普通小麦大赖草易位系T3AS·3AL-7Lr#1S的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1 200 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(M_1)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基因组专化探针Oligo-pAs1-2和B基因组专化探针Oligo-pSc119.2-2,进一步鉴定出该普通小麦-大赖草易位系为T3AS·3AL-7Lr#1S,且筛选出了可追踪该易位系的3个EST-STS分子标记BE591127、BQ168298和BE591737。该易位系的育成为小麦赤霉病遗传改良提供了新种质。
Leymus racemosus is highly resistant to wheat scab(Fusarum head blight, FHB). The transfer of scab resistant gene from L. racemosus to Triticum aestivum is of great significance for broadening the base of wheat resistance. In the present study, the pollen of T. aestivum-L. racemosus disomic addition line DA7 Lr with scab resistance was irradiated by~(60)Co-γ-rays 1 200 R(100 R·min~(-1)) prior to pollinating to emasculated T. aestivum cv. Chinese Spring. One plant with a translocated chromosome was detected in the M_1 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 ring bivalent was observed, indicating that the plant with two translocation chromosomes was one translocation homozygote. C-banding and sequential GISH/FISH analysis, using Oligo-pAs1-2 and Oligo-pSc119.2-2 as probe, translocation line T3 AS·3 AL-7 Lr#1 S was confirmed. Three molecular markers of EST-STS such as BE591127, BQ168298 and BE591737 were screened, and these markers can be used to track the translocation line. The translocation line had good resistance to wheat scab and feasibility to be used as a new germplasm in wheat breeding resistant to scab disease.
Leymus racemosus is highly resistant to wheat scab(Fusarum head blight, FHB). The transfer of scab resistant gene from L. racemosus to Triticum aestivum is of great significance for broadening the base of wheat resistance. In the present study, the pollen of T. aestivum-L. racemosus disomic addition line DA7 Lr with scab resistance was irradiated by~(60)Co-γ-rays 1 200 R(100 R·min~(-1)) prior to pollinating to emasculated T. aestivum cv. Chinese Spring. One plant with a translocated chromosome was detected in the M_1 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 ring bivalent was observed, indicating that the plant with two translocation chromosomes was one translocation homozygote. C-banding and sequential GISH/FISH analysis, using Oligo-pAs1-2 and Oligo-pSc119.2-2 as probe, translocation line T3 AS·3 AL-7 Lr#1 S was confirmed. Three molecular markers of EST-STS such as BE591127, BQ168298 and BE591737 were screened, and these markers can be used to track the translocation line. The translocation line had good resistance to wheat scab and feasibility to be used as a new germplasm in wheat breeding resistant to scab disease.
引文
[1] 李韬,李嫒嫒,李磊. 小麦赤霉病:从表型鉴定到抗性改良[J]. 科技导报,2016,34(22):75-80.LI T, LI A A, LI L. Fusarium head blight in wheat: From phenotyping to resistance improvement[J]. Science Technology Review, 2016, 34(22):75-80. (in Chinese)
[2] BAI G H, SHANER G. Management and resistance in wheat and barley to fusarium head blight[J]. Annual Review of Phytopathology, 2004,42(1): 135-161.
[3] PESTKA J J. Deoxynivalenol: mechanisms of action, human exposure,and toxicological relevance[J]. Archives of Toxicology, 2010, 84(9): 663-679.
[4] ZHANG J B, WANG J H, GONG A D, et al. Natural occurrence of fusarium head blight, mycotoxins and mycotoxin-producing isolates of Fusariumin commercial fields of wheat in Hubei[J]. Plant Pathology, 2013, 62(1): 92-102.
[5] 刘大钧. 小麦抗赤霉病育种一个世界性难题[M]//中国农学会. 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: 中国农业科技出版社, 2001: 4-12.
[6] MCGUIRE P E, DV?RáK J. High salt-tolerance potential in wheatgrasses[J]. Crop Science, 1981, 21(5): 702-705.
[7] MUJEEB-KAZI A, BEKELE G T, MIRAND J L. Incorporation of alien genetic information from Elymus giganteus into Triticum aestivum[C]// Proceedings of the 6th International Wheat Genetics Symposium. Kyoto, Japan, 1983. 223-231.
