普通小麦–大赖草易位系T5AS-7LrL·7LrS分子细胞遗传学鉴定
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摘要
大赖草对赤霉病具有较好的抗性,将大赖草赤霉病抗性基因转入普通小麦,对拓宽小麦赤霉病抗性基础有重要意义。本研究在获得抗赤霉病普通小麦–大赖草异附加系基础上,采用~(60)Co-γ射线(1200Rad,剂量率100Rad min~(-1))处理小麦–大赖草二体附加系DA7Lr,并用处理后的花粉授给去雄的普通小麦中国春,对其M_1代种子根尖细胞有丝分裂中期染色体进行GISH分析,获得1株具有一条普通小麦–大赖草易位染色体的植株,对其自交后代中具有2条易位染色体植株的花粉母细胞减数分裂中期I观察发现,2条易位染色体形成了稳定的环状二价体,表明该植株为纯合体。利用顺序GISH–双色FISH分析,结合C-分带、小麦D组专化探针Oligo-pAs1-2和B组专化探针Oligo-pSc119.2-2,进一步鉴定出该易位系为T5AS-7LrL·7LrS,同时筛选出可追踪该易位系的3个EST-STS分子标记,即BE591127、BQ168298和BE591737。该易位系的育成也为小麦赤霉病遗改良提供了新种质。
Leymus racemosus is highly resistant to wheat scab. The transfer of resistance genes from L. racemosus to common wheat(Triticum aestivum) is important for broadening the resistant sources against scab in common wheat. In this study, the pollen of DA7 Lr, a T. aestivum–L. racemosus disomic addition line with scab resistance, was irradiated with ~(60)Co-γ ray at 1200 Rad(100 Rad min~(-1)) before pollinating to emasculated T. aestivum cv. Chinese Spring. One plant with one translocation chromosome was detected in the M_1 generation by GISH. This plant was then self-pollinated and the pollen mother cells(PMCs) of the offspring plants with two translocation chromosomes were cytologically observed, and one ring bivalent was found at meiotic metaphase I, indicating that the plant with two translocation chromosomes was one translocation homozygote. The translocation line was proved to be T5 AS-7 LrL·7 LrS by C-banding, and sequential GISH–FISH using Oligo-pAs1-2 and Oligo-pSc119.2-2 as probes. Three EST-STS markers(BE591127, BQ168298, and BE591737) were identified to be able to track the T5 AS-7 LrL·7 LrS line. The translocation line also serves as an resistant source against wheat scab in wheat breeding programs.
Leymus racemosus is highly resistant to wheat scab. The transfer of resistance genes from L. racemosus to common wheat(Triticum aestivum) is important for broadening the resistant sources against scab in common wheat. In this study, the pollen of DA7 Lr, a T. aestivum–L. racemosus disomic addition line with scab resistance, was irradiated with ~(60)Co-γ ray at 1200 Rad(100 Rad min~(-1)) before pollinating to emasculated T. aestivum cv. Chinese Spring. One plant with one translocation chromosome was detected in the M_1 generation by GISH. This plant was then self-pollinated and the pollen mother cells(PMCs) of the offspring plants with two translocation chromosomes were cytologically observed, and one ring bivalent was found at meiotic metaphase I, indicating that the plant with two translocation chromosomes was one translocation homozygote. The translocation line was proved to be T5 AS-7 LrL·7 LrS by C-banding, and sequential GISH–FISH using Oligo-pAs1-2 and Oligo-pSc119.2-2 as probes. Three EST-STS markers(BE591127, BQ168298, and BE591737) were identified to be able to track the T5 AS-7 LrL·7 LrS line. The translocation line also serves as an resistant source against wheat scab in wheat breeding programs.
