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烟草中α-氨基膦酸酯衍生物Q-R与Harpin结合蛋白1的相互作用机制研究

Research on the Interaction Mechanism Between α Mino-Phosphonate Derivative Q-R and Harpin-Binding Protein 1 in Tobacco () Plants.

作者信息

Huang Maoxi, Yan Yunlong, Wang Li, Chen Jun, Liu Tao, Xie Xin, Li Xiangyang

机构信息

State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Guiyang, China.

College of Agriculture, Guizhou University, Guiyang, China.

出版信息

Front Microbiol. 2021 Mar 23;12:621875. doi: 10.3389/fmicb.2021.621875. eCollection 2021.

Abstract

Amino-phosphonate derivative R-diphenyl-1-(4-methylbenzothiazole-2-amino)-1-(thiphene-2-yl)-methylphosphonate (Q-R) has a high protective anti-tobacco mosaic virus (TMV) activity. However, the mechanism responsible for Q-R's effect on TMV infection is largely unknown. Here, we studied the expression levels of harpin-binding protein 1 (HrBP1) and pathogenesis-related protein-1a (PR-1a) in TMV-infected tobacco plants by using reverse transcription quantitative real-time PCR. Then, we verified the interactions between Q-R and the HrBP1 protein from using isothermal titration calorimetry and studied the Q-R-associated assembly of HrBP1 using size-exclusion chromatography. The results showed that the expression levels of HrBP1 and PR-1a genes were significantly increased by Q-R at the transcriptional level in TMV-infected tobacco plants, and the -expressed HrBP1 protein was assembled into oligomers by Q-R via binding to HrBP1 with a dissociation constant of 1.19 μM. We, therefore, concluded that Q-R activated the HrBP1 and PR-1a genes and enhanced the ability of HrBP1 to assemble in tobacco plants.

摘要

氨基膦酸酯衍生物R-二苯基-1-(4-甲基苯并噻唑-2-氨基)-1-(噻吩-2-基)甲基膦酸酯(Q-R)具有很高的抗烟草花叶病毒(TMV)保护活性。然而,Q-R对TMV感染的作用机制很大程度上尚不清楚。在此,我们通过逆转录定量实时PCR研究了TMV感染的烟草植株中harpin结合蛋白1(HrBP1)和病程相关蛋白1a(PR-1a)的表达水平。然后,我们用等温滴定量热法验证了Q-R与HrBP1蛋白之间的相互作用,并用尺寸排阻色谱法研究了Q-R相关的HrBP1组装。结果表明,在TMV感染的烟草植株中,Q-R在转录水平上显著提高了HrBP1和PR-1a基因的表达水平,且表达的HrBP1蛋白通过与HrBP1结合被Q-R组装成寡聚体,解离常数为1.19 μM。因此,我们得出结论,Q-R激活了HrBP1和PR-1a基因,并增强了HrBP1在烟草植株中的组装能力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/40e8/8044911/de8ce80ffee9/fmicb-12-621875-g001.jpg

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