Identification of the Potential Virulence Factors and RNA Silencing Suppressors of Mulberry Mosaic Dwarf-Associated Geminivirus

Yang, Xiuling; Ren, Yanxiang; Sun, Shaoshuang; Wang, Dongxue; Zhang, Fanfan; Li, Dawei; Li, Shifang; Zhou, Xueping

doi:10.3390/v10090472

Cited by 27 publications

(43 citation statements)

References 59 publications

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“…At 14 dpi, the PVX-C1-inoculated plants showed severe dwarfing symptoms (the overlapped C4 gene in the C1 gene has been mutated, so the symptom is caused only by the C1 protein). Previous research also showed that the C1 protein of Mulberry mosaic dwarf-associated virus can cause similar symptoms in host plants (Yang et al, 2018). Furthermore, Wezel et al found that PVX-driven expression of C1+C4 causes local and systemic necrosis in N. benthamiana, and that C4 is necessary for systemic necrosis induction (using a C1 mC4 construct) (Wezel et al, 2002).…”

Section: Discussionmentioning

confidence: 94%

Analysis of Pathogenicity and Virulence Factors of Ageratum leaf curl Sichuan virus

Jing

Ren

et al. 2020

Front. Plant Sci.

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Ageratum leaf curl Sichuan virus (ALCScV) is a novel monopartite begomovirus, which was identified from Ageratum conyzoides plants in Sichuan Province, China. In this study, we showed that ALCScV can induce typical dwarf and downward leaf-curling symptoms in Ageratum conyzoides, Helianthus annuus, and Nicotiana benthamiana plants and that the noncognate betasatellite can enhance disease symptoms and increase viral accumulation. Expression of the ALCScV-encoded V2, C1, and C4 proteins through a Potato virus X (PVX) vector caused severe symptoms in N. benthamiana. Further study revealed no symptoms in N. benthamiana plants inoculated with infectious ALCScV clones lacking the C4 protein and that the relative viral DNA accumulation levels significantly decreased when compared with ALCScV-inoculated plants. Thus, our mutational analyses demonstrated that C4 is a pathogenicity determinant that plays key roles in symptom formation and virus accumulation. Furthermore, we also demonstrated that the second glycine of C4 was critical for ALCScV pathogenicity.

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Section: Discussionmentioning

confidence: 94%

Analysis of Pathogenicity and Virulence Factors of Ageratum leaf curl Sichuan virus

Jing

Ren

et al. 2020

Front. Plant Sci.

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“…In African cassava mosaic virus (ACMV), the suppressor of silencing is C4 like Tomato leaf curl New Delhi virus (ToLCNDV) [26,59], while in CLCuDV both TrAP and C4 have suppressor functions [24]. Another protein associated with suppression is V2 from Mulberry mosaic dwarf-associated virus (MMDAV), Apple geminivirus (AGV), Tomato yellow leaf curl virus (TYLCV), and Beet curly top virus (BCTV) [29,48,[60][61][62]. Therefore, despite belonging to the same viral family each virus has different suppressor silencing proteins, so this function cannot be generalized in all geminiviruses and it is necessary to analyze each individual virus.…”

Section: Discussionmentioning

confidence: 99%

Superinfection by PHYVV Alters the Recovery Process in PepGMV-Infected Pepper Plants

Rodríguez-Gandarilla

Rodríguez-Negrete

Rivera-Bustamante

2020

Viruses

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Geminiviruses are important plant pathogens that affect crops around the world. In some geminivirus–host interactions, infected plants show recovery, a phenomenon characterized by symptom disappearance in newly emerging leaves. In pepper–Pepper golden mosaic virus (PepGMV) interaction, the host recovery process involves a silencing mechanism that includes both post-transcriptional (PTGS) and transcriptional (TGS) gene silencing pathways. Under field conditions, PepGMV is frequently found in mixed infections with Pepper huasteco yellow vein virus (PHYVV), another bipartite begomovirus. Mixed infected plants generally show a synergetic phenomenon and do not present recovery. Little is known about the molecular mechanism of this interaction. In the present study, we explored the effect of superinfection by PHYVV on a PepGMV-infected pepper plant showing recovery. Superinfection with PHYVV led to (a) the appearance of severe symptoms, (b) an increase of the levels of PepGMV DNA accumulation, (c) a decrease of the relative methylation levels of PepGMV DNA, and (d) an increase of chromatin activation marks present in viral minichromosomes. Finally, using heterologous expression and silencing suppression reporter systems, we found that PHYVV REn presents TGS silencing suppressor activity, whereas similar experiments suggest that Rep might be involved in suppressing PTGS.

