieu dfii re ciia cac ddng cho thfi'y cfic ddng deu cfl chieu dfii re tren 30cm (trfl ddng TP14 r l rfi't ngfin = 23,81cm); trong dd, cfic ddng TPl, TP3, TP6, TP7, TP15, TP18 vfi TP 23 re ngfin hdn ddi chflng, cac ddng cdn l^i r l dlu dfii hdn ho§c gdn tfldng dfldng doi chflng.
Ty 1$ RDW/SDW c6 tfldng quan thufin vdi kha nfing chiu hgn ciia cfiy. K^t qua thi nghiam cho thfi'y hdu het cac ddng nghien cflu chi tilu nfiy d l u dat tfldng dfldng ho$c vugt dli chflng, nhieu ddng cd ty 1§ nfiy khfi cao nhfl TP3 (0,78), TP4 (0,7), TP12 (0,74). TPl3 (1,03) vfi TP17 (0,83).
Tfldng quan gifla nfing sulTt cfi th^ ciia cac ddng ngd vdi m0t BI" tinh trgng tgo thfinh nfing sufi't vfi mgt s l tinh trang chiu ban cho thfi'y nfing sufi't cfi t h i tfldng quan chfit vdi so bfip/cay
Bang 3. Nang sufi't va yeu to'tao t r o n g vu Thu Dong
t h a n h n a n g su£ft ciia cfic dong nghign ciJfu nfim 2012 tai Gia Lfim, Ha N9i
STT 1 2 3 4 5 6 7 8 9 10 11 12 13 I t 15 16 17 18 19 20 21 22 23 24 25 26 27 26 29
Ky hi$u dflng T P l TP2 TP3 TP4 TP5 TP6 TP7 TP8 TP9 TPIO T P l l TP12 TP13 TP14 TP15 TP16 T P l 7 T P l 8 T P l 9 TP20 TP21 TP22 TP23 TP24 TP25 TP26 TP27 TP28 LCH9 (fl/c) LSDos CV%
S i b^p/cfiy 1,00 1,10 1,00 0,95 1,10 0,95 1,00 1,00 0,95 0,95 1,00 0,92 0,92 0,85 1,00 1,10 1,10 1,05 1,00 1,00 1,00 0,90 1,00 1,00 1,00 0,85 0,95 1,00 1,10
S i hdng hpt/b^p
14,15 13,95 13,05 12,55 13,35 14,78 12,50 13,76 13,64 13,10 12,88 14,33 14,45 12,88 12,88 12,74 12,80 12.75 12,97 12,50 13,50 13,58 13,56 12,49 14,39 12,26 15,55 12,81 14,85 0,96 3,5
S i hgt/hdng 28,75 27,65 22,76 24,33 25,71 23,08 23,95 25,83 24,42 24.40 22,98 24.27 25,90 24,32 25,25 24,56 26,44 22,25 27.00 22,25 24.74 22,53 24,93 23,18 25,46 23,09 27,47 27,90 26,20 2,27 4,5
P.oaor*(g) 218,98 209,32 221,13 211,65 219.83 210,64 227,26 220,08 221.38 222.92 229,65 240.84 241,50 230,91 231,70 228,03 225,38 221,75 228.86 227,53 210,43 203,60 216,03 240,85 224,94 231,59 214,01 232,87 249,51 23,65 5,2
Nfing su ill c&
thi(g) 125,88 132,05 125,75 122,72 127,47 121,28 127,80 125,67 117,52 115,47 126,20 126,16 116,32 119,19 135,89 149,53 140,40 125,92 124.52 122,40 124.93 111,48 123,61 122,34 122.91 111,66 126.12 126,91 136,30 10,44 4,1
188
Phan Oi>c Thjnh, Hofing Thj Thuy, Phgm Quang Tufin, VO Thj Bich Hgnh, Nguyln Thj Hfin, VO Vfin Lilt
Bang 4. Tinh trang cay cua 28 dong khi trong trong chgu va gay han nhan tao
STT
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29
Ky hi$u dflng
T P l TP2 TP3 TP4 TP5 TP6 TP7 TP8 TP9 T P I O T P l l T P l 2 T P l 3 T P l 4 TP15 T P 1 6 TP17 TP18 T P l 9 TP20 TP21 TP22 TP23 TP24 TP25 TP26 TP27 TP28 LGH9 (O/C) L S D M
cv%
Silfi
2,51 3,53 3,14 2.83 3,34 2.71 3,08 3,42 3,22 4,09 4.17 4,74 4,62 3,82 3,76 3,18 2.92 3,74 3,26 3,93 3,15 3,08 3,80 4,17 4,22 4,12 3.74 3.21 4,10 0,19 2.8
Dien tich la/cfly (cm^) 119,83 162,24 126,35 189,69 235,79 168.28 206,41 164,43 151,73 172,49 161,84 244.47 238,67 178,77 133,21 181,99 222,88 163,19 116,56 160,70 202,19 128,98 193,10 195,11 216,16 135,43 167,98 184,37 198,72 9,34
