Protein expression analysis in N. benthamiana plants agroinfiltrated with pBBWV1-Wt or pBBWV1-G492C compared with non-infected plants.
The effect of BBWV-1 infection on protein expression of N. benthamiana plants was analyzed by LC-MS/MS in plants agroinfiltrated with A. tumefaciens cells harboring the 35S-driven full-length cDNA infectious clone corresponding to BBWV-1 RNA1 and RNA2 wild type (pBBWV1-Wt). The role of BBWV-1 protein VP37 in the accumulation of N. benthamiana proteins was also studied by agroinfiltration of a BBWV-1 cDNA infectious clone mutant knocking out VP37 (pBBWV1-G4922C) (Carpino et al., 2020). As healthy controls, N. benthamiana plants were mock-infiltrated by using infiltration medium without A. tumefaciens cells. All N. benthamiana plants agroinfiltrated with pBBWV-Wt developed identical symptoms to those induced by BBWV-1 such as plant stunting and severe leaf mosaic symptoms in the new emergent leaves at 10–21 days post infiltration (dpi), whereas the plants agroinfiltrated with pBBWV1-G492C showed only slight mosaic symptoms. The healthy mock-infiltrated control plants showed no symptoms in the new emergent leaves and remained symptomless after the 21 dpi (Fig. 1). All plants showed some necrotic spots in the infiltrated leaves due to the mechanical injuries produced during the infiltration process. These results are identical to those previously reported that described severe symptoms in N. benthamiana plants agroinfiltrated with pBBWV1-Wt or mechanically infected with BBWV-1 and slight symptoms in equivalent plants agroinfiltrated with pBBWV1-G492C (Carpino et al., 2020a; Medina et al., 2022).
Samples of symptomatic apical leaves of N. benthamiana plants agroinfiltrated with BBWV-Wt or pBBWV1-G492C were collected at 15 dpi, when symptoms were completely developed. At the same time, samples of asymptomatic apical leaves of mock-infiltrated plants were collected and used as healthy controls. All the samples were processed, and the protein extracts corresponding to three individual plants per each group of plants (pBBWV1-Wt, pBBWV1-G4922C, or mock-infiltrated) were pooled to obtain one biological replicate. Then, four biological replicates for each group of plants were analyzed by LC-MS/MS. Statistical PCA and DA analysis of dimensionality reduction were performed to assess the homogeneity of results showing an acceptable aggrupation of the biological replicates in each plant group (Fig. 2). The LC-MS/MS output of spectral library identified 1186 proteins (FDR < 0.01). Analysis by using an Elastic-Net regression model of the individual biological replicates revealed a total of 46 proteins (including the two viral polyproteins) with differential expression (FDR < 0.01) and accumulation ratio between + 1.5 and − 2 that could be related to BBWV-1 infection (Fig. 3). In the N. benthamiana plants agroinfiltrated with pBBWV-Wt, 46 proteins (including the two viral polyproteins) were differentially expressed respect the healthy control plants (22 host proteins were overexpressed and 22 host proteins were underexpressed). Twenty out of 22 overexpressed proteins were also overexpressed in the N. bentamiana plants agroinfiltrated with pBBWV1-G4922C; one protein was overexpressed only in plants agroinfiltrated with pBBWV1-Wt, and other protein had high expression level in plants agroinfiltrated with pBBWV1-wt and healthy control plants.
Finally, it was detected 24 proteins with differential expression in N. benthamiana plants agroinfiltrated with pBBWV1-Wt in comparison to those agroinfiltrated with BBWV1-G4922C (not expressing BBWV-1 VP37 protein): two proteins were overexpressed, and 22 were underexpressed. The differential expression of host proteins in absence of VP37 could be related with the function of this viral protein that is a determinant of pathogenicity exacerbating symptoms, acting as suppressor of gene silencing and producing important subcellular alterations (Carpino et al., 2020a; Carpino et al., 2020b; Medina et al., 2022)
Proteomic analysis employing different platforms has been successfully used for the study of plant-virus interactions for several viruses (Jain et al., 2021; Souza et al., 2019). However, the detection of host proteins showing differential expression depended on many factors such as the virus-plant model studied, the proteomic technique employed (including restriction levels implemented by statistical analysis) and the infection period. It was reported that CPSMV infection, from the family Secoviridae, induced the differential expression of 2033 proteins in cowpea. Over these 2033 proteins, 1078 were detected at 2 dpi, whereas the number decreased to 955 proteins at 6 dpi (Varela et al., 2017). Pea (Pisum sativum) plants infected with pea seed-borne mosaic virus (PSbMV) showed 116 proteins with differential expression in comparison to healthy plants (Cerna et al., 2017). Tobacco mosaic virus (TMV) infection rendered different results, in different tobacco (Nicotiana tabacum) cultivars: the NC89 hypersensitive cultivar showed 260 proteins with differential expression, whereas in the Yuyan8 tolerant cultivar, the number decreased to 183 proteins (Wang et al., 2016).
