Tap clli Khoa hoc Tru&ng Dgi hgc Cdn Tha Phdn B: Ndng nghiep. Thiiy san ^ a Cong nghi Sinh hgc: 26 (2013). 104-111
Tap chl Khoa hpc Tru'dng Dai hpc Can Thd website: sj.ctu.edu.vn
TUYEN CHON VA TAI SINH MQT SO GIONG LUA CO KH.\ NANG CHIU MAN THICH ITNG VCtt BIEN DOI K H I HAU 6 DONG BANG SONG CtJU LONG Nguyen Thi Thanh "niao, Trin Thi Xuan Mai, Dd Tan Khang' va Trdn Nhan Diing'
' Viin Nghien cuu vd Phdt trien Cdng ngh? Sinh hoc, Tnrdng Dgi hoc Cdn Tho Thdng tin chung:Ngdy nhdn: 20/12/2012 Ngdy chdp nhgn: 20/06/2013 Title:
Selecting and regenerating sah tolerant rice varieties collected from Mekong Delta Tie khda:
Chdng chiu man. RAd206, RA(8094. md seo, tdi sinh choi
Keywords:
Salt tolerance. RM206.
RM8094, callus, regeneration
ABSTRACT
Eighteen rice varieties collected from .Mekong Delta M'ere used lo evaluate the salt tolerance using hydroponic system containing Yoshida solution with additional NaCl concentrations (0%o, 4%o. 6%o) The results showed that increasing salt levels, the survival rate, shoot height, root length, shoot and root dry weight of the tested rice plants was greatly reduced M'hile the chlorophyll concentration was stable. Four SSR markers including RM206, RM223, RM10745 and RM8094 were used lo identify the salt tolerant genotypes. Analysis of PCR products, only RM206 marker was associated with salt tolerant gene. Three high yielding rice varieties MTL480, MTL687 and ST20 were used for studying of induced mutation in vitro in order to select genetic variability in sah tolerance the calli ofhfTL480 and MTL687 showed high potential for regeneration (46,02% and 45,63%. respectively) in MS medium containing 5%o NaCl. However when NaCl concentration was increased to 10%o. there was only 30.67% calli of MTL480 regenerated. Tlie plantlets were transplanted in soil in the greenhouse lo test their response to sah tolerance, the result revealed that 100% plants were survive on 6%a NaCl concentration.
TOM
T A TMudi tdm gidng lua thu thdp dugc tir cdc tinh ddng bdng song Ciru Long dugc ddnh gid khd ndng chdng chiu man bdng cdch sir dung dung dich Yoshida bd sung 0%o, 4%o, 6%o NaCL Kit qud cho thdy ti li cdy sdng.
chiiu cao thdn. chiiu ddi ri. trong lugng klid thdn lua diu gidm mgnh khi ndng do mgn tdng lin trong khi hdm lirang chlorophyll thay ddi khdng ddng ki Bdn marker RM206. RS4223, R.\48094 vd RMI0745 dd dugc su dung de ddnh gid su liin kit vdi gen chiu mgn ciia cdc gidng thi nghiim Phdn tich kit qud PCR cho thdy r6ng chi co RM206 cho thdy su liin kit vdi kiiu gen chiu man. Ba gidng cao sdn MTL480. MTL687 vd ST20 dugc cbgn de nghiin ciru md dot biin trong mdi tnrang mdn. Kit qud nghiin cuu nuoi cdy md cho thdy hai gidng MTL480 va MTL687 cd kha ndng tdi sinh choi cao (46,02% vd 45.63%) khi bS sung 5%o NaCl vao mdi trudng nuoi cdy cd Khi ndng do NaCl tdng Hn 10%o thi chi cd 30.6 % md seo ciia giong MTL480 cd khd ndng tdi smh. Cav con duac chuyen sang nhd ludi di ddnh gid khd ndng chiu man, kit qud ghi nhdn lOrncdy con tdi sinh diu sdng sot sau 30 ngdy trong diiu kiin mgn 6%o.
Tgp chi Khoa hgc Tni&ng Dgi hgc Can Tha Pbdn B: Nong nghiep. Thm- san vd Cdng nghe Sinh hgc: 26 (2013): 104-111 1 DAT VAN DE
Trong nhieu nam qua, cac nhd khoa hgc da cd gang cai tien nhilu gidng lua cd tinh chdng chju man va da thanh cdng budc dau (Akbar et ai, 1986; Lang, 2000). Qua dd phuang phap lai tao va chgn lpc truyin thdng mit rit nhilu thai gian nhung hieu qua chua cao. Chgn gidng nhd marker phan lu lien kit vdi linh trang muc tieu (marker assisted selection MAS) la phuang phap dugc thi gidi ung hd manh me ttr nam 1995 cd t h i cho kit qua sau ba the he chgn lgc (Tanksley et ai, 1996).
