Characterization and Action Mechanism Analysis of VvmiR156b/c/d-VvSPL9 Module Responding to Multiple-Hormone Signals in the Modulation of Grape Berry Color Formation
Abstract
:1. Introduction
2. Materials and Methods
2.1. Plant Materials and Hormone Treatments
2.2. Ultra-Performance Liquid Chromatography (UPLC) Analysis
2.3. Gas Chromatography-Mass Spectrometer (GC-MS) Analysis
2.4. Total RNA and DNA Isolation, cDNA Synthesis
2.5. Cis-Element Analysis of The Promoters from VvMIR156b/c/d
2.6. Construction of the VvMIR156b/c/d Promoter Vector and Agrobacterium-Mediated Tobacco Transient Transformation
2.7. β-Glucuronidase (GUS) Staining and Activity Detection
2.8. Construction of vvmiR156b/c/d and VvSPL9 Overexpression Vectors and Instantaneous Injection of Tomato
2.9. Gene Expression Analysis
2.10. Statistical Analysis
3. Results
3.1. The Effect of Different Hormones on the Color and Quality Traits of Grape Berry
3.1.1. Effects of Different Hormones on Grape Phenotypic Characteristics
3.1.2. Effects of Different Hormones on Grape Sugar Content
3.1.3. Effects of Different Hormones on Grape Coloring
3.1.4. Effects of Different Hormones on Grape Volatile Compound
3.2. Cis-element Analysis and GUS Activity Detection of VvMIR156b/c/d Promoters in Response to Exogenous Hormones
3.3. The Expression Characteristics of VvmiR156b/c/d and VvSPL9 Responsive to Various Hormones in the Modulation of Grape Color Formation
3.3.1. Short-Term Response of VvmiR156b/c/d and VvSPL9 to Exogenous Hormones in ‘Wink’ Leaves
3.3.2. Long-Term Response of VvmiR156b/c/d and VvSPL9 to Exogenous Hormones in ‘Wink’ Berry Skin
3.4. Expression Modes of Genes Related to Anthocyanin Synthesis by Different Hormone Treatments
3.5. Interaction Mode Variation of VvmiR156b/c/d-VvSPL9 Modules in Response to Hormone Signal in the Regulation of the Grape Color Formation
3.6. Functional Verification of Overexpression of VvMIR156b/c/d and VvSPL9 in Tomatoes
4. Discussion
4.1. Regulatory Genes for Anthocyanin Biosynthesis
4.2. Regulatory Mechanism of Hormones on Anthocyanin Synthesis
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Physiological Indexes | Different Treatment Periods | |||||
---|---|---|---|---|---|---|
Treatment | 7DAT | 14DAT | 21DAT | 28DAT | 35DAT | |
transverse diameter (mm) | CK | 20.53 ± 0.59 b | 21.35 ± 0.61 bc | 20.57 ± 0.58 b | 21.75 ± 0.73 b | 21.54 ± 0.79 b |
GA | 19.87 ± 0.63 c | 21.19 ± 0.74 c | 20.34 ± 0.36 b | 20.98 ± 0.63 c | 21.31 ± 0.82 b | |
ABA | 21.01 ± 0.34 a | 21.96 ± 0.58 a | 21.54 ± 0.41 a | 22.12 ± 0.25 b | 22.27 ± 0.29 a | |
MeJA | 20.94 ± 0.64 a | 21.73 ± 0.62 ab | 21.23 ± 0.76 a | 22.67 ± 0.59 a | 22.16 ± 0.31 a | |
