拟南芥和大豆中用于RNA研究的细胞核制备和优化  

Preparation and optimization of nuclei from Arabidopsis and soybean for RNA research

作　　者：何思豪 苏京 董志诚 刘敏 朱家富 HE Sihao;SU Jing;DONG Zhicheng;LIU Min;ZHU Jiafu(Guangdong Provincial Key Laboratory of Plant Adaptation and Molecular Design,Guangzhou Key Laboratory of Crop Gene Editing,Innovative Center of Molecular Genetics and Evolution,School of Life Sciences,Guangzhou University,Guangzhou 510006,China)

机构地区：[1]广东省植物适应性与分子设计重点实验室,广州市作物基因编辑重点实验室,分子遗传与进化创新中心,广州大学生命科学学院,广州510006

出　　处：《植物生理学报》2024年第5期823-836,共14页Plant Physiology Journal

基　　金：国家自然科学基金(32101739)。

摘　　要：本研究旨在优化植物细胞核提取方法,以支持对核RNA完整性要求严格的转录组研究,如新生RNA检测方法、单细胞核RNA测序及其他组学技术。核心实验基于改进的Honda提取液配方,通过结合甘油、蔗糖和己二醇等三种渗压剂,并在此基础上添加或不添加聚蔗糖和葡聚糖T-40(Ficoll&Dextran T-40),形成六种不同的细胞核提取液。这些提取液被用于评估其在拟南芥和大豆样本中提取细胞核的效果。结果表明,含有己二醇的提取液在提取细胞核RNA及蛋白质方面表现出最高的得率,此外己二醇结合Ficoll&Dextran T-40能显著增强核RNA的纯度和完整性。此项研究为植物科学领域的组学研究提供了一种高效的细胞核提取方法,具有重要的参考价值。This study aims to optimize plant nuclear extraction methods to support transcriptomic studies where nuclear RNA integrity is strictly required,such as nascent RNA-sequencing,single-nucleus RNA sequencing,and other omics techniques.The core experiments are based on an improved Honda extraction solution formula,combining three types of osmotic agents:glycerol,sucrose,and hexylene glycol,with or without the addition of Ficoll and Dextran T-40,to create six different extraction solutions.These solutions were used to evaluate their effectiveness in extracting cell nuclei from Arabidopsis and soybean samples.The results indicate that the extraction solution containing hexylene glycol achieved the highest yield of nuclear RNA and proteins,and the combination of hexylene glycol with Ficoll&Dextran T-40 significantly enhanced the purity and integrity of nuclear RNA.This study provides an efficient method of nuclear extraction for omics research in the field of plant science and has important reference value.

关 键 词：细胞核RNA 拟南芥 大豆 核提取缓冲液 己二醇 

分 类 号：Q943.2[生物学—植物学] S565.1[农业科学—作物学]