一种提升流式细胞术分析效果的前处理方法  

A Plant Sample Optimal Pretreatment for Flow Cytometric Analysis

作　　者：张晋丹 冯旻[1] Jindan Zhang;Min Feng(State Key Laboratory of Systematic and Evolutionary Botany,Institute of Botany,Chinese Academy of Sciences,Beijing 100093,China)

机构地区：[1]中国科学院植物研究所,系统与进化植物学国家重点实验室,北京100093

出　　处：《植物学报》2023年第2期285-297,共13页Chinese Bulletin of Botany

基　　金：国家重点实验室专项经费(No.Y7106F1001)。

摘　　要：提取完整植物细胞核是流式细胞术检测前处理的关键步骤。分别以不同种属植物的新鲜叶片和大量硅胶快速干燥的植物叶片为研究材料,比较新优化的细胞核提取液PVPK12-mGB_(2)与其它2种常见提取液(LB01和CyStain^(■)PIAbsoluteP)的提取效果。结果表明,新优化的PVPK12-mGB_(2)细胞核提取液在测试多种富含干扰性次生代谢物的植物类群新鲜材料时效果最佳,提取的细胞核质量基本能满足流式细胞术分析的要求。此外,实验结果证实该细胞核提取液能够提升大量硅胶快速干燥保存植物材料的提取效果,测试结果明显优于其它2种细胞核提取液。研究建立了用于DNA流式细胞术的植物样品前处理方法,为野外采样分析提供了参考方法。Flow cytometry requires preparation of suspensions of intact nuclei,which is crucial step for analysis.A newly formulated buffer,PVPK12-mGB_(2)compared with two common buffers(LB01 and CyStain^(■)PI Absolute P),were used to isolate nuclei from fresh or silica gel desiccated leaf tissues of different species.PVPK12-mGB_(2)exhibited the best performance on all fresh leaves tested of plant species,of which the interfering secondary metabolites highly accumulated,the quality of extracted nuclei were able to satisfy demands for flow cytometric analysis.Moreover,results of the present study substantiate the enhanced effectiveness of PVPK12-mGB_(2),for silica gel desiccated plant tissue,compared to other buffers tested.The study established an optimal pretreatment of plants for DNA flow cytometry,which provided reference method for sampling and analysis in remote regions.

关 键 词：细胞核提取液 硅胶干燥 复杂植物次生代谢物 流式细胞术 

分 类 号：Q94-33[生物学—植物学]