应用分子标记辅助选育洋葱不育系及其杂交应用
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摘要
利用分子标记辅助筛选洋葱不育系,配制杂交组合,并利用分子标记检测杂交种的育性。结果表明,选育出101A、A1、D-9、A3和T共计5套不育系,101A为黄皮圆球形晚熟CMS-S型不育系,其他4个均为CMS-T型不育系,A1和D-9是利用不育分子标记orf725辅助筛选出的早熟黄皮圆球形不育系,A3和T是利用保持系分子标记jnurf13辅助分别筛选的早熟黄皮圆球形和晚熟红皮扁球形不育系;通过不育系A1和D-9配制64份杂交组合,筛选出23、39、40、72、73和84号共6份候选杂交组合,其中23号为全不育型杂交组合;另外,利用orf725分子标记检测市场上主栽的16个杂交品种的育性,其中帝皇、红将军、A1×3C和T×912 4份均为不育株,首次提出洋葱全不育型杂交种,为洋葱杂交育种提供新思路。所以,利用分子标记辅助筛选洋葱不育系、保持系和全不育型杂交种对生产具有重要作用。
By marker-assisted selection(MAS), we conducted the following research such as screening the onion sterile producing hybridization combination, detecting the fertility of the hybrid. The results showed that a total of 5 sterile lines of 101A, A1, D-9, A3 and T were selected. 101A was a yellow-skin-shaped late-maturing CMS-S type sterile line, and the other four were CMS-T-type sterile lines. A1 and D-9 were the early-maturing yellow-skin sterility lines which were selected by using molecular marker orf725. A3 and T were the early-ripening yellow-skin spherical and late-maturing red-skin spheroid sterile lines which were selected by using molecular marker jnurf13. Sixty-four hybrid combinations were prepared between sterile lines A1 and D-9, and 6 candidate hybrid combinations of 23, 39, 40, 72, 73 and 84 were screened. 23 was a complete sterile hybrid combination. In addition, the molecular marker orf725 was used to determine the fertility of the 16 popular hybrids on the market. The "Huangdi", the "Hongjiangjun", the "A1×3C" and the "T×912" were all infertile plants, and the complete sterile hybrid onion was first proposed, which provide fresh ideas for onion hybrid breeding. Therefore, the marker-assisted selection was an important role to screen onion male sterile lines, maintainer lines and complete sterile hybrids.
By marker-assisted selection(MAS), we conducted the following research such as screening the onion sterile producing hybridization combination, detecting the fertility of the hybrid. The results showed that a total of 5 sterile lines of 101A, A1, D-9, A3 and T were selected. 101A was a yellow-skin-shaped late-maturing CMS-S type sterile line, and the other four were CMS-T-type sterile lines. A1 and D-9 were the early-maturing yellow-skin sterility lines which were selected by using molecular marker orf725. A3 and T were the early-ripening yellow-skin spherical and late-maturing red-skin spheroid sterile lines which were selected by using molecular marker jnurf13. Sixty-four hybrid combinations were prepared between sterile lines A1 and D-9, and 6 candidate hybrid combinations of 23, 39, 40, 72, 73 and 84 were screened. 23 was a complete sterile hybrid combination. In addition, the molecular marker orf725 was used to determine the fertility of the 16 popular hybrids on the market. The "Huangdi", the "Hongjiangjun", the "A1×3C" and the "T×912" were all infertile plants, and the complete sterile hybrid onion was first proposed, which provide fresh ideas for onion hybrid breeding. Therefore, the marker-assisted selection was an important role to screen onion male sterile lines, maintainer lines and complete sterile hybrids.
引文
[1] Berninger E.Contribution à l’étude de la sterilité male del’oignon (Allium cepa L.) [J].Annals of Amélior Plantes,1965(15):183-199.
[2] Ones H,Emsweller S.A male-sterile onion [J].Proceedings of American Socirty for Horticultural Science,1936,34:582-585.
[3] Sato Y.PCR amplification of CMS-specific mitochondrial nucleotide sequences to identify cytoplasmic genotypes of onion (Allium cepa L.) [J].Theoretical and Applied Genetics,1998,96(3/4):367-370.
[4] Engeike T,Terefe D,Tatlioglu T.A PCR-based marker system monitoring CMS-(S),CMS-(T) and (N)-cytoplasm in the onion (Allium cepa L.) [J].Theoretical and Applied Genetics,2003,107:162-167.
[5] Havey M J.Identification of cytoplasms using the polymerase chain reaction to aid in the extraction of maintainer lines from open-pollinated populations of onion [J].Theoretical and Applied Genetics,1995,90(2):263-268.
[6] Kim S,Yoon M.Comparison of mitochondrial and chloroplast genome segments from three onion (Allium cepa L.) cytoplasm types and identification of a trans-splicing intron of cox2 [J].Current Genetics,2010,56:177-188.