引文
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[2]Mujeeb-Kazi A,Bekele G T,Mirand J L.Incorporation of alien genetic information from Elymus giganteus into Triticum aestivum.In:Proc the 6th Int Wheat Genet Symp,Kyoto,Japan,1983.pp 223-231
[3]Mujeeb-Kazi A,Rodriguez R.An intergeneric hybrid of Triticum aestivum L.×Elymus giganteus.J Hered,1981,72:253-256
[4]陈佩度,王兆悌,王苏玲,黄俐,王裕中,刘大钧.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,1995,22:206-210Chen P D,Wang Z T,Wang S L,Huang L,Wang Y Z,Liu D J.Transfer of useful germplasm from Leymus racemosus Lam.to common wheat:III.Development of addition lines with wheat scab resistance.Acta Genet Sin,1995,22:206-210(in Chinese with English abstract)
[5]Qi L L,Wang S L,Chen P D.Molecular cytogenetic analysis of Leymus racemosus chromosomes added to wheat.Theor Appl Genet,1997,95:1084-1091
[6]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,1999,41:463-467Liu W X,Chen P D,Liu D J.Development of Triticum aestivum-Leymus racemosus translocation lines by irradiating adult plants at meiosis.Acta Bot Sin,1999,41:463-467(in Chinese with English abstract)
[7]刘文轩,陈佩度,刘大钧.利用花粉辐射诱发普通小麦与大赖草染色体易位的研究.遗传学报,2000,27:44-49Liu W X,Chen P D,Liu D J.Studies of the development of Triticum aestivum-Leymus racemosus translocation lines by pollen irradiation.Acta Genet Sin,2000,27:44-49(in Chinese with English abstract)
[8]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,2000,26:305-309Liu W X,Chen P D,Liu D J.Selection,breeding and identification of T01:a Triticum aestivum-Leymus racemosus translocation line.Acta Agron Sin,2000,26:305-309(in Chinese with English abstract)
[9]杨宝军,窦全文,刘文轩,周波,陈佩度.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,2002,29:350-354Yang B J,Dou Q W,Liu W X,Zhou B,Chen P D.Development of Triticum aestivum-Leymus racemosus translocation lines NAU601 and NAU618 and their test-cross analysis with double ditelosomic.Acta Genet Sin,2002,29:350-354(in Chinese with English abstract)
[10]Yuan J H,Chen P D,Liu D J.Development of Triticum aestivumLeymus racemosus translocation lines using gametocidal chromosomes.Sci China Ser C:Life Sci,2003,46:522-530
[11]Wang L S,Chen P D,Wang X E.Molecular cytogenetic analysis of Triticum aestivum-Leymus racemosus reciprocal chromosomal translocation T7DS·5LrL/T5LrS·7DL.Chin Sci Bull,2010,55:1026-1031
[12]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,2013,39:191-197Cui C Q,Wang L S,Chen P D.Molecular and cytogenetic identification of Triticum aestivum-Leymus racemosus translocation lines T7BS·7Lr#1S and T2AS·2AL-7Lr#1S.Acta Agron Sin,2013,39:191-197(in Chinese with English abstract)
[13]Wang L S,Chen P D.Development of Triticum aestivum-Leymus racemosus sub.7Lr#1S(7A)with resistance to wheat scab and their analysis with meiosis.Chin Sci Bull,2008,53:3522-3529
[14]Qi L L,Pumphrey M O,Friebe B,Gill B S.Molecular cytogenetic characterization of alien introgressions with gene Fhb3 for resistance to Fusarium head blight disease of wheat.Theor Appl Genet,2008,117:1155-1166(in Chinese with English abstract)
[15]Gill B S,Friebe B.Standard karyotype and system for description of chromosome bands and structural aberrations in wheat(Triticum aestivum).Genome,1991,34:830-839
[16]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,1993,36:489-494
[17]Zhang P,Li W L,Friebe B,Gill B S.Simultaneous painting of three genomes in hexaploid wheat by BACK-FISH.Genome,2004,47:979-878
[18]Pedersen C,Linde-Laursen I.Chromosomal locations of four minor rDNA loci and a marker microsatellite sequence in barley.Chromos Res,1994,2:65-71
[19]Tang Z X,Yang Z J,Fu S L.Oligonucleotides replacing the roles of repetitive sequences pAs1,pSc119.2,pTa-535,pTa71,CCS1,and pAWRC.1 for FISH analysis.J Appl Genet,2014,55:313-318
[20]Sharp P J,Cao S,Desai S,Gale M D.The isolation characterization and application in the Triticae of a set of wheat RFLP probes identifying each homoeologous chromosome arm.Theor Appl Genet,1989,78:342-348
[21]王裕中,杨新宁,尚庆璞.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,1982,23(5):66-67Wang Y Z,Yang X N,Shang Q P.The improvement of identification technique of scab(Gibberella zeae Petch.)resistance of wheat and the development of resistant sources.Sci Agric Sin,1982,23(5):66-67(in Chinese with English abstract)
