Induction and Suspension Culture of Panax japonicus Callus Tissue for the Production of Secondary Metabolic Active Substances
Abstract
:1. Introduction
2. Results
2.1. Healing Callus Induction in Panax japonicus Roots
2.1.1. Disinfection Method of Panax japonicus Root
2.1.2. Screening of Induction Scheme for Root Callus of Panax japonicus
2.2. Proliferation Culture of Root Callus of Panax japonicus and Screening of High-Yield Cell Lines
2.2.1. Effects of NAA and 6-BA on Callus Proliferation of Panax japonicus
2.2.2. Screening of High-Yielding Cell Lines of Panax japonicus
2.2.3. Effect of Curing Agents on the Proliferation of Panax japonicus Callus Tissues
2.3. Establishment of Suspension Culture System of Panax japonicus
2.3.1. Effect of Carbon Source on Cell Suspension Culture
2.3.2. Effect of Initial Inoculum Size on Suspension Culture
2.3.3. Effect of Shaking Speed on Suspension Culture
2.3.4. Effect of Plant Growth Regulators on Suspension Culture of Callus Tissues
2.3.5. Effect of Culture Days on Suspension Culture
2.4. Effects of Suspension Culture Conditions on the Content of Total Saponins in Cells of Panax japonicus
2.4.1. The Effect of Carbon Source on the Content of Triterpenoid Glycosides in Cells
2.4.2. Effect of Initial Inoculation on Total Saponins Content
2.4.3. Effect of Shaking Speed on the Content of Triterpenoid Glycosides
2.4.4. Effects of Plant Growth Regulators on the Content of Triterpenoid Glycosides
2.4.5. The Effect of Culture Time on the Content of Triterpenoid Glycosides
3. Discussion
3.1. Callus Induction of Aseptic Roots of Panax japonicus
3.2. Callus Culture and High-Yield Cell Line Screening of Panax japonicus
3.3. Panax japonicus Cell Suspension Culture System
4. Materials and Methods
4.1. Materials
4.2. Methods
4.2.1. Callus Induction of Panax japonicus
Methods of Root Disinfection
Callus Induction Culture
Computing Formula
4.2.2. Proliferation Culture of Root Callus of Panax japonicus and Screening of High-Yield Cell Lines
Proliferation Culture of Loose Callus Tissue
Culture Cycle of Callus
Screening of High-Yield Cell Lines
Screening of Curing Agent
Computing Formula
4.3. Establishment of Cell Suspension Culture System of Panax japonicus
4.3.1. Screening of Suspension Culture Conditions
4.3.2. Carbon Source Screening
4.3.3. Screening of Initial Inoculation Amount
4.3.4. Cell Rotation Speed Screening
4.3.5. Hormone Concentration Screening
4.3.6. Screening of Incubation Time
4.3.7. Methods for Determination of Cell Growth Index
Determination of Cell Viability and Density
Determination of Settled Cell Volume
Determination of Biomass
4.3.8. Data Analysis Methods
4.4. Detection of Total Saponins Content in Suspension-Cultured Cells
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Scheme | Medium | Proliferation Rate/% | State of the Callus |
---|---|---|---|
1 | MS + 16.11 μmol·L−1 NAA + 0.44 μmol·L−1 6-BA + 30.0 g·L−1 sucrose + 3.8 g·L−1 gellan gum | 43.33 | Yellow and dense structure and not agglomerated (Figure S2a) |
2 | MS + 16.11 μmol·L−1 NAA +4.44 μmol·L−1 6-BA + 30.0 g·L−1 sucrose + 3.8 L−1 gellan gum | 126.55 | White and transparent; the structure is relatively dense (Figure S2b) |
3 | MS + 13.32 μmol·L−1 6-BA + 30.0 g·L−1 sucrose + 3.8 g·L−1 gellan gum | 57.53 | White and transparent; relatively loose structure (Figure S2c) |
4 | MS + 4.64 μmol·L−1 NAA + 13.32 μmol·L−1 6-BA + 30.0 g·L−1 sucrose + 3.8 g·L−1 gellan gum | 148.49 | White and transparent; loose structure (Figure S2d) |
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Lv, S.; Ding, F.; Zhang, S.; Nosov, A.M.; Kitashov, A.V.; Yang, L. Induction and Suspension Culture of Panax japonicus Callus Tissue for the Production of Secondary Metabolic Active Substances. Plants 2024, 13, 2480. https://doi.org/10.3390/plants13172480
Lv S, Ding F, Zhang S, Nosov AM, Kitashov AV, Yang L. Induction and Suspension Culture of Panax japonicus Callus Tissue for the Production of Secondary Metabolic Active Substances. Plants. 2024; 13(17):2480. https://doi.org/10.3390/plants13172480
Chicago/Turabian StyleLv, Siqin, Fan Ding, Shaopeng Zhang, Alexander M. Nosov, Andery V. Kitashov, and Ling Yang. 2024. "Induction and Suspension Culture of Panax japonicus Callus Tissue for the Production of Secondary Metabolic Active Substances" Plants 13, no. 17: 2480. https://doi.org/10.3390/plants13172480