[7] Kim S,Lee E,Cho D Y,et al.Identification of a novel chimeric gene,orf725,and its use in development of a molecular marker for distinguishing three cytoplasm types in onion (Allium cepa L.) [J].Theoretical and Applied Genetics,2009,118(2):433-441.
[8] Kims S,Parkb J Y,Yangb T.Comparative analysis of the complete chloroplast genome sequences of a normal male-fertile cytoplasm and two different cytoplasms conferring cytoplasmic male sterility in onion (Allium cepa L.) [J].The Journal of Horticultural Science and Biotechnology,2015,90(4):459-468.
[9] Kim S.A codominant molecular marker in linkage disequilibrium with a restorer-of-fertility gene (Ms) and its application in reevaluation of inheritance of fertility restoration in onions [J].Molecular Breeding,2014,34(3):769-778.
[10] Huo Y M,Liu B J,Yang Y Y,et al.AcSKP1,a multiplex PCR-based co-dominant marker in complete linkage disequilibrium with the male-fertility restoration (Ms) locus,and its application in open-pollinated populations of onion [J].Euphytica,2015,204 (3) :711-722.
[11] Yang Y Y,Huo Y M,Miao J,et al.Identification of two SCAR markers co-segregated with the dominant Ms and recessive ms alleles in onion ( Allium cepa L.) [J].Euphytica,2013,190 (2):267-277.
[12] Bang H,Cho D Y,Yoo K S,et al.Development of simple PCR-based markers linked to the Ms locus,a restorer-of-fertility gene in onion (Allium cepa L.) [J].Euphytica,2011,179 (3):439-449.
[13] 潘美红,杨海峰,惠林冲,等.洋葱CMS-T型育性分子标记的筛选与鉴定[J].西北农业学报,2018,27(4):576-585.
[14] 陈振泰,缪美华,薛萍,等.大直径黄皮洋葱新品种连葱096的选育[J].长江蔬菜,2011(9):50-51.
[15] 李峰,曹丽蓉,刘培园,等.小麦品种渝麦13号细胞质类型的分子标记鉴定[J].中国农业大学学报,2018,23(10):1-7.
[16] 张俊英,蒋美艳,江建霞,等.甘蓝型油菜显性核不育系的分子标记辅助选择[J].核农学报,2018,32(11):2119-2125.
[17] 张宏根,仲崇元,司华,等.分子标记辅助选择改良C418对红莲型粳稻不育系的恢复力[J].中国水稻科学,2018,32(5):445-452.
[18] 宋伟,苏爱国,邢锦丰,等.京724玉米自交系S型细胞质雄性不育系分子标记辅助选育研究[J].玉米科学,2016,24(1):33-36,42.
[19] Bang H,Kim S,Park S O,et al.Development of a codominant CAPS marker linked to the Ms locus controlling fertility restoration in onion (Allium cepa L.) [J].Scientia Horticulturae,2013,153(4):42-49.
[20] Park J,Bang H,Cho D Y,et al.Construction of high-resolution linkage map of the Ms locus,a restorer-of-fertility gene in onion (Allium cepa L.) [J].Euphytica,2013,192 (2):267-278.
[21] Kim S,Kim C W,Minkyu P,et al.Identification of candidate genes associated with fertility restoration of cytoplasmic male-sterility in onion (Allium cepa L.) using a combination of bulked segregant analysis and RNA-seq [J].Theoretical and Applied Genetics,2015,128 (11):2289-2299.
[22] Gokce A F,Havey M J.Linkage equilibrium among tightly linked RFLPs and the Ms locus in open-pollinated onion populations [J].Journal of the American Society for Horticultural Science,2002,127 (6):944-946.
[23] Gokce A F,Mccallum J,Sato Y,et al.Molecular tagging of the Ms locus in onion [J].Journal of the American Society for Horticultural Science,2002,127 (4):576-582.
[24] 陈振泰,薛萍,缪美华,等.紫皮洋葱新品种连葱11号的选育[J].长江蔬菜,2016(20):52-54.
[25] 倪西源,黄吉祥,柳寒,等.应用分子标记辅助选育甘蓝型油菜杂交种的可行性[J].浙江大学学报:农业与生命科学版,2017,43(2):173-182.
[2] Ones H,Emsweller S.A male-sterile onion [J].Proceedings of American Socirty for Horticultural Science,1936,34:582-585.
[3] Sato Y.PCR amplification of CMS-specific mitochondrial nucleotide sequences to identify cytoplasmic genotypes of onion (Allium cepa L.) [J].Theoretical and Applied Genetics,1998,96(3/4):367-370.
[4] Engeike T,Terefe D,Tatlioglu T.A PCR-based marker system monitoring CMS-(S),CMS-(T) and (N)-cytoplasm in the onion (Allium cepa L.) [J].Theoretical and Applied Genetics,2003,107:162-167.