[8] 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[J]. 遗传学报, 1995, 22(3): 206-210.CHEN P D, WANG Z T, WANG S L, et al. Transfer of useful germplasm from Leymus racemosus Lam. to common wheat. Ⅲ. Development of addition lines with wheat scab resistance[J]. Acta Genetica Sinica, 1995, 22(3): 206-210. (in Chinese with English abstract)
[9] WANG L S, CHEN P D. Development of Triticum aestivum-Leymus racemosus ditelosomic substitution line 7Lr#1S(7A) with resistance to wheat scab and its meiotic behavior analysis[J]. Chinese Science Bulletin, 2008, 53(22): 3522-3529.
[10] QI L L, PUMPHREY M O, FRIEBE B, et al. Molecular cytogenetic characterization of alien introgressions with gene Fhb3 for resistance to Fusarium head blight disease of wheat[J]. Theoretical and Applied Genetics, 2008, 117(7): 1155-1166.
[11] 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[J]. 植物学报,1999, 41(5):463-467.LIU W X, CHEN P D, LIU D J. Development of Triticum aestivum-Leymus racemosus translocation lines by irradiating adult plants at meiosis[J]. Acta Botanica Sinica, 1999, 41(5): 463-467. (in Chinese with English abstract)
[12] WANG L S, CHEN P D,WANG X E. Molecular cytogenetic analysis of Triticum aestivum-Leymus racemosus reciprocal chromosomal translocation T7DS·5LrL/T5LrS·7DL[J]. Chinese Science Bulletin, 2010, 55(11): 1026-1031.
[13] 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[J]. 作物学报, 2013,39(2):191-197.CUI 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[J]. Acta Agronomica Sinica, 2013, 39(2):191-197. (in Chinese with English abstract)
[14] GILL B S, FRIEBE B, ENDO T R. Standard karyotype and nomenclature system for description of chromosome bands and structural aberrations in wheat (Triticum aestivum)[J]. Genome, 1991, 34(5): 830-839.
[15] 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[J]. Genome, 1993, 36(3): 489-494.
[16] ZHANG P, LI W L, FRIEBE B, et al. Simultaneous painting of three genomes in hexaploid wheat by BAC-FISH[J]. Genome, 2004, 47(5): 979-987.
[17] 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]. Journal of Applied Genetics, 2014, 55(3): 313-318.
[18] SHARP P J, 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[J]. Theoretical and Applied Genetics, 1989, 78(3): 342-348.
[19] 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[J]. 中国农业科学,1982(5):66-67. WANG Y Z, YANG X N, XIAO Q P. The improvement of identification technique of scab (Gibberella zeae Petch.) resistance of wheat and the development of resistant sources[J]. Scientia Agricultura Sinica, 1982(5): 66-67.
[20] MA H X, YAO J B, ZHOU M P, et al. Molecular breeding for wheat Fusarium head blight resistance in China[J]. Cereal Research Communications, 2008, 36(Suppl):203-212.
[21] ZHANG J, ZHANG J P, LIU W H, et al. An intercalary translocation from Agropyron cristatum 6P chromosome into common wheat confers enhanced kernel number per spike[J]. Planta, 2016, 244(4): 853-864.
[22] SONG L, LU Y, ZHANG J, et al. Physical mapping of Agropyron cristatum chromosome 6P using deletion lines in common wheat background[J]. Theoretical and Applied Genetics,2016 ,129(5):1023-1034.
[2] BAI G H, SHANER G. Management and resistance in wheat and barley to fusarium head blight[J]. Annual Review of Phytopathology, 2004,42(1): 135-161.
[3] PESTKA J J. Deoxynivalenol: mechanisms of action, human exposure,and toxicological relevance[J]. Archives of Toxicology, 2010, 84(9): 663-679.
[4] ZHANG J B, WANG J H, GONG A D, et al. Natural occurrence of fusarium head blight, mycotoxins and mycotoxin-producing isolates of Fusariumin commercial fields of wheat in Hubei[J]. Plant Pathology, 2013, 62(1): 92-102.