Nhieu nghien ctru cho thay gen chii lire dilu khien tinh frang chdng chiu man cua cay liia nim fr§n nhiem sac the sd 1 (saltol). mgl so QTL dinh vi uen nhiem sic ihl sd 3, 4. 8. 10, 12 cung dugc ghi nhan Id co quan he vdi khd nang chdng chiu man (Teng, 1994).
Tren ca sd dd, de tai ''Tuyin chgn va tdi sinh mpt so giong Itia co khd ndng chiu mgn thich Ang v&i bien ddi khi hau of dong bdng song Ciru Long " dugc thuc hien nham thu thdp, tuyen chgn dugc gidng Ida chju man tu Bang 1: Cac giong Ilia thanfa loc trong nfaa ludi
cac gidng dang dugc frdng d cac viing cd dat ngap man ciia ddng bang sdng Cuu Long (DBSCL) qua dd lam ca sd cho chgn gidng.
2 PHlTONG PHAP NGHIEN C f U Cac gidng liia dugc thu thap tir cdc vung cd dat nhilm man d DBSCL nhu Bdng 1, 2.1 Danh gia kha nang chju man dira tren
cac dap ung sinh ly
Khdo sdt khd ndng chiu man trong nhd ludi:
Thanh Igc theo quy uinh thanh lgc man ciia IRRI (1997) cd cdi lien. Cay ma dugc danh gid c ^ diem theo tieu chuan SES (Standard E\'aluating Score) de phan biet tu mdn cam den khang, Khi cho man vao dua vao trieu chung ciia cay Ida ma danh gia tinh chdng chiu theo lieu chuin cua IRRI (1997).
Tieu chuan ddnh gia muc do chdng chiu man d giai doan tang trudng va phat trien theo Bang 2.
STT Ten giong Ncri lay mau STT Ten giong Noi lay mau I DH2
DH3 OM4900 CL8 DH4 DH5 8 OM1348 9 OM1350
Trutmg Long Hoa, Duyen Hai, Tra Vinh
Dan Thanh. Duyen Hai, Tra Vinh Hiep My, Cau Ngang, Tra Vinh Ngu Lac. Duyen Hai, Tra Vinh Long VTnh. Duyen Hai, Tra Vinh Ngu Lac. Duyen Hai. Tra Vinh Long Khanh. Duyen Hai. Tra Vinh
Phu Long, huyen Binh Dai. Ben Tre
xa Lpc Thuan. huyen Binh Dai,
Ben Tre ^ ^ ^
TC2 Giong lon, Dai An. Tra Cii, Tra Vinh
CN 1 Hiep Hoa, Cau Ngang. Tra Vinh Mpt bui do Lpc Ninh, Hong Dan, Bac Lieu OM576 Lpc Ninh. Hong Dan. Bac Lieu Thanh Phii. huj'en Cai Nudc, Ca Mau
Long Phu, Soc Trang OM6677
OM6976
MTL504 Long Phu, Sdc Trang 17 Pokkali* Vien Iiia DBSCL
Vien Ilia DBSCL Ghi chd • Pokkali lam chudn khdng. IR29 ldm chudn nhiim
Bang 2: Tieu chuan danfa gia mux dp kfaang man giai doan tang tnrdng
c i p Quan sat Muc chong chiu
I TSng tnrdng binh thudng, khong co trieu chiing tren la 3 Tang trudng gan nhu binh thudng nhung chop la hoac phan nua 5 TSng trudng cham lai, hau het la bj ction, chi cd vai la con co the mpc
dai ra
7 TSng trudng bi ngung hoan loan, hau hit nhimg la khd di. mot vai choi bi chet
9 Tit ca cay bj chet hojc kho
Chong chiu tot Chong chiu Chdng chiu trung binh Nhiem man
Tap cbi Khoa hgc Tiirang Dai lioc Cdn Tha Phdn B Ndng ngbHp Thuy san vd Cdng nghe Smh bgc: 26 (2013): 104-111
Ddnh gid tang tiirang cdc giong lua trong dieu kien man
Ghi nhan cdc chi lieu lang trudng cua cay gdm:
- Chilu dai re (cm): Tinh tu cd rl din dinh sinh trudng cua chdp re dai nhit.
Chieu dai than la (cm): Tinh tu cd rl den vimg la non cao nhat cdn xanh tdt.
Trgng Iugng khd rl (g): Say khd re ciia
TL trong m5n - TL trong doi chung
6 cay ma/gidng (d 80°C trong 2 ngay, sau do li Uong desiccators trudc khi do frgng lugng khd) can va tinh tnmg binh.
