生物技术通报2024，Vol.40Issue(2)：48-54,7.DOI:10.13560/j.cnki.biotech.bull.1985.2023-0844
基于流式细胞仪鉴定番石榴倍性方法的建立及应用
Establishment and Application of Ploidy Method for the Identification of Psidium guajava by Flow Cytometry
摘要
Abstract
[Objective]The flow cytometry was used to identify the ploidy of Psidium guajava,which may lay the foundation for ploidy breeding and hybrid breeding of P.guajava.[Method]Using P.guajava as the material,the effects of sample parts,centrifugation treatment,and storage methods on ploidy detection results were compared.[Result]P.guajava petals had the highest number of collected nuclear DNA,with a CV value of 1.96%and the best peak shape.When preparing nuclear suspension,the best detection effect was achieved by directly staining and analyzing without centrifugation after filtration.The nuclear DNA content obtained from fresh P.guajava petals was significantly higher than that of refrigerated and other frozen treatment groups,with fewer background fragments and a clear main peak.The-80℃freezing treatment caused the least damage to the samples,and the detection effect was second only to that of fresh P.guajava petals.The established flow cytometry method for ploidy analysis of P.guajava involved taking 0.50-1.00 cm2 of pomegranate petals,mixing them with 1 mL of mGb lysis buffer,filtering,and adding 30 μL of PI staining for 1 min before analysis.[Conclusion]Using this established flow cytometry method,ploidy identification was performed on 33 P.guajava germplasm resources,resulting in the detection of 32 diploids and 1 hexaploid.This method is simple,efficient,and accurate,and may provide an effective approach for ploidy identification of P.guajava germplasm resources.关键词
番石榴/花瓣/倍性鉴定/流式细胞术Key words
Psidium guajava/petal/ploidy identification/flow cytometry引用本文复制引用
邵雪花,李桂兰,肖维强,赖多,庄庆礼,秦健..基于流式细胞仪鉴定番石榴倍性方法的建立及应用[J].生物技术通报,2024,40(2):48-54,7.基金项目
乡村振兴战略专项资金(2022-NPY-01-005),广东省农业科学院横向科技项目(2022-22141) （2022-NPY-01-005）
