Light- and Temperature-Induced Expression of an R2R3-MYB Gene Regulates Anthocyanin Biosynthesis in Red-Fleshed Kiwifruit
Abstract
:1. Introduction
2. Results and Discussion
2.1. Identification, Genomic Location, Gene Structure, and Motif Composition of R2R3-MYB Genes in Kiwifruit
2.2. Phylogenetic Analysis of AccR2R3-MYB TFs
2.3. Expression Preference of AccR2R3-MYBs in Kiwifruit
2.4. Identification of Cis-Acting Regulatory Elements of Anthocyanin-Related AccR2R3-MYBs
2.5. Light and Low-Temperature Synergistically Induced the Red Pigmentation in the Callus of A. chinensis cv. Hongyang
2.6. Sequence Analysis, Transcriptional Activation Ability, and Expression Characteristic of Acc00493 (AcMYB10)
2.7. Gene Functional Study by Overexpression of AcMYB10 in A. chinensis cv. Hongyang and Using the VIGS (Virus-Induced Gene Silencing) System in A. chinensis cv. Hongshi No. 2
2.8. Subcellular Localization Analysis
2.9. Protein–Protein Interactions between AcMYB10 and AcbHLH42
2.10. AcMYB10 and AcbHLH42 Regulate the Promoter Activities of AcLDOX and AcF3GT
2.11. AcMYB10 Interacts with AcbHLH42 to Activate Anthocyanin Biosynthesis in N. tabacum Leafs and A. arguta Fruit
3. Materials and Methods
3.1. Plant Materials, Treatment, and Nucleic Acid Extraction
3.2. Genome-Wide Analysis and Expression Profile of Kiwifruit R2R3-MYB Genes
3.3. Quantitative Real-Time RT-PCR
3.4. Determining the Concentration of Anthocyanin, Chlorophyll, and Flavonoid Metabolites
3.5. Isolation and Sequence Alignments and Transcriptional Activation Analysis of AcMYB10
3.6. Dual Luciferase Assay of Transiently Transformed Tobacco Leaves
3.7. Gene Functional Study Using the VIGS System in Kiwifruit and Over-Expression of AcMYB10 in A. chinensis, A. arguta, and N. tabacum
3.8. Subcellular Localization Analysis
3.9. Yeast Two-Hybrid (Y2H) Assays
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
HMM | Hidden Markov Model |
CDS | coding sequences |
FPKM | Fragments per kilobase of transcript per million mapped reads |
VIGS | Virus-induced gene silencing |
qRT-PCR | Quantitative real-time PCR |
DAP | days after pollination |
YFP | yellow fluorescent protein |
RFP | red fluorescent protein |
3AT | 3-amino-1,2,4-triazole |
CRE | Cis-acting regulatory element |
LTR | Low-temperature stress |
MRE | MYB-recognizing element |
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Yu, M.; Man, Y.; Wang, Y. Light- and Temperature-Induced Expression of an R2R3-MYB Gene Regulates Anthocyanin Biosynthesis in Red-Fleshed Kiwifruit. Int. J. Mol. Sci. 2019, 20, 5228. https://doi.org/10.3390/ijms20205228
Yu M, Man Y, Wang Y. Light- and Temperature-Induced Expression of an R2R3-MYB Gene Regulates Anthocyanin Biosynthesis in Red-Fleshed Kiwifruit. International Journal of Molecular Sciences. 2019; 20(20):5228. https://doi.org/10.3390/ijms20205228
Chicago/Turabian StyleYu, Min, Yuping Man, and Yanchang Wang. 2019. "Light- and Temperature-Induced Expression of an R2R3-MYB Gene Regulates Anthocyanin Biosynthesis in Red-Fleshed Kiwifruit" International Journal of Molecular Sciences 20, no. 20: 5228. https://doi.org/10.3390/ijms20205228