Exposure of Lycopersicon Esculentum to Microcystin-LR: Effects in the Leaf Proteome and Toxin Translocation from Water to Leaves and Fruits
Abstract
:1. Introduction
2. Results
2.1. Photosynthetic Efficiency
2.2. Protein Expression in Tomato Leaves
Biological Function | Protein Name/Accession (NCBI) | SSP | C | MCE | MCP | Stat. | |||
---|---|---|---|---|---|---|---|---|---|
Mean | SD | Mean | SD | Mean | SD | ||||
ATP synthesis | ATP synthase CF1 epsilon subunit/gi|89280641 | 3,113 | 2,161.17 | 523.64 | 1,083.10 | 67.11 | 1,614.57 | 289.78 | a↑ |
ATP synthase CF1 alpha chain/gi|89280620 | 4,713 | 16,331.87 | 415.97 | 9,004.00 | 3,492.91 | 8,123.90 | 4,598.80 | a↓, b↓ | |
ATP synthase CF1 alpha subunit/gi|28261702 | 4,723 | 1,784.37 | 416.06 | 892.93 | 96.02 | 1,144.53 | 739.62 | a↓ | |
ATP synthase delta chain, chloroplastic/gi|416681 | 6,209 | 2,544.37 | 344.27 | 1,369.07 | 225.35 | 1,968.50 | 505.96 | a↓ | |
Carbon fixation | Ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit/gi|77798370 | 7,704 | 265.55 | 197.49 | 503.30 | 0.00 | 650.77 | 158.04 | b↑ |
Ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit/gi|21634137 | 7,735 | 526.30 | 570.78 | 906.40 | 704.28 | 1331.10 | 293.83 | a↓ | |
Photosynthesis | Cytochrome b6-f complex iron-sulfur subunit, chloroplastic/gi|68566175 | 3,120 | 2,037.23 | 437.05 | 780.53 | 432.47 | 1,609.97 | 76.46 | b↓ |
Oxygen-evolving enhancer protein 1, chloroplastic/gi|12644171 | 3,339 | 17,411.43 | 4,066.45 | 21,763.37 | 11,695.21 | 9,877.70 | 688.13 | a↓, b↓ | |
Oxygen-evolving enhancer protein 2, chloroplastic/gi|350538909 | 4,107 | 29,152.87 | 1,140.43 | 9,704.93 | 4,910.78 | 18,299.77 | 2,863.23 | a↓ | |
Carbohydrate | Ribose-5-phosphate isomerase/gi|225451299 | 5,316 | 4,453.07 | 361.92 | 2,821.67 | 319.86 | 3,672.17 | 362.55 | a↓ |
Metabolism | Ribose-5-phosphate isomerase/gi|225451299 | 5,350 | 1,201.00 | 247.53 | 610.03 | 42.48 | 899.83 | 239.29 | b↑ |
Proteolysis | ATP-dependent Clp protease ATP-binding subunit clpA homolog CD4B, chloroplastic/gi|399213 | 3,869 | 316.30 | 0.00 | 451.53 | 262.70 | 686.13 | 279.57 | a↓, b↓ |
Embryogenesis and stress response | 24K germin like protein/gi|58244966 | 220 | 18,919.30 | 5,892.38 | 6,637.10 | 948.09 | 7,986.73 | 2,663.81 | b↓ |
Uncharacterized protein | Heme-binding protein 2/ gi|225470739 | 1210 | 1,193.80 | 332.38 | 711.40 | 223.81 | 576.47 | 187.06 | b↑ |
Cytoskeleton | Actin/gi|32186906 | 3,632 | 384.07 | 92.56 | 1,043.73 | 1,034.54 | 1,606.87 | 731.04 | a↑ |
2.3. Toxin Quantification in Tomato Tissues
Tissue | Treatment | MC-LR (μg/kg FW Tissue) | |
---|---|---|---|
Week 1 | Week 2 | ||
Root | C | ND | <LD |
MCE | ND | 1635.21 ± 941.11 | |
MCP | ND | <LD | |
Leaves | C | <LD | <LD |
MCE | 12,298.18 ± 8962.03 | nd | |
MCP | ND | <LD | |
Green tomato | C | <LD | <LD |
MCE | 5.41 ± 0.49 | <LD | |
MCP | 5.15 ± 0.93 | <LD | |
Mature tomato | C | <LD | <LD |
MCE | 10.52 ± 6.48 | <LD | |
MCP | 10.83 ± 0.94 | <LD |
3. Discussion
3.1. Impact of Cyanobacterial Compounds and MC-LR in Plant Metabolism
3.2. Bioaccumulation of MC-LR and Toxin Translocation from Roots to Leaves and Fruits
4. Experimental Section
4.1. Biological Material
4.2. Cyanobacterial Cell Extracts and Quantification of MC-LR
4.3. Experimental Exposure
4.4. MC-LR Extraction, Clean-up and Quantification in Plant Tissues
4.5. Chlorophyll Fluorescence
4.6. Protein Extraction for 2DE
4.7. Two-Dimensional Gel Electrophoresis (2DE)
4.8. Gel Image Acquisition and Protein Expression Analysis
4.9. Protein Identification
4.10. Statistics
5. Conclusions
Supplementary Files
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Gutiérrez-Praena, D.; Campos, A.; Azevedo, J.; Neves, J.; Freitas, M.; Guzmán-Guillén, R.; Cameán, A.M.; Renaut, J.; Vasconcelos, V. Exposure of Lycopersicon Esculentum to Microcystin-LR: Effects in the Leaf Proteome and Toxin Translocation from Water to Leaves and Fruits. Toxins 2014, 6, 1837-1854. https://doi.org/10.3390/toxins6061837
Gutiérrez-Praena D, Campos A, Azevedo J, Neves J, Freitas M, Guzmán-Guillén R, Cameán AM, Renaut J, Vasconcelos V. Exposure of Lycopersicon Esculentum to Microcystin-LR: Effects in the Leaf Proteome and Toxin Translocation from Water to Leaves and Fruits. Toxins. 2014; 6(6):1837-1854. https://doi.org/10.3390/toxins6061837
Chicago/Turabian StyleGutiérrez-Praena, Daniel, Alexandre Campos, Joana Azevedo, Joana Neves, Marisa Freitas, Remédios Guzmán-Guillén, Ana María Cameán, Jenny Renaut, and Vitor Vasconcelos. 2014. "Exposure of Lycopersicon Esculentum to Microcystin-LR: Effects in the Leaf Proteome and Toxin Translocation from Water to Leaves and Fruits" Toxins 6, no. 6: 1837-1854. https://doi.org/10.3390/toxins6061837