普通小麦-大赖草易位系T6DL·7LrS的分子细胞遗传学鉴定
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摘要
大赖草高抗小麦赤霉病,将大赖草抗性基因导入普通小麦,对创新小麦赤霉病抗性种质有重要意义。为了获得普通小麦-大赖草抗赤霉病易位系,采用12 00 R~(60)Co-γ射线处理小麦-大赖草二体附加系DA7Lr花粉,授予已去雄的普通小麦中国春,对其后代(M1)种子根尖细胞有丝分裂中期染色体进行GISH分析,获得了1株具有1条普通小麦-大赖草易位染色体的植株,让其自交,对自交后代中具有2条易位染色体植株的花粉母细胞减数分裂中期I进行观察,发现2条易位染色体形成了稳定的棒状二价体,表明该植株为纯合体。利用顺序GISH-双色FISH分析,结合小麦D组专化探针Oligo-pAs1-2和B组专化探针Oligo-pSc119.2-2,进一步鉴定出该普通小麦-大赖草易位系为T6DL·7LrS,该易位系的育成也为小麦赤霉病遗传改良提供了新种质。
Leymus racemosus had a high resistant capacity to wheat scab(Fusarum head blight). The transfer of scab resistant gene from L. racemosus to Triticum aestivum is of great significance for broadening the germplasm of wheat resistance. To obtain Triticum aestivum-Leymus racemosus translocation line with scab resistance, we irradiated the pollen of T. aestivum-L. racemosus disomic addition line DA7 Lr by ~(60)Co-γ-rays 1 200 R(100 R/min) prior to pollinating to emasculation T. aestivum cv. Chinese Spring. One plant with one translocation chromosome was detected in the M1 by GISH. The plant with one translocation chromosome was self-pollinated, and at meiotic metaphase I its progenies with two translocation chromosomes were analyzed for chromosome pairing behavior in their pollen mother cells(PMCs). One rod bivalent was observed at meiotic metaphase I, indicating that the plant with two translocation chromosomes was one translocation homozygote. Sequential GISH-FISH analysis, using Oligo-pAs1-2 and Oligo-pSc119.2-2 as probe, translocation line was confirmed as T6 DL·7 LrS. The translocation line had higher resistance to wheat scab and feasibility to be used as a new source in wheat breeding resistant to scab disease.
Leymus racemosus had a high resistant capacity to wheat scab(Fusarum head blight). The transfer of scab resistant gene from L. racemosus to Triticum aestivum is of great significance for broadening the germplasm of wheat resistance. To obtain Triticum aestivum-Leymus racemosus translocation line with scab resistance, we irradiated the pollen of T. aestivum-L. racemosus disomic addition line DA7 Lr by ~(60)Co-γ-rays 1 200 R(100 R/min) prior to pollinating to emasculation T. aestivum cv. Chinese Spring. One plant with one translocation chromosome was detected in the M1 by GISH. The plant with one translocation chromosome was self-pollinated, and at meiotic metaphase I its progenies with two translocation chromosomes were analyzed for chromosome pairing behavior in their pollen mother cells(PMCs). One rod bivalent was observed at meiotic metaphase I, indicating that the plant with two translocation chromosomes was one translocation homozygote. Sequential GISH-FISH analysis, using Oligo-pAs1-2 and Oligo-pSc119.2-2 as probe, translocation line was confirmed as T6 DL·7 LrS. The translocation line had higher resistance to wheat scab and feasibility to be used as a new source in wheat breeding resistant to scab disease.
引文
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[2]Yin GM,Zhao JH,Zhang YJ,et al.Adaption strategy about Leymus racemosus under different grazing patterns.Anim Husband Feed Sci,2013,34(11):33-36,39(in Chinese).殷国梅,赵金花,张英俊,等.大赖草对不同放牧方式的适应策略.畜牧与饲料科学,2013,34(11):33-36,39.
[3]Dewey DR.The genomic system of classification as a guide to intergeneric hybridization with the perennial Triticeae//Gustafson JP,ed.Gene Manipulation in Plant Improvement.New York:Plenum Press,1984,209-279.
[4]McGuire PE,Dv?rák J.High salt-tolerance potential in wheatgrasses.Crop Sci,1981,21(5):702-705.
[5]Mujeeb-Kazi A,Bekele GT,Mirand JL.Incorporation of alien genetic information from Elymus giganteus into Triticum aestivum//Sakamoto S,ed.Proceedings of the 6th International Wheat Genet Symposium.Maruzen,Kyoto,Japan,1983:223-231.
