黄化苗测交法检测玉米单倍体诱导率
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摘要
以本课题组选育出的纯合黄化自交系Z52作母本,与课题组选育的6份改良诱导系杂交,进行诱导能力鉴定,评价人工遗传标记筛选的准确率,并通过观察叶片单位面积气孔数进行验证。结果表明6份改良诱导系的诱导率为2.02%～5.62%,平均为3.66%。人工筛选的平均假阳性率为7.4%,平均假阴性率为0.92%。单位面积气孔数的观察结果表明,黄化单倍体单位面积平均气孔数为36.56个,极显著高于对照的黄化二倍体(21.78个)以及杂交二倍体(18.56个)。本研究期望为玉米诱导系诱导率的准确测定提供一定参考,并为玉米单倍体育种工作提供科学依据。
The pure and yel low inbred line Z52, which was bred by our research group, was used as female parent. It was crossbred with 6 improved induction lines which was bred by our group to identify the ability of induction. At the same time, the accuracy of artificial genetic marker screening was verificated by observing the stomatal number per unit area of leaves. The results showed that the induction rate of 6 improved lines was 2.02%～5.62%, with an average of 3.66%. The average false positive rate and false negative rate of artificial screening were7.4% and 0.92%, respectively. The detection of stomatal per unit area indicated that the average stomatal number per unit area of yellow haploid was 36.56, which was significantly higher than that of yellow diploid(21.78) and hybrid diploid(18.56). This study was expected to provide scientific basis for accurate identification of the induction rate of maize haploid inducer, and provide scientific reference for maize haploid breeding.
The pure and yel low inbred line Z52, which was bred by our research group, was used as female parent. It was crossbred with 6 improved induction lines which was bred by our group to identify the ability of induction. At the same time, the accuracy of artificial genetic marker screening was verificated by observing the stomatal number per unit area of leaves. The results showed that the induction rate of 6 improved lines was 2.02%～5.62%, with an average of 3.66%. The average false positive rate and false negative rate of artificial screening were7.4% and 0.92%, respectively. The detection of stomatal per unit area indicated that the average stomatal number per unit area of yellow haploid was 36.56, which was significantly higher than that of yellow diploid(21.78) and hybrid diploid(18.56). This study was expected to provide scientific basis for accurate identification of the induction rate of maize haploid inducer, and provide scientific reference for maize haploid breeding.
引文
Cai Y.,2011,Creation and genetic analysis of the different ploidy maize-Z.perennis hybrids,Thesis for M.S.,College of A-gronomy,Sichuan Agricultural University,Supervisor:Tang Q.L.,pp.40-42(蔡毅,2011,玉米与四倍体多年生类玉米不同倍性杂种的创制与遗传分析,硕士学位论文,四川农业大学农学院,导师:唐祈林,pp.40-42)
Chen S.J.,and Song T.M.,2003,Identification haploid with high oil xenia effect in maize,Zuowu Xuebao(Acta Agronomica Sinica),29(4):587-590(陈绍江,宋同明,2003,利用高油分的花粉直感效应鉴别玉米单倍体,作物学报,29(4):587-590)
Du H.W.,Dai J.R.,and Li J.S.,2010,Study proceeding in haploid breeding of maize,Yumi Kexue(Journal of Maize Sciences),18(6):1-7(杜何为,戴景瑞,李建生,2010,玉米单倍体育种研究进展,玉米科学,18(6):1-7)
Li L.,Xu X.W.,Jin W.W.,and Chen S.J.,2009,Morphological and molecular evidences for DNA introgression in haploid induction via a high oil inducer CAUHOI in maize,Planta,230(2):367-376
