High-throughput Sample Preparation Method for the Identification of Potato Ploidy Using Flow Cytometry
[Objective]Chromosome ploidy identification is an important part of the evaluation of potato germplasm resources.The establishment of a high-throughput sample preparation method for the identification of potato chromosome ploidy based on flow cytometry would lay a foundation for the subsequent large-scale identification of potato ploidy.[Method]The 30 parthenogenetic offspring of potato were used to compare the effects of low-temperature steel-bullet beating,liquid nitrogen grinding and blade shredding to prepare nuclear suspensions.The ploidy of diploid potato IVP101 and tetraploid potato HTJ349-3 was identified.[Result]The suspension of cell nuclei prepared by the low-temperature steel-bullet beating method had obvious fluorescence signal,sufficient cell lysis and few impurities.Due to the simple and easy operation of the low-temperature steel-bullet beating method,the sample preparation time was greatly shortened,and the identification efficiency was 45-60 times higher than that of the liquid nitrogen grinding method,and 105-180 times higher than that of the blade shredding method.The chromosome ploidy of potato IVP101 and HTJ349-3 was accurately detected by this low-temperature steel-bullet beating method.[Conclusion]The results by the low-temperature steel-bullet beating method are as accurate as that by the liquid nitrogen grinding method and the blade cutting method,which can achieve high-throughput identification of potato chromosome ploidy.
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