[6]Mujeeb-Kazi A,Rodriguez R.An intergeneric hybrid of Triticum aestivum L.X Elymus giganteus.J Hered,1981,72:253-256.
[7]Chen PD,Wang ZT,Wang SL,et al.Transfer of useful germplasm from Leymus racemosus Lam.to common wheat.III.Development of addition lines with wheat scab resistance.Acta Genet Sin,1995,22(3):206-210(in Chinese).陈佩度,王兆悌,王苏玲,等.将大赖草种质转移给普通小麦的研究:III.抗赤霉病异附加系选育.遗传学报,1995,22(3):206-210.
[8]Qi LL,Wang SL,Chen PD,et al.Molecular cytogenetic analysis of Leymus racemosus chromosomes added to wheat.Theoret Appl Genet,1997,95(7):1084-1091.
[9]Liu WX,Chen PD,Liu DJ.Development of Triticum aestivum-Leymus racemosus translocation lines by irradiating adult plants at meiosis.Acta Bot Sin,1999,41(5):463-467(in Chinese).刘文轩,陈佩度,刘大钧.利用减数分裂期成株电离辐射选育小麦-大赖草易位系的研究.植物学报,1999,41(5):463-467.
[10]Wang LS,Chen PD,Wang XE.Molecular cytogenetic analysis of Triticum aestivum-Leymus racemosus reciprocal chromosomal translocation T7DS·5LrL/T5LrS·7DL.Chin Sci Bull,2010,55(11):1026-1031.
[11]Cui CQ,Wang LS,Chen PD.Molecular and cytogenetic identification of Triticum aestivum-Leymus racemo-sus translocation lines T7BS·7Lr#1S and T2AS·2AL-7Lr#1S.Acta Agron Sin,2013,39(2):191-197(in Chinese).崔承齐,王林生,陈佩度.普通小麦-大赖草易位系T7BS·7Lr#1S和T2AS·2AL-7Lr#1S的分子细胞遗传学鉴定.作物学报,2013,39(2):191-197.
[12]Gill BS,Friebe B,Endo TR.Standard karyotype and nomenclature system for description of chromosome bands and structural aberrations in wheat(Triticum aestivum).Genome,1991,34(5):830-839.
[13]Mukai Y,Nakahara Y,Yamamoto M.Simultaneous discrimination of the three genomes in hexaploid wheat by multicolor fluorescence in situ hybridization using total genomic and highly repeated DNA probes.Genome,1993,36(3):489-494
[14]Zhang P,Li W L,Friebe B,et al.Simultaneous painting of three genomes in hexaploid wheat by BACK-FISH.Genome,2004,47(5):979-987.
[15]Tang ZX,Yang ZJ,Fu SL.Oligonucleotides replacing the roles of repetitive sequences pAs1,pSc119.2,pTa-535,pTa71,CCS1,and pAWRC.1 for FISH analysis.J Appl Genet,2014,55(3):313-318.
[16]Sharp PJ,Chao S,Desai S,et al.The isolation characterization and application in the Triticeae of a set of wheat RFLP probes identifying each homoeologous chromosome arm.Theor Appl Genet,1989,78(3):342-348.
[17]Wang YZ,Yang XN,Xiao QP.The improvement of identification technique of SCAB(Gibberella zeae Petch)resistance of wheat and the development of resistant sources.Sci Agric Sin,1982,15(5):66-67(in Chinese).王裕中,杨新宁,肖庆璞.小麦赤霉病抗性鉴定技术的改进及其抗源的开拓.中国农业科学,1982,15(5):66-67.
[18]Liu DJ.Breeding wheat for scab resistance-a worldwide hard nut to crack//Proceedings of International Conference on Wheat Genetics and Breeding-Perspectives of the 21st Century for Wheat Genetics and Breeding.Beijing:Agriculture Publisher,2001:4-7(in Chinese).刘大钧.小麦抗赤霉病育种--一个世界性难题//21世纪小麦遗传育种展望--小麦遗传育种国际学术讨论会文集.北京:农业出版社,2001:4-7.
[19]Liu WX,Chen PD,Liu DJ.Selection,breeding and identification of T01-a Triticum aestivum-Leymus racemosus translocation line.Acta Agron Sin,2000,26(3):305-309(in Chinese).刘文轩,陈佩度,刘大钧.一个普通小麦_大赖草易位系T01的选育与鉴定.作物学报,2000,26(3):305-309.
