Characterization and Functional Analysis of Chalcone Synthase Genes in Highbush Blueberry (Vaccinium corymbosum)
Abstract
:1. Introduction
2. Results
2.1. Identification and Physiochemical Property Analysis of Blueberry CHSs
2.2. Gene Duplication and Synteny Analysis of Blueberry CHS Genes
2.3. Conserved Motifs of VcCHSs and Gene Structures of Their Corresponding Genes
2.4. Promoter Analysis Results of Blueberry CHS Genes
2.5. Expression Analysis of VcCHS Genes in Blueberry Fruits
2.6. Effects of Transient Overexpression of VcCHS8, VcCHS13 and VcCHS21 on Flavonoids and Anthocyanin Accumulations in Blueberry Leaves
2.7. Effects of Transient Overexpression of VcCHS8, VcCHS13 and VcCHS21 on Flavonoids and Anthocyanin Accumulations in Apple Fruits
2.8. Binding Ability of the Anthocyanin Biosynthesis Regulatory bHLH Transcription Factors to the VcCHS21 Promoter
3. Discussion
3.1. The Expansion of Blueberry CHS Gene Family Is Mainly Caused by Segmental Duplications
3.2. The Subfamily I CHS Members Are Closely Related to Flavonoid and Anthocyanin Accumulation in Blueberry Fruits
3.3. The Expression of the Blueberry CHSs Is Regulated by Transcription Factors
4. Materials and Methods
4.1. Identification of the Blueberry CHS Genes
4.2. Bioinformatic Analysis of Blueberry CHSs and Their Encoded Proteins
4.3. Phylogenetic Analysis of Blueberry CHS Proteins
4.4. Synteny Analysis of Blueberry CHS Genes
4.5. Gene Structure Analysis of VcCHSs and Conserved Motif analysis of Their Encoded Proteins
4.6. Promoter Analysis
4.7. Gene Expression Analysis of VcCHSs
4.8. Gene Cloning and Vector Construction
4.9. Transient Overexpression Analysis of VcCHS8, VcCHS13 and VcCHS21 in Blueberry Leaves and Apple Fruits
4.10. Yeast One-Hybrid (Y1H) Assay
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Gene Name | Gene ID | Number of AA | Molecular Weight/Da | pI | Instability Index | Aliphatic Index | GRAVY | Subcellular Localization |
---|---|---|---|---|---|---|---|---|
VcCHS1 | VaccDscaff12-processed-gene-318.4 | 475 | 52,294.14 | 5.40 | 39.3 | 95.22 | −0.132 | Cytoplasm |
VcCHS2 | VaccDscaff12-snap-gene-319.33 | 395 | 43,328.65 | 5.55 | 38.54 | 96.53 | −0.129 | Cytoplasm |
VcCHS3 | VaccDscaff13-augustus-gene-46.32 | 398 | 43,199.79 | 5.92 | 38.68 | 90.63 | −0.084 | Cytoplasm |
VcCHS4 | VaccDscaff14-processed-gene-382.0 | 385 | 42,193.76 | 6.19 | 39.09 | 93.27 | −0.143 | Cytoplasm |
VcCHS5 | VaccDscaff23-processed-gene-74.2 | 557 | 61,129.82 | 8.26 | 47.69 | 87.34 | −0.253 | Cytoplasm |
VcCHS6 | VaccDscaff23-snap-gene-74.34 | 403 | 44,634.05 | 5.44 | 36.25 | 93.13 | −0.163 | Cytoplasm |