Identification of the host proteins showing differential expression in N. benthamiana plants agroinfiltrated with pBBWV1-Wt or pBBWV1-G492C
The host proteins showing differential expression among the three groups of N. benthamiana (agroinfiltrated with pBBWV1-Wt or BBWV1-G4922C and mock-infiltrated control plants) were analyzed by using the UniprotKB (https://www.uniprot.org/) and Gen Ontology (GO, http://geneontology.org/) databases. All information obtained in these analyses such as the detection of homologous proteins in different hosts, potential functions, and subcellular localizations are shown in Table 1.
Table 1
Database analysis (UniprotKB and Gen ontology (GO) of the 44 Nicotiana benthamiana proteins showing significant differences (FDR < 0.01) in expression level among plants agroinfiltrated with pBBWV1-Wt, pBBWV1-G492C and mock-infiltrated used as healthy controls.
Accession number | Name | Subcellular localization | Organism | Function |
sp|Q42971|ENO_ORYSJ | Enolase | Cytoplasmic | Oryza sativa | Glycolysis |
sp|Q9M4S8|TPIC_FRAAN | Triosephosphate isomerase | Chloroplastic | Fragaria ananassa | Calvin cycle |
sp|Q9SVD7|UEV1D_ARATH | Ubiquitin-conjugating enzyme E2 variant 1D | Nucleus | Arabidopsis thaliana | Ubiquitination |
sp|Q10AZ4|ACT3_ORYSJ | Actin-3 | Cytoplasmic | Oryza sativa | Cell cytoskeleton, cytoplasmic streaming, cell shape and division, organelle movement |
sp|P36183|ENPL_HORVU | Endoplasmin homolog | Endoplasmic reticulum | Hordeum vulgare | Chaperone (protein folding) |
sp|O22263|PDI21_ARATH | Protein disulfide-isomerase like 2 − 1 | Endoplasmic reticulum | Arabidopsis thaliana | Rearrangement of -S-S- bonds in proteins: Plant fertility and embryo development. |
sp|P35016|ENPL_CATRO | Endoplasmin homolog | Endoplasmic reticulum | Catharanthus roseus | Chaperone (protein folding) |
sp|Q9FGX1|ACLB2_ARATH | ATP-citrate synthase beta chain protein 2 | Cytoplasmic | Arabidopsis thaliana | Lipid synthesis |
sp|P50218|IDHC_TOBAC | Isocitrate dehydrogenase [NADP] | Cytoplasmic | Nicotiana tabacum | Glutamate synthase pathway |
sp|P29196|CAPP_SOLTU | Phosphoenolpyruvate carboxylase | Chloroplastic | Solanum tuberosum | Calvin cycle |
sp|P46225|TPIC_SECCE | Triosephosphate isomerase | Chloroplastic | Secale cereale | Calvin cycle |
sp|Q9LEC8|DHEB_NICPL | Glutamate dehydrogenase B | Cytoplasmic | Nicotiana plumbaginifolia | Glutamate pathway |
sp|Q42876|AMPL2_SOLLC | Leucine aminopeptidase 2 | Chloroplastic | Solanum lycopersicum | N-terminal amino acid release of proteins |
sp|Q9ZUC2|BCA3_ARATH | Beta carbonic anhydrase 3 | Cytoplasmic | Arabidopsis thaliana | Reversible hydration of carbon dioxide. |
sp|Q6YZX6|ACOC_ORYSJ | Putative aconitate hydratase | Cytoplasmic | Oryza sativa | Glyoxylate and dicarboxylate metabolism |
sp|P37225|MAON_SOLTU | NAD-dependent malic enzyme 59 kDa isoform | Mitochondrial | Solanum tuberosum | Malate metabolism an amino acid biosynthesis |
sp|Q9FGT9|GPDL6_ARATH | Glycerophosphodiester phosphodiesterase GDPDL6 | Membrane | Arabidopsis thaliana | Lipid and glicerol metabolism |
sp|Q9T0G2|CYC2_ARATH | Cytochrome c-2 | Mitochondrial | Arabidopsis thaliana | Electron transfer activity, ATP synthesis |
sp|P29696|LEU3_SOLTU | 3-isopropylmalate dehydrogenase | Chloroplastic | Solanum tuberosum | L-leucine biosynthesis |
sp|P17614|ATPBM_NICPL | ATP synthase subunit beta | Mitochondrial | Nicotiana plumbaginifolia | ATP synthesis |