[20]Yang BJ,Dou QW,Liu WX,et al.Development of Triticum aestivum-Leymus racemosus translocation lines NAU601 and NAU618 and their test-cross analysis with double ditelosomic.Acta Genet Sin,2002,29(4):350-354,377(in Chinese).杨宝军,窦全文,刘文轩,等.普通小麦-大赖草易位系NAU601和NAU618的选育及双端二体测交分析.遗传学报,2002,29(4):350-354,377.
[21]Yuan JH,Chen PD,Liu DJ.Development of Triticum aestivum-Leymus racemosus translocation lines using gametocidal chromosomes.Sci China Ser C:Life Sci,2003,46(5):522-530.
[22]Sears ER.The transfer of leaf-rust resistance from Aegilops umbellulata to wheat.Brookhaven Symp Biol,1956,9:1-22.
[23]Sharma D,Knott DR.The transfer of leaf-rust resistance from Agropyron to Triticum by irradiation.Genome,1966,8(1):137-143.
[24]Sears ER,Gustafson JP.Use of radiation to transfer alien chromosome segments to wheat.Crop Sci,1993,33(5):897-901.
[25]Friebe B,Jiang JM,Raupp WJ,et al.Characterization of wheat-alien translocations conferring resistance to diseases and pests:Current status.Euphytica,1996,91(1):59-87.
[26]Luan Y,Wang XG,Liu WH,et al.Production and identification of wheat-Agropyron cristatum 6Ptranslocation lines.Planta,2010,232(2):501-510.
[27]Zhang J,Liu WP,Wu XH,et al.An intercalary translocation from Agropyron cristatum 6Pchromosome into common wheat confers enhanced kernel number per spike.Planta,2016,244(4):853-864.
[28]Song LQ,Lu YQ,Zhang JP,et al.Physical mapping of Agropyron cristatum chromosome 6P using deletion lines in common wheat background.Theor Appl Genet,2016,129(5):1023-1034.
[2]Yin GM,Zhao JH,Zhang YJ,et al.Adaption strategy about Leymus racemosus under different grazing patterns.Anim Husband Feed Sci,2013,34(11):33-36,39(in Chinese).殷国梅,赵金花,张英俊,等.大赖草对不同放牧方式的适应策略.畜牧与饲料科学,2013,34(11):33-36,39.
[3]Dewey DR.The genomic system of classification as a guide to intergeneric hybridization with the perennial Triticeae//Gustafson JP,ed.Gene Manipulation in Plant Improvement.New York:Plenum Press,1984,209-279.
[4]McGuire PE,Dv?rák J.High salt-tolerance potential in wheatgrasses.Crop Sci,1981,21(5):702-705.
[5]Mujeeb-Kazi A,Bekele GT,Mirand JL.Incorporation of alien genetic information from Elymus giganteus into Triticum aestivum//Sakamoto S,ed.Proceedings of the 6th International Wheat Genet Symposium.Maruzen,Kyoto,Japan,1983:223-231.
[6]Mujeeb-Kazi A,Rodriguez R.An intergeneric hybrid of Triticum aestivum L.X Elymus giganteus.J Hered,1981,72:253-256.
[7]Chen PD,Wang ZT,Wang SL,et al.Transfer of useful germplasm from Leymus racemosus Lam.to common wheat.III.Development of addition lines with wheat scab resistance.Acta Genet Sin,1995,22(3):206-210(in Chinese).陈佩度,王兆悌,王苏玲,等.将大赖草种质转移给普通小麦的研究:III.抗赤霉病异附加系选育.遗传学报,1995,22(3):206-210.
[8]Qi LL,Wang SL,Chen PD,et al.Molecular cytogenetic analysis of Leymus racemosus chromosomes added to wheat.Theoret Appl Genet,1997,95(7):1084-1091.
[9]Liu WX,Chen PD,Liu DJ.Development of Triticum aestivum-Leymus racemosus translocation lines by irradiating adult plants at meiosis.Acta Bot Sin,1999,41(5):463-467(in Chinese).刘文轩,陈佩度,刘大钧.利用减数分裂期成株电离辐射选育小麦-大赖草易位系的研究.植物学报,1999,41(5):463-467.
[10]Wang LS,Chen PD,Wang XE.Molecular cytogenetic analysis of Triticum aestivum-Leymus racemosus reciprocal chromosomal translocation T7DS·5LrL/T5LrS·7DL.Chin Sci Bull,2010,55(11):1026-1031.
[11]Cui CQ,Wang LS,Chen PD.Molecular and cytogenetic identification of Triticum aestivum-Leymus racemo-sus translocation lines T7BS·7Lr#1S and T2AS·2AL-7Lr#1S.Acta Agron Sin,2013,39(2):191-197(in Chinese).崔承齐,王林生,陈佩度.普通小麦-大赖草易位系T7BS·7Lr#1S和T2AS·2AL-7Lr#1S的分子细胞遗传学鉴定.作物学报,2013,39(2):191-197.
