中国农业科学2025，Vol.58Issue(14)：2739-2750,12.DOI:10.3864/j.issn.0578-1752.2025.14.003
GmSZFP互作蛋白的筛选及GmERF7在大豆抵抗SMV侵染过程中的功能分析
Identification of GmSZFP-Interacting Proteins and Functional Analysis of GmERF7 in Soybean Resistance to SMV Infection
摘要
Abstract
[Objective]Soybean,an important economic and oil crop,is frequently threatened by soybean mosaic virus(SMV)disease,which is caused by the soybean mosaic virus(SMV).Previous studies identified a differentially expressed C2H2 single zinc finger protein gene,GmSZFP,which positively regulated soybean resistance to SMV infection.Using a yeast two-hybrid library,we screened for proteins that interact with GmSZFP and explored their functions in the soybean-SMV interaction.This research provides a theoretical basis for further elucidating the regulatory network of transcription factors involved in soybean resistance to SMV infection.[Method]In this study,the compatible(Jidou 7 and SMV strain SC-8)and incompatible(Jidou 7 and SMV strain N3)combinations were used to screen the potential interaction proteins of GmSZFP by yeast two-hybrid library.the protein-protein interactions were validated through yeast two-hybrid system(Y2H)and bimolecular fluorescence complementation(BiFC).Real-time quantitative PCR(qPCR)and Tobacco rattle virus(TRV)-based virus-induced gene silencing(VIGS)were employed to analyze the transcriptional expression levels and functions of the identified interaction proteinin the soybean-SMV interaction.[Result]We identified GmERF7 as apotential target protein of GmSZFP through yeast two-hybrid system.The interaction between GmSZFP and GmERF7 was confirmed by Y2H and BiFC.GmERF7 is a 393-amino acid protein containing an AP2/ERF domain and two nuclear localization signals(NLS).qPCR results showed that the expression levels of GmERF7 were significantly higher in the compatible combination than those in the incompatible combination at 4,12,and 24 hours after infection,peaking at 24 hours.VIGS-mediated silencing of GmERF7 in Jidou 7 plants infected with SC-8(compatible combination)results in increased callose deposition at the inoculation site compared to the control.At the 14 days after inoculation,the expression of SMV coat protein CP gene was undetectable in the upper leaves of the GmERF7-silenced plants,and no SMV infection symptoms were observed.In contrast,the CP gene was detected in the control plants,which exhibited typical susceptibility symptoms such as mosaic and green-loss.These results indicate silencing GmERF7 weakened virus transport and spread between cells,thereby enhancing plant resistance to SMV.This suggest that GmERF7 negatively regulates soybean resistance to SMV.[Conclusion]The ERF transcription factor GmERF7 interacts with the zinc finger protein GmSZFP,and negatively regulates soybean resistance to SMV.关键词
大豆花叶病毒/GmSZFP/GmERF7/互作蛋白/酵母双杂交文库/双分子荧光互补试验Key words
Soybean mosaic virus/GmSZFP/GmERF7/interacting proteins/yeast two-hybrid system/BiFC引用本文复制引用
齐梦楠,赵玎玲,张雪妍,张玉洁,王荣纳,刘兵强,闫龙,张洁,王冬梅..GmSZFP互作蛋白的筛选及GmERF7在大豆抵抗SMV侵染过程中的功能分析[J].中国农业科学,2025,58(14):2739-2750,12.基金项目
国家自然科学基金(32472111)、河北省自然科学基金(C2023204093)、河北省现代农业产业技术体系创新团队建设项目(HBCT2023040101,HBCT2023040202)、大豆资源精准鉴定与优异种质创新利用(21326313D-1) （32472111）