3,3
C h i i u cao cfiy (cm) 32,95 40,69 46,22 47,22 56,19 50.25 55,44 59,92 48,28 46,82 45,97 48,75 51,14 52,13 39,13 47,14 51,28 50,60 49,17 45,71 48,61 47,15 50,87 52,82 50,97 44,28 38,05 53,14 53,01 1,79 2,3
K h i l lu'dng thdn kha (g) 0,92 2.25 1,86 1,49 1,68 2,31 1,76 2,56 2,00 1,38 2,25 1,97 1,14 2,29 1,88 2,87 1,79 1,65 2,97 2,17 1,84 2,61 1,98 2,13 3,36 1,97 2,61 2,24 0,91
(0,638), sfl' hat/hang (0,709). chieu dfii re chieu dfii re (0,768), khd'i Ifldng 1000 hat vdi (0,578). khdi Ifldng r l (0,630), khoi Ifldng r l khfl chilu dfii re (0.570). Ket qua nghien cflu chi ra (0,664). Ket qua nghien cflu cua chung tdi phu rSng cd the ap dung phUdng phap danh gia gay hdp vdi nghien cflu cua cac tac gia Camacho vfi ban nhan tao trdng trong chau vai va cac chi so' cs. 1994, Pierangelo Landi va cs., 2010). Tfldng chiu ban d l nghien cflu thanh lgc vat lieu di quan gifla cac yl'u to tao thfinh nfing sufi't vdi truyen chiu ban trong qua trinh chon tao gid'ng tinh trang re cung hilu hidn qua so bap/cay vdi ngd cho vung khd khan ve nfldc tUdi.
Chon loc d6ng ngS c6 k h i nSng chju h?n iv^ trSn ki4u hinh v4 marker phSn tl>
Bang 5. Tlnh t r a n g r l c u a 28 dong ngo tif phoi t r o n g t h i n g h i $ m c h ^ u plastic n t m 2012
STT
1 2 3 4 5 6 7
a
9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29
K^ hi$u ddng
T P l TP2 TP3 TP4 TP5 TP6 TP7 TP8 TP9 TPIO TP11 TP12 T P l 3 TP14 TP15 TP16 TP17 T P l 8 T P l 9 TP20 TP21 TP22 TP23 TP24 TP25 TP26 TP27 TP28 LCH9 (D/C) LSDo.M
CV%
T h i tich rfi (ml) 1,15 3,06 2,02 2,24 2,24 1,96 1,92 2.18 2,85 2,84 3,80 3,52 5,10 0,79 2,00 2,02 4,12 2,75 2,54 1,48 2,18 3,43 2,12 3,50 2,91 4.75 1,66 2,77 2,40 0,57 14,4
C h i i u ddi ih (cm)
31,47 39,03 33,62 50,08 38.37 35,45 34,52 43,51 48,60 44.39 42.08 48,79 56,93 23,81 30,25 37,39 '46,70 32,51 46,12 43,33 48,52 38,69 35,66 42,58 42,91 47,50 44,71 37,29 43,00 3,23 5.2
K h i l lu-gng r i tircKg)
1,05 3,64 1,74 1,68 2,01 1,49 1,30 2,50 3,40 2.06 1,99 3,25 4,18 3,13 1,90 1,90 3,93 2,63 2,73 1,58 2,03 1,87 1,56 3,64 2,48 2,98 1,74 2,54 2.89
K h i l l u g n g rS k h 6 ( g )
0,60 1,14 1,44 1,04 0,90 0,98 0,73 1,01 1,08 0,77 0,78 1,45 1,17 1,15 0,81 1,06 1.48 1,05 1,18 0,78 0,82 0,76 0.70 1,26 1,36 0,88 1,16 1,02 0,66
T ^ l $ RDW/SDW
0,65 0,51 0,78 0,70 0,54 0,43 0,42 0,40 0,54 0,56 0,35 0,74 1,03 0,51 0.43 0,37 0,83 0,63 0,40 0,36 0,45 0,29 0,35 0.59 0.41 0,45 0,44 0,46 0,73 0.06 6.1
3.3. Nhan biet QTL chiu h a n t r o n g 28 dong condition), vfi cfip moi nc 133 dd tim QTL dieu ngo nghien ciJfu bang m a r k e r p h a n tijf SSR khien di truyen kha nfing chong chiu bfit thu§n nUdc CrOL. tolerance); c^p mli umc2359 dd tim QTL dieu kien chi so chd'ng chiu bfi't thufin nfldc trin cd sd tham khao k^t qua nghien cflu cua cua Phan tich xac dinh QTL chiu ban ciia 28
ddng vfit lieu la cac ddng tu phoi tfl t h i he S3 den S12 vfi doi chflng la giong lai LCH 9 (doi chflng).
