Novel DnaJ Protein Facilitates Thermotolerance of Transgenic Tomatoes
Abstract
:1. Introduction
2. Results
2.1. Identification and Bioinformatics Analysis of SlDnaJ20
2.2. Subcellular Localization of SlDnaJ20
2.3. Expression Analysis of SlDnaJ20 in Tomato
2.4. Identification of Transgenic Plants
2.5. SlDnaJ20 Overexpression Enhanced Heat Stress Resistance
2.6. SlDnaJ20 Overexpression Alleviates ROS Accumulation by Maintaining High Levels of SOD and APX Activities
2.7. SlDnaJ20 Overexpression Alleviates Photoinhibition of Photosystem II (PSII) under Heat Stress
2.8. SlDnaJ20 Overexpression Promotes Expression of HSFs under Heat Stress
2.9. Interaction between SlDnaJ20 and Chloroplast Hsp70 (cpHsp70)
3. Discussion
4. Materials and Methods
4.1. Plant Materials, Growth Conditions, and Stress Treatments
4.2. Isolating and Sequencing of SlDnaJ20
4.3. Subcellular Localization of SlDnaJ20
4.4. Tomato Genetic Transformation and Identification
4.5. Real-Time Quantitative PCR (qPCR) Analysis
4.6. Antibody Production, Protein Acquisition, and Protein Level Analysis
4.7. Measurement of Net Photosynthetic Rate (Pn) and Chlorophyll Fluorescence
4.8. Histochemical Staining and Measurements of H2O2 and O2•−
4.9. Measurements of Chlorophyll Content and Antioxidative Enzyme Activities
4.10. Yeast Two-Hybrid Assays
4.11. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
APX | ascorbate peroxidase |
CaMV35 S | cauliflower mosaic virus 35 S |
DAB | 3,3′-diaminobenzidine |
EGFP | enhanced green fluorescent protein |
H2O2 | hydrogen peroxide |
HSF | heat-shock transcription factor |
HSP | heat-shock protein |
NBT | nitroblue tetrazolium |
O2•− | superoxide anion radical |
PSII | photosystem II |
qPCR | quantitative real-time polymerase chain reaction |
ROS | reactive oxygen species |
sHSP | small heat-shock protein |
SlDnaJ20 | Solanum lycopersicum DnaJ protein 20 |
SOD | superoxide dismutase |
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Wang, G.; Cai, G.; Xu, N.; Zhang, L.; Sun, X.; Guan, J.; Meng, Q. Novel DnaJ Protein Facilitates Thermotolerance of Transgenic Tomatoes. Int. J. Mol. Sci. 2019, 20, 367. https://doi.org/10.3390/ijms20020367
Wang G, Cai G, Xu N, Zhang L, Sun X, Guan J, Meng Q. Novel DnaJ Protein Facilitates Thermotolerance of Transgenic Tomatoes. International Journal of Molecular Sciences. 2019; 20(2):367. https://doi.org/10.3390/ijms20020367
Chicago/Turabian StyleWang, Guodong, Guohua Cai, Na Xu, Litao Zhang, Xiuling Sun, Jing Guan, and Qingwei Meng. 2019. "Novel DnaJ Protein Facilitates Thermotolerance of Transgenic Tomatoes" International Journal of Molecular Sciences 20, no. 2: 367. https://doi.org/10.3390/ijms20020367