Trgng lugng khd thM (g): Say khd than ciia 6 cay ma/gidng, can va tinh trung binh vao cudi thi nghiem.
Tinh ti le bien ddng trgng lugng (TL) khd than, r l cua nghiem thuc man so vdi nghiem thuc doi chung:
Ti le faien ddng (%)=
TL kho than trong man xlOO
Ddnh gid hdm lircmg chlorophyl trong Id Nong dp chlorophyll a (Chia), chlorophyll b (Chlb) va chlorophyll tdng so dugc xac dinh theo cac phuong phap ciia Shabala vao giai doan giong chuin nhilm da chet.
2.2 Khao sat kha nang tai sinh ciia md lua trong dieu kien man
Ba gidng cao san MTL480, MTL687 va ST20 dugc chpn de nghien cuu md dot bien frong mdi trudng man.
Chgn nhiing hat cdn nguyen ven, bdc bo vd, cho vao binh tam giac de tiln hanh khu trung bang con 70°C (20 giay), Javel, 1-2 gipt Tween 20 tren may khuay trong 25 phiit, rua Iai vdi nudc cat vo trung 7-10 Ian. Hal gao khu triing dugc cay vao mdi trudng MS b6 sung 3 %
So mo seo sdng Ti le mo seo sdng sdt/dia (%) = — ; :
Sd md cay vao
maltose (CIM 3 % maltose), 20 hat/dta, u trong toi vdi nhiet dg 32,4°C. Cay chuyen cac mo seo CO nhilu ciu tnic bi, mau vang sang sang dTa moi trudng mdi de nhan sd lugng mo seo sau moi 3 tuan.
Cac mo seo tdt cua cac gi6ng thi nghiera dugc chuyen vao mdi trudng CIM 3 % maltose cd bd sung 0%o (doi chung), 5%o, 10%o va 20%o NaCl, dat trong tdi 32,4°C, liy so lieu vl khd nang sdng sdt, ldc do sinh trudng va cay chuyin vao mdi trudng tuong dng sau 3 tuin.
Nhiing md sdng sdt cd mau trang nga, kich thudc md seo tang. Nhiing md chit cd mau den hoac frang, kfch thudc khdng thay doi. Sau 1 lan cay chuyen cac md dugc chuyen sang mdi trudng tai sinh choi.
Cac mo seo song dugc trong moi frudng mdn dugc cay chuyen sang mdi trudng SIM de tdi sinh ra chdi, ti trong toi d nhiet dp 32,4'^C mdt luan, sau do dem ra sang vdi quang ki 18 gid sang - 6 gid toi.
Khi cay con 3 la. r l dai 3 - 5 cm, dung kep lay cay ra, riia agar va Uong vao chau dit. Sau khi cay da khoe lien hanh tudi dung dich Yoshida + NaCl 6%o de so sdnh kha nang sinh trudng khac nhau giiJa cay liia ldi sinh trong
dieu kien man vdi cay Iua tdi sinh trong dieu kien binh thudng. Ghi nhan kit qua sau 3 tuan chung man.
2.3 Danh gia s u lien ket cua mgt so marker phan tiV den gen chiu man
Trinh ty cac mdi dugc mieu ta d Bang 3.
Ngoai cac gi6ng Ida d Bdng 1, trong phan danh gia marker phan tur, DNA ciia gidng liia 1R28 dugc su dung nhu la giong chuin nhilm.
T^ chi Khoa hgc Tru&ng Dai hoc Cdn Tha PhdnB. Ndng nghiep. Thin-san vd Cdng nghe Sinh hgc- 26l20I3l: 104-111 Bang 3: Trinfa tijr cac mm dung tfai ngfaiem
Ten cfai tfai T a c g
For 5' AAGTTTGTACACATCGTATACA 3"
Rev 5" CGCGACCAGTACTACTACTA 3' For5TGACGAATTGACACACCGAGTACG 3 Rev 5' ACTTCACCGTCGGCAACATGG 3' For 5" CCC ATG CGT TTA ACT ATT CT 3"
Rev 5' CGT TCC ATC GAT CCG TAT GG 3' For 5' AGT GAG CTT GGG CTG AAA C 3"
Rev 5' GAA GGC AAG TCT TGG CAC TG 3'
Nejadera/., 2008
Nejadera/., 2008
Lang era/.. 2002
Rahman et al..
