栽培稻-药用野生稻杂种F_1及回交后代的基因组原位杂交鉴定  

Identification of Oryza sativa×Oryza officinalis F1 and BackorossProgenies Using Genomic in situ Hybridization

作　　者：颜辉煌[1,2] 颜辉煌[1,2] 程祝宽[1] 刘国庆[1] 陈纯贤[1] 闵绍楷[2] 朱立煌 

机构地区：[1]中国科学院遗传研究所,北京100101 [2]中国水稻研究所,杭州310006

出　　处：《Acta Genetica Sinica》1999年第2期157-162,共6页

基　　金：国家科委水稻基因组计划项目;国家自然科学基金

摘　　要：以ｄｉｇｏｘｉｇｅｎｉｎ标记的药用野生稻（Ｏｒｙｚａ　ｏｆｆｉｃｉｎaｌｉｓ）总ＤＮＡ作探针，未标记的栽培稻（Ｏｒｙｚａ　ｓａｔｉｖａ）总ＤＮＡ封阻（１：１６），对栽培稻一药用野生稻Ｆｉ及回交后代材料根尖体细胞染色体制片进行基因组原位杂交，鉴定出３个双单体异源附加系和４个单体异源附加系。当标记探针用ａｎｔｉ－ｄｉｇｏｘｉｇｅｎｉｎ－ＰＯＤ和ＤＡＢ检测时，药用野生稻染色体显棕色，栽培稻染色体则因Ｇｉｅｍｓａ对染而显浅蓝色；标记探针用ａｎｔｉ－ｄｉｇｏｘｉｇｅｎｉｎ－ＦＩＴＣ检测时，药用野生稻染色体发出黄绿色荧光，而栽培稻染色体则因ＰＩ对染而发出红色荧光。在异源附加系的有丝分裂间期细胞中，药用野生稻染色质所占区域也很明显。由此看来，有可能利用ＧＩＳＨ技术来区分Ａ与稻属的其他基因组。ith digoxigenin-labelled total genomic DNA from O.officinalis as Frobe and 16-fold unlabelled total genomic DNA from O,sative as blocking,genomic in situhybridization was applied to somatic chromosome preparations of Oryza sativa×Oryzaofficinali, F1 hybrid and tackcross progenies, which led to the confirmation of threedouble and four monosomic alien addition lines.When anti-digoxigenin-POD and DABwere used for detection.the color of officinalis chromosome was brown, against lightbluc sativa chromosome from Giemas counterstain. When anti-digoxigenin-FITC wasused instead, officinalis chromosome appeared greenish- yellow while sativachromosome fluoresced red with PI counterstain.In the interphase of alien addition line officinalis chromatins were visealized as distinct areas. Hence it is likely to be alsofeasibie for discriminating genome A from other oryza genomes.

关 键 词：栽培稻 药用野生稻 异源附加系 基因组原位杂交 

分 类 号：S511.035.1[农业科学—作物学]