利用Indel标记鉴定‘豫甘3号’结球甘蓝种子纯度
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摘要
本研究利用InDel分子标记技术对结球甘蓝"豫甘3号"杂交种进行了纯度快速鉴定。从395对InDel引物中筛选出6对具有双亲互补条带、差异明显的引物,分别是BrID101201,BrID90436,BrID90043,BrID101009,BrID10229和BrID101085。用该6个标记对126株‘豫甘3号’杂交种进行纯度分析,结果表明,杂交种纯度分别为94.4%、96.8%、96%、96%、96%和96%,其中122株的鉴定结果完全相同,鉴定结果一致率达到96.8%。因此,筛选出的6个InDel标记均可用于结球甘蓝‘豫甘3号’种子纯度的快速鉴定。
In this research,the InDel marker was used to quickly identify the purity of cabbage hybrid 'Yugan No.3'.Six primer pairs,namely,BrID101201,BrID90436,BrID90043,BrID101009,BrID10229 and BrID101085,with obvious different bands between parents and with codominant bands in hybrid‘Yugan No.3',were screened from 395 pairs of InDel primers.Then,the six InDel markers were chosen to test the purity of 126 'Yugan No.3'hybrids.The results showed that the seed purity were 94.4% 、96.8% 、96% 、96% 、96% and 96%,respectively,of which the genotype of 122 seedlings tested by the six InDel markers were totally identical,and the concordance of molecular test was high to 96.8%.Therefore,the screened six InDel markers could be used to quickly identify the seed purity of cabbage‘Yugan No.3'.
In this research,the InDel marker was used to quickly identify the purity of cabbage hybrid 'Yugan No.3'.Six primer pairs,namely,BrID101201,BrID90436,BrID90043,BrID101009,BrID10229 and BrID101085,with obvious different bands between parents and with codominant bands in hybrid‘Yugan No.3',were screened from 395 pairs of InDel primers.Then,the six InDel markers were chosen to test the purity of 126 'Yugan No.3'hybrids.The results showed that the seed purity were 94.4% 、96.8% 、96% 、96% 、96% and 96%,respectively,of which the genotype of 122 seedlings tested by the six InDel markers were totally identical,and the concordance of molecular test was high to 96.8%.Therefore,the screened six InDel markers could be used to quickly identify the seed purity of cabbage‘Yugan No.3'.
引文
Chen C.,Zhang X.T.,Cheng F.,Zhuang M.,Liu Y.M.,Yang L.M.,Zhang Y.Y.,and Fang Z.Y.,2011,Establishment of SSRfingerprinting on autumn Cabbage hybrids and their parents,Yuanyi Xuebao(Acta Horticulturae Sinica),38(1):159-164(陈琛,张兴桃,程斐,庄木,刘玉梅,杨丽梅,张扬勇,方智远,2011,秋甘蓝品种的SSR指纹图谱的构建,园艺学报,38(1):159-164)
Chu Z.Z.,Guo H.B.,Zeng D.C.,and Liu Y.G.,2016,Development of 295 In Del markers for Indica and Japanica rice,Zuowu Xuebao(Acta Agronomica Sinica),42(6):932-941(初志战,郭海滨,曾栋昌,刘耀光,2016,籼粳稻基因组295个In Del标记的开发,作物学报,42(6):932-941)
Fang S.G.,Zeng X.L.,Chen W.H.,and Chen X.,2009,Identification of seed purit y of hybrid Chinese cabbage Zhongshu No.5 by RAPD markers,Changjiang Shucai(Journal of Changjiang Vegetables),42(6):19-21(方淑桂,曾小玲,陈文辉,陈铣,2009,大白菜杂交种中熟5号纯度的RAPD鉴定,长江蔬菜,42(6):19-21)
Feng F.J.,Luo L.J.,Li Y.,Zhou L.G.,Xu X.Y.,Wu J.H.,Cheng H.W.,Chen L.,and Mei H.W.,2005,Comparative analysis of polymorphism of In Del and SSR markers in rice,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),3(5):725-730(冯芳君,罗利军,李荧,周立国,徐小艳,吴金红,陈宏伟,陈亮,梅捍卫,2005,水稻In Del和SSR标记多态性的比较分析,分子植物育种,3(5):725-730)
