Two viral proteins translated from one open reading frame target different layers of plant defense

Multilayered defense responses are activated upon pathogen attack.Viruses utilize a number of strategies to maximize the coding capacity of their small genomes and produce viral proteins for infection,including suppression of host defense.Here,we reveal translation leakage as one of these strategies:two viral effec-tors encoded by tomato golden mosaic virus,chloroplast-localized C4(cC4)and membrane-associated C4(mC4),are translated from two in-frame start codons and function cooperatively to suppress defense.cC4 localizes in chloroplasts,to which it recruits NbPUB4 to induce ubiquitination of the outer membrane;as a result,this organelle is degraded,and chloroplast-mediated defenses are abrogated.However,chloroplast-localized cC4 induces the production of singlet oxygen(1O2),which in turn promotes translo-cation of the 1O2 sensor NbMBS1 from the cytosol to the nucleus,where it activates expression of the CERK1 gene.Importantly,an antiviral effect exerted by CERK1 is countered by mC4,localized at the plasma membrane.mC4,like cC4,recruits NbPUB4 and promotes the ubiquitination and subsequent degradation of CERK1,suppressing membrane-based,receptor-like kinase-dependent defenses.Importantly,this translation leakage strategy seems to be conserved in multiple viral species and is related to host range.Thisfinding suggests that stacking of different cellular antiviral responses could be an effective way to abrogate viral infection and engineer sustainable resistance to major crop viral diseases in thefield.

Tomato leaf curl New Delhi virus:an emerging plant begomovirus threatening cucurbit production

Tomato leaf curl New Delhi virus(ToLCNDV),a bipartite begomovirus,was first reported to infect tomato and has recently spread rapidly as an emerging disease to Cucurbitaceae crops.To date,the virus has been reported to infect more than 11 cucurbit crops,in 16 countries and regions,causing severe yield losses.In autumn 2022,ToLCNDV was first isolated from cucurbit plants in Southeastern coastal areas of China.Phylogenetic analysis established that these isolates belong to the Asian ToLCNDV clade,and shared high nucleotide identity and closest genetic relationship with the DNA-A sequence from the Chinese tomato-infecting ToLCNDV isolate(Accession no.OP356207)and the tomato New Delhi ToLCNDV-Severe isolate(Accession no.HM159454).In this review,we summarize the occurrence and distribution,host range,detection and diagnosis,control strategies,and genetic resistance of ToLCNDV in the Cucurbitaceae.We then summarize pathways that could be undertaken to improve our understanding of this emerging disease,with the objective to develop ToLCNDV-resistant cucurbit cultivars.

Molecular characterization and pathogenicity of an infectious clone of tomato leaf curl New Delhi virus isolate infecting Cucumis melo

Tomato leaf curl New Delhi virus(ToLCNDV)is a member of the genus Begomovirus,and causes devastating disease in the world.In recent years,ToLCNDV was rapidly spreading in China and induces severe economic losses in agriculture.In this study,we sequenced and characterized the complete genome of ToLCNDV isolates from melon plants showing leaf curling and stunting symptoms in Jiangsu Province of China.We constructed a full-length infectious cDNA clone of ToLCNDV,which could induce systemic infection with typical symptoms in Nicotiana benthamiana,Cit-rullus melo,and Citrullus lanatus plants through agrobacterium-mediated inoculation.Further experimental evidence demonstrated that the virions produced in plants infected with the infectious clone of ToLCNDV are biologically active and sap-transmissible.We also evaluated the resistance of commercial melon cultivars to ToLCNDV and found all testing melon cultivars were susceptible to ToLCNDV.Collectively,the reverse genetic system developed herein will facilitate further research on biological functions of proteins encoded by ToLCNDV and plant-ToLCNDV interactions,which might provide new insights into breeding resistance germplasm in crops.

Nucieocytoplasmic Shuttling of Geminivirus C4 Protein Mediated by Phosphorylation and Myristoylation Is Critical for Viral Pathogenicity

Many geminivirus C4 proteins induce severe developmental abnormalities in plants.We previously demon- strated that Tomato leaf curl Yunnan virus (TLCYnV)C4 induces plant developmental abnormalities at least partically by decreasing the accumulation of NbSKη,an ortholog of Arabidopsis BIN2 kinase involved in the brassinosteroid signaling pathway,in the nucleus through directing it to the plasma membrane.However, the molecular mechanism by which the membrane-associated C4 modifies the localization of NbSKη in the host cell remains unclear.Here,we show that TLCYnV C4 is a nucleocytoplasmic shuttle protein,and that C4 shuttling is accompanied by nuclear export of NbSKTI.TLCYnV C4 is phosphorylated by NbSKη in the nucleus,which promotes myristoylation of the viral protein.Myristoylation of phosphorylated C4 favors its interaction with exportin-α(XPO I);which in turn facilitates nuclear export of the C4/NbSKTI complex. Supporting this model,chemical inhibition of N-myristoyltransferases or exportin-α enhanced nuclear retention of C4,and mutations of the putative phosphorylation or myristoylation sites in C4 resulted in increased nuclear retention ofrC4 and thus decreased severity of C4-induced developmental abnormalities. The impact of C4 on development is also lessened when a nuclear localization signal or a nuclear export signal is added to its C-terminus,restricting it to a specific cellular niche and therefore impairing nucleocytoplasmic shuttling.Taken together,our results suggest that nucleocytoplasmic shuttling of TLCYnV C4,enabled by phosphorylation by NbSKη,myristoylation,and interaction with exportin-α is critical for its function as a pathogenicity factor.