o'., J 1, - -- a- T. - 1- -.n^^ J- .^ Mohammadreza Shiri (2011).
Su dung ba cap moi dac hieu la umc 1862 do tim
QTL dilu khiln nang sua't ngo dudi dilu kien bfi't Marker umcl862 do tim QTL dieu khien thufin nfldc (YS, yield of a Unes in water stressed nang sufi't dfldi dieu kien bfi't thufin nfldc cho 190
Phan Oirc Thjnh. Hofing Thj Thuy, Phgm Quang TuSn, VO Thj B(ch Hgnh, Nguyln Thj Hfin, VO Vfin Uet
th^y ca 28 dong vfi dli chflng deu xufit hi^n band vdi kich thudc n&m trong pham vi 100 din 200hp, hfiu h i t kfch thudc khoang 150 bp nSm tren nhilm sac the s^ 1 va bin 11. Mflc da hinh cao vfi giai thich 30% phfldng sai kieu hinh, do vfiy cho phip kl't lufin cac dong nghien cflu deu mang QTL dilu khien nfing sufi't dfldi dilu kien ban. Kit qua phii hdp vdi ke't qua ciia Mohammadreza Shiri da cdng bo nfim 2011.
marker umcl862 lien kl't vdi nfing sufi't hat dudi dilu kien bfi't thu^n, Ribaut vfi cs. (1997) cung nhfin biet 2 locus nam tren NST sd' 1 vfi s6' 9 giai thich 21% phUdng sai kilu hinh nfing sua't dudi dieu kiln hgn.
Chi s l chong chiu bfi't thufin (STI) lien quan den cd c h i sing sflt cua cfiy khi gap bfi't thufin.
Marker ncl33 lien ket vdi gen. QTL dilu khien TOL, marker nfiy cung Hen ke't vdi chi so' mfin cam bfi't thufin (stress susceptibility index - SSI). Marker ncl33 dUdc dung de dd tim QTL nay tren NST so' 2 ciia 28 ddng nghien cflu cung da nhfin biet 28 ddng vat lieu mang QTL chong chiu ban (TOL). Kich thfldc cac QTL trong khoang 100 den 180 bp, kich thudc cac band sai
khfic nhau khfi ro ret. QTL nfiy n^m tren NST s l 2 vfi bin 05 cfl mflc da binh rfi't cao chflng td rSng marker lien kit ch$t vdi chIng chiu bfi't thufin vfi cfl kha nfing phan anh dung kieu hinh ciia cac ddng ngd nghien cflu. Kl't qua nghien cflu phfl hgp vdi kit qua nghien cflu cua cac tfic gia khfic nhfl Quarrie & cs, (1999); Shiri (2011);
Jiufeng Guol cs. (2008), Gieng so 21 xufi't hien 2 band, nhfl vfiy ddng tfl phm TP13 mang ca 2 QTL chIng chiu bfi't thufin vfi chi so' mfin cam bfi't thufin (SSI). Marker umc 2359 ciing da dd tim dugc QTL lien quan de'n nfing sufi't dfldi dieu ki^n bfi't thufin vdi 14 ddng vfi doi chflng LCH 9 (Hinh 2).
Marker umc 2359 cdn dd tim QTL dieu khien nfing sufi't trung binh (mean productivity - MP) dfldi dieu ki&n ban. Kit qua dfi xac dinh ddng mang QTL chiu ban bo sung cho kit lufin vl kieu hinh dam bao do tin cfiy. Nhfl vfiy, cd thi sii dung phUdng phap chgn Igc nhd marker phan tfl (MAS) trong chgn tao gid'ng ngd chiu han vdi cac ngudn vfit lieu khac nhau de khuyin cao nhflng ddng flu tu cfl dfic diem nong sinh hgc phu hop va mang QTL cho'ng chiu ban.
