Characterization of the Role of a Non-GPCR Membrane-Bound CFEM Protein in the Pathogenicity and Germination of Botrytis cinerea

Arya, Gulab Chand; Srivastava, Dhruv Aditya; Pandaranayaka, Eswari P. J.; Manasherova, Ekaterina; Prusky, D.; Elad, Y.; Frenkel, Omer; Dvir, Hay; Harel, Amnon

doi:10.3390/microorganisms8071043

Cited by 14 publications

(13 citation statements)

References 47 publications

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“…Recent studies have revealed that necrotrophic pathogens also secrete small proteinaceous effectors that can function as pathogenicity factors. Some prominent examples of proteinaceous necrotrophic effectors that induce necrotic cell death include PtrToxA, SnToxA, BcCFEM1, BcXYG1, and the NEP-like proteins in various necrotrophs ( Ciuffetti et al, 2010 ; Dallal Bashi et al, 2010 ; Arya et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

Functional Characterization of the Nep1-Like Protein Effectors of the Necrotrophic Pathogen – Alternaria brassicae

et al. 2021

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Alternaria brassicae is an important necrotrophic pathogen that infects the Brassicaceae family. A. brassicae, like other necrotrophs, also secretes various proteinaceous effectors and metabolites that cause cell death to establish itself in the host. However, there has been no systematic study of A. brassicae effectors and their roles in pathogenesis. The availability of the genome sequence of A. brassicae in public domain has enabled the search for effectors and their functional characterization. Nep1-like proteins (NLPs) are a superfamily of proteins that induce necrosis and ethylene biosynthesis. They have been reported from a variety of microbes including bacteria, fungi, and oomycetes. In this study, we identified two NLPs from A. brassicae viz. AbrNLP1 and AbrNLP2 and functionally characterized them. Although both AbrNLPs were found to be secretory in nature, they localized differentially inside the plant. AbrNLP2 was found to induce necrosis in both host and non-host species, while AbrNLP1 could not induce necrosis in both species. Additionally, AbrNLP2 was shown to induce pathogen-associated molecular pattern (PAMP)-triggered immunity in both host and non-host species. Overall, our study indicates that AbrNLPs are functionally and spatially (subcellular location) distinct and may play different but important roles during the pathogenesis of A. brassicae.

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Section: Introductionmentioning

confidence: 99%

Functional Characterization of the Nep1-Like Protein Effectors of the Necrotrophic Pathogen – Alternaria brassicae

et al. 2021

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“…In B. Cinerea , the CFEM protein Bcin07g03260 contains only one transmembrane domain, which differs from the G-protein-coupled receptor. When the Bcin07g03260 gene was knockout, the fungus would significantly reduce spore germination and bud tube elongation resulting decreased infection rate on leaves [ 44 ]. In this study, three CFEM proteins from M. anisopliae have multiple transmembrane regions (CFEM82 and CFEM91 with six transmembrane regions, CFEM80 with seven transmembrane regions), and one (CFEM83) has a single transmembrane region ( Figure 2 ).…”

Section: Discussionmentioning

confidence: 99%

Bioinformatics Analysis and Functional Characterization of the CFEM Proteins of Metarhizium anisopliae

Cai

Liu

Yan

et al. 2022

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The entomopathogen Metarhizium anisopliae is a facultative rhizosphere or endophytic fungus available for managing pests and improving plant growth. The CFEM (common in fungal extracellular membrane) proteins form a unique group in fungi but are rarely reported in entomopathogens. In this study, we cloned and identified 13 CFEM genes from M. anisopliae (MaCFEMs). Sequence alignment and WebLogo analysis showed that eight cysteines were the most conserved amino acids in their CFEM domain. Phylogenic analysis suggested that these 13 proteins could be divided into 4 clades based on the presence of the transmembrane region and the position of CFEM domain in the whole sequence. Six MaCFEM proteins with a signal peptide and without a transmembrane domain were considered candidate effector proteins. According to Phyre2 analysis, the MaCFEM88 and MaCFEM85 have the most homologous to Csa2 in Candida albicans. Subcellular localization analysis revealed that five effectors were located in the plasma membrane, while MaCFEM88 may locate in both plasma membrane and nucleus in the treated Nicotiana benthamiana. Expression pattern analysis showed that MaCFEM81, 85, 88, and 89 expression level was significantly higher in the sporulation stage compared to other growth stages. Furthermore, the yeast secretion assay showed that six candidate effectors were able to secrete out of the cell. All of the MaCFEMs couldn’t affect INF1-induced programmed cell death (PCD), but MaCFEM85 and 88 could trigger a slight hypersensitive response both when applied separately or in combination with INF1 in N. benthamiana leaves. These findings showed that six MaCFEM potential effectors with various structures and subcellular localizations in host cells might be used to illustrate the roles of MaCFEM proteins during M. anisopliae-plant interactions.

