甘蓝rDNA及C_0t-1 DNA荧光原位杂交及其核型分析  

Karyotyping of Brassica oleracea L.Based on rDNA and C_0t-1 DNA Fluorescence in situ Hybridization

作　　者：王太霞[1] 吴春红[1] 黄进勇[2] 魏文辉[3] 

机构地区：[1]河南师范大学生命科学学院,河南新乡453007 [2]郑州大学生命科学学院,河南郑州450001 [3]中国农业科学院油料作物研究所,湖北武汉430062

出　　处：《武汉大学学报（理学版）》2006年第2期230-234,共5页Journal of Wuhan University:Natural Science Edition

基　　金：中国农业科学院油料作物研究所所长基金资助项目(200301);湖北省杰出青年基金资助项目(2005ABB028);武汉市青年科技晨光计划资助项目(20045006071-37)

摘　　要：为了探索一种高效可靠的芸薹属植物核型分析方法,本文以甘蓝品种黑叶小平头为实验材料,从其基因组DNA中分离出C0t-1 DNA并用生物素标记作探针,对有丝分裂中期相染色体进行原位杂交,每对染色体上均显示出了特定的荧光原位杂交带型.将植物25S和5S rDNA分别用地高辛和生物素标记作探针,单色荧光原位杂交结果显示黑叶小平头2对染色体具有25S rDNA基因座,1对染色体具有5S rDNA基因座.生物素标记的C0t-1 DNA与地高辛标记的25S rDNA等量混合作探针,双色荧光原位杂交证实了C0t-1 DNA与25S rDNA二者具有一致的染色体位置特征,表明基于rDNA及C0t-1 DNA荧光原位杂交的核型分析技术,优于目前普遍采用的只基于rDNA荧光原位杂交的核型分析方法.结合已报道的rDNA染色体定位结果,及C0t-1 DNA荧光原位杂交带型与染色体形态,更准确地构建了甘蓝的核型.To explore an effective and reliable karyotyping method in Brassica crop plants, C0t-1 DNA was isolated from Brassica oleracea genome, labeled as probe with Biotin-Nick Translation Mix kit, in situ hybridized to mitotic spreads, specific fluorescent bands were showed on each chromosome pairs. 25S and 5S rDNA were labeled as probes with DIG-Nick Translation Mix kit and Biotin Nick Translation Mix kit, respectively, in situ hybridized to mitotic preparations, 25S rDNA could be detected on two chromosome pairs, and 5S rDNA only one. C0t-1 DNA contains rDNA,chromosome sites identity between C0t-1 DNA and 25S rDNA was determined by dual-colour fluorescence in situ hybridization. All these showed that the karyotyping technique based on a combination of rDNA and C0t-1 DNA chromosome landmarks is superior to alone one. A more exact karyotype of B. oleracea have been analysed based on a combination of rDNA sites, C0-1 DNA fluorescent bands, chromosome lengths and arm ratios.

关 键 词：甘蓝 RDNA C0T-1 DNA 荧光原位杂交 核型分析 

分 类 号：Q321.3[生物学—遗传学]