粳稻BT型细胞质雄性不育恢复基因功能标记的开发与应用
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摘要
为提高粳稻BT型细胞质雄性不育恢复基因的选择效率,在前人克隆的基础上,根据其恢复系与不育系在Rf1a位点574bp的碱基插入/缺失,设计出功能标记InDel-Rf1a。利用该标记对不同地区来源的72份籼、粳稻材料进行检测,结果表明所有的常规籼稻品种、恢复系及保持系在Rf1a位点并不存在缺失,其基因型为Rf1aRf1a。这些材料对粳稻BT型细胞质雄性不育系具有恢复育性的作用;而绝大多数常规粳稻品种(爱知106和伊粳12号除外)在该位点存在缺失,其基因型为rf1arf1a,它们对BT型细胞质雄性不育系具有保持不育的能力。同时,为进一步验证该标记对不同基因型的检测效果,利用其对粳稻恢复系、不育系、杂交组合以及863A/宁恢8号F2分离群体的DNA进行扩增,根据其电泳带型可准确区分出Rf1a位点的3种基因型。
To improve the selection efficiency of restoring gene for BT-type cytoplasmic male sterility(CMS) in japonica rice,we designed a functional marker InDel-Rf1a based on a 574 bp insertion/deletion(InDel) in Rf1a locus between japonica restorer line and CMS line reported by previous studies and used it to identify the genotypes of Rf1a locus in 72 indica or japonica rice materials.The results showed that there was no 574 bp deletion in indica conventional rice varieties,restorer and maintain lines and they could restore the fertility of BT-type CMS in japonica rice,with genotypes of Rf1aRf1a;However,the same deletion was detected in most of japonica conventional rice varieties,except in Aichi 106 and Yijing 12,and they had the ability to maintain sterility of BT-type CMS lines,with genotypes of Rf1aRf1a.For further verification of the detection effect with this marker for different genotypes in this locus,we used it to amplify the DNA of different japonica rice restorer lines,CMS lines,hybrid F1 and the F2 population derived from a cross between 863A and Ninghui 8,and three genotypes in Rf1a locus could be distinguished distinctly by gel electrophoresis.
To improve the selection efficiency of restoring gene for BT-type cytoplasmic male sterility(CMS) in japonica rice,we designed a functional marker InDel-Rf1a based on a 574 bp insertion/deletion(InDel) in Rf1a locus between japonica restorer line and CMS line reported by previous studies and used it to identify the genotypes of Rf1a locus in 72 indica or japonica rice materials.The results showed that there was no 574 bp deletion in indica conventional rice varieties,restorer and maintain lines and they could restore the fertility of BT-type CMS in japonica rice,with genotypes of Rf1aRf1a;However,the same deletion was detected in most of japonica conventional rice varieties,except in Aichi 106 and Yijing 12,and they had the ability to maintain sterility of BT-type CMS lines,with genotypes of Rf1aRf1a.For further verification of the detection effect with this marker for different genotypes in this locus,we used it to amplify the DNA of different japonica rice restorer lines,CMS lines,hybrid F1 and the F2 population derived from a cross between 863A and Ninghui 8,and three genotypes in Rf1a locus could be distinguished distinctly by gel electrophoresis.
引文
[1]张峭,赵俊晔.中国稻米供需分析与展望.农业展望,2007(1):9-14.
[2]吴乐,邹文涛.我国稻谷消费中长期趋势分析.农业技术经济,2011,5:87-96.
[3]邓华风,何强,舒服,等.中国杂交粳稻研究现状与对策.杂交水稻,2006,21(1):1-6.
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[5]袁隆平,陈洪新.杂交水稻育种栽培学.长沙:湖南科学技术出版社,1988:83-84.
[6]李铮友.滇型杂交水稻育种.昆明:云南科技出版社,2000:1-15.
[7]贺和初.滇一型和BT型杂交稻育性遗传和不育机理研究.云南农业大学学报,1988,3(1):54-68.
[8]王才林,汤玉庚.我国杂交粳稻育种的现状与展望.中国农业科学,1989,22(5):8-13.
[9]Akagi H,Yokozeki Y,Inagaki A,et al.A codominant DNAmarker closely linked to the rice nuclear restorer gene,Rf-1,identified with inter-SSR fingerprinting.Genome,1996,39(6):1205-1209.
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[11]Komori T,Yamamoto T,Takemori N,et al.Fine geneticmapping of the nuclear gene,Rf-1,that restores the BT-typecytoplasmic male sterility in rice(Oryza sativa L.)by PCR-based markers.Euphytica,2003,129(2):241-247.
[12]Akagi H,Nakamura A,Yokozeki-Misono Y,et al.Positionalcloning of the rice Rf-1 gene,a restorer of BT-type cytoplas-mic male sterility that encodes a mitochondria-targeting PPRprotein.Theor Appl Genet,2004,108(8):1449-1457.