Huang J.D.,Shao G.R.,Chen W.H.,Fang S.G.,and Zhong K.Q.,2012,Genetic purity test of Cabbage Xiahua No.2 using SRAP markers,Zhongguo Guacai,25(2):15-17(黄建都,邵贵荣,陈文辉,方淑桂,钟开勤,2012,SRAP标记在夏华2号甘蓝种子纯度检测中的应用,中国瓜菜,25(2):15-17)
Li X.J.,Wang Y.H.,Xuan S.X.,Zhao J.J.,and Gu A.X.,2013,Screening of specific SSR markers on different linkage groups of Chinese Cabbage compared with Cabbage,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),14(4):694-698(李雪姣,王彦华,轩淑欣,赵建军,顾爱侠,2013,大白菜相对于结球甘蓝连锁群特异SSR标记的筛选,植物遗传资源学报,14(4):694-698)
Liang B.P.,Yuan Y.X.,Piao F.Z.,Zhang X.W.,Jiang W.S.,Liang S.,Yao Q.J.,Zhang Q.,and Zhao Y.Y.,2012,The optimization of technique of native polyacrylamide gel electrophyresis(Native-PAGE),Henan Nongye Kexue(Journal of Henan Agricultural Sciences),41(5):129-132(梁宝萍,原玉香,朴凤植,张晓伟,蒋武生,梁爽,姚秋菊,张强,赵艳艳,2012,大白菜非变性聚丙烯酰胺凝胶电泳技术的优化,河南农业科学,41(5):129-132)
Liang S.,Yuan Y.X.,Zhang X.W.,Xue Y.G.,Yao Q.J.,Jiang W.S.,Tian B.M.,Zhang Q.,and Zhao Y.Y.,2012,Optimization of In Del-PCR reaction system for Chinese Cabbage(Brassica rapa L.ssp.pekinensis),Henan Nongye Kexue(Journal of Henan Agricultural Sciences),41(9):110-113(梁爽,原玉香,张晓伟,薛银鸽,姚秋菊,蒋武生,田保明,张强,赵艳艳,2012,大白菜In Del-PCR反应体系的优化,河南农业科学,41(9):110-113)
Liu S.Y.,Liu Y.M.,Yang X.H.,Tong C.B.,Edwards D.,Parkin I.A.P.,Zhao M.X.,Ma J.X.,Yu J.Y.,Huang S.M.,Wang X.Y.,Wang J.Y.,and Lu K.,2014,The Brassica oleracea genome reveals the asymmetrical evolution of polyploid genomes,Nature Communications,5(9):3930-3934
Miao S.P.,2017,Application of SSR molecular marker technique in hybrid purity identification of violet Cabbage of Zigan NO.1,Xinyang Nonglin Xueyuan Xuebao(Journal of Xinyang College of Agriculture and Forestry),27(2):88-90(苗素平,2017,SSR标记技术在“紫甘1号”紫甘蓝杂交种纯度鉴定中的应用,信阳农林学院学报,27(2):88-90)
Qie L.J.,Shen S.X.,Xuan S.X.,Wang Y.H.,Chen X.P.,Zhang C.H.,Li X.F.,and Luo S.X.,2007,Karyotype analysis of Chinese Cabbage and Cabbage by genome in situ hybridization,Yuanyi Xuebao(Acta Horticulturae Sinica),34(6):1459-1464(郄丽娟,申书兴,轩淑欣,王彦华,陈雪平,张成合,李晓峰,罗双霞,2007,大白菜和结球甘蓝基因组原位杂交及核型分析,园艺学报,34(6):1459-1464)
Tian L.,Peng R.D.,Wang H.,and Jia X.H.,2010,Research on Chinese cabbage varieties purity identification using SSRmarker,Beijing Nongye(Beijing Agricultural),9(4):23-28(田雷,彭瑞迪,王辉,贾希海,2010,应用SSR技术进行白菜品种纯度鉴定的研究,北京农业,9(4):23-28)
Wang T.T.,Zhang S.J.,Zhang S.F.,Li F.,Zhang H.,and Sun R.F.,2012,Mapping of clubroot resistance gene in Chinese Cabbage,Zhongguo Shucai(China Vegetables),1(14):31-35(王彤彤,张淑江,章时蕃,李菲,张慧,孙日飞,2012,大白菜根肿病抗性基因的标记和定位,中国蔬菜,1(14):31-35)
Wang Y.,Sun S.L.,Liu B..,Wang H.,Deng J.,Liao Y.C.,Wang41(8):1699-1706(朱东旭,王彦华,赵建军,刘博,李晓峰,轩淑欣,申书兴,2014,结球甘蓝相对于大白菜连锁群特异In Del标记的建立及应用,园艺学报,41(8):1699-1706)
Chu Z.Z.,Guo H.B.,Zeng D.C.,and Liu Y.G.,2016,Development of 295 In Del markers for Indica and Japanica rice,Zuowu Xuebao(Acta Agronomica Sinica),42(6):932-941(初志战,郭海滨,曾栋昌,刘耀光,2016,籼粳稻基因组295个In Del标记的开发,作物学报,42(6):932-941)
Fang S.G.,Zeng X.L.,Chen W.H.,and Chen X.,2009,Identification of seed purit y of hybrid Chinese cabbage Zhongshu No.5 by RAPD markers,Changjiang Shucai(Journal of Changjiang Vegetables),42(6):19-21(方淑桂,曾小玲,陈文辉,陈铣,2009,大白菜杂交种中熟5号纯度的RAPD鉴定,长江蔬菜,42(6):19-21)