sp|Q5E924|G3PP2_ARATH | Glyceraldehyde-3-phosphate dehydrogenase GAPCP2 | Chloroplastic | Arabidopsis thaliana | Glycolysis |
sp|Q40412|ABA2_NICPL | Zeaxanthin epoxidase | Chloroplastic | Nicotiana plumbaginifolia | Abscisate biosynthesis |
sp|P68173|SAHH_TOBAC | Adenosylhomocysteinase | Cytoplasmic | Nicotiana tabacum | L-homocysteine biosynthesis |
sp|P37222|MAOC_SOLLC | NADP-dependent malic enzyme | Chloroplastic | Solanum lycopersicum | C4 acid pathway |
sp|Q93ZN9|DAPAT_ARATH | LL-diaminopimelate aminotransferase | Chloroplastic | Arabidopsis thaliana | Lysine biosynthesis |
sp|O82802|SIR1_TOBAC | Sulfite reductase 1 [ferredoxin] | Chloroplastic | Nicotiana tabacum | Assimilatory sulphate reduction pathway involved in development and growth |
sp|O81155|CYSKP_SOLTU | Cysteine synthase | Chloroplastic/Chromoplastic | Solanum tuberosum | L-cysteine biosynthesis |
sp|Q43848|TKTC_SOLTU | Transketolase | Chloroplastic | Solanum tuberosum | Calvin cycle |
sp|P25851|F16P1_ARATH | Fructose-1,6-bisphosphatase | Chloroplastic | Arabidopsis thaliana | Calvin cycle |
sp|Q9SYI0|SECA1_ARATH | Protein translocase subunit SECA1 | Chloroplastic | Arabidopsis thaliana | Protein signalling and translocation |
sp|P14671|TRPB1_ARATH | Tryptophan synthase beta chain 1 | Chloroplastic | Arabidopsis thaliana | L-tryptophan biosynthesis |
sp|Q42601|CARB_ARATH | Carbamoyl-phosphate synthase large chain | Chloroplastic | Arabidopsis thaliana | L-arginine biosynthesis |
sp|Q94A94|DCDA2_ARATH | Diaminopimelate decarboxylase 2 | Chloroplastic | Arabidopsis thaliana | L-lysine biosynthesis |
sp|Q9LMM0|GPAT4_ARATH | Glycerol-3-phosphate 2-O-acyltransferase 4 | Membrane | Arabidopsis thaliana | Esterifies acyl-group |
sp|Q9SIM4|RL141_ARATH | 60S ribosomal protein L14-1 | Ribosomal | Arabidopsis thaliana | Ribosomal large subunit biogenesis |
sp|O23254|GLYC4_ARATH | Serine hydroxymethyltransferase 4 | Cytoplasmic | Arabidopsis thaliana | one-carbon (C1) pathways by catalyzing the reversible conversions of L-serine to glycine and tetrahydrofolate (THF) to 5,10-methylene THF. |
sp|Q43794|SYE_TOBAC | Glutamate–tRNA ligase | Chloroplastic/Mitochondrial | Nicotiana tabacum | Attachment of glutamate to tRNA |
sp|Q9MAP3|RK11_ARATH | 50S ribosomal protein L11 | Chloroplastic | Arabidopsis thaliana | Ribosomal component |
sp|O24133|CHLD_TOBAC | Magnesium-chelatase subunit ChlD | Chloroplastic | Nicotiana tabacum | Chlorophyll biosynthesis |
sp|P21239|RUB1_BRANA | RuBisCO large subunit-binding protein subunit alpha | Chloroplastic | Brassica napus | RuBisCO small and large subunits binding and assembly of the enzyme oligomer |
sp|Q42694|RUBA_CHLRE | RuBisCO large subunit-binding protein subunit alpha | Chloroplastic | Chlamydomonas reinhardtii | RuBisCO small and large subunits binding and assembly of the enzyme oligomer |
sp|P93841|ISPE_SOLLC | 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase | Chloroplastic/Chromoplastic | Solanum lycopersicum | Isopentenyl diphosphate biosynthesis |
sp|O24163|PPOC_TOBAC | Protoporphyrinogen oxidase | Chloroplastic | Nicotiana tabacum | Chlorophyll biosynthesis |
sp|P46299|RS4_GOSHI | 40S ribosomal protein S4 | Cytoplasmic | Gossypium hirsutum | Ribosomal component |