[13]Tan X L,Tan Y L,Zhao Y H,et al.Identification of the Rfgene conferring fertility restoration of the CMS Dian-type 1inrice by using simple sequence repeat markers and advanced in-bred lines of restorer and maintainer.Plant Breeding,2004,123(4):338-341.
[14]程宝山,洪德林,万志兵,等.与粳稻BT型细胞质育性恢复基因连锁的实用SSR标记的筛选.南京农业大学学报,2011,34(1):1-7.
[15]Wang Z H,Zou Y J,Li X Y,et al.Cytoplasmic male sterilityof rice with Boro II cytoplasm is caused by a cytotoxic peptideand is restored by two related PPR motif genes via distinctmodes of mRNA silencing.Plant Cell,2006,18(3):676-687.
[16]Dellaporta S L,Wood J,Hicks J B.A plant DNA miniprepa-ration:VersionⅡ.Plant Mol Biol Rep,1983,1(1):19-21.
[17]Chen X,Temnykh S,Xu Y,et al.Development of a microsat-ellite framework map providing genome-wide coverage in rice(Oryza sativa L.).Theor Appl Genet,1997,95(4):553-567.
[18]汤述翥,张宏根,朱正斌,等.红莲型不育细胞质应用于粳稻杂种优势的思考与初探.西南农业学报,2009,22(4):19-20.
[19]罗盛财,杨聚宝.BT型光身不育系—包珍兰A选育初报.杂交水稻,1989(4):18-19.
[20]张群宇,刘耀光,张桂权,等.野败型水稻细胞质雄性不育恢复基因Rf-4的分子标记定位.遗传学报,2002,29(4):1001-1004.
[21]Fujii S,Toriyama K.Molecular mapping of the fertility re-storer gene for ms-CW-type cytoplasmic male sterility of rice.Theor Appl Genet,2005,111(4):696-701.
[22]李亮杰,周海鹏,占小登,等.水稻印尼水田谷型细胞质雄性不育恢复系R68的恢复基因初步定位.中国水稻科学,2007,21(5):547-549.
[23]Guo J X,Liu Y G.The genetic and molecular basis of cyto-plasmic male sterility and fertility restoration in rice.Chin SciBull,2009,54(14):2404-2409.
[24]何小平,刘永柱,郭涛,等.水稻细胞质雄性不育恢复基因定位及相互关系研究进展.中国农学通报,2011,27(15):1-5.
[25]Hu J,Wang K,Huang W C,et al.The rice pentatricopeptiderepeat protein RF5restores fertility in Hong-lian cytoplasmicmale-sterile lines via a complex with the glycine-rich proteinGRP162.Plant Cell,2012,24(1):109-122.
[26]Sugiura N,Tsuji T,Fujii K,et al.Molecular marker-assistedselection in a recurrent backcross breeding for the incorpora-tion of resistance to rice stripe virus and panicle blast in rice(Oryza sativa L.).Breeding Res,2004,6(3):143-148.
[27]吐尔干江,陈疆,沙达尔,等.高产优质水稻新品种—伊粳12号.新疆农垦科技,2006(1):40-41.
[28]李建红,洪德林.新选粳稻BT型同质恢复系农艺和品质性状的配合力研究.作物学报,2005,31(7):851-857.
[29]闫双勇,马忠友,范俊山,等.我国杂交粳稻恢复系的遗传多样性.贵州农业科学,2011,39(10):1-4.
[30]李爱宏,戴正元,季红娟,等.不同基因型水稻种质对黑条矮缩病抗性的初步分析.扬州大学学报:农业与生命科学版,2008,29(3):18-22.
[31]王才林.江苏省杂交粳稻育种的现状、问题与对策.西南农业学报,2009,22(4):1165-1169.
[32]姚姝,张亚东,朱镇,等.粳稻恢复系条纹叶病抗性的定向改良.杂交水稻,2010(S1):264-267.
[2]吴乐,邹文涛.我国稻谷消费中长期趋势分析.农业技术经济,2011,5:87-96.
[3]邓华风,何强,舒服,等.中国杂交粳稻研究现状与对策.杂交水稻,2006,21(1):1-6.
[4]邓华凤.中国杂交粳稻.北京:中国农业出版社,2008:11-12.
[5]袁隆平,陈洪新.杂交水稻育种栽培学.长沙:湖南科学技术出版社,1988:83-84.
[6]李铮友.滇型杂交水稻育种.昆明:云南科技出版社,2000:1-15.
[7]贺和初.滇一型和BT型杂交稻育性遗传和不育机理研究.云南农业大学学报,1988,3(1):54-68.
[8]王才林,汤玉庚.我国杂交粳稻育种的现状与展望.中国农业科学,1989,22(5):8-13.