VcCHS7 | VaccDscaff25-augustus-gene-173.26 | 389 | 42,577.31 | 6.04 | 37.78 | 90.49 | −0.082 | Cytoplasm |
VcCHS8 | VaccDscaff25-augustus-gene-174.22 | 389 | 42,603.35 | 5.97 | 37.3 | 90.75 | −0.078 | Cytoplasm |
VcCHS9 | VaccDscaff2-augustus-gene-67.14 | 389 | 42,505.25 | 6.18 | 37.33 | 90.75 | −0.068 | Cytoplasm |
VcCHS10 | VaccDscaff2-processed-gene-435.12 | 407 | 44,652.54 | 5.97 | 44.56 | 93.02 | −0.109 | Cytoplasm |
VcCHS11 | VaccDscaff303-processed-gene-0.17 | 407 | 44,676.58 | 5.97 | 41.41 | 91.11 | −0.116 | Cytoplasm |
VcCHS12 | VaccDscaff30-augustus-gene-303.29 | 398 | 43,215.86 | 5.92 | 38.12 | 90.63 | −0.076 | Cytoplasm |
VcCHS13 | VaccDscaff32-augustus-gene-318.21 | 398 | 43,199.79 | 5.92 | 38.68 | 90.63 | −0.084 | Cytoplasm |
VcCHS14 | VaccDscaff35-augustus-gene-1.21 | 389 | 42,538.25 | 6.04 | 35.09 | 92.49 | −0.045 | Cytoplasm |
VcCHS15 | VaccDscaff36-augustus-gene-240.30 | 389 | 42,538.25 | 6.04 | 35.09 | 92.49 | −0.045 | Cytoplasm |
VcCHS16 | VaccDscaff3-augustus-gene-334.17 | 389 | 42,591.34 | 6.04 | 37.78 | 90.75 | −0.083 | Cytoplasm |
VcCHS17 | VaccDscaff40-snap-gene-213.40 | 383 | 42,048.07 | 5.56 | 38.31 | 92.95 | −0.197 | Cytoplasm |
VcCHS18 | VaccDscaff41-snap-gene-198.34 | 403 | 44,647.09 | 5.53 | 36.46 | 93.37 | −0.162 | Cytoplasm |
VcCHS19 | VaccDscaff41-snap-gene-198.39 | 386 | 42,379.59 | 5.3 | 41.41 | 104.33 | −0.097 | Cytoplasm |
VcCHS20 | VaccDscaff42-augustus-gene-14.30 | 398 | 43,227.81 | 5.92 | 40.12 | 90.63 | −0.086 | Cytoplasm |
VcCHS21 | VaccDscaff4-processed-gene-104.3 | 389 | 42,538.25 | 6.04 | 35.09 | 92.49 | −0.045 | Cytoplasm |
VcCHS22 | VaccDscaff9-processed-gene-64.0 | 389 | 42,538.25 | 6.04 | 35.09 | 92.49 | −0.045 | Cytoplasm |
Gene ID | Gene Name | Gene ID | Gene Name | Ka | Ks | Ka_Ks | Duplication Date/Mya | Duplication Type |
---|---|---|---|---|---|---|---|---|
23-processed-gene-74.2 | VcCHS5 | 23-snap-gene-74.34 | VcCHS6 | 0.0413 | 0.1090 | 0.3788 | 4.19 | Tandem duplication |
12-processed-gene-318.4 | VcCHS1 | 23-processed-gene-74.2 | VcCHS5 | 0.1050 | 0.2063 | 0.5090 | 7.93 | Segmental duplication |
2-processed-gene-435.12 | VcCHS10 | 303-processed-gene-0.17 | VcCHS11 | 0.0065 | 0.0423 | 0.1534 | 1.63 | Segmental duplication |
30-augustus-gene-303.29 | VcCHS12 | 32-augustus-gene-318.21 | VcCHS13 | 0.0011 | 0.0388 | 0.0286 | 1.49 | Segmental duplication |
30-augustus-gene-303.29 | VcCHS12 | 42-augustus-gene-14.30 | VcCHS20 | 0.0022 | 0.0570 | 0.0390 | 2.19 | Segmental duplication |
32-augustus-gene-318.21 | VcCHS13 | 42-augustus-gene-14.30 | VcCHS20 | 0.0011 | 0.0533 | 0.0208 | 2.05 | Segmental duplication |
35-augustus-gene-1.21 | VcCHS14 | 36-augustus-gene-240.30 | VcCHS15 | 0 | 0.0569 | 0 | 2.19 | Segmental duplication |
