生物技术通报2026，Vol.42Issue(6)：77-86,10.DOI:10.13560/j.cnki.biotech.bull.1985.2026-0247
流式细胞术的优化及其在甘薯倍性鉴定中的应用
Optimization of Flow Cytometry and Its Application in Ploidy Identification of Sweetpotato
摘要
Abstract
[Objective]Chromosome ploidy identification is an important step in evaluating sweetpotato germplasm resources.To overcome the limitations of traditional methods in sweetpotato sampling,this study optimized the application of flow cytometry for ploidy identification,established an efficient method,and provided a foundation for subsequent large-scale identification of sweetpotato materials with different ploidy levels.[Method]Using'Xuzishu 8'as the experimental material,this study systematically compared the effects of the liquid nitrogen-ground leaf method and the blade-chopped root tip method in preparing cell nucleus suspensions.The comparison focused on nucleus yield,sample preparation time,and the quality of flow cytometry profiles.To address the difficulty of sampling precious materials,we explored the feasibility of ploidy identification using fully expanded mature sweetpotato leaves by adjusting the concentrations of MgSO ₄·7H₂O,DTT,and Triton X-100 in the lysis buffer.The method was validated by individual and mixed detection of materials with different ploidy levels.[Result]The liquid nitrogen-ground leaf method significantly improved preparation efficiency while maintaining detection accuracy,and both the crushing step and the on-machine detection time per sample were markedly reduced.For mature leaf identification,doubling the concentrations of DTT and Triton X-100 in the lysis buffer remarkably increased the release of intact nuclei,successfully overcoming the difficulty of detecting mature leaves.This method showed excellent resolution in both individual and mixed detection of diploid,tetraploid,and hexaploid materials,and the observed fluorescence intensity ratio was highly consistent with the theoretical ploidy ratio(1:2:3),confirming its stability and universality.[Conclusion]For sweetpotato ploidy identification,the flow cytometry method improved by the liquid nitrogen-ground leaf approach is not restricted by sampling limitations such as root tip dependence,and can rapidly and accurately determine the ploidy of sweetpotato materials and facilitate material screening.关键词
甘薯/流式细胞术/倍性鉴定/液氮研磨叶片法Key words
sweetpotato/flow cytometry/ploidy identification/liquid nitrogen-ground leaf method引用本文复制引用
李建功,邓逸桐,赵路宽,王珧,曹清河..流式细胞术的优化及其在甘薯倍性鉴定中的应用[J].生物技术通报,2026,42(6):77-86,10.基金项目
国家重点研发计划(2023YFD1202702),现代农业产业技术体系专项(CARS-10),江苏省种业振兴"揭榜挂帅"项目(JBGS(2021)010) （2023YFD1202702）
