The Expression Profiles of the Salvia miltiorrhiza 3-Hydroxy-3-methylglutaryl-coenzyme A Reductase 4 Gene and Its Influence on the Biosynthesis of Tanshinones
Abstract
:1. Introduction
2. Results
2.1. Organ-Specific Expression of S. miltiorrhiza HMGR4 Gene
2.2. Potential Regulators of S. miltiorrhiza HMGR4 Gene Expression
2.3. Effect of GA3, IAA, SA on S. miltiorrhiza HMGR4 Gene Expression
2.4. Impact of pRI201-AN-HMGR4 Transformation on S. miltiorrhiza HMGR4 Gene Expression
2.5. Influence of HMGR4 Overexpression on the Biosynthesis of Tanshinones in S. miltiorrhiza
2.6. Organ-Dependent Accumulation of Tanshinones in S. miltiorrhiza
2.7. Impact of Growth Environment on the Biosynthesis of Tanshinones in S. miltiorrhiza
2.8. Effect of GA3 and IAA on the Biosynthesis of Tanshinones in S. miltiorrhiza
3. Discussion
4. Materials and Methods
4.1. Establishment of S. miltiorrhiza Culture and Treatments
4.2. Selection of Potential Regulators of S. miltiorrhiza HMGR4 Gene Expression
4.3. Preparation of pRI201-AN-HMGR4 Overexpression Construct
4.4. Transformation, Selection, Regeneration and Treatments of S. miltiorrhiza Culture
4.5. RNA Isolation, Reverse Transcription and Quantitative Real-Time PCR
4.6. Quantitative Analysis of Tanshinones
4.7. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Sample Availability
References
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TF Family Name | TF Gene Name and Locus | TFBSs Number |
---|---|---|
GA3 | ||
AT-Hook | AHL25; At4g35390 | 6 |
MYB-related | CCA1; At2g46830 | 1 |
RVE8; At3g09600 | 2 | |
RVE4; At5g02840 | ||
Homeodomain; HD-ZIP | ATHB-23; At1g26960 | 1 |
bHLH | PIF3; At1g09530 | 2 |
GATA | GATA22; At4g26150 | 62 |
GATA21; At5g56860 | ||
MADS box; MIKC | SOC1; At2g45660 | 8 |
AGL24; At4g24540 | 15 | |
MADS box | AGL71; At5g51870 | 13 |
AGL72; At5g51860 | 14 | |
AGL42; At5g62165 | ||
NF-YC | NFYC3; At1g54830 | 48 |
IAA | ||
MYB-related | CCA1; At2g46830 | 1 |
RVE 8; At3g09600 | 2 | |
Homeodomain; HD-ZIP | HAT2; At5g47370 | 2 |
ATHB-20; At3g01220 | 3 | |
CAMTA | CAMTA1; At5g09410 | 2 |
MADS box; MIKC | AGL14; At4g11880 | 17 |
SA | ||
NAC; NAM | NAC081; At5g08790 | 1 |
NAC062; At3g49530 | 2 | |
MYB | MYB46; At5g12870 | 4 |
Myb/SANT; MYB | MYB3; At1g22640 | 1 |
MYB-related | CCA1; At2g46830 | 1 |
RVE8; At3g09600 | 2 | |
RVE4; At5g02840 | ||
Dof | DOF1.1; At1g07640 | 34 |
DOF3.4; At3g50410 | ||
bZIP | TGA2; At5g06950 | 32 |
bHLH | LRL1; At2g24260 | 2 |
CG-1; CAMTA | CAMTA2; At5g64220 | 3 |
CAMTA | CAMTA4; At1g67310 | 2 |
CAMTA6; At3g16940 | ||
CSD | CSP2; At4g38680 | 2 |
WRKY | WRKY6; At1g62300 | 9 |
WRKY40; At1g80840 | ||
WRKY4; At1g13960 | 8 | |
WRKY60; At2g25000 | ||
WRKY21; At2g30590 | ||
WRKY54; At2g40750 | ||
WRKY70; At3g56400 | ||
WRKY53; At4g23810 | ||
WRKY18; At4g31800 | ||
WRKY26; At5g07100 | ||
WRKY38; At5g22570 | ||
WRKY30; At5g24110 |
Primer Name | Sequence | Product Size [bp] |
---|---|---|
Confirmation of Transformation | ||
Kanamycin_F | TGATCTCGTCGTGACCCAT | 234 |
Kanamycin_R | AGAAGGCGATAGAAGGCGA | |
Real-Time PCR | ||
HMGR4_F | CTCAACCTGCTTGGCGTAA | 185 |
HMGR4_R | AGTCTCGTGATGTCCCTGCT | |
ACT7_F | TCCGTCTTGATCTTGCTGGT | 170 |
ACT7_R | CGTCTTTGCAGTTTCGAGCT |
Analyte | Retention Time [min] | Standard Curve | R2 |
---|---|---|---|
DHTI | 6.025 | y = 190.163x − 16.690 | 0.99973 |
CT | 10.009 | y = 146.106x − 10.926 | 0.99992 |
TI | 10.938 | y = 241.276x − 21.816 | 0.99989 |
TIIA | 14.808 | y = 327.209x − 20.014 | 0.99996 |
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Majewska, M.; Szymczyk, P.; Gomulski, J.; Jeleń, A.; Grąbkowska, R.; Balcerczak, E.; Kuźma, Ł. The Expression Profiles of the Salvia miltiorrhiza 3-Hydroxy-3-methylglutaryl-coenzyme A Reductase 4 Gene and Its Influence on the Biosynthesis of Tanshinones. Molecules 2022, 27, 4354. https://doi.org/10.3390/molecules27144354
Majewska M, Szymczyk P, Gomulski J, Jeleń A, Grąbkowska R, Balcerczak E, Kuźma Ł. The Expression Profiles of the Salvia miltiorrhiza 3-Hydroxy-3-methylglutaryl-coenzyme A Reductase 4 Gene and Its Influence on the Biosynthesis of Tanshinones. Molecules. 2022; 27(14):4354. https://doi.org/10.3390/molecules27144354
Chicago/Turabian StyleMajewska, Małgorzata, Piotr Szymczyk, Jan Gomulski, Agnieszka Jeleń, Renata Grąbkowska, Ewa Balcerczak, and Łukasz Kuźma. 2022. "The Expression Profiles of the Salvia miltiorrhiza 3-Hydroxy-3-methylglutaryl-coenzyme A Reductase 4 Gene and Its Influence on the Biosynthesis of Tanshinones" Molecules 27, no. 14: 4354. https://doi.org/10.3390/molecules27144354