Liu Ying-Li, Guo Ying-Hua, Song Xue-Qin, Hu Meng-Xuan, Zhao Shu-Tang
State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing, China.
National Center for Protein Sciences at Peking University, Beijing, China.
Front Plant Sci. 2023 Jun 9;14:1196618. doi: 10.3389/fpls.2023.1196618. eCollection 2023.
Programmed cell death (PCD) is a genetically regulated developmental process leading to the death of specific types of plant cells, which plays important roles in plant development and growth such as wood formation. However, an efficient method needs to be established to study PCD in woody plants. Flow cytometry is widely utilized to evaluate apoptosis in mammalian cells, while it is rarely used to detect PCD in plants, especially in woody plants. Here, we reported that the xylem cell protoplasts from poplar stem were stained with a combination of fluorescein annexin V-FITC and propidium iodide (PI) and then sorted by flow cytometry. As expected, living cells (annexin V-FITC negative/PI negative), early PCD cells (annexin V-FITC positive/PI negative), and late PCD cells (annexin V-FITC positive/PI positive) could be finely distinguished through this method and then subjected for quantitative analysis. The expression of cell-type- and developmental stages-specific marker genes was consistent with the cell morphological observation. Therefore, the newly developed fluorescence-activated cell sorting (FACS) method can be used to study PCD in woody plants, which will be beneficial for studying the molecular mechanisms of wood formation.
程序性细胞死亡（PCD）是一种由基因调控的发育过程，导致特定类型的植物细胞死亡，它在植物发育和生长（如木材形成）中发挥着重要作用。然而，需要建立一种有效的方法来研究木本植物中的PCD。流式细胞术被广泛用于评估哺乳动物细胞中的凋亡，而很少用于检测植物中的PCD，尤其是木本植物中的PCD。在此，我们报道了用荧光素膜联蛋白V-FITC和碘化丙啶（PI）对杨树茎的木质部细胞原生质体进行染色，然后通过流式细胞术进行分选。正如预期的那样，通过这种方法可以精细地区分活细胞（膜联蛋白V-FITC阴性/PI阴性）、早期PCD细胞（膜联蛋白V-FITC阳性/PI阴性）和晚期PCD细胞（膜联蛋白V-FITC阳性/PI阳性），然后进行定量分析。细胞类型和发育阶段特异性标记基因的表达与细胞形态观察结果一致。因此，新开发的荧光激活细胞分选（FACS）方法可用于研究木本植物中的PCD，这将有助于研究木材形成的分子机制。
