西南农业学报2025，Vol.38Issue(5)：885-892,8.DOI:10.16213/j.cnki.scjas.2025.5.001
梨褪绿叶斑伴随病毒P5蛋白与NbWRKY1的互作研究
Interaction between P5 protein encoded by pear chlorotic leaf spot-associated virus and NbWRKY1
摘要
Abstract
[Objective]Previous studies have confirmed that the protein P5 of pear chlorotic leaf spot-associated virus(PCLSaV)was a silen-cing suppressor and pathogenic factor.The study aimed to explore the effect of NbWRKY1 to the biological function of PCLSaV P5,and to pro-vide insights into the mechanism for biological functions of PCLSaV P5.[Method]In the present study,the interaction between NbWRKY 1 and P5 were analyzed using bimolecular fluorescence complementation(BiFC)and co-immunoprecipitation(Co-IP)assays.Additionally,the biological effects of NbWRKY1 on P5 were examined through Agrobacterium tumefaciens-mediated transient expression and green fluorescent protein(GFP)-based RNA silencing induction experiments.[Result]NbWRKY1 was found to interact with P5 and co-localize in the nucle-us,forming uniform nuclear body particles that excluded nucleoli.This distribution pattern was similar to that of NbWRKY1 in Nicotiana benthamiana cells.Furthermore,P5 was shown to induce the up-regulation of NbWRKY1 gene expression,with a significant 4.2-fold increase at 2 days,which slightly decreased by 4 days but remained higher than the control.Virus-induced gene silencing(VIGS)revealed that the si-lencing efficiency of NbWRKY1 in wild-type N.benthamiana plants followed a dynamic trend,increasing initially and then decreasing,with the highest silencing efficiency at 14 days.And the NbWRKY1-silened plants exhibited phenotypes similar to those of wild-type plants.Nota-bly,the degree of cell death caused by P5 on the leaves of NbWRKY1-silenced plants was significantly reduced compared to that in wild-type plants.It was determined that down-regulation of NbWRKY1 effectively alleviated the cell death caused by PCLSaV P5.The RNA silencing suppression activities of NbWRKY1 and P5 were detected.It was found that NbWRKY1 had the ability to directly suppress RNA silencing,and there was a superposition effect between the RNA silencing suppression ability of P5 and NbWRKY1.When the silencing suppression ac-tivity of the two proteins were combined,resulting in stronger RNA silencing suppression activity than that of P5 protein alone.[Clonclusion]The study demonstrated that there was interaction relationship between NbWRKY1 and PCLSaV P5.NbWRKY1 could directly inhibit RNA silencing and its down-regulation could also alleviate cell death caused by PCLSaV P5.Moreover,there was a synergistic effect between the RNA silencing inhibitory activity of NbWRKY1 and PCLSaV P5.These results provide a new theoretical basis for analyzing the biological function of PCLSaV P5.关键词
梨褪绿叶斑伴随病毒/特异蛋白P5/NbWRKY1/蛋白互作/RNA沉默抑制Key words
Pear chlorotic leaf spot-associated virus/Specific protein P5/NbWRKY1/Protein-protein interaction/RNA silencing suppression分类
农业科技引用本文复制引用
高玉洁,任秋婷,张永乐,张樾,张红艺,邓惠方,王先洪,洪霓,王国平..梨褪绿叶斑伴随病毒P5蛋白与NbWRKY1的互作研究[J].西南农业学报,2025,38(5):885-892,8.基金项目
国家重点研发计划项目(2019YFD1001800) （2019YFD1001800）