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“…Disruption of this gene results in decreased virulence and increased sensitivity to osmotic and cell wall stress, indicating that BcCFEM1 is required for virulence and plays a key role in stress resistance [ 25 ]. The recent work by Arya et al [ 26 ] illustrates a potential new role for a non-GPCR membrane CFEM in pathogenic fungi to control virulence in the fungus B. cinerea .…”

Section: Discussionmentioning

confidence: 99%

“…Target deletion of BcCFEM1 in Botrytis cinerea suggested that the gene contributes to conidial production and stress tolerance, and most importantly the gene disrupt mutants resulted in decreased virulence on French bean ( Phaseolus vulgaris ) leaves, indicated that it was involved in pathogenicity [ 25 ]. Recently, the study by Arya et al indicated that Bcin07g03260 (a non-GPCR membrane-bound CFEM protein) deletion mutants of B. cinerea also showed significantly reduced progression of a necrotic lesion on tomato ( Solanum lycopersicum ) leaves [ 26 ].…”

Section: Introductionmentioning

confidence: 99%

Bioinformatics and Transcriptome Analysis of CFEM Proteins in Fusarium graminearum

Chen

Wang

Yang

et al. 2021

JoF

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Fusarium blight of wheat is usually caused by Fusarium graminearum, and the pathogenic fungi will secrete effectors into the host plant tissue to affect its normal physiological process, so as to make it pathogenic. The CFEM (Common in Fungal Extracellular Membrane) protein domain is unique to fungi, but it is not found in all fungi. The CFEM protein contained in F. graminearum may be closely related to pathogenicity. In this study, 23 FgCFEM proteins were identified from the F. graminearum genome. Then, features of these proteins, such as signal peptide, subcellular localization, and transmembrane domains, etc., were analyzed and candidate effectors were screened out. Sequence alignment results revealed that each FgCFEM protein contains one CFEM domain. The amino acids of the CFEM domain are highly conserved and contain eight spaced cysteines, with the exception that FgCFEM8, 9, and 15 lack two cysteines and three cysteines were missed in FgCFEM18 and FgCFEM22. A recently identified CFEM_DR motif was detected in 11 FgCFEMs, and importantly we identified two new conserved motifs containing about 29 and 18 amino acids (CFEM_WR and CFEM_KF), respectively, in some of FgCFEM proteins. Transcriptome analysis of the genes encoding CFEM proteins indicated that all the CFEM-containing genes were expressed during wheat infection, with seven and six genes significantly up- and down-regulated, respectively, compared with in planta and in vitro. Based on the above analysis, FgCFEM11 and FgCFEM23 were predicted to be F. graminearum effectors. This study provides the basis for future functional analyses of CFEM proteins in F. graminearum.

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Characterization of the Role of a Non-GPCR Membrane-Bound CFEM Protein in the Pathogenicity and Germination of Botrytis cinerea

Cited by 14 publications

References 47 publications

Functional Characterization of the Nep1-Like Protein Effectors of the Necrotrophic Pathogen – Alternaria brassicae

Functional Characterization of the Nep1-Like Protein Effectors of the Necrotrophic Pathogen – Alternaria brassicae

Bioinformatics Analysis and Functional Characterization of the CFEM Proteins of Metarhizium anisopliae

Bioinformatics and Transcriptome Analysis of CFEM Proteins in Fusarium graminearum

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