[22]Sears E R.The transfer of leaf rust resistance from Aegilops umbellulata to wheat.Brookhaven Symp Biol,1956,9:1-22
[23]Sharma D,Knott D R.The transfer of leaf-rust resistance from Agropyron to Triticum by irradiation.Genome,1966,8:137-143
[24]Sears E,Gustafson J.Use of radiation to transfer alien chromosome segments to wheat.Crop Sci,1993,33:897-901
[25]Friebe B,Jiang J M,Raupp W J,McIntosh R A,Gill B S.Characterization of wheat-alien translocations conferring resistance to diseases and pets:current status.Euphytica,1996,91:59-87
[26]Luan Y,Wang X,Liu W,Li C,Zhang J,Gao A,Wang Y,Yang X,Li L.Production and identification of wheat-Agropyron cristatum6P translocation lines.Planta,2010,232:501-510
[27]Zhang J,Liu W,Wu X,Yang X,Li X,Lu Y,Li L.An intercalary translocation from Agropyron cristatum 6P chromosome into common wheat confers enhanced kernel number per spike.Planta,2016,244:853-864
[28]Song L,Lu Y,Zhang J,Pan C,Yang X,Li X,Liu W,Li L.Physical mapping of Agropyron cristatum chromosome 6P using deletion lines in common wheat background.Theor Appl Genet,2016,129:1023-1034
[29]Chen P D,Qi L L,Zhou B,Zhang S Z,Liu D J.Development and molecular cytogenetic analysis of wheat-Haynaldia villosa6VS/6A:I.Translocation lines specifying resistance to powdery mildew.Theor Appl Genet,1995,91:1125-1128
[30]李桂萍,陈佩度,张守忠,赵和.小麦-簇毛麦6VS/6AL易位染色体对小麦农艺性状的影响.植物遗传资源报,2011,12:744-749Li G P,Chen P D,Zhang S Z,Zhao H.Effects of the 6VS/6ALtranslocation chromosome on agronomic characteristics of wheat.J Plant Genet Resour,2011,12:744-749(in Chinese with English abstract)
[31]Subbarao G V,Ban T,Masahiro K,Osamu I,Samejima H,Wang H Y,Pearse S J,Gopalakrishnan S,Nakahara K,Zakir Hossain AK M,Tsujimoto H,Berry W L.Can biological nitrification inhibition(BNI)genes from perennial Leymus racemosus(Triticeae)combat nitrification in wheat farming?Plant&Soil,2007,299:55-64
[32]Subbarao G V,Sahrawat K L,Nakahara K,Rao I M,Ishitani M,Hash C T,Kishii M,Bonnett D G,Berry W L,Lata J C.A paradigm shift towards low-nitrifying production systems:the role of biological nitrification inhibition(BNI).Ann Bot,2013,112:297-316
[2]Mujeeb-Kazi A,Bekele G T,Mirand J L.Incorporation of alien genetic information from Elymus giganteus into Triticum aestivum.In:Proc the 6th Int Wheat Genet Symp,Kyoto,Japan,1983.pp 223-231
[3]Mujeeb-Kazi A,Rodriguez R.An intergeneric hybrid of Triticum aestivum L.×Elymus giganteus.J Hered,1981,72:253-256
[4]陈佩度,王兆悌,王苏玲,黄俐,王裕中,刘大钧.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,1995,22:206-210Chen P D,Wang Z T,Wang S L,Huang L,Wang Y Z,Liu D J.Transfer of useful germplasm from Leymus racemosus Lam.to common wheat:III.Development of addition lines with wheat scab resistance.Acta Genet Sin,1995,22:206-210(in Chinese with English abstract)
[5]Qi L L,Wang S L,Chen P D.Molecular cytogenetic analysis of Leymus racemosus chromosomes added to wheat.Theor Appl Genet,1997,95:1084-1091
[6]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,1999,41:463-467Liu W X,Chen P D,Liu D J.Development of Triticum aestivum-Leymus racemosus translocation lines by irradiating adult plants at meiosis.Acta Bot Sin,1999,41:463-467(in Chinese with English abstract)
[7]刘文轩,陈佩度,刘大钧.利用花粉辐射诱发普通小麦与大赖草染色体易位的研究.遗传学报,2000,27:44-49Liu W X,Chen P D,Liu D J.Studies of the development of Triticum aestivum-Leymus racemosus translocation lines by pollen irradiation.Acta Genet Sin,2000,27:44-49(in Chinese with English abstract)
[8]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,2000,26:305-309Liu W X,Chen P D,Liu D J.Selection,breeding and identification of T01:a Triticum aestivum-Leymus racemosus translocation line.Acta Agron Sin,2000,26:305-309(in Chinese with English abstract)
[9]杨宝军,窦全文,刘文轩,周波,陈佩度.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,2002,29:350-354Yang B J,Dou Q W,Liu W X,Zhou B,Chen P D.Development of Triticum aestivum-Leymus racemosus translocation lines NAU601 and NAU618 and their test-cross analysis with double ditelosomic.Acta Genet Sin,2002,29:350-354(in Chinese with English abstract)
[10]Yuan J H,Chen P D,Liu D J.Development of Triticum aestivumLeymus racemosus translocation lines using gametocidal chromosomes.Sci China Ser C:Life Sci,2003,46:522-530