NAA | 19.28 ± 0.58 d | 19.43 ± 0.54 d | 19.31 ± 0.49 c | 19.84 ± 0.62 d | 20.06 ± 0.76 c | |
longitudinal diameter (mm) | CK | 28.12 ± 0.62 b | 28.91 ± 0.77 b | 29.14 ± 0.59 ab | 29.11 ± 0.99 ab | 29.42 ± 0.89 bc |
GA | 27.36 ± 0.86 c | 28.20 ± 0.94 c | 28.85 ± 0.63 b | 28.82 ± 0.83 b | 29.17 ± 0.68 c | |
ABA | 28.58 ± 0.74 a | 29.31 ± 0.96 a | 29.45 ± 0.74 a | 29.50 ± 0.39 a | 29.90 ± 0.43 a | |
MeJA | 28.49 ± 0.57 ab | 29.15 ± 0.62 ab | 29.32 ± 0.92 a | 29.31 ± 0.65 a | 29.61 ± 1.03 ab | |
NAA | 26.82 ± 0.94 d | 27.31 ± 0.54 d | 27.89 ± 0.91 c | 28.33 ± 0.78 c | 28.62 ± 0.73 d | |
berry weigh (g) | CK | 6.13 ± 0.16 a | 6.81 ± 0.14 bc | 8.06 ± 0.09 ab | 8.31 ± 0.15 a | 8.53 ± 0.14 b |
GA | 6.05 ± 0.16 a | 6.56 ± 0.22 c | 7.78 ± 0.35 b | 8.23 ± 0.11 ab | 8.38 ± 0.19 b | |
ABA | 6.23 ± 0.19 a | 7.72 ± 0.21 a | 8.39 ± 0.38 a | 8.50 ± 0.22 a | 8.92 ± 0.29 a | |
MeJA | 6.19 ± 0.12 a | 6.97 ± 0.23 b | 8.18 ± 0.23 a | 8.41 ± 0.27 a | 8.61 ± 0.26 ab | |
NAA | 6.10 ± 0.18 a | 6.57 ± 0.15 c | 7.32 ± 0.17 c | 7.84 ± 0.14 b | 7.97 ± 0.13 c | |
soluble solid (%) | CK | 12.57 ± 0.22 c | 15.61 ± 0.21 b | 17.05 ± 0.39 c | 18.12 ± 0.47 c | 21.21 ± 0.73 c |
GA | 12.25 ± 0.37 c | 12.57 ± 0.22 c | 15.41 ± 0.21 d | 17.58 ± 0.39 d | 19.09 ± 0.61 d | |
ABA | 14.63 ± 0.41 a | 16.46 ± 0.52 a | 18.81 ± 0.52 a | 20.13 ± 0.52 a | 22.71 ± 0.67 a | |
MeJA | 13.84 ± 0.52 b | 15.89 ± 0.34 b | 17.93 ± 0.62 b | 19.44 ± 0.69 b | 21.82 ± 0.59 b | |
NAA | 11.39 ± 0.23 d | 12.46 ± 0.31 c | 14.82 ± 0.39 e | 16.37 ± 0.43 e | 17.54 ± 0.41 e |
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Su, Z.; Wang, X.; Xuan, X.; Sheng, Z.; Jia, H.; Emal, N.; Liu, Z.; Zheng, T.; Wang, C.; Fang, J. Characterization and Action Mechanism Analysis of VvmiR156b/c/d-VvSPL9 Module Responding to Multiple-Hormone Signals in the Modulation of Grape Berry Color Formation. Foods 2021, 10, 896. https://doi.org/10.3390/foods10040896
Su Z, Wang X, Xuan X, Sheng Z, Jia H, Emal N, Liu Z, Zheng T, Wang C, Fang J. Characterization and Action Mechanism Analysis of VvmiR156b/c/d-VvSPL9 Module Responding to Multiple-Hormone Signals in the Modulation of Grape Berry Color Formation. Foods. 2021; 10(4):896. https://doi.org/10.3390/foods10040896
Chicago/Turabian StyleSu, Ziwen, Xicheng Wang, Xuxian Xuan, Zilu Sheng, Haoran Jia, Naseri Emal, Zhongjie Liu, Ting Zheng, Chen Wang, and Jinggui Fang. 2021. "Characterization and Action Mechanism Analysis of VvmiR156b/c/d-VvSPL9 Module Responding to Multiple-Hormone Signals in the Modulation of Grape Berry Color Formation" Foods 10, no. 4: 896. https://doi.org/10.3390/foods10040896