[12]Gill BS,Friebe B,Endo TR.Standard karyotype and nomenclature system for description of chromosome bands and structural aberrations in wheat(Triticum aestivum).Genome,1991,34(5):830-839.
[13]Mukai Y,Nakahara Y,Yamamoto M.Simultaneous discrimination of the three genomes in hexaploid wheat by multicolor fluorescence in situ hybridization using total genomic and highly repeated DNA probes.Genome,1993,36(3):489-494
[14]Zhang P,Li W L,Friebe B,et al.Simultaneous painting of three genomes in hexaploid wheat by BACK-FISH.Genome,2004,47(5):979-987.
[15]Tang ZX,Yang ZJ,Fu SL.Oligonucleotides replacing the roles of repetitive sequences pAs1,pSc119.2,pTa-535,pTa71,CCS1,and pAWRC.1 for FISH analysis.J Appl Genet,2014,55(3):313-318.
[16]Sharp PJ,Chao S,Desai S,et al.The isolation characterization and application in the Triticeae of a set of wheat RFLP probes identifying each homoeologous chromosome arm.Theor Appl Genet,1989,78(3):342-348.
[17]Wang YZ,Yang XN,Xiao QP.The improvement of identification technique of SCAB(Gibberella zeae Petch)resistance of wheat and the development of resistant sources.Sci Agric Sin,1982,15(5):66-67(in Chinese).王裕中,杨新宁,肖庆璞.小麦赤霉病抗性鉴定技术的改进及其抗源的开拓.中国农业科学,1982,15(5):66-67.
[18]Liu DJ.Breeding wheat for scab resistance-a worldwide hard nut to crack//Proceedings of International Conference on Wheat Genetics and Breeding-Perspectives of the 21st Century for Wheat Genetics and Breeding.Beijing:Agriculture Publisher,2001:4-7(in Chinese).刘大钧.小麦抗赤霉病育种--一个世界性难题//21世纪小麦遗传育种展望--小麦遗传育种国际学术讨论会文集.北京:农业出版社,2001:4-7.
[19]Liu WX,Chen PD,Liu DJ.Selection,breeding and identification of T01-a Triticum aestivum-Leymus racemosus translocation line.Acta Agron Sin,2000,26(3):305-309(in Chinese).刘文轩,陈佩度,刘大钧.一个普通小麦_大赖草易位系T01的选育与鉴定.作物学报,2000,26(3):305-309.
[20]Yang BJ,Dou QW,Liu WX,et al.Development of Triticum aestivum-Leymus racemosus translocation lines NAU601 and NAU618 and their test-cross analysis with double ditelosomic.Acta Genet Sin,2002,29(4):350-354,377(in Chinese).杨宝军,窦全文,刘文轩,等.普通小麦-大赖草易位系NAU601和NAU618的选育及双端二体测交分析.遗传学报,2002,29(4):350-354,377.
[21]Yuan JH,Chen PD,Liu DJ.Development of Triticum aestivum-Leymus racemosus translocation lines using gametocidal chromosomes.Sci China Ser C:Life Sci,2003,46(5):522-530.
[22]Sears ER.The transfer of leaf-rust resistance from Aegilops umbellulata to wheat.Brookhaven Symp Biol,1956,9:1-22.
[23]Sharma D,Knott DR.The transfer of leaf-rust resistance from Agropyron to Triticum by irradiation.Genome,1966,8(1):137-143.
[24]Sears ER,Gustafson JP.Use of radiation to transfer alien chromosome segments to wheat.Crop Sci,1993,33(5):897-901.
[25]Friebe B,Jiang JM,Raupp WJ,et al.Characterization of wheat-alien translocations conferring resistance to diseases and pests:Current status.Euphytica,1996,91(1):59-87.
[26]Luan Y,Wang XG,Liu WH,et al.Production and identification of wheat-Agropyron cristatum 6Ptranslocation lines.Planta,2010,232(2):501-510.
[27]Zhang J,Liu WP,Wu XH,et al.An intercalary translocation from Agropyron cristatum 6Pchromosome into common wheat confers enhanced kernel number per spike.Planta,2016,244(4):853-864.
[28]Song LQ,Lu YQ,Zhang JP,et al.Physical mapping of Agropyron cristatum chromosome 6P using deletion lines in common wheat background.Theor Appl Genet,2016,129(5):1023-1034.