Hinh 1, San pham PCR cua 28 dong tfl phoi va do'i chiing v6i marker UMC 1862 t r i n gel Agarose 2%
M 16 17 IS 19 20 21 22 23 24 25 26 27 28 29 30
Hinh 2. San pham PCR cua 14 dong tfl phoi va doi chufng vdi marker umc 2359 t r i n gel Agarose 2%
Chgn Igc ddng ngfl cfl khd nfing chju hgn dg'a trdn kilu htnh vfi marker phfin t u
B a n g 6 . NhuTng d a c d i l ' m c d b a n c u a c&c d 6 n g c h o n lfim v a t l i e u c h o c h p n g i o n g n g o c h i u h a n
Dflng TP17 TP12 TP2 TP5 TP24
TGST (ngfiy) 115 104 110 102 107
CCG (cm) 197 215 190 195 217
S i blp/cfiy SHH SH/H KLH
(gam) ASI (ngfiy)
NSCT (g/cay) 1,10 12.80 26,44 225,38
0,95 14,33 24,27 240,84 1.00 13,95 27,65 209,32 1.1 13,35 25,71 219,83 1,00 12,49 23,18 240,85
140,4 126.2 132,1 127,4 122,3 Ghi chii: TGST: thai gian ainh tritdng; CCC: chiiu cao c&y; SHH: t
h^t. ASI: chdnh l^ch tri cd phun r&u, NSCT n&ngsuitc6 thi
Mdt s o ' t i n h t r a n g cd b a n nhfl t h d i g i a n s i n h trfldng, c h i e u c a o cay, c h i e u dfii r e , k h o i lUdng re, c h e n h lech t r o c d - p h u n r a u vfi nfing sufi't ca t h i dugc dUa vfio c h g n d d n g vfit l i l u . Ke't q u a sfl d u n g chUdng t r i n h chi so c h g n lgc vdi c u d n g d o chgn lgc 2 0 % d a c h g n dUdc 5 d d n g u u t u nhfi't ( B a n g 6). Nhflng d d n g c h g n d e u m a n g Q T L c h i u b a n q u a k e t q u a d o t i m bfing m a r k e r p h a n tfl.
4. K E T L U A N
Cac d o n g ngd tU p h o i n g h i e n cflu cd t h d i gian s i n h t r u d n g tfl 9 7 ngfiy (TP27) dl'n 115 n g a y (TP2 vfi T P 2 2 ) , t h u g c n h d m c h i n t r u n g b i n h . Cac d d n g cd d a c d i e m n d n g s i n h hgc nhfl chieu cao cay, c h i e u cao d d n g bfip, n a n g sufi't vfi y e u to' t a o t h a n h nfing sufi't p h u h d p cho p h a t t r i e n giong ngd flu t h e lai.
Cac chi t i e u h i n h t h a i cay v a bg r e cd tfldng q u a n vdi nfing sufi't ciia d d n g tfl phoi t r o n g d i e u k i l n gay b a n n h a n t a o b a n g chfiu vai. D u a t r i n cac dSc diem p h a t t r i e n c u a r e v a t h a n la d a nhfin bie't dfldc 7 d d n g cd k h a nfing c h i u b a n Ifi T P l l , T P 1 2 , T P 1 3 , T P 1 7 , T P 2 4 , T P 2 5 , T P 2 6 . .
Sfl d i i n g m a r k e r p h a n tfl dd t i m Q T L l i l n q u a n de'n c h i u b a n d a nhfin b i e t 2 8 d d n g cd Q T L dieu k h i e n nfing sufi't n g d dfldi d i e u kidn bfi't thufin n u d c (YS) v a Q T L chi so c h o n g c h i u bfi't thufin (TSI), 14 d d n g m a n g Q T L c h o n g c h i u vdi dieu k i e n bfi't thufin,
D u a t r e n d a n h gia k i e u h i n h vfi m a r k e r p h a n tfl d a c h g n dUdc 5 d d n g T P 1 7 , T P 1 2 , T P 2 . T P 5 va T P 2 4 Uu t u d l u l a n h f l n g d d n g p h a t
^Ang h^t/bip; SH/H: aih^t/hdng; KIM: khSi lu^g 1000
t r i e n tfl gid'ng n g d d i a p h f l d n g Vifit N a m cd t h i sfl d u n g d c lai thfl k h a nfing k e t h d p chgn tao gid'ng ngd l a i c h i u b a n .
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