The 44 host proteins showing differential expression between N. benthamiana plants agroinfiltrated with pBBWV1-Wt and healthy controls were related with a broad spectrum of biochemical and physiological processes such as cytoskeleton formation, amino acid, carbohydrate or fatty metabolism, photosynthesis, and photorespiration, synthesis of proteins and plant hormones, ubiquitination, intercellular communication, and plant development. Each protein is also involved in different biochemical and biological pathways, showing a complex interaction of BBWV-1 with its host. These proteins were mainly located in the chloroplast (24 proteins), whereas the rest of them had a cytoplasmic (10 proteins), mitochondrial (four proteins), reticulum endoplasmic (three proteins), membranous (two proteins), ribosomal (one protein), chromoplast (one protein) and nuclear (one protein) localization. Two chloroplastic proteins, the glutamate-tRNA ligase and the cysteine synthase, are also in the mitochondria and chromoplast, respectively.
Following the UniprotKB and Gen Ontology database, the 20 overexpressed proteins in both N. benthamiana plants agroinfiltrated with pBBWV1-Wt or BBWV1-G4922C respect to the healthy control plants were related with: i) amino acid metabolism (isocitrate dehydrogenase, 3-isopropylmalate dehydrogenase, glutamate dehydrogenase B, leucine aminopeptidase 2 and NAD-dependent malic enzyme 59 kDa isoform), ii) carbon fixation in the Calvin cycle during the photosynthesis (triosephosphate isomerase, phosphoenolpyruvate carboxylase and triosephosphate isomerase), iii) carbohydrate metabolism (enolase and putative aconitate hydratase), iv) fatty metabolism (ATP-citrate synthase beta chain protein and glycerophosphodiester phosphodiesterase GDPDL6), v) protein folding (two chaperone endoplasmic homologs), vi) ATP synthesis (cytochrome C-2 and ATP synthase subunit beta), vii) embryo development and plant fertility (protein -isomerase like 2 − 1), viii) component of the cellular cytoskeleton (actin-3), ix) reversible hydration of carbon dioxide (beta carbonic anhydrase 3), and multiple regulatory functions such as protein degradation, cellular growth, nucleic acid reparation, and stress response (ubiquitin-conjugating enzyme E2 variant 1D). These proteins seem to be related to the BBWV-1 infection independently of VP37 expression.
On the other hand, the 22 proteins underexpressed in the N. benthamiana plants agroinfiltrated with pBBWV1-Wt respect to those agroinfiltrated with pBBWV1-G492C or healthy control plants were related with: i) amino acid metabolism (adenosylhomocysteinase, LL-diaminopimelate aminotransferase, cysteine synthase, tryptophan synthase beta chain 1, carbamoyl-phosphate synthase large chain and Diaminopimelate decarboxylase 2), ii) photosynthesis (NADP-dependent malic enzyme, transketolase, fructose-1,6-bisphosphatase, magnesium-chelatase subunit ChlD and protoporphyrinogen oxidase), iii) protein synthesis (60S ribosomal protein L14-1, glutamate–tRNA ligase, 50S ribosomal protein L11 and 40S ribosomal protein S4), iv) fatty metabolism (glycerol-3-phosphate 2-O-acyltransferase 4 and 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase), v) protein folding (two annotations of RuBisCO large subunit-binding protein subunit alpha, in different microorganisms), vi) one-carbon (C1) pathway involved in photorespiration and stress responses (serine hydroxymethyltransferase 4), vii) protein signalling and translocation (protein translocase subunit SECA1) and viii) development and growth (sulfite reductase 1).