[5] 刘大钧. 小麦抗赤霉病育种一个世界性难题[M]//中国农学会. 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: 中国农业科技出版社, 2001: 4-12.
[6] MCGUIRE P E, DV?RáK J. High salt-tolerance potential in wheatgrasses[J]. Crop Science, 1981, 21(5): 702-705.
[7] MUJEEB-KAZI A, BEKELE G T, MIRAND J L. Incorporation of alien genetic information from Elymus giganteus into Triticum aestivum[C]// Proceedings of the 6th International Wheat Genetics Symposium. Kyoto, Japan, 1983. 223-231.
[8] 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[J]. 遗传学报, 1995, 22(3): 206-210.CHEN P D, WANG Z T, WANG S L, et al. Transfer of useful germplasm from Leymus racemosus Lam. to common wheat. Ⅲ. Development of addition lines with wheat scab resistance[J]. Acta Genetica Sinica, 1995, 22(3): 206-210. (in Chinese with English abstract)
[9] WANG L S, CHEN P D. Development of Triticum aestivum-Leymus racemosus ditelosomic substitution line 7Lr#1S(7A) with resistance to wheat scab and its meiotic behavior analysis[J]. Chinese Science Bulletin, 2008, 53(22): 3522-3529.
[10] QI L L, PUMPHREY M O, FRIEBE B, et al. Molecular cytogenetic characterization of alien introgressions with gene Fhb3 for resistance to Fusarium head blight disease of wheat[J]. Theoretical and Applied Genetics, 2008, 117(7): 1155-1166.
[11] 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[J]. 植物学报,1999, 41(5):463-467.LIU W X, CHEN P D, LIU D J. Development of Triticum aestivum-Leymus racemosus translocation lines by irradiating adult plants at meiosis[J]. Acta Botanica Sinica, 1999, 41(5): 463-467. (in Chinese with English abstract)
[12] WANG L S, CHEN P D,WANG X E. Molecular cytogenetic analysis of Triticum aestivum-Leymus racemosus reciprocal chromosomal translocation T7DS·5LrL/T5LrS·7DL[J]. Chinese Science Bulletin, 2010, 55(11): 1026-1031.
[13] 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[J]. 作物学报, 2013,39(2):191-197.CUI 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[J]. Acta Agronomica Sinica, 2013, 39(2):191-197. (in Chinese with English abstract)
[14] GILL B S, FRIEBE B, ENDO T R. Standard karyotype and nomenclature system for description of chromosome bands and structural aberrations in wheat (Triticum aestivum)[J]. Genome, 1991, 34(5): 830-839.
[15] 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[J]. Genome, 1993, 36(3): 489-494.
[16] ZHANG P, LI W L, FRIEBE B, et al. Simultaneous painting of three genomes in hexaploid wheat by BAC-FISH[J]. Genome, 2004, 47(5): 979-987.
[17] 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]. Journal of Applied Genetics, 2014, 55(3): 313-318.
[18] SHARP P J, 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[J]. Theoretical and Applied Genetics, 1989, 78(3): 342-348.
[19] 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[J]. 中国农业科学,1982(5):66-67. WANG Y Z, YANG X N, XIAO Q P. The improvement of identification technique of scab (Gibberella zeae Petch.) resistance of wheat and the development of resistant sources[J]. Scientia Agricultura Sinica, 1982(5): 66-67.
[20] MA H X, YAO J B, ZHOU M P, et al. Molecular breeding for wheat Fusarium head blight resistance in China[J]. Cereal Research Communications, 2008, 36(Suppl):203-212.
[21] ZHANG J, ZHANG J P, LIU W H, et al. An intercalary translocation from Agropyron cristatum 6P chromosome into common wheat confers enhanced kernel number per spike[J]. Planta, 2016, 244(4): 853-864.
[22] SONG L, LU Y, ZHANG J, et al. Physical mapping of Agropyron cristatum chromosome 6P using deletion lines in common wheat background[J]. Theoretical and Applied Genetics,2016 ,129(5):1023-1034.