Profiling the Proteome of Cyst Nematode-Induced Syncytia on Tomato Roots
Abstract
:1. Introduction
2. Results
2.1. Optimization of the Fixation Method, Tissue Embedding, and Sectioning Procedure
2.2. Tissue Analysis and LCM
2.3. Protein Isolation
2.4. Mass Spectrometry Results
3. Discussion
4. Materials and Methods
4.1. Plant Material and Specimen Preparation
4.2. Laser-Capture Microdissection and Protein Sample Preparation
4.3. Mass Spectrometry
4.4. RNA Isolation, cDNA Synthesis, and qPCR Reaction
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Collection Buffer | Tissue Homogenization | Protein Sample Preparation | Polymer Purification | Sample Size |
---|---|---|---|---|
8 M urea, 1× protease inhibitors cocktail, 1× phosphatase inhibitors cocktail | Homogenizer and glass beads | PAGE-separated gel sections | Not included | 4 mm2 |
8M urea, 1× protease inhibitors cocktail, 1× phosphatase inhibitors cocktail, 100 mM Tris, 150 mM NaCl, 5 mM EDTA, 10 mM DTT, 0.5% (v/v) Triton X-100 | Pestle and Eppendorf tube | Methanol–chloroform precipitate | Included | 15 mm2 |
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Filipecki, M.; Żurczak, M.; Matuszkiewicz, M.; Święcicka, M.; Kurek, W.; Olszewski, J.; Koter, M.D.; Lamont, D.; Sobczak, M. Profiling the Proteome of Cyst Nematode-Induced Syncytia on Tomato Roots. Int. J. Mol. Sci. 2021, 22, 12147. https://doi.org/10.3390/ijms222212147
Filipecki M, Żurczak M, Matuszkiewicz M, Święcicka M, Kurek W, Olszewski J, Koter MD, Lamont D, Sobczak M. Profiling the Proteome of Cyst Nematode-Induced Syncytia on Tomato Roots. International Journal of Molecular Sciences. 2021; 22(22):12147. https://doi.org/10.3390/ijms222212147
Chicago/Turabian StyleFilipecki, Marcin, Marek Żurczak, Mateusz Matuszkiewicz, Magdalena Święcicka, Wojciech Kurek, Jarosław Olszewski, Marek Daniel Koter, Douglas Lamont, and Mirosław Sobczak. 2021. "Profiling the Proteome of Cyst Nematode-Induced Syncytia on Tomato Roots" International Journal of Molecular Sciences 22, no. 22: 12147. https://doi.org/10.3390/ijms222212147