In addition to the mentioned host proteins, there was one protein, the glyceraldehyde-3-phosphate dehydrogenase (GAPCP2), that was only overexpressed in N. benthamiana plants agroinfiltrated with pBBWV1-Wt but not in those agroinfiltrated with pBBWV1-G4922C or healthy control plants. Another host protein, the zeaxanthin epoxidase, was overexpressed in N. benthamiana plants agroinfiltrated with pBBWV1-Wt and in control plants in comparison with those agroinfiltrated with BBWV1-G4922C.
During infection, viruses produce different subcellular and molecular changes in the host to allow their replication, movement, and spread. These host changes can also include modifications in energy sources, development, and reproductive stage of infected plants (Souza et al., 2019). For example, rice stripe virus (RSV) alters the biosynthesis of the chlorophyll pigment (Wang et al., 2015), affecting the photosynthesis in infected rice (Oryza sativa) plants. In this work, it was found the alteration of the accumulation of eight N. benthamiana proteins involved in photosynthesis that could be associated with BBWV-1 infection (triosephosphate isomerase, phosphoenolpyruvate carboxylase, triosephosphate isomerase, NADP-dependent malic enzyme, transketolase, fructose-1,6-bisphosphatase, magnesium-chelatase subunit ChlD, and protoporphyrinogen oxidase). Five of these proteins were underexpressed in plants agroinfiltrated with pBBWV1-Wt compared with those agroinfiltrated with pBBWV1-G4922C or healthy control plants, and therefore, they could be related to VP37 expression and the increase of virus pathogenicity. Among these proteins is the fructose-1,6-biphosphatase, which modulates the CO2 fixation in the Calvin Cycle. Cowpea infected with CPSMV and beet infected with beet curly top virus (BCTV), showed a deregulation of the photosynthesis associated with changes in the accumulations of this protein (Varela et al., 2017).
Other host proteins with altered expression during BBWV-1 infection are the actin-3 and cytochrome C-2. The first protein is an essential component of the cell cytoskeleton responsible for cytoplasmic streaming, cell shape determination, cell division, organelle movement, and growth. The actin can interact with some plant viruses such as TMV, potato virus X (PVX), or tomato bushy stunt virus (TBSV), allowing the viral movement to adjacent cells through plasmodesmata structures (Harries et al., 2009; Varela et al., 2017). Cytochrome C-2 is a protein with electron transfer activity and ATP synthesis and changes in its accumulation were reported for some plant viruses as mungbean yellow mosaic India virus (MYMIV) in Vigna mungo (Souza et al., 2019).
Amino acid biosynthesis and protein formation are also important metabolic pathways affected by plant virus infection (Zeier, 2013). Plant viruses optimize the translation of their own proteins modulating the tRNAs and the synthesis of host ribosomal proteins (Albers et al., 2016; Bahir et al., 2009). In this work, 14 host proteins related to the biosynthesis of different amino acids such as glutamate, leucine, or lysine, and the ribosomal proteins L14-1, L11, and S4 showed differential accumulation in N. benthamiana plants infected with BBWV-1. Nine of these proteins were related to BBWV-1 VP37 protein, including the glutamate-tRNA ligase and all the ribosomal proteins. Other important proteins that could be related to VP37 are the sulfite reductase 1, serine hydroxymethyltransferase 4, and GAPCP2. Sulfite reductase is coupled with the ferredoxin in the chloroplast photosystems, and it is also involved in plant development and growth. Some viral products such as the CP of ToMV or the HC-Pro of sugarcane mosaic virus (SCMV), can modulate the function of this protein (Yang et al., 2020). Serine hydroxymethyltransferase 4 is involved in one-carbon (C1) pathway, and changes in its accumulation can affect the photorespiratory pathway and stress responses (Zhang et al., 2019). GAPCP2 plays an important role in different pathways such as glycolysis, starch metabolism, generation of primary metabolites for fatty acid and amino acid synthesis and pollen formation. Also, this protein contributes to the generation of signalling molecules in the abscisic acid pathway (Agut et al., 2016; Cho and Yoo, 2011). The abscisic acid is a plant hormone with an important role in the development process including seed and bud dormancy, and its biosynthesis is usually activated for both abiotic and biotic stresses (Chan, 2012). In this work, it was determined that GAPCP2 was overexpressed in N. benthamiana plants agroinfiltrated with pBBWV1-Wt in comparison with those agroinfiltrated with pBBWV1-G4922C and the healthy control plants. Overexpression of GAPCP2 protein was reported in plants infected with different viruses such as RSV, TMV, and CPSMV (Varela et al., 2019; Wang et al., 2015; Wang et al., 2016). Finally, the zeaxanthin epoxidase, which is involved also in the abscisic acid biosynthesis, was underexpressed in N. benthamiana plants agroinfiltrated with pBBWV1-G4922C in comparison with those agroinfiltrated with pBBWV1-Wt and healthy control plants, that showed similar levels of accumulation. Curiously, the lack of viral VP37 protein reduce the expression of zeaxanthin epoxidase.
