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Transcriptomic and proteomic analysis of a compatible tomato-aphid interaction reveals a predominant salicylic acid-dependent plant response.

Coppola V, Coppola M, Rocco M, Digilio MC, D'Ambrosio C, Renzone G, Martinelli R, Scaloni A, Pennacchio F, Rao R, Corrado G - BMC Genomics (2013)

Bottom Line: Among them, the SA-signaling pathway and stress-responsive SA-dependent genes play a dominant role.Furthermore, tomato response is characterized by a reduced accumulation of photosynthetic proteins and a modification of the expression of various cell wall related genes.Considering the rapid advancement of tomato genomics, this information will be important for the development of new protection strategies.

View Article: PubMed Central - HTML - PubMed

Affiliation: Dipartimento di Agraria, Università degli Studi di Napoli Federico II, 80055 Portici, NA, Italy.

ABSTRACT

Background: Aphids are among the most destructive pests in temperate climates, causing significant damage on several crops including tomato. We carried out a transcriptomic and proteomic study to get insights into the molecular mechanisms and dynamics of the tomato response to the Macrosyphum euphorbiae aphid.

Results: The time course analysis of aphid infestation indicated a complex, dynamic pattern of gene expression. Several biological functions were affected and genes related to the stress and defence response were the most represented. The Gene Ontology categories of the differentially expressed genes (899) and identified proteins (57) indicated that the tomato response is characterized by an increased oxidative stress accompanied by the production of proteins involved in the detoxification of oxygen radicals. Aphids elicit a defense reaction based on the cross-communication of different hormone-related signaling pathways such as those related to the salicylic acid (SA), jasmonic acid (JA), ethylene and brassinosteroids. Among them, the SA-signaling pathway and stress-responsive SA-dependent genes play a dominant role. Furthermore, tomato response is characterized by a reduced accumulation of photosynthetic proteins and a modification of the expression of various cell wall related genes.

Conclusions: Our work allowed a more comprehensive understanding of the signaling events and the defense dynamics of the tomato response to aphids in a compatible interaction and, based on experimental data, a model of the tomato-aphid molecular interaction was proposed. Considering the rapid advancement of tomato genomics, this information will be important for the development of new protection strategies.

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2-DE proteomic map of tomato leaves from non-infested tomato plants. Protein extracts were analyzed in first dimension (pH 4–7 linear IPG, 18 cm); second dimension was performed on a vertical slab (12%T) gel. Protein detection was achieved by using colloidal Coomassie staining. Numbering refers to differentially-represented protein spots in the M. euphorbiae-infested plants, which were then excised, digested and identified by MS procedures as reported in Additional file1: Table S5.
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Figure 5: 2-DE proteomic map of tomato leaves from non-infested tomato plants. Protein extracts were analyzed in first dimension (pH 4–7 linear IPG, 18 cm); second dimension was performed on a vertical slab (12%T) gel. Protein detection was achieved by using colloidal Coomassie staining. Numbering refers to differentially-represented protein spots in the M. euphorbiae-infested plants, which were then excised, digested and identified by MS procedures as reported in Additional file1: Table S5.

