一种利用流式细胞术鉴定梨倍性的方法

流式细胞术鉴定植物染色体倍性的方法广泛应用于果树多倍体育种中，然而相关流式细胞术鉴定梨染色体倍性的方法还没有系统的研究。以梨树叶片为试材系统优化了各个流式细胞术关键实验节点，结果表明：实验材料选取当年春季新梢第3~5 片梨树嫩叶，尤其是大棚新梢嫩叶最好；叶片前处理时用蒸馏水、去离子水依次洗净叶片表面后，再用去离子水浸泡10 min；使用WPB解离液对梨叶片细胞核进行提取后，用500 目滤膜过滤2 次，离心1 次后用流式细胞仪进行检测可得到理想的实验结果。该方法步骤简单、结果可靠，是一种快速、高效的梨染色体倍性鉴定的方法，可广泛用于梨树多倍体鉴定。

A Method of Ploidy Identification in Pear Using Flow Cytometry

Although it has been widely used in the polyploid breeding of fruit trees, ploidy identification by flow cytometry facilitated in pear trees has not been well reported. In this study, key links in flow cytometry experiment were systemically optimized using pear leaves as materials. The results showed that the young leaves from the third-to-fifth leaf of the spring shoots, especially grew in greenhouse, were better for flow cytometry test. Before xperiment, successively washing young leaves with water and distilled water, and then immersing in distilled water for 10 min was also benefit for the test. In addition, the processes using WPB buffer to extract cell nucleus, twice filtrating the solution with 500 mesh filter membrane and once centrifugation of cell lysis solution were optimal for the ploidy identification by flow cytometry. In total, a simple, rapid and efficient method of flow cytometry suitable for pear ploidy identification was established and could be generally used for pear polyploid identification.