BBWV-1 infection induces strong symptoms and subcellular alterations in N. benthamiana plants, and the role played by the viral VP37 protein has been studied (Carpino et al., 2020b; Medina et al., 2022). Severe leaf mosaic symptoms and subcellular changes in mesophilic cells such as large starch and plastoglobuli accumulation in chloroplasts, mitochondrial proliferation, cytoplasmic vesiculation, and the presence of plasmalemmasomes were reported in N. benthamiana plants agroinfiltrated with pBBWV1-Wt. However, equivalent plants agroinfiltrated with pBBWV1-G492C knocking out VP37 showed slight leaf mosaic symptoms, and the subcellular alterations were restricted to the presence of plastoglobuli in chloroplasts and cytoplasmic vesiculation.
Plant symptoms and subcellular alterations observed in N. benthamiana plants agroinfiltrated with pBBWV1-Wt could be related to the differential expression of 44 plant proteins reported here. These proteins (up to 24) were localized in chloroplasts, and most of them (up to 19) were related to the viral VP37 protein. Therefore, deregulation of these chloroplastic proteins may be associated with the high accumulations of starch observed in N. benthamiana plants agroinfiltrated with pBBWV1-Wt. One of these proteins is the GAPCP2, that is directly involved in starch metabolism. GAPCP2 is overexpressed in N. bentamiana agroinfiltrated with pBBWV1-Wt in comparison with those agroinfiltrated with pBBWV1-G4922C or healthy control plants. Starch is synthesized in the chloroplasts from ADP-Glucose (primary product of photosynthesis) and functions as a transient reserve of carbohydrates metabolized during night conditions (Zeeman et al., 2010). Abiotic and/or biotic stresses such as infection by plant viruses, alter the starch metabolism inducing its accumulation (Ali Fayez and Younis Mahmoud, n.d.; Zechmann et al., n.d.; Zhan et al., 2021).
Plasmalemmasomes or paramural bodies are membranous structures showing continuity of the plasmatic membrane. The function of these structures has been related to the virus movement between cells (Wan and Laliberté, 2015). We found two proteins with membranous localization: i) the glycerophosphodiester phosphodiesterase GDPDL6 overexpressed in both N. benthamiana plants agroinfiltrated with pBBWV1-Wt and pBBWV1-G4922C, and ii) the glycerol-3-phosphate 2-O-acyltransferase 4 that is underexpressed only in N. benthamiana plants agroinfiltrated with pBBWV1-Wt. For these two host proteins, only the underexpression of the last one was associated with the viral VP37 protein expression and plasmalemmasomes formation. Other membranous alterations reported previously in N. benthamiana plants agroinfiltrated with pBBWV1-Wt and pBBWV1-G4922C were the increase of cytoplasmic vesicles (Medina et al., 2022). In this case, cytoplasmic vesiculation did not depend on viral VP37 protein, and could be consequence of overexpression of GDPDL6 in both groups of plants.
In conclusion, LC-MS/MS analysis revealed the differential expression of 44 host proteins related to BBWV-1 infection, plant symptom development, and subcellular alterations in mesophilic cells. Predicted functions and localizations of the proteins showing differential expression revealed an alteration in the principal metabolic and physiological processes of N. benthamiana plants infected with BBWV-1. These proteins were localized in different cellular organelles but mainly in chloroplast, which is affected by the virus infection inducing the accumulation of starch and plastoglobuli (Medina et al., 2022).
This work is the first analysis of the host protein expression profile during BBWV-1 infection. Results presented here complement previous studies conducted to understand the pathological process during BBWV-1 infection and virus-host interactions (Carpino et al., 2020a,b; Medina et al, 2022). Moreover, provide new data about the putative role of BBWV-1 VP37 to modulate plant symptoms and subcellular alterations during BBWV-1 infection.