Mentions: We also carried out a proteomic analysis of tomato leaves after M. euphorbiae damage. The leaf tissue of control and infested plants at 48 hours following infestation was used for protein extraction. Proteins were subjected to 2-DE analysis and a representative Coomassie-stained gel from control leaves is shown in Figure 5. Peptide spots showing qualitative and statistically different quantitative differences between infested and control plants were further analyzed. Eighty-seven spots were selected as differentially expressed in tomato after aphid damage with a cut-off of a twofold change compared to the control. A database search with data from peptide mass fingerprinting using MALDI-TOF-MS experiments allowed the identification of the protein uniquely present in 45 spots; the remaining ones were analyzed by nanoLC-ESI-LIT-MS/MS, which identified 12 additional unique components. In the residual 30 spots, we detected multiple polypeptide species, which did not allow a quantitative evaluation of the protein expression level. The list of all the identified proteins is reported in Additional file1: Table S5, together with the corresponding quantitative variations. The annotation of their protein coding genes indicated that the most represented biological process was “stress and defence response”, followed by “primary metabolism” (Figure 6). Among the differentially represented proteins after aphid attack, those involved in the photosynthesis included the oxygen-evolving enhancer protein 1 (OEE1) (spots 35, 36, 37, 41 and 83), oxygen-evolving enhancer protein 2 (OEE2) (spots 61, 62, 65 and 78), photosystem II oxygen-evolving complex protein 3 (spot 73), ATP synthase subunit beta (spots 7, 8 and 9), ATP synthase (spot 42 and 46) and cytochrome f (spot 31). A similar trend was also observed for enzymes of the photorespiration system, such as the RuBisCO activase (spots 17, 20, 21 and 59), RuBisCO decarboxylase small chain (spot 86), aminomethyltransferase (spot 26 and 27) and glycine/serine hydroxymethyltransferase (spot 11 and 13). As a result of concomitant multiple spot changes often with a negative quantitative trend, some of them showed variations that were suggestive of the occurrence of post-translational modifications. All proteins occurring in multiple spots have been already reported to be phosphorylated on other plant species (http://phosphat.mpimp-golm.mpg.de and http://www.p3db.org). A down-representation of proteins involved in carbohydrate metabolism, namely glycosyl hydrolase family 3 protein (spot 4) and triosephosphate isomerase (spots 44, 49 and 53), was also observed. Among the proteins related to the “transport” category, the mitochondrial outer membrane protein porin (VDAC) (spot 33) and ferredoxin-1, chloroplastic (spot 66 and 74) were up-regulated. Ferredoxin also participates in other reactions in the chloroplast (e.g. redox regulation)[39] and is strongly up-regulated after pathogen attack[40,41]. Many proteins correlated to defense and stress response were down-regulated in infested plants, except for the Nodulin-related protein (spot 67). Among the down-regulated proteins two were involved in oxidative stress, such as thioredoxin peroxidase 1 (spot 64) and oxidoreductase (spot 23). The category “protein metabolism” included proteins involved in translation, complex assembly, proteolysis and folding (spots 1, 2, 3, 6, 14, 15, 19, 28, 43 and 87). All these proteins, except for the chaperonin 20 (spots 50 and 51), were down-represented, suggesting that protein synthesis and secretion patterns are significantly affected in infested tomato. Finally, glycine-rich RNA-binding protein and RNA recognition motif (RRM)-containing protein were down-regulated, as also observed in rice leaf sheaths in response to infestation by the brown planthopper (Nilaparvata lugens)[42].


Transcriptomic and proteomic analysis of a compatible tomato-aphid interaction reveals a predominant salicylic acid-dependent plant response.

Coppola V, Coppola M, Rocco M, Digilio MC, D'Ambrosio C, Renzone G, Martinelli R, Scaloni A, Pennacchio F, Rao R, Corrado G - BMC Genomics (2013)

2-DE proteomic map of tomato leaves from non-infested tomato plants. Protein extracts were analyzed in first dimension (pH 4–7 linear IPG, 18 cm); second dimension was performed on a vertical slab (12%T) gel. Protein detection was achieved by using colloidal Coomassie staining. Numbering refers to differentially-represented protein spots in the M. euphorbiae-infested plants, which were then excised, digested and identified by MS procedures as reported in Additional file1: Table S5.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3733717&req=5