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“…The RepA, V2, and V3 genes from MMDaV (GenBank accession number KP303687) (Ma et al, 2015), the V2 dm61−77aa from pGEM-T-V2 dm61−77aa (Yang et al, 2018) and the coding sequences of the fibrillarin 2 gene of N. benthamiana cDNA (GenBank accession number AM269909, NbFib2) were cloned into pENTR-D/TOPO entry vector without stop codon by standard protocols (Invitrogen, Beijing, China) to yield pENTR-RepA, pENTR-V2, pENTR-V3, pENTR-V2 dm61−77aa , and pENTR-NbFib2, respectively. For subcellular localization analysis, NbFib2 and RepA was gateway-cloned into plant binary vectors pEarleyGate102 [N-terminal cyan fluorescent protein (CFP) fusion] and pGWB454 (N-terminal RFP fusion) to yield Fib2-RFP and RepA-CFP, respectively (Earley et al, 2006;Nakagawa et al, 2007).…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…Grapevine red blotch-associated virus (GRBaV) V2 localizes to the nucleoplasm, Cajal bodies, and cytoplasm, and is redirected to the nucleolus upon coexpression with the nucleus and Cajal body-associated protein fibrillarin 2 (Fib2) (Guo et al, 2015). The V2 protein of a distinct monopartite geminivirus, Mulberry mosaic dwarf-associated virus (MMDaV), was shown to localize to both subnuclear foci and cytoplasm of the epidermal cells of Nicotiana benthamiana plants (Yang et al, 2018). Most of these experiments, however, have studied the subcellular localization of V2 in the absence of virus infection.…”

Section: Introductionmentioning

confidence: 99%

RepA Promotes the Nucleolar Exclusion of the V2 Protein of Mulberry Mosaic Dwarf-Associated Virus

et al. 2020

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Plant viruses have limited coding capacities so that they rely heavily on the expression of multifunctional viral proteins to achieve a successful infection. The functional specification of viral proteins is often related to their differential interaction with plant and viral components and somewhat depends on their localization to various subcellular compartments. In this study, we analyzed the intracellular localization of the V2 protein of Mulberry mosaic dwarf-associated virus (MMDaV), an unsigned species of the family Geminiviridae. We show that the V2 protein colocalizes with the nucleolar protein fibrillarin (NbFib2) in the nucleolus upon transient expression in the epidermal cells of Nicotiana benthamiana. A yeast-two hybrid assay, followed by bimolecular fluorescence complementation assays, demonstrated the specific interaction between V2 and NbFib2. Intriguingly, we find that the presence of MMDaV excludes the V2 protein from the nucleolus to nucleoplasm. We present evidence that the replication-associated protein A (RepA) protein of MMDaV interacts with V2 and enables the nucleolar exclusion of V2. We also show that, while V2 interacts with itself primarily in the nucleolus, the presence of RepA redirects the site of V2-V2 interaction from the nucleolus to the nucleoplasm. We further reveal that RepA promotes V2 out of the nucleolus presumably by directing the NbFib2-V2 complex from the nucleolus to the nucleoplasm. Considering the critical role of the nucleolus in plant virus infection, this RepA-dependent modulation of V2 nucleolar localization would be crucial for understanding the involvement of this subcellular compartment in plant-virus interactions.

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Identification of the Potential Virulence Factors and RNA Silencing Suppressors of Mulberry Mosaic Dwarf-Associated Geminivirus

Cited by 27 publications

References 59 publications

Analysis of Pathogenicity and Virulence Factors of Ageratum leaf curl Sichuan virus

Analysis of Pathogenicity and Virulence Factors of Ageratum leaf curl Sichuan virus

Superinfection by PHYVV Alters the Recovery Process in PepGMV-Infected Pepper Plants

RepA Promotes the Nucleolar Exclusion of the V2 Protein of Mulberry Mosaic Dwarf-Associated Virus

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