2010 2.4 Xir ly sd lieu
Sd lieu thi nghiem dugc XTJ K' bang phan mem Excel va phan tich thdng ke bang chuang Uinh SPSS Version 16,
3 K E T QUA VA THAO LUAN 3.1 Danh gia kha nang chiu man ciia cac
giong lua dura tren dap ung sinh ty i . /. I Ty le song sot
Kel qua ghi nhan kha nang sdng sdt cho thay cay lua d nghiem thuc ddi chung sdng 100% den khi ket thuc thi nghiem. 6 dieu kien nhiem man 4%t» giong IR29 chet sau 19 ngdy
\ u ly man. gidng OM6677, Pokkali sdng ban 70% va cd 8 gidng sdng uen 50% sau 21 ngay la: CL8. DH4^ DH5. OM1348, TC2. CNl, Mdt bui dd. OM6976. CJ dieu kien nhilm man 6%o gidng IR29 chet sau 9 ngay xu ly man, sau 21 ngay da sd cdc gidng deu chet chi cdn Pokkali.
DH4, Mdt bui dd, va OM6677 cdn sdng sdt nhtmg ti le sdng deu thap hon 50%, Nhu \ a \ . cd mdt sd giong cd the sdng frong dieu kien man 4%o nhung lai chet khi ndng do man tang len 6%o. Tu kit qud na> cho thay cac gidng cd khd nang sdng sdt lau vdi ti le cao nhu: CL8.
DH4. DH5. OM1348. TC2, CNI, Mgi bui dd
\ a OM6976 cd thi xem la cac gidng tiem nang cho cdng tac chgn Igc, lai tao gidng chiu man.
3.1.2 Mirc do chong chiu man
Su sinh trudng cua cac gidng lua thi nghiem cho thiy uong dieu kien man 0%o cay ma tana trudng binh thudng khdng cd frieu
chung nhiem man. Trong dieu kien man 49oo ca\ ma bit dau hi dnh hudng \ a muc do dnh hudng tang dan theo thdi gian. Sau 7 ngay xu
|\' cac gidng deu chua hi anh hudng chi 1R29 va OM4900 anh hudng den cap 3: sau 14 ngay phan Idn cac gidng hi anh hudng d cap 3 - 5 rieng IR29 va OM4900 nhiem cap den 7; sau 21 ngay gidng pokkali. CL8, OM6677 the hien khang man tdt (cap 3). 8 giong Ida the hien khang dimg binh la: DH2. DH3, DH4, DH5, OMr348. Mgt bui dd. OM6976. MTL504 (dnh hudng d cdp 5), gidng IR29. 1R50404 va OM4900 bieu hien rat nhilm va da chet hoan loan (cap 9). cdc gidng cdn lai cd bieu hien nhiem trung binh.
Bieu hien cua cac gidng Ida d mdi trudng man 6%o cung tuong tu nhu d mdi trudng man 4%o nhung muc do dnh hudng nang ban. Sau 7 ngay xu ly man gidng IR29 cd bieu hien nhiem d cap 7. gidng Pokkali, DH4, DH5, OM1348.
OM6677, MTL504 cd bieu hien d cap 3. cac gidng Ida khac bieu hien cap 5 - 7 , Sau 14 ngay xu ly man, gidng IR29 va OM4900 chet hoan loan (nhiem cap 9), gidng Pokkali bieu hien cap 5 cimg vdi 6 gidng Ida cd bieu hien tuong duang la: CL8, DH4, OM1348.
OM6677, OM6976, MTL504. Sau 21 ngay xu K man hau het cac gidng lua da chit, chi cd 4 gidng cdn lai vai cay nhiem cap 5 Id: Pokkali.
bH4"! Mdt bui dd' OM6677. Nhu \ d \ . d nghiem thuc man 4%o da quan sat dugc 2 giong chiu man kha, 8 gidng chju man trung binh va 8 gidng nhiem sau 21 ngav chung man, Trong khi d nghiem thuc man 6%o tai
Tgp chi Khoa hoc Tnr&ng Dgi hgc Cdn Tlia Pbdn B. Nong nghiip. Thiry san vd Cong nghe Smh hoc: 26 (2013): 104-111 thdi diim giong chuin nhilm IR29 chet co 6
gidng ch6ng chiu trung binh nhung den mpt tuin sau do (21 ngay sau chung man) chi cdn 1 giong chdng chiu trung binh Id Pokkali. Ket qua thi nghiem cho thiy gidng Pokkali, DH4, Mdt bui dd, OM6677 cd khd nang chju man tdt nhit, gidng DH2, DH3, CL8, DH5, OM1348, OM6976, MTL504 bilu hien khdng d nong do man 4%o nhung bieu hien nhiem frong ndng do man 6%o.
