Establishment of a novel virus‐induced virulence effector assay for the identification of virulence effectors of plant pathogens using a PVX‐based expression vector

Shi, Jinxia; Zhu, Yuanhong; Li, Ming; Ma, Yonggang; Hua-Rong, Liu; Zhang, Peng; Fang, Di; Guo, Yushuang; Xu, Ping; Qiao, Yongli

doi:10.1111/mpp.13000

Cited by 12 publications

(19 citation statements)

References 39 publications

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“…Based on previously obtained B. tabaci transcriptomic data (Lee et al, 2018; van Kleeff et al 2022, under revision) G4 is highly expressed in the salivary glands of adult whiteflies, while not being expressed in the nymphal stage. Last, G4 was potentially identified as a virulence effector using the VIVE assay, although this paper contains some uncertainties (Shi et al, 2020).…”

Section: Resultsmentioning

confidence: 99%

Whitefly effector G4 interacts with tomato proteins of which MIPDB141 affects whitefly performance

Naalden

Dermauw

Ilias

et al. 2023

Preprint

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The phloem-feeding insect Bemisia tabaci is an important pest, responsible for the transmission of several crop-threatening virus species. While feeding, the insect secretes a cocktail of effectors to modulate defense responses. Here, we present a set of proteins that was identified in artificial diet on which B. tabaci was salivating. We studied whether these candidate effectors can play a role in plant immune suppression. Effector G4 was the most robust suppressor of the flg22-induced ROS response when transiently expressed in Nicotiana benthamiana. In addition, G4 was able to supress ROS in Solanum lycopersicum (tomato) and Capsicum annuum (pepper). Fused to a fluorescence tag, G4 localized in the cytoplasm in N. benthamiana. A yeast two-hybrid screen combined with a luciferase bimolecular complementation and co-localization assays resulted in the identification of two target proteins in tomato: REF-like stress related protein 1 (RSP1) and meloidogyne-induced giant cell protein DB141 (MIPDB141). Silencing of MIPDB141 in tomato, using virus-induced gene silencing, reduced whitefly fecundity up to 40% demonstrating that the protein is involved in susceptibility to B. tabaci. Together our data demonstrate that effector G4 impairs tomato immunity to whiteflies by interfering with the ROS production and via a direct interaction with tomato susceptibility protein MIPDB141.

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Section: Resultsmentioning

confidence: 99%

Whitefly effector G4 interacts with tomato proteins of which MIPDB141 affects whitefly performance

Naalden

Dermauw

Ilias

et al. 2023

Preprint

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“…The extracted DNA was also quantified by Nanodrop and then diluted into the same concentration for biomass assay. Primers of GmCYP2 ( Hu et al, 2009 ) from soybean and PsActin from P. sojae were used for biomass assay ( Shi et al, 2020 ; Supplementary Table 5 ).…”

Section: Methodsmentioning

confidence: 99%

Genome-Wide Analysis of Soybean Lateral Organ Boundaries Domain Gene Family Reveals the Role in Phytophthora Root and Stem Rot

Feng

Shi

et al. 2022

Front. Plant Sci.

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The plant-specific lateral organ boundaries (LOB) domain (LBD) proteins, a family of transcription factors, play important roles in plant growth and development, as well as in responses to various stresses. However, little is known about the functions of LBD genes in soybean (Glycine max). In this study, we investigated the evolution and classification of the LBD family in soybean by a phylogenetic tree of the LBD gene family from 16 species. Phylogenetic analysis categorized these proteins into two classes (Class I and Class II) with seven subgroups. Moreover, we found that all the 18 LBD ancestors in angiosperm were kept in soybean, common bean genomes, and genome-wide duplication, suggesting the main force for the expansion of LBD from common bean to soybean. Analysis of gene expression profiling data indicated that 16 GmLBD genes were significantly induced at different time points after inoculation of soybean plants (cv. Huachun 6) with Phytophthora sojae (P. sojae). We further assessed the role of four highly upregulated genes, GmLBD9, GmLBD16, GmLBD23, and GmLBD88, in plant defense in soybean hairy roots using the transient overexpression and knockdown assays. The results showed that GmLBD9 and GmLBD23 negatively regulate plant immunity against P. sojae, whereas GmLBD16 and GmLBD88 positively manipulate plant immunity against P. sojae. Collectively, our findings expand our knowledge of the origin and evolution of the GmLBD gene family in soybean and promote the potential application of these genes in soybean genetic improvement.

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“…2018b; Shi et al . 2020). Restriction endonucleases used for digesting pBI121 were BamH I and Sac I; for pSUC2 we used EcoR I and Xho I; and for pGR107 we used Cla I and Sac I (Takara).…”

Section: Methodsmentioning

confidence: 99%

Identification of RNA silencing suppressor encoded by wheat blue dwarf (WBD) phytoplasma

Wang

Chen

et al. 2021

Plant Biol J

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Plants possess an innate immune system for defence against pathogens. In turn, pathogens have various strategies to overcome complex plant defences. Among diverse pathogens, phytoplasmas are associated with serious diseases in a range of species. RNA silencing serves as an efficient defence system against pathogens in eukaryotes but can be interrupted by RNA silencing suppressors (RSSs) encoded by pathogens. Currently, many RSSs have been identified in viruses, bacteria, oomycetes and fungi. Phytoplasmas are pathogens in several hundred plant species. In this research, 37 candidate effectors of wheat blue dwarf (WBD) phytoplasma were screened for presence of RSS. Agro‐infiltration assay, yeast expression system, floral‐dip method for constructing transgenic A. thaliana, Western blotting and RT‐qPCR were used for identification of RNA silencing suppressors. SWP16 encoded by WBD phytoplasma was found to be a secretory protein that inhibited accumulation of GFP siRNA and led to the accumulation of GPF mRNA in systemic N. benthamiana 16c. Furthermore, in A. thaliana SWP16 inhibited production of miRNAs, which are components of RNA silencing. SWP16 also promoted infection of potato virus X. We conclude that SWP16 encoded by WBD phytoplasma was an RSS, suppressing systemic RNA silencing. This is the first evidence that a phytoplasma encodes an RSS and provides a theoretical basis for research on the interaction mechanisms between pathogens and plants.

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Establishment of a novel virus‐induced virulence effector assay for the identification of virulence effectors of plant pathogens using a PVX‐based expression vector

Cited by 12 publications

References 39 publications

Whitefly effector G4 interacts with tomato proteins of which MIPDB141 affects whitefly performance

Whitefly effector G4 interacts with tomato proteins of which MIPDB141 affects whitefly performance

Genome-Wide Analysis of Soybean Lateral Organ Boundaries Domain Gene Family Reveals the Role in Phytophthora Root and Stem Rot

Identification of RNA silencing suppressor encoded by wheat blue dwarf (WBD) phytoplasma

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