College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, China.
National Engineering Research Center of Tree breeding and Ecological Restoration, College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, China.
Int J Mol Sci. 2022 Jul 13;23(14):7718. doi: 10.3390/ijms23147718.
Jasmonic acid (JA) is an important hormone that functions in plant defense. and mutants were more resistant to than in the wild-type (WT) Arabidopsis group. In addition, JA concentration in and mutants was higher compared with the WT group. To explore how these two proteins affect the resistance of Arabidopsis plants, we used a yeast two-hybrid assay, firefly luciferase complementation imaging assay and in vitro pull-down assay confirming that calmodulin 1 (CAM1) interacted with WRKY53. However, these two proteins separate when calcium concentration increases in Arabidopsis leaf cells. Then, electrophoretic mobility shift assay and luciferase activation assay were used to verify that WRKY53 could bind to lipoxygenases 3 (LOX3) and lipoxygenases 4 (LOX4) gene promoters and negatively regulate gene expression. This study reveals that CAM1 and WRKY53 negatively regulate plant resistance to herbivory by regulating the JA biosynthesis pathway via the dissociation of CAM1-WRKY53, then the released WRKY53 binds to the promoters to negatively regulate gene expression. This study reveals WRKY53's mechanism in insect resistance, a new light on the function of WRKY53.
茉莉酸（JA）是一种在植物防御中起重要作用的激素。和突变体比野生型（WT）拟南芥群体更能抵抗。此外，和突变体中的 JA 浓度比 WT 组更高。为了探究这两种蛋白质如何影响拟南芥植物的抗性，我们使用酵母双杂交测定、萤火虫荧光素酶互补成像测定和体外下拉测定证实钙调素 1（CAM1）与 WRKY53 相互作用。然而，当钙浓度在拟南芥叶细胞中增加时，这两种蛋白质会分离。然后，电泳迁移率变动分析和荧光素酶激活分析用于验证 WRKY53 可以结合到脂氧合酶 3（LOX3）和脂氧合酶 4（LOX4）基因启动子上，并负调控基因表达。本研究表明，CAM1 和 WRKY53 通过 CAM1-WRKY53 的解离负调控 JA 生物合成途径，从而负调控植物对草食性的抗性，然后释放的 WRKY53 结合到启动子上，负调控基因表达。本研究揭示了 WRKY53 在昆虫抗性中的机制，为 WRKY53 的功能提供了新的认识。