Kit qua nghien cuu cua Nguyin Trung Tiln (2009) cho thiy ti Ie song sdt ciia cay lua giam khi ndng do man tang len ddng thai nhtrng gidng cd bilu hien khang man thi deu cd thai gian song sdt lau uong mdi frudng man. Kit qua cua thi nghiem nay rat phu hgp vdi kit qua nghien cuu Uen cua Nguyen Trung Tien va Phan Vinh Quang,
3.1.3 Anh huong cua man den chieu cao thdn Id cda cdc giong Ida thanh lgc Chilu cao thdn la Ida d cac ndng dp mudi khac nhau thi rat khdc nhau. Cdy lua Uong dieu ki?n man 0%o cd chilu cao than la keo dai lien luc frong qud Uinh thi nghiem. Theo dd sau 7 ngay chung man cdc gidng deu cd the keo dai than la ngoai trir gidng 1R29 va OM4900 d nghiem thuc man 6%o. Sau 14 ngay chung man, d dp man 4%o giong 1R29 giam 37,69% chilu cao than Id, 2 giong DH3 va OM4900 cung cd chieu cao than la giam dang ke; d nong dd man 6%o gidng chuan nhilm IR29 chit hoan toan, OM4900 giam 59,37%, da sd cdc gidng cd chieu cao than Id gidm manh dac biet chi OM1348 va Pokkali cd chieu cao tSng. Nhu vay. cac gidng DH2, DH3, CL8, IR50504, DH5. OM6677 khdng gidm chieu cao d ndng do man 4%o nhung lai gidm manh d nfing do man 6%o. Giai doan 21 ngay sau khi xd ly man, d ndng dp man 4%o giong chuan nhilm IR29 chet hoan loan, da so cdc gidng deu cd chilu cao than Id gidm ngoai tru DH4, TC2, Mot bui dd. Pokkali thay ddi khdng ddng ke. OM6677 va MTL504 tang cd y nghTa thdng ke; d ndng do man 6%o chi Pokkali, DH4, Mpt bui do, OM6677 cdn lai vai cay nhung chilu cao cung gidm dang ke. cac gidng khac deu da chit.
Ket qua nghien cuu cua Islam et al, 2007 da ghi nhan chieu cao cay Ida ti \h nghich vdi ndng dp man; Akita va Cabuslay (1988) thay ring dudi dnh hudng cua man cac ia gia chet sdm hon Id non. Ket qud cua thi nghiem nay i^t tuang dong vdi ket qud cua cac nghien cuu ndi tren.
3.1.4 Anh hu&ng man den chieu dai re cua cdc giong Ilia thanh loc
Chilu dai re liia da giam khi ndng do mudi tang len nhung muc do giam khdng cao nhu sy gidm d than. Dilu nay phii hgp vdi ghi nhan cda Akbar et al. (1972) ring khi bi man anh hudng thi su sinh frudng d nggn Iua gidm nhilu ban su sinh trudng d re.
3.1.5 Anh hu&ng man den trong luang kho thdn, re cda cdc giong lua thanh lpc Trgng lugng khd than, re cua da sd cac giong Iua thi nghiem deu giam khi nong dp man tang ISn va khdi Iugng khd than gidm nhilu han khdi lugng khd re. Sau 21 ngay chiing man, trgng lugng khd than gidng OM4900, IR29, OM576, OM1350, DH2, DH3, IR50404 giam hem 50%; trgng lugng re 1R29 gidm cao nhit 47,65%, tilp din OM4900 giam 44,75%, cdc gidng cdn Iai deu cd ti le gidm thip han 23%, gidng Pokkali gidm 2,67%. Dac biel gidng CL8, DH4, DH5, TC2 cd khdi lugng vat chit khd tang nhe so vdi ddi chung. Nam 1986. Akita ghi n h ^ la: khi bi dnh hudng cua man, trgng lugng khd cd xu hudng tang len frong mdt thdi gian, sau do gidm nghiem trgng do suy giam dien tich la;
trong dieu kien thiet hai nang ban, trpng lugng khd cua chdi va cua re suy giam luang ung vdi muc do thiet hai. Nhu vay kit qua cua thi nghiem nay rat phu hgp vdi ghi nhan cua Akita (1986) va cdc gidng OM4900, IR29, OM576, OM1350, DH2. DH3, IR50404 la cac gidng cd khd nang chiu man kem nen da giam manh khdi lugng khd sau 21 ngay nhiem man, cdc gi6ng CO kha nSng chiu man khd hem nen ti le gidm thip, gidng c6 khdi lugng than r l khdng gidm nhu CL8. DH4, DH5, TC2 Id nhOng gidng cd tiem nang chiu man tdt.
Tgp chi Khoa hgc Tru&ng Dgi hgc Cdn Tha Phdn B: Ndng nghiip. Tbu\- s d Cdng nghe Smh hgc: 26 i2013r !f'4-lll 3.1.6 Bdnh gid dnh hu&ng ciia man len ham
Itrong chlorophyll trong Id
0 ndng do man 4%o gidng 1R29 cd chlorophyll tdng la 1,683 mg/g thip ban chlorophyll tdng ciia Pokkali (2,122 mg/g).
