Studies on Cold Tolerance of Rice, Oryza sativa L.

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1 ACTA AGRONOMICA SINICA Vol. 29, No. 5 pp Sept., , Ξ ( 1, ; 2, ; 3, , ) F 2, 159 RFLP SSR STATISTIC, ( r) ; ( ) 43, , 7 9, 7 8 ; 9 11, ; ; ; ; : S511 : A Studies on Cold Tolerance of Rice, Oryza sativa L.. Molecular Basis for Special Fertility Percentage as Evaluation Increterior of Cold Toleance DAI Lu2Yuan 1,2 LIN Xing2Hua 2 YE Chang2Rong 1 KATO Akira 3 SAITO Koji 3 YU Teng2Qiong 1 XU Fu2Rong 1 ZHANG Duan2Pin 2 ( 1 Crop Genetic Resources Institute, Yunnan Academy of Agricultural Sciences, Kunming, Yunnan ; 2 Huazhong Agricultural University, Wuhan, Hubei, , China ; 3 National Agriculture Research Center for the Hokkaido Region, Sapporo, Hokkaido, , Japan) Abstract Here is the third part of a series papers to describe the studies on cold tolerance of rice. This part describes the results obtained by using the parents, Kunmingxiaobaigu, on indigenous variety with cold2tolerance from Yunnan, and Tow2 ada, a cold2sensitive variety bred in Japan, as well as their F 2 progenies as tested materials, combining by using 159 RFLP and SSR markers and STATISTIC software. The total fertility percentage ( TFP) was used as indicator of cold tolerance. The coefficient of relationship between TFP and special fertility percentage( SFP) of single plant was markers were related to cold tolerance based on the TFP as its indicator, distributed in eleven regions on eight chromosomes while 34 ones related to SFP over nine regions on seven chromosomes. The seven chromosomes related to SFP belonged to the eight ones related to TFP. The nine regions related to SFP belonged to the eleven regions related to TFP. The above results showed that SFP can be used to represent TFP in evaluation of cold tolerance. The issue over SFP to represent TFP in other fields was discussed. Key words Rice ( Oryza sativa L. ) ; Criterion of cold tolerance ; Special fertility percentage(sfp) ; Molecular marker ; Yunnan : : ( ),,,,, Tel : ,Fax : Received( ) : , Accepted( ) :

2 5 :. 709, 1, [1,2 ],, [2 4 ] [2, ] [2,5 ] ( 1916 m), F 1 F 2,,5 7,, 10 cm 17 cm,, 4 ( ), 4 [6 8 ],, Satake et al. [9,10 ] , F 3, F 3 30, , 9 [2,7 ] DNA DNA Liu et al. [15 ], ;, I (tetrad) ( the first contraction stage) BamH Dra EcoR (young microspore stage), EcoRV Hind, 018 % (Sigma ; ), 012 molπl, HCl 10 min, DNA 014 molπl NaOH ( Hybond (leptotene) N), Southern DNA, 10 ml 65 [11 13 ], (5 SSC,50 mmolπl Phosphate buffer, 215 mmolπl EDTA, 5 Denhardts, 5 % Dextran Sulfate, [2 4,14 ] 011 % SDS,100 gπml ssdna), 65, 6 12 h, : DNA 100 ng,, 2 32 P2dCTP, 1 SSC 011 %, SDS ( 65 ) 65 2, 15 min, X, d PCR 10 L, 012,molΠL DNA 10 ng dntps 100 molπl 50 mmolπl KCl 10 mmolπl Tris2Cl (ph 813) 115 mmolπl MgCl Gold DNA polymerase PCR

3 min, 45, s s s, min 6 L PCR % Nusieve 3 1 Agarose ( FMC, Rockland, Marine, 2 F 2 SA), 16 V 1, 50 min, 2. 2 F 2 F 2 X, 2, ( r) ( n = RFLP SSR 249),,, 1, 2, 3, 2. 3 F 2, 1, 4 ;, F 2, 43 2, 5,, -, MAPMAKERΠEXP. Version 3. 0 F 2 F 2 RFLP STATISTIC, 14 20, 1 ( )

4 5 :. 711 Fig ( ) F 2 Linkage map of the F 2 population between Kunmingxiaobaigu and Towada Fig. 2 2 F 2 Correlation between total fertility percentage and special fertility percentage in the F 2 population G1128), 3 G1128 ( F = ),, F, RM232 F = ( P = ) XNPB358 F = ( P = ) 2. 4 F 2 F 2 8, 7 8, 7 1 F 2, 12 34, 31 (, 8, 8 ), % ; 3 ( RM232 XNPB358

5 Table 1 1 F 2 F Markers affecting significantly total fertile spikelet percentage and special fertile spikelet percentage in F 2 population and their F values Marker F 2 Total fertile spikelet percentage in F 2 Chro No. F F value F 2 Special fertile spikelet percentage in F 2 Marker Chro R R C RM RM R RZ RZ C C RG RG C C No. F F value RM C C R R RG RG RG RG G G R R C C RM RM RM RM RZ RZ RM RM R R R R RM C C RG RG RG RG RG Y2668A Y2668A R R RM RM RM RM G G RG G XNPB RM RM G G G G G C G G R XNPB G RM RM

