鉴定病毒诱导的兰花 Argonaute 5b 启动子及其在病毒感染过程中的功能特征。

Characterization of Virus-Inducible Orchid Argonaute 5b Promoter and Its Functional Characterization in during Virus Infection.

机构信息

Graduate Institute of Biotechnology, National Chung Hsing University, Taichung 40227, Taiwan.

Advanced Plant Biotechnology Center, National Chung Hsing University, Taichung 40227, Taiwan.

出版信息

Int J Mol Sci. 2022 Aug 29;23(17):9825. doi: 10.3390/ijms23179825.

DOI:10.3390/ijms23179825

PMID:36077222

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9456093/

Abstract

Plant ARGONAUTES (AGOs) play a significant role in the defense against viral infection. Previously, we have demonstrated that AGO5s encoded in subsp. (PaAGO5s) took an indispensable part in defense against major viruses. To understand the underlying defense mechanism, we cloned PaAGO5s promoters (PaAGO5s) and analyzed their activity in transgenic using β-glucuronidase (GUS) as a reporter gene. GUS activity analyses revealed that during (CymMV) and (ORSV) infections, PaAGO5b activity was significantly increased compared to PaAGO5a and PaAGO5c. Analysis of PaAGO5b 5'-deletion revealed that PaAGO5b_941 has higher activity during virus infection. Further, yeast one-hybrid analysis showed that the transcription factor NbMYB30 physically interacted with PaAGO5b_941 to enhance its activity. Overexpression and silencing of resulted in up- and downregulation of expression, respectively. Exogenous application and endogenous measurement of phytohormones have shown that methyl jasmonate and salicylic acid respond to viral infections. NbMYB30 overexpression and its closest related protein, PaMYB30, in subsp. reduced CymMV accumulation in subsp. . Based on these discoveries, this study uncovers the interaction between virus-responsive promoter and the corresponding transcription factor in plants.

摘要

植物 ARGONAUTES（AGOs）在抗病毒感染中发挥重要作用。以前，我们已经证明，亚属中编码的 AGO5s（PaAGO5s）在防御主要病毒方面起着不可或缺的作用。为了了解潜在的防御机制，我们克隆了 PaAGO5s 启动子（PaAGO5s），并使用β-葡萄糖醛酸酶（GUS）作为报告基因分析其在转基因中的活性。GUS 活性分析表明，在（CymMV）和（ORSV）感染期间，与 PaAGO5a 和 PaAGO5c 相比，PaAGO5b 的活性显著增加。对 PaAGO5b 5'-缺失的分析表明，PaAGO5b_941 在病毒感染期间具有更高的活性。此外，酵母单杂交分析表明，转录因子 NbMYB30 与 PaAGO5b_941 物理相互作用以增强其活性。NbMYB30 的过表达和沉默分别导致表达的上调和下调。外源应用和内源性测定植物激素表明，茉莉酸甲酯和水杨酸响应病毒感染。NbMYB30 的过表达及其在亚属中的最接近相关蛋白 PaMYB30 降低了 CymMV 在亚属中的积累。基于这些发现，本研究揭示了植物中病毒响应启动子与相应转录因子之间的相互作用。