Gong Junqing, Wang Zhaoyuan, Guo Zhujuan, Yao Lijuan, Zhao Chuanfang, Lin Sheng, Ma Songling, Shen Yingbai
National Engineering Research Center of Tree breeding and Ecological Restoration, College of Biological Sciences and Technology, Beijing Forestry University, No. 35, Qinghua East Road, Beijing 100083, PR China.
National Engineering Research Center of Tree breeding and Ecological Restoration, College of Biological Sciences and Technology, Beijing Forestry University, No. 35, Qinghua East Road, Beijing 100083, PR China.
Int J Biol Macromol. 2023 Mar 15;231:123503. doi: 10.1016/j.ijbiomac.2023.123503. Epub 2023 Feb 1.
Evk (ethyl vinyl ketone) is a signal substance for plant defense, but little is known about how evk mediates stomatal closure. Through stomatal biology experiments, we found that evk can mediate stomatal closure, and stomatal closure is weakened when DORN1 (DOES NOT RESPOND TO NUCLEOTIDES 1) and GORK (GATED OUTWARDLY-RECTIFYING K+ CHANNEL) are mutated. In addition, it was found by non-invasive micro-test technology (NMT) that the K efflux mediated by evk was significantly weakened when DORN and GORK were mutated. Yeast two-hybrid (Y2H), firefly luciferase complementation imaging (LCI), and in vitro pull-down assays demonstrated that DORN1 and GORK could interact in vitro and in vivo. It was found by molecular docking that evk could combine with MRP (Multidrug Resistance-associated Protein), thus affecting ATP transport, promoting eATP (extracellular ATP) concentration increase and realizing downstream signal transduction. Through inoculation of botrytis cinerea, it was found that evk improved the antibacterial activity of Arabidopsis thaliana. As revealed by reverse transcription quantitative PCR (RT-qPCR), the expression of defense related genes was enhanced by evk treatment. Evk is a potential green antibacterial drug.
乙基乙烯基酮（Evk）是一种植物防御信号物质，但关于Evk如何介导气孔关闭知之甚少。通过气孔生物学实验，我们发现Evk可以介导气孔关闭，而当DORN1（不响应核苷酸1）和GORK（向外整流钾离子通道）发生突变时，气孔关闭会减弱。此外，通过非损伤微测技术（NMT）发现，当DORN和GORK发生突变时，Evk介导的钾离子外流会显著减弱。酵母双杂交（Y2H）、萤火虫荧光素酶互补成像（LCI）和体外下拉试验表明，DORN1和GORK可以在体外和体内相互作用。通过分子对接发现，Evk可以与多药耐药相关蛋白（MRP）结合，从而影响ATP转运，促进细胞外ATP（eATP）浓度升高并实现下游信号转导。通过接种灰霉病菌发现，Evk提高了拟南芥的抗菌活性。逆转录定量PCR（RT-qPCR）结果显示，Evk处理增强了防御相关基因的表达。Evk是一种潜在的绿色抗菌药物。