Figure 5: 2-DE proteomic map of tomato leaves from non-infested tomato plants. Protein extracts were analyzed in first dimension (pH 4–7 linear IPG, 18 cm); second dimension was performed on a vertical slab (12%T) gel. Protein detection was achieved by using colloidal Coomassie staining. Numbering refers to differentially-represented protein spots in the M. euphorbiae-infested plants, which were then excised, digested and identified by MS procedures as reported in Additional file1: Table S5.
Mentions: We also carried out a proteomic analysis of tomato leaves after M. euphorbiae damage. The leaf tissue of control and infested plants at 48 hours following infestation was used for protein extraction. Proteins were subjected to 2-DE analysis and a representative Coomassie-stained gel from control leaves is shown in Figure 5. Peptide spots showing qualitative and statistically different quantitative differences between infested and control plants were further analyzed. Eighty-seven spots were selected as differentially expressed in tomato after aphid damage with a cut-off of a twofold change compared to the control. A database search with data from peptide mass fingerprinting using MALDI-TOF-MS experiments allowed the identification of the protein uniquely present in 45 spots; the remaining ones were analyzed by nanoLC-ESI-LIT-MS/MS, which identified 12 additional unique components. In the residual 30 spots, we detected multiple polypeptide species, which did not allow a quantitative evaluation of the protein expression level. The list of all the identified proteins is reported in Additional file1: Table S5, together with the corresponding quantitative variations. The annotation of their protein coding genes indicated that the most represented biological process was “stress and defence response”, followed by “primary metabolism” (Figure 6). Among the differentially represented proteins after aphid attack, those involved in the photosynthesis included the oxygen-evolving enhancer protein 1 (OEE1) (spots 35, 36, 37, 41 and 83), oxygen-evolving enhancer protein 2 (OEE2) (spots 61, 62, 65 and 78), photosystem II oxygen-evolving complex protein 3 (spot 73), ATP synthase subunit beta (spots 7, 8 and 9), ATP synthase (spot 42 and 46) and cytochrome f (spot 31). A similar trend was also observed for enzymes of the photorespiration system, such as the RuBisCO activase (spots 17, 20, 21 and 59), RuBisCO decarboxylase small chain (spot 86), aminomethyltransferase (spot 26 and 27) and glycine/serine hydroxymethyltransferase (spot 11 and 13). As a result of concomitant multiple spot changes often with a negative quantitative trend, some of them showed variations that were suggestive of the occurrence of post-translational modifications. All proteins occurring in multiple spots have been already reported to be phosphorylated on other plant species (http://phosphat.mpimp-golm.mpg.de and http://www.p3db.org). A down-representation of proteins involved in carbohydrate metabolism, namely glycosyl hydrolase family 3 protein (spot 4) and triosephosphate isomerase (spots 44, 49 and 53), was also observed. Among the proteins related to the “transport” category, the mitochondrial outer membrane protein porin (VDAC) (spot 33) and ferredoxin-1, chloroplastic (spot 66 and 74) were up-regulated. Ferredoxin also participates in other reactions in the chloroplast (e.g. redox regulation)[39] and is strongly up-regulated after pathogen attack[40,41]. Many proteins correlated to defense and stress response were down-regulated in infested plants, except for the Nodulin-related protein (spot 67). Among the down-regulated proteins two were involved in oxidative stress, such as thioredoxin peroxidase 1 (spot 64) and oxidoreductase (spot 23). The category “protein metabolism” included proteins involved in translation, complex assembly, proteolysis and folding (spots 1, 2, 3, 6, 14, 15, 19, 28, 43 and 87). All these proteins, except for the chaperonin 20 (spots 50 and 51), were down-represented, suggesting that protein synthesis and secretion patterns are significantly affected in infested tomato. Finally, glycine-rich RNA-binding protein and RNA recognition motif (RRM)-containing protein were down-regulated, as also observed in rice leaf sheaths in response to infestation by the brown planthopper (Nilaparvata lugens)[42].

Bottom Line: Among them, the SA-signaling pathway and stress-responsive SA-dependent genes play a dominant role.Furthermore, tomato response is characterized by a reduced accumulation of photosynthetic proteins and a modification of the expression of various cell wall related genes.Considering the rapid advancement of tomato genomics, this information will be important for the development of new protection strategies.

View Article: PubMed Central - HTML - PubMed

Affiliation: Dipartimento di Agraria, Università degli Studi di Napoli Federico II, 80055 Portici, NA, Italy.

ABSTRACT

Background: Aphids are among the most destructive pests in temperate climates, causing significant damage on several crops including tomato. We carried out a transcriptomic and proteomic study to get insights into the molecular mechanisms and dynamics of the tomato response to the Macrosyphum euphorbiae aphid.

Results: The time course analysis of aphid infestation indicated a complex, dynamic pattern of gene expression. Several biological functions were affected and genes related to the stress and defence response were the most represented. The Gene Ontology categories of the differentially expressed genes (899) and identified proteins (57) indicated that the tomato response is characterized by an increased oxidative stress accompanied by the production of proteins involved in the detoxification of oxygen radicals. Aphids elicit a defense reaction based on the cross-communication of different hormone-related signaling pathways such as those related to the salicylic acid (SA), jasmonic acid (JA), ethylene and brassinosteroids. Among them, the SA-signaling pathway and stress-responsive SA-dependent genes play a dominant role. Furthermore, tomato response is characterized by a reduced accumulation of photosynthetic proteins and a modification of the expression of various cell wall related genes.

Conclusions: Our work allowed a more comprehensive understanding of the signaling events and the defense dynamics of the tomato response to aphids in a compatible interaction and, based on experimental data, a model of the tomato-aphid molecular interaction was proposed. Considering the rapid advancement of tomato genomics, this information will be important for the development of new protection strategies.

Show MeSH
Related in: MedlinePlus