[11]Wang L S,Chen P D,Wang X E.Molecular cytogenetic analysis of Triticum aestivum-Leymus racemosus reciprocal chromosomal translocation T7DS·5LrL/T5LrS·7DL.Chin Sci Bull,2010,55:1026-1031
[12]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,2013,39:191-197Cui C Q,Wang L S,Chen P D.Molecular and cytogenetic identification of Triticum aestivum-Leymus racemosus translocation lines T7BS·7Lr#1S and T2AS·2AL-7Lr#1S.Acta Agron Sin,2013,39:191-197(in Chinese with English abstract)
[13]Wang L S,Chen P D.Development of Triticum aestivum-Leymus racemosus sub.7Lr#1S(7A)with resistance to wheat scab and their analysis with meiosis.Chin Sci Bull,2008,53:3522-3529
[14]Qi L L,Pumphrey M O,Friebe B,Gill B S.Molecular cytogenetic characterization of alien introgressions with gene Fhb3 for resistance to Fusarium head blight disease of wheat.Theor Appl Genet,2008,117:1155-1166(in Chinese with English abstract)
[15]Gill B S,Friebe B.Standard karyotype and system for description of chromosome bands and structural aberrations in wheat(Triticum aestivum).Genome,1991,34:830-839
[16]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,1993,36:489-494
[17]Zhang P,Li W L,Friebe B,Gill B S.Simultaneous painting of three genomes in hexaploid wheat by BACK-FISH.Genome,2004,47:979-878
[18]Pedersen C,Linde-Laursen I.Chromosomal locations of four minor rDNA loci and a marker microsatellite sequence in barley.Chromos Res,1994,2:65-71
[19]Tang Z X,Yang Z J,Fu S L.Oligonucleotides replacing the roles of repetitive sequences pAs1,pSc119.2,pTa-535,pTa71,CCS1,and pAWRC.1 for FISH analysis.J Appl Genet,2014,55:313-318
[20]Sharp P J,Cao S,Desai S,Gale M D.The isolation characterization and application in the Triticae of a set of wheat RFLP probes identifying each homoeologous chromosome arm.Theor Appl Genet,1989,78:342-348
[21]王裕中,杨新宁,尚庆璞.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,1982,23(5):66-67Wang Y Z,Yang X N,Shang Q P.The improvement of identification technique of scab(Gibberella zeae Petch.)resistance of wheat and the development of resistant sources.Sci Agric Sin,1982,23(5):66-67(in Chinese with English abstract)
[22]Sears E R.The transfer of leaf rust resistance from Aegilops umbellulata to wheat.Brookhaven Symp Biol,1956,9:1-22
[23]Sharma D,Knott D R.The transfer of leaf-rust resistance from Agropyron to Triticum by irradiation.Genome,1966,8:137-143
[24]Sears E,Gustafson J.Use of radiation to transfer alien chromosome segments to wheat.Crop Sci,1993,33:897-901
[25]Friebe B,Jiang J M,Raupp W J,McIntosh R A,Gill B S.Characterization of wheat-alien translocations conferring resistance to diseases and pets:current status.Euphytica,1996,91:59-87
[26]Luan Y,Wang X,Liu W,Li C,Zhang J,Gao A,Wang Y,Yang X,Li L.Production and identification of wheat-Agropyron cristatum6P translocation lines.Planta,2010,232:501-510
[27]Zhang J,Liu W,Wu X,Yang X,Li X,Lu Y,Li L.An intercalary translocation from Agropyron cristatum 6P chromosome into common wheat confers enhanced kernel number per spike.Planta,2016,244:853-864
[28]Song L,Lu Y,Zhang J,Pan C,Yang X,Li X,Liu W,Li L.Physical mapping of Agropyron cristatum chromosome 6P using deletion lines in common wheat background.Theor Appl Genet,2016,129:1023-1034
[29]Chen P D,Qi L L,Zhou B,Zhang S Z,Liu D J.Development and molecular cytogenetic analysis of wheat-Haynaldia villosa6VS/6A:I.Translocation lines specifying resistance to powdery mildew.Theor Appl Genet,1995,91:1125-1128
[30]李桂萍,陈佩度,张守忠,赵和.小麦-簇毛麦6VS/6AL易位染色体对小麦农艺性状的影响.植物遗传资源报,2011,12:744-749Li G P,Chen P D,Zhang S Z,Zhao H.Effects of the 6VS/6ALtranslocation chromosome on agronomic characteristics of wheat.J Plant Genet Resour,2011,12:744-749(in Chinese with English abstract)
[31]Subbarao G V,Ban T,Masahiro K,Osamu I,Samejima H,Wang H Y,Pearse S J,Gopalakrishnan S,Nakahara K,Zakir Hossain AK M,Tsujimoto H,Berry W L.Can biological nitrification inhibition(BNI)genes from perennial Leymus racemosus(Triticeae)combat nitrification in wheat farming?Plant&Soil,2007,299:55-64
[32]Subbarao G V,Sahrawat K L,Nakahara K,Rao I M,Ishitani M,Hash C T,Kishii M,Bonnett D G,Berry W L,Lata J C.A paradigm shift towards low-nitrifying production systems:the role of biological nitrification inhibition(BNI).Ann Bot,2013,112:297-316