6 5 :. 713 F 2 43, 1, 9 F 2 43, 3, (RZ288 Y2668A G179), 40, 8 ( ), 6 3, 13, ; 1 7, [1,6 ], 5 ; 8, 1,, [4,10,11,15 ] 11 F 2, 6, , , DNA, F 2 (RZ288 Y2668A), 32, 6 10, 4 6, 7 5, 3 10, 4, 1 2, 12, 1,, 9 F 2, 1 6 2,, , F 2 9 F 2 (, ) 11, 8 F 2 1, ; 6, 3, 1 9, 8 F 2, G126, G126 RM223 RM210 F 2,,, 8 F 2, 8, F 2 8 G126,, F 2 F 2, 7 ; F 2 10, F 2 References 9 1, 6 [1 ] Dai L2Y( ),Ye C2R( ),Yu T2Q( ),Xu F2R(

7 ). Studies on cold tolerance of rice, Oryza sativa L. Descrip2 tion on types of cold injury and classifications of evaluation methods on cold tolerance of rice. Southwest China Journal of Agricultural Sciences ( ),2002,15(1) :41 44 [2 ] Dai L2Y( ), Kariya K( ), Ye C2R( ),,,Yu T2Q( ),Xu F2R( ),Ma C2W ( ). Studies on cold tolerance of rice, Oryza sativa L. Evaluation on cold tolerance of Yunnan rice genetic resoureces. Southwest China Journal of Agricultural Sciences ( ),2002,15 (3) :47 52 [3 ] Saito K, Miura K, Nagano K, Hayano2Saito Y, Araki H, Kato A. Identification of two closely linked quantitative trait loci for cold toler2 ance on chromosome 4 of rice and their association with anther length. Theor Appl Genet, 2001, 103 : [4 ] Tanno H, Kiuchi H,Hirayama Y,Kikuchi H. Development of a simple testing method for cool weather tolerance at the flowering stage of rice using an air2conditioned room. Japan Jour Crop Sci, 2000, 69 :43 48 [5 ] Dai L2Y( ), Ye C2R( ), Xiong J2H( ), Wang H2Y( ). Methods for evaluating cold tolerance in rice. Chinese Journal of Rice Science ( ),1999,13(1) : 62 [ 6 ] Nishiyama I, Satake T. Male sterility caused by cooling treatment at the young microspore stage in rice plants. XIX. The difference in suscepti2 bility to coolness among spikelets on a panicle. Japan Jour Crop Sci, 1979,48(2) : [ 7 ] Nishiyama I. Male sterility caused by cooling treatment at the young mi2 crospore stage in rice plants. XXIII. Anther length,pollen number and the difference in susceptibility to coolness among spikelets on the pani2 cle. Japan Jour Crop Sci. 1982, 51(4) : [8 ] Shimizu M, Ujiie S. Sterile type cool injury in Miyagi prefecture in Tohoku no Nogyo Kisho, 1957, 2 : [9 ] Satake T, Hayase H. Male sterility caused by cooling treatment at the young microspore stage in rice plants. V. Estimations of pollen devel2 opmental stage and the most sensitive stage to coolness. Soc Japan, 1970, 39(4) : Proc Crop Sci [10 ] Satake T, Hayase H. Male sterility caused by cooling treatment at the young microspore stage in rice plants. X. A seconary sensitive stage at the beginning of meiosis. Proc Crop Sci Soc Japan, 1974, 43 (1) : [11 ] Nishiyama I. Male sterility caused by cooling treatment at the young microspore stage in rice plants. XI. Effects of some substances on ste2 rility. Proc Crop Sci Soc Japan, 1975, 44(4) : [12 ] Bertin P, Bouhaumont J. Use of somaclonal variation and in vitro se2 lection for chilling tolerance improvement in rice. Euphytica, 1997, 96 : [13 ] Bertin P, Bouhaumont J, Kinet J M. Somaclonal variation and im2 provement in chilling tolerance in rice : changes in chilling2induced electrolyte leakage. Plant Breeding, 1996, 115 : [ 14 ] Satake T. Determination of the most sensitive stage to sterile2cool inju2 ry in rice plants. Research Bulletin No. 113 of the Hokkaido National Agricultural Experiment Station, [15 ] Liu K D, Wang J, Li H B, et al. A genome2wide analysis of wide compatibility on rice and the precise location of the S 5 locus in the mo2 lecular map. Theor Appl Genet, 1997, 95 : , ( ) ( ),, ( ), 10, 16,120, : CN ΠS, : ISSN , : : ( 399 ), Q4380 :12, 48, : 33 :712100, : :, (7), : E2mail net. cn

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