Enhanced Tolerance against a Fungal Pathogen and Insect Resistance in Transgenic Tobacco Plants Overexpressing an Endochitinase Gene from Serratia marcescens
Abstract
:1. Introduction
2. Results
2.1. Bacteria Strain Identification and Phylogenetic Analysis
2.2. Cloning of Chitinase Genes
2.3. Tobacco Transformation and Molecular Analysis
2.4. Western Blot Analysis
2.5. Endochitinase Activity
2.6. Antifungal Inhibition Activity Assays on Crude Protein Extracts
2.7. Botrytis Cinerea Resistance Assays on Detached Leaves
2.8. Insect Toxicity Assay
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Screening of Bacterial Isolate for Chitinolytic Aactivity
4.3. Bacterial Identification
4.4. Chitinase Gene Isolation
4.5. Tobacco Transformations
4.6. Molecular Analyses of Tobacco Transgenic Lines
4.7. Progeny Segregation
4.8. Preparation of Crude Protein Extracts from Transgenic Tobacco Lines
4.9. Western Blotting
4.10. Endochitinase Assay
4.11. Botrytis cinerea Spore Harvest for Antifungal Inhibition Assays
4.12. Assays of Antifungal Inhibition Activity on Crude Protein Extracts
4.13. Resistance Assays of Botrytis cinerea on Detached Leaves
4.14. Bioassays for Toxicity to Insects
4.15. Analysis
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
PR- | Pathogenesis-related |
GlcNAc | β-1,4-N-acetylglucosamine |
S. marcescens | Serratia marcescens |
B. cinerea | Botrytis cinerea |
S. frugiperda | Spodoptera frugiperda |
B. thuringiensis | Bacillus thuringiensis |
PM | Peritrophic matrix |
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Navarro-González, S.S.; Ramírez-Trujillo, J.A.; Peña-Chora, G.; Gaytán, P.; Roldán-Salgado, A.; Corzo, G.; Lina-García, L.P.; Hernández-Velázquez, V.M.; Suárez-Rodríguez, R. Enhanced Tolerance against a Fungal Pathogen and Insect Resistance in Transgenic Tobacco Plants Overexpressing an Endochitinase Gene from Serratia marcescens. Int. J. Mol. Sci. 2019, 20, 3482. https://doi.org/10.3390/ijms20143482
Navarro-González SS, Ramírez-Trujillo JA, Peña-Chora G, Gaytán P, Roldán-Salgado A, Corzo G, Lina-García LP, Hernández-Velázquez VM, Suárez-Rodríguez R. Enhanced Tolerance against a Fungal Pathogen and Insect Resistance in Transgenic Tobacco Plants Overexpressing an Endochitinase Gene from Serratia marcescens. International Journal of Molecular Sciences. 2019; 20(14):3482. https://doi.org/10.3390/ijms20143482
Chicago/Turabian StyleNavarro-González, Samantha Sarai, José Augusto Ramírez-Trujillo, Guadalupe Peña-Chora, Paul Gaytán, Abigail Roldán-Salgado, Gerardo Corzo, Laura Patricia Lina-García, Víctor Manuel Hernández-Velázquez, and Ramón Suárez-Rodríguez. 2019. "Enhanced Tolerance against a Fungal Pathogen and Insect Resistance in Transgenic Tobacco Plants Overexpressing an Endochitinase Gene from Serratia marcescens" International Journal of Molecular Sciences 20, no. 14: 3482. https://doi.org/10.3390/ijms20143482