35-augustus-gene-1.21 | VcCHS14 | 4-processed-gene-104.3 | VcCHS21 | 0 | 0.0336 | 0 | 1.29 | Segmental duplication |
35-augustus-gene-1.21 | VcCHS14 | 9-processed-gene-64.0 | VcCHS22 | 0 | 0.0569 | 0 | 2.19 | Segmental duplication |
36-augustus-gene-240.30 | VcCHS15 | 4-processed-gene-104.3 | VcCHS21 | 0 | 0.0688 | 0 | 2.65 | Segmental duplication |
36-augustus-gene-240.30 | VcCHS15 | 9-processed-gene-64.0 | VcCHS22 | 0 | 0.0530 | 0 | 2.04 | Segmental duplication |
4-processed-gene-104.3 | VcCHS21 | 9-processed-gene-64.0 | VcCHS22 | 0 | 0.0688 | 0 | 2.65 | Segmental duplication |
13-augustus-gene-46.32 | VcCHS3 | 30-augustus-gene-303.29 | VcCHS12 | 0.0011 | 0.0571 | 0.0194 | 2.19 | Segmental duplication |
13-augustus-gene-46.32 | VcCHS3 | 32-augustus-gene-318.21 | VcCHS13 | 0 | 0.0174 | 0 | 0.67 | Segmental duplication |
13-augustus-gene-46.32 | VcCHS3 | 42-augustus-gene-14.30 | VcCHS20 | 0.0011 | 0.0460 | 0.0241 | 1.77 | Segmental duplication |
14-processed-gene-382.0 | VcCHS4 | 2-processed-gene-435.12 | VcCHS10 | 0.0046 | 0.0449 | 0.1017 | 1.73 | Segmental duplication |
14-processed-gene-382.0 | VcCHS4 | 303-processed-gene-0.17 | VcCHS11 | 0 | 0.0221 | 0 | 0.85 | Segmental duplication |
23-processed-gene-74.2 | VcCHS5 | 41-snap-gene-198.34 | VcCHS18 | 0.0390 | 0.1091 | 0.3575 | 4.20 | Segmental duplication |
25-augustus-gene-173.26 | VcCHS7 | 3-augustus-gene-334.17 | VcCHS16 | 0.0011 | 0.0298 | 0.0376 | 1.15 | Segmental duplication |
2-augustus-gene-67.14 | VcCHS9 | 25-augustus-gene-173.26 | VcCHS7 | 0.0034 | 0.0336 | 0.1003 | 1.29 | Segmental duplication |
2-augustus-gene-67.14 | VcCHS9 | 3-augustus-gene-334.17 | VcCHS16 | 0.0045 | 0.0185 | 0.2434 | 0.71 | Segmental duplication |
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Zhang, Z.; Qu, P.; Hao, S.; Li, R.; Zhang, Y.; Zhao, Q.; Wen, P.; Cheng, C. Characterization and Functional Analysis of Chalcone Synthase Genes in Highbush Blueberry (Vaccinium corymbosum). Int. J. Mol. Sci. 2023, 24, 13882. https://doi.org/10.3390/ijms241813882
Zhang Z, Qu P, Hao S, Li R, Zhang Y, Zhao Q, Wen P, Cheng C. Characterization and Functional Analysis of Chalcone Synthase Genes in Highbush Blueberry (Vaccinium corymbosum). International Journal of Molecular Sciences. 2023; 24(18):13882. https://doi.org/10.3390/ijms241813882
Chicago/Turabian StyleZhang, Zening, Pengyan Qu, Siyi Hao, Ruide Li, Yongyan Zhang, Qi Zhao, Pengfei Wen, and Chunzhen Cheng. 2023. "Characterization and Functional Analysis of Chalcone Synthase Genes in Highbush Blueberry (Vaccinium corymbosum)" International Journal of Molecular Sciences 24, no. 18: 13882. https://doi.org/10.3390/ijms241813882