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Journal of Integrative Agriculture
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A bacterial protein Rhp-PSP inhibits plant viral proliferation through endoribonuclease activity
Lijie Chen1, 2, Qianze Peng2, Xiaohua Du2, Weixing Zhang2, Ju’e Cheng2, Shu’e Sun2, Deyong Zhang1, 2, Pin Su1, 2#, Yong Liu1, 2#

1 Longping Branch, College of Biology, Hunan University, Changsha 410082, China

2 State Key Laboratory of Hybrid Rice and Institute of Plant Protection, Hunan Academy of Agricultural Sciences, Changsha 410125, China

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摘要  植物病毒病的发生在全球范围内造成巨大的作物损失。然而,目前没有非常有效的措施来控制植物病毒病。此前,我们鉴定了一种由沼泽红假单胞菌JSC-3b菌株产生的抗病毒蛋白Rhp-PSP。本研究中,我们发现Rhp-PSP的抗病毒活性依赖于其核酸内切酶活性。将129位的精氨酸(R)残基转化为丙氨酸(A),降低了对烟草花叶病毒(TMV)外壳蛋白(CPRNA的核糖核酸内切酶活性,从而损害了Rhp-PSP的抗病毒活性。进一步的研究表明,突变体Rhp-PSPR129A不能像野生型那样形成同源三聚体,这说明了四级连接对核糖核酸内酶活性的重要性Rhp-PSP在本氏烟草中的过表达显著增强了幼苗对TMV的抗性,而Rhp-PSPR129A过表达则没有,这证实了核糖核酸内切酶活性是Rhp-PSP抗病毒活性的原因。此外,Rhp-PSP喷施在番茄和辣椒叶面上,显著降低了病毒性疾病的发病指数,表明Rhp-PSP在实践中显示出开发抗病毒药物的潜力。

Abstract  Plant virus causes massive crop losses globally. However, there is currently no effective measure to control plant viral disease. Previously, we identify an antiviral protein Rhp-PSP, produced by the bacterial Rhodopseudomonas palustris strain JSC-3b. In this study, we discover that the antiviral activity of Rhp-PSP relies on its endoribonuclease activity. Convert the arginine (R) residue at position 129 onto alanine (A) abolish its endoribonuclease activity on coat protein (CP) RNA of tobacco mosaic virus (TMV), consequentially, compromised the antiviral activity of Rhp-PSP. Further investigation demonstrates that, the mutant Rhp-PSPR129A is unable to form the homotrimer as the wild type, indicating the importance of quaternary junction for the endoribonuclease activity. Overexpression of Rhp-PSP in Nicotiana benthamiana significantly enhances the resistance against TMV of seedlings, while expression of Rhp-PSPR129A did not, confirming that endoribonuclease activity is responsible for the antiviral activity of Rhp-PSP. In addition, foliar spray of Rhp-PSP solution on tomato and pepper plants significantly reduces the disease index of viral diseases, indicating that Rhp-PSP shows potential to develop antiviral agent in practice.
Keywords:  Rhodopseudomonas palustris              Rhp-PSP              Tobacco mosaic virus              endoribonuclease              antiviral activity  
Online: 16 April 2024  
Fund: This research was supported by National Key R&D Program of China (2022YFD1400700); the key Research and Development program of Hunan Province (2022NK2014); Hunan Natural Science Foundation (2022JJ40234); Agricultural Science and Technology Innovation Fund Project of Hunan Province (2022CX1) and Changsha Natural Science Foundation (kq2202338). The grant agencies had no role in study design, data collection and analysis, the decision to publish, or the preparation of this manuscript. We are thankful to the Institute of Plant Protection, Hunan Academy of Agricultural Sciences for their support.
About author:  #Correspondence Pin Su, E-mail: supin102@163.com; Yong Liu, E-mail: liuyong@hunaas.cn, haoasliu@163.com

Cite this article: 

Lijie Chen, Qianze Peng, Xiaohua Du, Weixing Zhang, Ju’e Cheng, Shu’e Sun, Deyong Zhang, Pin Su, Yong Liu. 2024. A bacterial protein Rhp-PSP inhibits plant viral proliferation through endoribonuclease activity. Journal of Integrative Agriculture, Doi:10.1016/j.jia.2024.03.051

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