植物寡核苷酸荧光原位杂交技术方法  

Protocals for Oligonucleotide Fluorescence in situ Hybridization in Plants

作　　者：张凡凡 邢新滢 石文清 沈懿[2] 程祝宽[2,3] Fanfan Zhang;Xinying Xing;Wenqing Shi;Yi Shen;Zhukuan Cheng(Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education,College of Life Sciences,Beijing Normal University,Beijing 100875,China;State Key Laboratory of Plant Genomics and Center for Plant Gene Research,Institute of Genetics and Developmental Biology,Chinese Academy of Sciences,Beijing 100101,China;Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops,Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Key Laboratory of Plant Functional Genomics of the Ministry of Education,College of Agriculture,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]北京师范大学生命科学学院,细胞增殖与调控生物学教育部重点实验室,北京100875 [2]中国科学院遗传与发育生物学研究所,植物基因组学国家重点实验室及植物基因研究中心,北京100101 [3]扬州大学农学院,江苏省作物基因组学与分子育种重点实验室/植物功能基因组学教育部重点实验室,江苏省粮食作物现代生产技术协同创新中心,扬州225009

出　　处：《植物学报》2023年第2期274-284,共11页Chinese Bulletin of Botany

基　　金：北京市自然科学基金(No.6214043);国家自然科学基金(No.32001522,No.32000368);中央高校基本科研业务费专项资金(No.2021NTST20)。

摘　　要：寡核苷酸荧光原位杂交技术是一种整合了生物信息学分析、DNA高通量合成以及荧光原位杂交实验的新兴技术。该技术适用于任何具有参考基因组的植物物种,并可根据研究需求设计靶向某个染色体区域、整条染色体或一组染色体的寡核苷酸探针,目前已成功应用于数种植物的核型分析、染色体变异、种群演化、同源染色体配对、单倍型分析和异源多倍体识别等研究。该文详述了植物寡核苷酸探针的设计、扩增和标记、染色体制备以及荧光原位杂交的具体操作流程,以期促进寡核苷酸荧光原位杂交技术在细胞遗传学研究中的应用。Oligonucleotide fluorescence in situ hybridization(Oligo-FISH)is a new technology that integrates bioinfor-matics analysis,high throughput DNA synthesis and fluorescence in situ hybridization experiments.It is applicable to plant species with a reference genome.Oligo probes are able design to a chromosomal region,an entire chromosome,and a set of chromosomes according to researchers’requirements.Oligo-FISH has been successfully applied to studies on karyotyping,chromosome variation,population evolution,homologous chromosome pairing,haplotype analysis,and heteropolyploid identification in several plant species.Here,we introduce the concrete operational processes of oligonu-cleotide-based probe design,amplification and labeling,chromosomes preparation,and fluorescence in situ hybridization in plants,so as to facilitate Oligo-FISH application in the cytogenetics research in the future.

关 键 词：荧光原位杂交 寡核苷酸探针 染色体 植物 

分 类 号：Q94-33[生物学—植物学]