Chen Xinru, Gao Zihan, Li Yangyang, Nie Xiaoqian, Xie Qiaoli, Chen Guoping, Hu Zongli
Laboratory of molecular biology of tomato, Bioengineering College, Chongqing University, Chongqing 400030, China.
Plants (Basel). 2024 Oct 6;13(19):2800. doi: 10.3390/plants13192800.
N6-methyladenosine (mA) is a widespread post-transcriptional modification in eukaryotic mRNAs. Proteins with the YTH structural domain act as mA-binding proteins by recognizing the mA modification and regulating mRNA through this recognition. In this study, , a prototypical mA -binding protein gene in the YTH family was expressed in various tissues, and subcellular localization analyses indicated that the SlYTHDF2 protein was localized in the nucleus and cytoplasm. knockout lines were obtained using CRISPR/Cas9 technology and showed the senesced leaves prematurely increased endogenous ABA accumulation compared with the wild type. Moreover, we found that dark promoted leaf senescence in knockout lines and exogenous ABA further accelerated leaf senescence under dark conditions. The qRT-PCR analysis revealed significant alterations in the expression of genes associated with the ABA pathway. Relative to the wild type, the plants exhibited reduced levels of photosynthetic pigments, higher accumulation of reactive oxygen species, and increased damage to cell membranes. Additionally, we discovered that SlYTHDF2 interacts with the chloroplast-binding protein SlRBCS3 through yeast two-hybrid and BiFC experiments. Overall, our data suggest the important role of in regulating tomato leaf senescence.
N6-甲基腺苷（mA）是真核生物mRNA中一种广泛存在的转录后修饰。具有YTH结构域的蛋白质通过识别mA修饰并通过这种识别来调节mRNA，从而充当mA结合蛋白。在本研究中，YTH家族中的一个典型的mA结合蛋白基因在各种组织中表达，亚细胞定位分析表明SlYTHDF2蛋白定位于细胞核和细胞质中。使用CRISPR/Cas9技术获得了敲除株系，与野生型相比，这些株系的衰老叶片过早地增加了内源脱落酸（ABA）的积累。此外，我们发现黑暗促进了敲除株系中的叶片衰老，并且外源ABA在黑暗条件下进一步加速了叶片衰老。qRT-PCR分析揭示了与ABA途径相关的基因表达的显著变化。相对于野生型，这些植株表现出光合色素水平降低、活性氧积累增加以及细胞膜损伤加剧。此外，我们通过酵母双杂交和双分子荧光互补实验发现SlYTHDF2与叶绿体结合蛋白SlRBCS3相互作用。总体而言，我们的数据表明其在调节番茄叶片衰老中具有重要作用。