Tuy nhien, xet cimg mdt giong thi ham lugng Chl hau nhu khdng thay ddi khi tang nong do xu ly man Mt 0%o len 4%o. Giong 1R29, DH2, DH3, TC2 cd ham lugng chlorophyll giam khi
xu ly man nhung su thay ddi ndy khdng cd y nghta. Ndng do man 496o hau nhu khdng anh hudng den ham lugng chlorophyl fren Id cua cac gidng liia thi nghiem, kit qua tucmg tu cung dugc Tang Thi Hanh et al. 2010 quan sat.
3.2 Danh gia kieu gen cac gidng thu t h a p dugc bang marker phan tir Marker RM 206
200bp
Hinfa 1: S a n pfaim P C R ciia chi thi R M 2 0 6 / : Ladder 2: DH2. 3 DH3, 4 OM4900. 5: DH5. 6: CL8, 7: OM6677. 8 CNl. 9: Mdt bui do.
10: Pokkali, IT. IR28, 12: IR29. 13: Ladder, 14: OM6976. IS. MTL504. 16: Nu&c Ket qua phan tich gel d hinh 1 cho thay
CL8, OM1348, Mpt bui dd, OM6677, OM6976, MTL504 cd bang tuang ung vdi Pokkali mang kieu gen chdng chiu man (170bp); cd sdu gidng co bang tuang ung vdi IR28, 1R29 mang gen nhiem man (I90bp) la:
DH2, DH3, OM4900. IR50464, OM1350, OM576. Qua dd co thi thdy RM206 lien kit khd chat vdi khd nang chong chiu man tren cay Ilia, kel qud cua thi nghiem nay tucmg ung vdi kit qua thi nghiem cua Nguyin Thj Lang et al, 2008. Thdng qua RM206 da nhan dien dugc giong cd mang gen chong chiu man la CL'S, OM1348, Mpt bui dd, OM6677, OM6976. MTL504.
Marker RM223 vdRMl0745 Trong nghien cuu nay cac doan DNA do RM233 va RM 10745 khulch dai dugc da khdng thi hien su khac biet giua gidng chuan nhiem IR28 vd giong khdng chuin Pokkali.
Nhu vay, trong nghien cuu nay marker RM223 va RMI0745 da khdng lien kit chat vdi kha
nang chiu man cua cac gidng lua, Nejad (2008) ciing cho rang RM 10745 lien ket chat vdi kha nang khdng man cua liia.
Ket qua kiem tra san pham PCR vdi marker RM8094 cho thay co bang khac nhau da dugc nhan dien lu cac gidng Ida Ihi nghiem, dac biet hai gidng IR28 va IR29 deu la gidng chudn nhiem nhung san pham PCR cung cho ra cdc bang khac nhau. Dieu nay cho thdy RM8049 khdng phu hop de phan tich gen khang man nhtmg cd the su dung frong phan lich da dang di iruyen. Tinh da hinh di iruyen cao giua cdc gidng Itia cung da dugc Nejad (2008) ghi nhan vdi RM8094. Ket qua cua thi nghiem nay rat phu hgp vdi ket qua ciia Nejad.
RM206 lien ket chat vdi gen khang man.
Nhu va\. trong 4 cap primer dugc su dung trong thi nghiem thi RM206 cd kha nang lien kit cao nhat vdi kha nSng khang man;
RM8094 the hien tinh da hinh di truyen rat cao giira cac gidng Iua, vi vay RM206 tdt cho phdn tich kha nang khang man trong khi RM8094
Tap clli Khoa Itgc Tnr&ng Dai hoc Cdn Tlia Phdn B: Ndng nghiep, Thuy s d Cdng nghi Sinh hgc. 26 (2013)- 104-111 tdt cho phan lich tinh da dang di truyen cua cac
gidng lua.