[5] Havey M J.Identification of cytoplasms using the polymerase chain reaction to aid in the extraction of maintainer lines from open-pollinated populations of onion [J].Theoretical and Applied Genetics,1995,90(2):263-268.
[6] Kim S,Yoon M.Comparison of mitochondrial and chloroplast genome segments from three onion (Allium cepa L.) cytoplasm types and identification of a trans-splicing intron of cox2 [J].Current Genetics,2010,56:177-188.
[7] Kim S,Lee E,Cho D Y,et al.Identification of a novel chimeric gene,orf725,and its use in development of a molecular marker for distinguishing three cytoplasm types in onion (Allium cepa L.) [J].Theoretical and Applied Genetics,2009,118(2):433-441.
[8] Kims S,Parkb J Y,Yangb T.Comparative analysis of the complete chloroplast genome sequences of a normal male-fertile cytoplasm and two different cytoplasms conferring cytoplasmic male sterility in onion (Allium cepa L.) [J].The Journal of Horticultural Science and Biotechnology,2015,90(4):459-468.
[9] Kim S.A codominant molecular marker in linkage disequilibrium with a restorer-of-fertility gene (Ms) and its application in reevaluation of inheritance of fertility restoration in onions [J].Molecular Breeding,2014,34(3):769-778.
[10] Huo Y M,Liu B J,Yang Y Y,et al.AcSKP1,a multiplex PCR-based co-dominant marker in complete linkage disequilibrium with the male-fertility restoration (Ms) locus,and its application in open-pollinated populations of onion [J].Euphytica,2015,204 (3) :711-722.
[11] Yang Y Y,Huo Y M,Miao J,et al.Identification of two SCAR markers co-segregated with the dominant Ms and recessive ms alleles in onion ( Allium cepa L.) [J].Euphytica,2013,190 (2):267-277.
[12] Bang H,Cho D Y,Yoo K S,et al.Development of simple PCR-based markers linked to the Ms locus,a restorer-of-fertility gene in onion (Allium cepa L.) [J].Euphytica,2011,179 (3):439-449.
[13] 潘美红,杨海峰,惠林冲,等.洋葱CMS-T型育性分子标记的筛选与鉴定[J].西北农业学报,2018,27(4):576-585.
[14] 陈振泰,缪美华,薛萍,等.大直径黄皮洋葱新品种连葱096的选育[J].长江蔬菜,2011(9):50-51.
[15] 李峰,曹丽蓉,刘培园,等.小麦品种渝麦13号细胞质类型的分子标记鉴定[J].中国农业大学学报,2018,23(10):1-7.
[16] 张俊英,蒋美艳,江建霞,等.甘蓝型油菜显性核不育系的分子标记辅助选择[J].核农学报,2018,32(11):2119-2125.
[17] 张宏根,仲崇元,司华,等.分子标记辅助选择改良C418对红莲型粳稻不育系的恢复力[J].中国水稻科学,2018,32(5):445-452.
[18] 宋伟,苏爱国,邢锦丰,等.京724玉米自交系S型细胞质雄性不育系分子标记辅助选育研究[J].玉米科学,2016,24(1):33-36,42.
[19] Bang H,Kim S,Park S O,et al.Development of a codominant CAPS marker linked to the Ms locus controlling fertility restoration in onion (Allium cepa L.) [J].Scientia Horticulturae,2013,153(4):42-49.
[20] Park J,Bang H,Cho D Y,et al.Construction of high-resolution linkage map of the Ms locus,a restorer-of-fertility gene in onion (Allium cepa L.) [J].Euphytica,2013,192 (2):267-278.
[21] Kim S,Kim C W,Minkyu P,et al.Identification of candidate genes associated with fertility restoration of cytoplasmic male-sterility in onion (Allium cepa L.) using a combination of bulked segregant analysis and RNA-seq [J].Theoretical and Applied Genetics,2015,128 (11):2289-2299.
[22] Gokce A F,Havey M J.Linkage equilibrium among tightly linked RFLPs and the Ms locus in open-pollinated onion populations [J].Journal of the American Society for Horticultural Science,2002,127 (6):944-946.
[23] Gokce A F,Mccallum J,Sato Y,et al.Molecular tagging of the Ms locus in onion [J].Journal of the American Society for Horticultural Science,2002,127 (4):576-582.
[24] 陈振泰,薛萍,缪美华,等.紫皮洋葱新品种连葱11号的选育[J].长江蔬菜,2016(20):52-54.
[25] 倪西源,黄吉祥,柳寒,等.应用分子标记辅助选育甘蓝型油菜杂交种的可行性[J].浙江大学学报:农业与生命科学版,2017,43(2):173-182.