Establishment and Application of Ploidy Method for the Identification of Psidium guajava by Flow Cytometry
[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.
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