中国农业大学学报2025，Vol.30Issue(7)：119-130,12.DOI:10.11841/j.issn.1007-4333.2025.07.10
菊花倍性流式细胞术快速检测方法的优化
Optimization of rapid chrysanthemum ploidy detection method by flow cytometry
摘要
Abstract
To establish a flow cytometry method for large-scale rapid identification of chrysanthemum ploidy,the diploid chrysanthemum indicum and the hexaploid chrysanthemum 'Qiuyan' were used as study materials.Three methods were adopted to prepare nuclear suspension.By comparing the advantages and disadvantages of the three methods in determining the ploidy,the grinder grinding method was selected as the optimal method.A total of five factors in the grinder grinding method were explored,including lysis buffer,the rotation speed of the grinder,lysis time,the mesh number of the filtering nylon mesh and staining time.The results showed that:1)All three sample preparation methods could accurately determine the ploidy.The sample prepared by the blade chopping method displayed a better peak shape,but it took the longest time.The sample prepared by the mortar grinding method had a medium peak shape and a moderate time consumption.The sample prepared by the grinder grinding method had a medium peak shape and the shortest time consumption.2)The grinder grinding method,using Otto lysis buffer with the rotation speed of 1 200 to 1 400 r/min,the lysis time of 0 min,200 to 500 mesh nylon mesh and thestaining time of 5 min,could process the samples more quickly and achieve a better detection effect.In conclusion,the grinder grinding method based on flow cytometry achieved the same accurate detection results as the mortar grinding method and the blade chopping method for sample preparation.It realized the rapid batch identification of chrysanthemums' chromosome ploidy,providing a quick and reliable technical method for the ploidy identification of chrysanthemums.关键词
菊花/倍性检测/流式细胞仪/方法优化/快速鉴定Key words
chrysanthemum/ploidy identification/flow cytometry/method optimization/rapid identification分类
农业科技引用本文复制引用
郭艳超,侯宇航,陈菊,徐彦杰,孟静柔,郑伶杰,姚玉涛,张国君..菊花倍性流式细胞术快速检测方法的优化[J].中国农业大学学报,2025,30(7):119-130,12.基金项目
河北省农林科学院科技创新专项(2022KJCXZX-BHS-5,2023KJCXZX-BHS-8) （2022KJCXZX-BHS-5,2023KJCXZX-BHS-8）
河北省现代农业产业技术体系建设专项(HBCT2024200409) （HBCT2024200409）