Liu J.,2015,The study of automatic identification of in vivo-induced haploid seeds and new haploid inducer lines selection in maize,Dissertation for Ph.D.,China Agricultural University,Supervisor:Chen S.J.,pp.11-14(刘金,2015,玉米单倍体自动化鉴别与新型诱导系选育研究,博士学位论文,中国农业大学,导师:陈绍江,pp.11-14)
Ren X.J.,Yang W.,Cui X.Y.,Yang W.G.,and Wu C.S.,2015Induction rate of maize haploid inducer JS9,Yumi Kexue(Journal of Maize Sciences),23(3):34-38(任雪娇,杨巍,崔学宇,杨伟光,吴春胜,2015,玉米单倍体诱导系JS9诱导率研究,玉米科学,23(3):34-38)
Rober F.K.,Gordillo G.A.,and Geiger H.H.,2005,In vivo haploid induction in maize-performance of new inducers and significance of doubles haploid lines in hybrid breeding Maydica,50(3):275-283
Rotarenco V.A.,Dicu G.,State D.,and Fuia S.,2010,New inducers of maternal haploids in maize,Maize Genet.Coop Newslett.,84:15
Shi X.F.,Peng J.G.,and Wang L.P.,2009,Identifying chromosome ploidy by the number of stomatal guard cell chloroplast in watermelon,Hunan Nongye Daxue Xuebao(Journa of Hunan Agricultural University),35(6):640-642,663(施先锋,彭金光,汪李平,2009,用西瓜叶片气孔保卫细胞叶绿体数鉴定西瓜染色体倍性,湖南农业大学学报,35(6)640-642,663)
Wang P.,Yu F.,Huang Y.J.,Chen S.Y.,and Deng Z.H.,2015Physical localization of 5S r DNA on the chromosomes of different ploidy Saccharum spontaneum,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(8)1761-1768(王平,余凡,黄永吉,陈胜烨,邓祖湖,2015,5Sr DNA在不同倍性割手密染色体上的物理定位,基因组学与应用生物学,34(8):1761-1768)
Wang X.Y.,Shan X.H.,Wu Y.,Su S.Z.,Li S.P.,Liu H.K.,Han JY.,Xue C.M.,and Yuan Y.P.,2016,i TRAQ-based quantitative proteomic analysis reveals new metabolic pathways responding to chilling stress in maize seedlings,J.Proteomics146:14-24
Xu G.L.,Dai Y.X.,Liu X.D.,Ren J.,Li S.H.,Yu M.Y.,and Cai Z.,2012,Research on the induced rate and the doubling rates of haploid in maize,Yumi Kexue(Journal of Maize Sciences),20(2):1-5(徐国良,代玉仙,刘晓丹,任军,李淑华,于明彦,才卓,2012,玉米单倍体诱导率和加倍率研究,玉米科学,20(2):1-5)
Yang G.,Zou H.D.,Wu Y.,Liu H.K.,and Yuan Y.P.,2011,Identification and characterization of candidate genes involved in chilling responses in maize(Zea mays L.),Plant Cell Tiss.Org.,106(1):127-141
Yang W.,Ren X.J.,Cui X.Y.,Ci J.B.,Ren Q.B.,Yang W.G.,and Wu C.S.,2015,Induced rate of maize haploid inducer JS6-2,Xibei Nonglin Keji Daxue Xuebao(Journal of Northwest A&F University),43(7):89-94(杨巍,任雪娇,崔学宇,慈佳宾,任启彪,杨伟光,吴春胜,2015,玉米单倍体诱导系JS6-2诱导效果的研究,西北农林科技大学学报,43(7):89-94)
Yue Y.H.,Lu M.,Zhang J.X.,Ma Y.J.,Zhou X.D.,Zhao W.Q.,Wang S.P.,Zhang Z.J.,and Liu W.G.,2017,The breeding of Jiyou 101 haploid inducer with high frequency parthenogenesis in maize,Zuowu Zazhi(Crops),(3):35-38(岳尧海,路明,张建新,马英杰,周旭东,赵万庆,王绍平,张志军,刘文国,2017,玉米单倍体高频诱导系吉诱101号的选育,作物杂志,(3):35-38)
Zhang X.F.,Geng S.S.,and Chen B.,2009,Relationship between chromosome ploidy and stomatal characters of leaf in pepper,Jiangsu Nongye Xuebao(Jiangsu Journal of Agricultural Sciences),25(2):339-342(张晓芬,耿三省,陈斌,2009,辣椒染色体倍性与气孔性状的相关性,江苏农业学报,25(2):339-342)
Chen S.J.,and Song T.M.,2003,Identification haploid with high oil xenia effect in maize,Zuowu Xuebao(Acta Agronomica Sinica),29(4):587-590(陈绍江,宋同明,2003,利用高油分的花粉直感效应鉴别玉米单倍体,作物学报,29(4):587-590)