[9]Akagi H,Yokozeki Y,Inagaki A,et al.A codominant DNAmarker closely linked to the rice nuclear restorer gene,Rf-1,identified with inter-SSR fingerprinting.Genome,1996,39(6):1205-1209.
[10]Ichikawa N,Kishimoto N,Inagaki A,et al.A rapid PCR-ai-ded selection of a rice line containing the Rf-1 gene which is in-volved in restoration of the cytoplasmic male sterility.MolBreeding,1997,3(3):195-202.
[11]Komori T,Yamamoto T,Takemori N,et al.Fine geneticmapping of the nuclear gene,Rf-1,that restores the BT-typecytoplasmic male sterility in rice(Oryza sativa L.)by PCR-based markers.Euphytica,2003,129(2):241-247.
[12]Akagi H,Nakamura A,Yokozeki-Misono Y,et al.Positionalcloning of the rice Rf-1 gene,a restorer of BT-type cytoplas-mic male sterility that encodes a mitochondria-targeting PPRprotein.Theor Appl Genet,2004,108(8):1449-1457.
[13]Tan X L,Tan Y L,Zhao Y H,et al.Identification of the Rfgene conferring fertility restoration of the CMS Dian-type 1inrice by using simple sequence repeat markers and advanced in-bred lines of restorer and maintainer.Plant Breeding,2004,123(4):338-341.
[14]程宝山,洪德林,万志兵,等.与粳稻BT型细胞质育性恢复基因连锁的实用SSR标记的筛选.南京农业大学学报,2011,34(1):1-7.
[15]Wang Z H,Zou Y J,Li X Y,et al.Cytoplasmic male sterilityof rice with Boro II cytoplasm is caused by a cytotoxic peptideand is restored by two related PPR motif genes via distinctmodes of mRNA silencing.Plant Cell,2006,18(3):676-687.
[16]Dellaporta S L,Wood J,Hicks J B.A plant DNA miniprepa-ration:VersionⅡ.Plant Mol Biol Rep,1983,1(1):19-21.
[17]Chen X,Temnykh S,Xu Y,et al.Development of a microsat-ellite framework map providing genome-wide coverage in rice(Oryza sativa L.).Theor Appl Genet,1997,95(4):553-567.
[18]汤述翥,张宏根,朱正斌,等.红莲型不育细胞质应用于粳稻杂种优势的思考与初探.西南农业学报,2009,22(4):19-20.
[19]罗盛财,杨聚宝.BT型光身不育系—包珍兰A选育初报.杂交水稻,1989(4):18-19.
[20]张群宇,刘耀光,张桂权,等.野败型水稻细胞质雄性不育恢复基因Rf-4的分子标记定位.遗传学报,2002,29(4):1001-1004.
[21]Fujii S,Toriyama K.Molecular mapping of the fertility re-storer gene for ms-CW-type cytoplasmic male sterility of rice.Theor Appl Genet,2005,111(4):696-701.
[22]李亮杰,周海鹏,占小登,等.水稻印尼水田谷型细胞质雄性不育恢复系R68的恢复基因初步定位.中国水稻科学,2007,21(5):547-549.
[23]Guo J X,Liu Y G.The genetic and molecular basis of cyto-plasmic male sterility and fertility restoration in rice.Chin SciBull,2009,54(14):2404-2409.
[24]何小平,刘永柱,郭涛,等.水稻细胞质雄性不育恢复基因定位及相互关系研究进展.中国农学通报,2011,27(15):1-5.
[25]Hu J,Wang K,Huang W C,et al.The rice pentatricopeptiderepeat protein RF5restores fertility in Hong-lian cytoplasmicmale-sterile lines via a complex with the glycine-rich proteinGRP162.Plant Cell,2012,24(1):109-122.
[26]Sugiura N,Tsuji T,Fujii K,et al.Molecular marker-assistedselection in a recurrent backcross breeding for the incorpora-tion of resistance to rice stripe virus and panicle blast in rice(Oryza sativa L.).Breeding Res,2004,6(3):143-148.
[27]吐尔干江,陈疆,沙达尔,等.高产优质水稻新品种—伊粳12号.新疆农垦科技,2006(1):40-41.
[28]李建红,洪德林.新选粳稻BT型同质恢复系农艺和品质性状的配合力研究.作物学报,2005,31(7):851-857.
[29]闫双勇,马忠友,范俊山,等.我国杂交粳稻恢复系的遗传多样性.贵州农业科学,2011,39(10):1-4.
[30]李爱宏,戴正元,季红娟,等.不同基因型水稻种质对黑条矮缩病抗性的初步分析.扬州大学学报:农业与生命科学版,2008,29(3):18-22.
[31]王才林.江苏省杂交粳稻育种的现状、问题与对策.西南农业学报,2009,22(4):1165-1169.
[32]姚姝,张亚东,朱镇,等.粳稻恢复系条纹叶病抗性的定向改良.杂交水稻,2010(S1):264-267.