《植物生理学报》 2014, 50(8): 1216-1222

白菜rDNA及C₀t-1 DNA的荧光原位杂交及其核型分析

吴春红¹, 范双莉², 王力军^3,*

¹河南理工大学图书馆, 河南焦作454000; ²济源职业技术学院护理系, 河南济源459000; ³中国农业科学院油料作物研究所/油料作物生物学与遗传改良农业部重点实验室, 武汉430062

通信作者：王力军；E-mail: wanglijun@caas.cn；Tel: 027-86711930

摘　要：

以早熟白菜苔为实验材料, 从其基因组DNA中分离出C₀t-1 DNA并用生物素标记作探针, 25S rDNA用地高辛标记作探针, 对有丝分裂中期相染色体进行双色荧光原位杂交。每对染色体上均显示出了特定的C₀t-1 DNA荧光原位杂交带型, 5对染色体上显示出了25S rDNA荧光原位杂交带型。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的荧光原位杂交核型分析技术, 优于目前普遍采用的只基于rDNA的荧光原位杂交核型分析方法。结合C₀t-1 DNA与25S rDNA的荧光原位杂交带型和传统的染色体的形态学标记分析方法及白菜已公布的基于rDNA分布的核型分析结果, 创建了一个精确的白菜核型。

关键词：白菜; rDNA; C₀t-1 DNA; 荧光原位杂交; 核型分析

收稿：2014-01-26   修定：2014-06-25

资助：国家自然科学基金面上项目(30671312)。

Fluorescence in situ Hybridization and Karyotyping Analysis of rDNA and C₀t-1 DNA in Brassica campestris

WU Chun-Hong¹, FAN Shuang-Li², WANG Li-Jun^3,*

¹Library of Henan Polytechnic University, Jiaozuo, Henan 454000, China; ²Nursing Department, Jiyuan Vocational and Technical College, Jiyuan, Henan 459000, China; ³Institute of Oil Crops, Chinese Academy of Agricultural Sciences/Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, China

Corresponding author: WANG Li-Jun; E-mail: wanglijun@caas.cn; Tel: 027-86711930

Abstract:

With Brassica campestris cv. Zaoshu baicaitai as experimental material, C₀t-1 DNA was isolated from the genomic DNA and labeled with Biotin-Nick as a probe, while 25S rDNA was labeled with DIG-Nick. Fluorescence in situ hybridized to the mitotic metaphase chromosomes of B. campestris showed the specific fluorescence bands of C₀t-1 rDNA on each chromosome pair. However, the specific fluorescence bands of 25S rDNA were detected on 5 chromosome pairs. The dual fluorescence in situ hybridization confirmed the C₀t-1 DNA and rDNA had identical chromosome sites. And also the karyotyping technique based on a combination of rDNA and C₀t-1 DNA chromosome landmarks was superior to alone one. Based on a combination of rDNA sites, C₀t-1 DNA fluorescent bands, chromosome lengths and arm ratios, a more exact karyotype idiogram of B. campestris was described.

Key words: Brassica campestris; rDNA; C₀t-1 DNA; fluorescence in situ hybridization; karyotyping