Du H.W.,Dai J.R.,and Li J.S.,2010,Study proceeding in haploid breeding of maize,Yumi Kexue(Journal of Maize Sciences),18(6):1-7(杜何为,戴景瑞,李建生,2010,玉米单倍体育种研究进展,玉米科学,18(6):1-7)
Li L.,Xu X.W.,Jin W.W.,and Chen S.J.,2009,Morphological and molecular evidences for DNA introgression in haploid induction via a high oil inducer CAUHOI in maize,Planta,230(2):367-376
Liu J.,2015,The study of automatic identification of in vivo-induced haploid seeds and new haploid inducer lines selection in maize,Dissertation for Ph.D.,China Agricultural University,Supervisor:Chen S.J.,pp.11-14(刘金,2015,玉米单倍体自动化鉴别与新型诱导系选育研究,博士学位论文,中国农业大学,导师:陈绍江,pp.11-14)
Ren X.J.,Yang W.,Cui X.Y.,Yang W.G.,and Wu C.S.,2015Induction rate of maize haploid inducer JS9,Yumi Kexue(Journal of Maize Sciences),23(3):34-38(任雪娇,杨巍,崔学宇,杨伟光,吴春胜,2015,玉米单倍体诱导系JS9诱导率研究,玉米科学,23(3):34-38)
Rober F.K.,Gordillo G.A.,and Geiger H.H.,2005,In vivo haploid induction in maize-performance of new inducers and significance of doubles haploid lines in hybrid breeding Maydica,50(3):275-283
Rotarenco V.A.,Dicu G.,State D.,and Fuia S.,2010,New inducers of maternal haploids in maize,Maize Genet.Coop Newslett.,84:15
Shi X.F.,Peng J.G.,and Wang L.P.,2009,Identifying chromosome ploidy by the number of stomatal guard cell chloroplast in watermelon,Hunan Nongye Daxue Xuebao(Journa of Hunan Agricultural University),35(6):640-642,663(施先锋,彭金光,汪李平,2009,用西瓜叶片气孔保卫细胞叶绿体数鉴定西瓜染色体倍性,湖南农业大学学报,35(6)640-642,663)
Wang P.,Yu F.,Huang Y.J.,Chen S.Y.,and Deng Z.H.,2015Physical localization of 5S r DNA on the chromosomes of different ploidy Saccharum spontaneum,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(8)1761-1768(王平,余凡,黄永吉,陈胜烨,邓祖湖,2015,5Sr DNA在不同倍性割手密染色体上的物理定位,基因组学与应用生物学,34(8):1761-1768)
Wang X.Y.,Shan X.H.,Wu Y.,Su S.Z.,Li S.P.,Liu H.K.,Han JY.,Xue C.M.,and Yuan Y.P.,2016,i TRAQ-based quantitative proteomic analysis reveals new metabolic pathways responding to chilling stress in maize seedlings,J.Proteomics146:14-24
Xu G.L.,Dai Y.X.,Liu X.D.,Ren J.,Li S.H.,Yu M.Y.,and Cai Z.,2012,Research on the induced rate and the doubling rates of haploid in maize,Yumi Kexue(Journal of Maize Sciences),20(2):1-5(徐国良,代玉仙,刘晓丹,任军,李淑华,于明彦,才卓,2012,玉米单倍体诱导率和加倍率研究,玉米科学,20(2):1-5)
Yang G.,Zou H.D.,Wu Y.,Liu H.K.,and Yuan Y.P.,2011,Identification and characterization of candidate genes involved in chilling responses in maize(Zea mays L.),Plant Cell Tiss.Org.,106(1):127-141
Yang W.,Ren X.J.,Cui X.Y.,Ci J.B.,Ren Q.B.,Yang W.G.,and Wu C.S.,2015,Induced rate of maize haploid inducer JS6-2,Xibei Nonglin Keji Daxue Xuebao(Journal of Northwest A&F University),43(7):89-94(杨巍,任雪娇,崔学宇,慈佳宾,任启彪,杨伟光,吴春胜,2015,玉米单倍体诱导系JS6-2诱导效果的研究,西北农林科技大学学报,43(7):89-94)
Yue Y.H.,Lu M.,Zhang J.X.,Ma Y.J.,Zhou X.D.,Zhao W.Q.,Wang S.P.,Zhang Z.J.,and Liu W.G.,2017,The breeding of Jiyou 101 haploid inducer with high frequency parthenogenesis in maize,Zuowu Zazhi(Crops),(3):35-38(岳尧海,路明,张建新,马英杰,周旭东,赵万庆,王绍平,张志军,刘文国,2017,玉米单倍体高频诱导系吉诱101号的选育,作物杂志,(3):35-38)
Zhang X.F.,Geng S.S.,and Chen B.,2009,Relationship between chromosome ploidy and stomatal characters of leaf in pepper,Jiangsu Nongye Xuebao(Jiangsu Journal of Agricultural Sciences),25(2):339-342(张晓芬,耿三省,陈斌,2009,辣椒染色体倍性与气孔性状的相关性,江苏农业学报,25(2):339-342)