Qua kilm fra cd 6 frong cac giong thu thap dugc cd cd kieu gen vd kieu hinh khdng man la: CL8, OM1348, Mgt Bui Dd, OM6677, OM6976, MTL504; mdt sd gidng cd kilu hinh khang nhtmg xuit hien bang la (150bp) khi phan t k h san phim PCR la DH4, DH5, TC2, CNl: Cac gidng nay cd the bd sung vao ngan hang cdc giong liia cd kha nang chju man nhim lam ngudn cho cac nghien cuu tiep theo 3.3 Khao sat kha uang tai sinh cua md liia
trong dieu kien man
3 3.1 Anh hu&ng cua NaCl den ti le song sot cua mo seo
Cd 3 gidng lua MTL480, MTL687 va ST20 deu cd khd nang lao md seo Uen mdi trudng C1M3% Maltose. Ket qua ddnh gid li le song sol cua mo seo Uong cdc ndng do man cua cac gidng liia cho thdy ndng dp man cang cao thi ty le sdng sdt ciia md seo cdng gidm, md seo cua cac gidng khac nhau co kha nang chju man rat khdc nhau. Trong dieu kien khdng chiing mdn, md seo cdc gidng deu cd li le song rdt cao (khoang 98%). Trong cdc mdi trudng chung mSn 5%o : 10%o : 20%o, ty le song sdt ciia md see MTL480 la cao nhit (94,64%, 43,33% va 8,88%); tilp theo Id md seo MTL687 (85,18%, 30,16% vd 0%); ty le sdng sdt ciia md seo ST20 la friip nhit (70.28%, 9.09% vd 0%).
Bao cao ciia Lang va Tam (2003) cho thiy mo s?o cOa mgl sd gidng cd khd nang sdng sot cao trong mdi frudng man 15%o nhu Sdc Nau, Trang Thai Lan. Doc Do. frong thi nghiem nay giong MTL480 co kha nang sdng dugc d nong dg man 20%o.
3.3.2 Khd ndng tdi sinb choi tir mo seo sau khi xir ly man
Ket qua thi nghiem cho thay: nong do man 20%o lam mo seo cua cd ba gidng thi nghiem deu mat khd nang tai sinh choi; sau khi xu ly man md seo ST20 hodn loan khdng co kha ndng tai sinh choi: ti le tai sinh chdi cua MTL480 va MTL687 gidm khi nong do man tdng. Giong MTL480 cd khd nang tai sinh choi cao nhit trong cac giong thi nghiem: d nghiem
thuc man 5%o md see MTL480 cd ti le tdi sinh chdi tuang duang d nghiem thdc ddi chirng; d nong do man 10%o md seo MTL480 cd ti le tao chdi kha cao (30,67%). Ddi vdi giong MTL687 cung cd ti le tai sinh chdi cao d nghiem thuc doi chung (60,42%) va man 5%o (45,63%), nhtmg dat ti le rit thap (1,7%) d nghiem thtrc 10%o. Nhu vdy, khd nang tai sinh chdi tu md seo ciia liia khdng nhirng chiu dnh hudng cua thanh phdn mdi trudng nudi cay (Lee et al.. 2003) ma cdn chiu anh hudng ciia kilu gen va nong do xir ly man.
3.3.3 Khd ndng tgo re cda cdc choi dugc tdi sinh tie moi tru&ng man
Cd 100% cdc chdi tai sinh tir cac nghiem thuc khi chuyen san^ mdi trudng tao re MS deu cd bg rl vung chac sau 2 tuan, cd 86% so cay con sdng tot Uong vudn sau 2 tuan. Cac cay con chet d giai doan uam cd the Id do bi sdc nhiet hoac do bg re chua hoat ddng tot.
3.3.4 Kiem tra khd ndng song sot cua cdc cdy lua tai sinh trong dieu kiin man Cac cay liia dugc tai sinh trong dieu kien man deu sdng sdt khi frdng frong chau dat tudi hoan loan bang dung dich Yoshida bo sung 6%o NaCl sau 30 ngay, Ket qua tren cho thdy cdc cay lua dugc lai sinh ttr mdi trudng man co kha nang sdng sdt cao frong mdi trudng hi nhiem man. Ket qud na\' luang tu vdi kit qua nghien ciiu ciia Le Trin Binh et al. (2007).
4 K E T LUAN
Cd 11 Uong 18 gidng thu dugc tu cac tinh ngdp man cua DBSCL cd bilu hien khdng man d giai doan ma, frong do cd 3 giong liia cd kha nang chiu man tot la: DH4, OM1348, Pokkali; 8 gidng chiu man trung binh la : CL8, DH5, TC2, C N l , Mdt bui dd, OM6677, OM6976 vd MTL504.
RM206 lien ket chat vdi gen chiu mdn, trong khi RM8094 t h i hien tinh da hinh di truyin cao giira cac giong Ida thi nghiem.
Chieu cao thdn. trgng lugng khd than Id giam manhkhi cdy liia bj nhiem mdn Uong khi chieu dai re, ham lugng chlorophyll trong la la chi tieu thay ddi it.
Tgp chi Kboa hgc Tnr&ng Dgi hgc Cdn Tba Phdn B Ndng nghiep. Tlmy san vd Cdng nghi Smb hgc. 26 (2013). 104-111 - Md seo cua ba giong MTL480,
MTL687, ST20 deu cd the sinh trudng uong mdi trudng CIM3% maltose bd sung 5%o va I0%o NaCL Tuy nhien, chi cd gidng MTL480 va MTL687 cd kha nang lai sinh choi sau khi xu ly man.