Feng F.J.,Luo L.J.,Li Y.,Zhou L.G.,Xu X.Y.,Wu J.H.,Cheng H.W.,Chen L.,and Mei H.W.,2005,Comparative analysis of polymorphism of In Del and SSR markers in rice,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),3(5):725-730(冯芳君,罗利军,李荧,周立国,徐小艳,吴金红,陈宏伟,陈亮,梅捍卫,2005,水稻In Del和SSR标记多态性的比较分析,分子植物育种,3(5):725-730)
Huang J.D.,Shao G.R.,Chen W.H.,Fang S.G.,and Zhong K.Q.,2012,Genetic purity test of Cabbage Xiahua No.2 using SRAP markers,Zhongguo Guacai,25(2):15-17(黄建都,邵贵荣,陈文辉,方淑桂,钟开勤,2012,SRAP标记在夏华2号甘蓝种子纯度检测中的应用,中国瓜菜,25(2):15-17)
Li X.J.,Wang Y.H.,Xuan S.X.,Zhao J.J.,and Gu A.X.,2013,Screening of specific SSR markers on different linkage groups of Chinese Cabbage compared with Cabbage,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),14(4):694-698(李雪姣,王彦华,轩淑欣,赵建军,顾爱侠,2013,大白菜相对于结球甘蓝连锁群特异SSR标记的筛选,植物遗传资源学报,14(4):694-698)
Liang B.P.,Yuan Y.X.,Piao F.Z.,Zhang X.W.,Jiang W.S.,Liang S.,Yao Q.J.,Zhang Q.,and Zhao Y.Y.,2012,The optimization of technique of native polyacrylamide gel electrophyresis(Native-PAGE),Henan Nongye Kexue(Journal of Henan Agricultural Sciences),41(5):129-132(梁宝萍,原玉香,朴凤植,张晓伟,蒋武生,梁爽,姚秋菊,张强,赵艳艳,2012,大白菜非变性聚丙烯酰胺凝胶电泳技术的优化,河南农业科学,41(5):129-132)
Liang S.,Yuan Y.X.,Zhang X.W.,Xue Y.G.,Yao Q.J.,Jiang W.S.,Tian B.M.,Zhang Q.,and Zhao Y.Y.,2012,Optimization of In Del-PCR reaction system for Chinese Cabbage(Brassica rapa L.ssp.pekinensis),Henan Nongye Kexue(Journal of Henan Agricultural Sciences),41(9):110-113(梁爽,原玉香,张晓伟,薛银鸽,姚秋菊,蒋武生,田保明,张强,赵艳艳,2012,大白菜In Del-PCR反应体系的优化,河南农业科学,41(9):110-113)
Liu S.Y.,Liu Y.M.,Yang X.H.,Tong C.B.,Edwards D.,Parkin I.A.P.,Zhao M.X.,Ma J.X.,Yu J.Y.,Huang S.M.,Wang X.Y.,Wang J.Y.,and Lu K.,2014,The Brassica oleracea genome reveals the asymmetrical evolution of polyploid genomes,Nature Communications,5(9):3930-3934
Miao S.P.,2017,Application of SSR molecular marker technique in hybrid purity identification of violet Cabbage of Zigan NO.1,Xinyang Nonglin Xueyuan Xuebao(Journal of Xinyang College of Agriculture and Forestry),27(2):88-90(苗素平,2017,SSR标记技术在“紫甘1号”紫甘蓝杂交种纯度鉴定中的应用,信阳农林学院学报,27(2):88-90)
Qie L.J.,Shen S.X.,Xuan S.X.,Wang Y.H.,Chen X.P.,Zhang C.H.,Li X.F.,and Luo S.X.,2007,Karyotype analysis of Chinese Cabbage and Cabbage by genome in situ hybridization,Yuanyi Xuebao(Acta Horticulturae Sinica),34(6):1459-1464(郄丽娟,申书兴,轩淑欣,王彦华,陈雪平,张成合,李晓峰,罗双霞,2007,大白菜和结球甘蓝基因组原位杂交及核型分析,园艺学报,34(6):1459-1464)
Tian L.,Peng R.D.,Wang H.,and Jia X.H.,2010,Research on Chinese cabbage varieties purity identification using SSRmarker,Beijing Nongye(Beijing Agricultural),9(4):23-28(田雷,彭瑞迪,王辉,贾希海,2010,应用SSR技术进行白菜品种纯度鉴定的研究,北京农业,9(4):23-28)
Wang T.T.,Zhang S.J.,Zhang S.F.,Li F.,Zhang H.,and Sun R.F.,2012,Mapping of clubroot resistance gene in Chinese Cabbage,Zhongguo Shucai(China Vegetables),1(14):31-35(王彤彤,张淑江,章时蕃,李菲,张慧,孙日飞,2012,大白菜根肿病抗性基因的标记和定位,中国蔬菜,1(14):31-35)
Wang Y.,Sun S.L.,Liu B..,Wang H.,Deng J.,Liao Y.C.,Wang41(8):1699-1706(朱东旭,王彦华,赵建军,刘博,李晓峰,轩淑欣,申书兴,2014,结球甘蓝相对于大白菜连锁群特异In Del标记的建立及应用,园艺学报,41(8):1699-1706)