Comparative Proteomic Analysis by iTRAQ Reveals that Plastid Pigment Metabolism Contributes to Leaf Color Changes in Tobacco (Nicotiana tabacum) during Curing
Abstract
:1. Introduction
2. Results
2.1. Color and Phenotypic Changes of Tobacco Leaves during Curing
2.2. Ultrastructural Observations of Tobacco Leaves during Curing
2.3. Physiological Attributes of Tobacco Leaves during Curing
2.4. Pigment Degradation Products Analysis in Tobacco Leaves during Curing
2.5. Protein Profile Analysis of Tobacco Leaves Using iTRAQ
2.6. DEPs Identified and Functional Analysis
2.7. DEPs Involved in Carotenoid and Chlorophyll Metabolism
2.8. Validation of iTRAQ Data by qRT-PCR
3. Discussion
3.1. Leaf Color Change is Determined by the Carotenoid and Chlorophyll Content
3.2. Effect of Cell Ultrastructure Damage on Pigment Metabolism and Leaf Color Change
3.3. Role of Physiological Parameters in Pigment Metabolism and Leaf Color Change
3.4. Role of DEPs in Carotenoid and Chlorophyll Metabolism and Color Change
4. Materials and Methods
4.1. Plant Material and Sampling
4.2. Color Analysis
4.3. Ultrastructural Observation
4.4. Physiological Measurements
4.5. Pigment Degradation Products Analysis
4.6. Protein Extraction
4.7. iTRAQ Labeling and Strong Cation Exchange (SCX) Fractionation
4.8. LC-ESI-MS/MS Analysis
4.9. iTRAQ protein Identification and Quantification
4.10. Bioinformatics Analysis
4.11. RNA Extraction and qRT-PCR Analysis
4.12. Data Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Concentration | Curing Time (h) | ||
---|---|---|---|
0 | 48 | 72 | |
β-carotene (μg·gDM‒1) | 275.37 ± 7.32 | 151.19 ± 3.92** | 70.63 ± 1.20** |
Lutein (μg·gDM‒1) | 384.29 ± 5.02 | 213.85 ±6.64** | 86.23 ± 1.93** |
Violaxanthin (μg·gDM‒1) | 99.73 ± 2.98 | 46.49 ± 2.55** | 26.22 ± 0.62** |
Neoxanthin (μg·gDM‒1) | 39.72 ± 1.53 | 14.85 ± 0.44** | 7.97 ± 0.28** |
Chlorophyll a (mg·gFM‒1) | 0.71 ± 0.02 | 0.10 ± 0.01** | 0.04 ± 0.01** |
Chlorophyll b (mg·gFM‒1) | 0.33 ± 0.01 | 0.07 ± 0.01** | 0.04 ± 0.01** |
SPAD value | 21.87 ± 0.63 | 5.36 ± 0.39** | 1.45 ± 0.23** |
Carotenoids/Chlorophylls | 0.29 ± 0.01 | 1.30 ± 0.06** | 2.22 ± 0.14** |
Xanthophylls/β-carotene | 1.91 ± 0.06 | 1.84 ± 0.07 | 1.71 ± 0.03* |
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Wu, S.; Guo, Y.; Adil, M.F.; Sehar, S.; Cai, B.; Xiang, Z.; Tu, Y.; Zhao, D.; Shamsi, I.H. Comparative Proteomic Analysis by iTRAQ Reveals that Plastid Pigment Metabolism Contributes to Leaf Color Changes in Tobacco (Nicotiana tabacum) during Curing. Int. J. Mol. Sci. 2020, 21, 2394. https://doi.org/10.3390/ijms21072394
Wu S, Guo Y, Adil MF, Sehar S, Cai B, Xiang Z, Tu Y, Zhao D, Shamsi IH. Comparative Proteomic Analysis by iTRAQ Reveals that Plastid Pigment Metabolism Contributes to Leaf Color Changes in Tobacco (Nicotiana tabacum) during Curing. International Journal of Molecular Sciences. 2020; 21(7):2394. https://doi.org/10.3390/ijms21072394
Chicago/Turabian StyleWu, Shengjiang, Yushuang Guo, Muhammad Faheem Adil, Shafaque Sehar, Bin Cai, Zhangmin Xiang, Yonggao Tu, Degang Zhao, and Imran Haider Shamsi. 2020. "Comparative Proteomic Analysis by iTRAQ Reveals that Plastid Pigment Metabolism Contributes to Leaf Color Changes in Tobacco (Nicotiana tabacum) during Curing" International Journal of Molecular Sciences 21, no. 7: 2394. https://doi.org/10.3390/ijms21072394