- Cay lua tai sinh d dilu kien man cua MTL480, MTL687 diu cd kha nang sdng sdt d nha ludi trong dieu kien man 6%o sau 30 ngay.
TAI LIEU THAM KHAO
1. Le Trin Binh, Le Thi Muoi (1998), Phan lap gen va chpn ddng chdng chiu ngoai canh bat lgi d cay liia, Nxb Dai hpc Quoc gia. Ha Ngi.
2. Nguyen Trung Tiln. 2009. Nghien cuu tinh chong chju man tren nhom giong liia miia dja phuong. Omonrice, Vol. 14, trang 39-43 3. TSng Thi Hanh, Duong Thi H6ng Mai, Trin
Van Luyen, Pham Van Cudng. Le Kha Tudng.
Phan Thi Nga. 2010. Nghien ciiu kha nang chiu man ciia mpt s6 nguin gen liia ltm giu tai ngan hang gen cay Uong quoc gia. Tgp chi Ndng nghiip vd Phdt triin Ndng thdn.
4. Akbar M.Yabuno Y.Nakao 5(1972).
Breeding for saline resistant varieties of rice. I.
Vari^ility for salt-tolerance among some rice varieties. Jpn. I Breed 22: 277-284.
5. Akbar M and F.N. Ponnamperuma. 1982.
Saline soils of Southeast Asia as potential land. IRRI, Los Banos, Philippines.
6. Akbar M, I.E. Gunawardena, F.N.
Ponnamperuma. 1986. Breeding for soil stress. Progress in rainfed lowland rice.
Intemational Rice Research Institute, Los Banos. Philippines. Pages 263-272.
7. Akagi HY, Yokozeki A. Inagaki T. Fujimura (1997). Highly polymorphic microsatellites of rice consist of AT repeats, and a classification of closely related cultivars with these microsatellite loci. Theor. Appl. Genet. 94:61-67.
8. Akita, S and Cabuslay, G S. 1988.
Physiological basis of differential response to salinity in rice cultivars. In: Proc. 3"". Intl.
Symp, Genet, Aspects Plant Mineral NuU. 19-24 June. 1988. Braunschweig, Germany, pp 37.
9. Gregorio GB, Senadhira D, Mendoza RD (1997). Screening rice for salinity tolerance, IRRI Discussion paper Series No.22.
International Rice Research Institute, Los Banos. Laguna, Philippines.
10. Islam M.Z., M. A. Basel Mia, M.R. Islam, and A. Akter (2007), Effect of different saline level on growth and yield attributes of mutant rice.
J.Soil .Nature . 1(2), pp 18-22.
11. Lang NT, Yanagihara S, Buu BC (2000).
Quantitative tr^l loci for salt tolerance in rice via molecular markers. Omonrice 8:37-48.
12. Nguyen Thi Lang, B.C. Buu and A. Ismail, 2008. Molecular mapping and marker- assisted selection for salt tolerence in rice {Oryza sativa L.). Omonrice, Vol. 16, pp50-56.
13. Nguyen Thi Lang, Nguyen Van Tao. Bui Thi Duong Khuyeu, Trinh Hoang Khai. Dang Minh Tam, Bui Xuan Ky. Hiroyuki Hiraoka.
Hiromi Kobayashi and Bui Chi Buu, 2003.
Genetic Variabilis of Salt Tolerance in Rice (Oryza sativa 1 L.).
www.ctu.edu.vn/institutes/mdi/jircas/JlRCAS.
14. Lee. I.S., D. S. Kim, D.Y. Hyun, S. J. Lee, H.
S. Song. Y. P. Lim. and Y. 1. Lee, 2003.
Isolation of gamma-inuced rice mutants with increased tolerance to salt by anther culture. J.
PlanlBiotechnology. 5(1): 51-57,2003.
15. Nejad, G.M., A. Arzani, A.M. Rezai, R.K.
Singh and G.B. Gregorio, 2008. Assessment of rice genotypes for salt tolerance using microsatellite markers associated with the saltol QTL. Afr. J, Biotech., 7: 730-736.
16. Tanksley, S.D., S. Grandillo, T.M. Fulton. D.
Zamir, Y. Eshed, V. Petiard, J. Lopez and T, Beck-Bunn. 1996. Advanced backcross QTL analysis in a cross between an elite processing line of tomato and its wild relative L.
pimpinellifolium. Theor Appl Genet 92: 213-224.
17. TengS. ]994. Gene tagging for salt tolerance in rice (Oryza sativar L.). PhD Thesis. The university ofthe Philippines, Los Banos, Laguna, Philippines, 118 p.
