LED Lights Affecting Morphogenesis and Isosteroidal Alkaloid Contents in Fritillaria cirrhosa D. Don—An Important Chinese Medicinal Herb
Abstract
:1. Introduction
2. Material and Methods
2.1. Callus Multiplication
2.2. Influence of Different Light Spectra on Morphogenesis in Embryogenic Calli and Contents of Isosteroidal Alkaloids
2.3. Development of Bulblets from Somatic Embryos
2.4. Chemicals and Other Materials
2.5. Preparation of Fe3O4@C18 Nanoparticle Composite
2.6. Extraction Procedure
2.7. LC-MS/MS Conditions
2.8. Statistical Analysis
3. Results and Discussion
3.1. Callus Proliferation
3.2. Influence of LED Lights on Morphogenesis of Embryogenic Calli (EC) of F. cirrhosa
3.3. Influence of LED Lights on the Contents of Isosteroidal Alkaloids in In Vitro Cultures of F. cirrhosa
3.4. Development of Bulblets in Somatic Embryos of F. cirrhosa
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Marker Compounds | tR a (min) | Mass/Charge (m/z) | Linearity and Range | Sensitivity c | |||
---|---|---|---|---|---|---|---|
Regression Equation b | Correlation Coefficient (r2) | Linear Range (μg/g) | LOD (ng/g) | LOQ (ng/g) | |||
Peimisine | 5.78 | 428.316 | y = 0.0396x + 0.0630 | 0.9923 | 0.1–40 | 0.01 | 0.04 |
Sipeimine | 8.22 | 430.332 | y = 0.5426x + 0.1531 | 0.9968 | 0.1–40 | 0.02 | 0.06 |
Peiminine | 10.72 | 430.332 | y = 0.0047x + 0.0010 | 0.9967 | 0.1–40 | 0.97 | 3.23 |
Peimine | 11.88 | 432.347 | y = 0.0067x + 0.0016 | 0.9977 | 0.1–40 | 0.56 | 1.88 |
LED Light Treatment | Av No. of Somatic Embryos/Bottle | Av No. Of Somatic Embryos with Cotyledonary Leaves/Bottle | Av Total Fresh Weight of Cultures/Bottle (g) | Morphological Features of Cultures (Somatic Embryos = SEs) |
---|---|---|---|---|
CW-5000K * | 63.7 ± 7.6 c ** | 12.0 ± 6.0 abc | 12.82 ± 1.09 c | SEs at all stages from globular to mature, SEs with long cotyledonary leaves. Color of cultures is light green. |
WW-2700K | 122.0 ± 65.0 bc | 22.3 ± 13.9 a | 13.64 ± 2.82 c | SE stages are similar to CW-5000K. Color of cultures is light green. |
7R1G1B | 108.0 ± 38.2 bc | 8.5 ± 3.5 bc | 12.99 ± 2.02 c | A majority of SEs in early stages, including globular shapes. Cotyledonary leaves absent. Color of cultures is light green to light brown. |
8R1B | 157.3 ± 9.3 ab | 16.7 ± 8.3 ab | 15.23 ± 0.97 abc | Stages similar to CW-5000K. Color of cultures is light green. |
9B | 169.0 ± 66.1 ab | 3.7 ± 2.3 c | 14.61 ± 1.02 bc | SE stages similar to CW-5000K, but cotyledonary leaves shorter in length. Color of cultures is light green to light yellow. |
9R | 223.7 ± 57.5 a | 5.3 ± 4.9 c | 17.92 ± 0.77 a | A majority of SEs in the early stages, including globular shapes. Cotyledonary leaves absent. Color of cultures is white. |
9IR | 231.3 ± 62.3 a | 4.7 ± 0.6 c | 16.67 ± 1.85 ab | SE stages and color of cultures similar to 9R. Only a few cotyledonary leaves are seen. Color of cultures is white. |
3R3B3IR | 230.7 ± 23.4 a | 4.3 ± 2.5 c | 17.56 ± 2.35 a | SE at all stages from globular to mature but cotyledonary leaves shorter in length. Color of cultures is dark green. |
LED Light Treatment | Plant Material | Isosteroidal Alkaloids (µg/g/dw) | Total of Four Alkaloids (µg/g/dw) | |||
---|---|---|---|---|---|---|
Peimisine | Sipeimine | Peimine | Peiminine | |||
CW-5000K | In vitro cultures | ND * | ND | ND | 0.12 ± 0.20 b ** | 0.12 ± 0.20 c ** |
WW-2700K | In vitro cultures | ND | ND | ND | 0.28 ± 0.27 b | 0.28 ± 0.27 c |
7R1G1B | In vitro cultures | ND | ND | ND | 0.19 ± 0.32 b | 0.19 ± 0.32 c |
8R1B | In vitro cultures | ND | ND | ND | ND | 0.00 ± 0.00 c |
9B | In vitro cultures | ND | ND | ND | 0.60 ± 0.43 b | 0.65 ± 0.45 c |
9R | In vitro cultures | 3.65 ± 1.68 b ** | ND | 0.38 ± 0.11 a | 2.40 ± 0.30 a | 6.42 ± 2.06 b |
9IR | In vitro cultures | 3.22 ± 3.28 b | ND | 0.05 ± 0.09 b | 2.21 ± 0.87 a | 5.48 ± 3.21 b |
3R3B3IR | In vitro cultures | ND | ND | ND | 0.26 ± 0.24 b | 0.26 ± 0.24 c |
Fluorescent tube | In vitro bulblets (3 months old) | 0.91 ± 0.97 b | ND | ND | 2.98 ± 1.09 a | 3.90 ± 1.51 bc |
Natural habitat | Commercial bulbs (wild type, 3 years old) | 68.4 ± 7.8 a | 0.6 ± 0.4 a | ND | ND | 69.0 ± 7.4 a |
Type of Somatic Embryo (SE) * | Percentage of Response (%) | Av No. of Bulblets/SE |
---|---|---|
Single embryo | 90.0 ± 10.0 a ** | 4.7 ± 1.3 a ** |
Cluster of five embryos | 86.7 ± 12.0 a | 3.3 ± 1.5 ab |
Embryo with the cotyledonary leaf | 43.6 ± 29.0 b | 1.1 ± 0.7 b |
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Chen, C.-C.; Lee, M.-R.; Wu, C.-R.; Ke, H.-J.; Xie, H.-M.; Tsay, H.-S.; Agrawal, D.C.; Chang, H.-C. LED Lights Affecting Morphogenesis and Isosteroidal Alkaloid Contents in Fritillaria cirrhosa D. Don—An Important Chinese Medicinal Herb. Plants 2020, 9, 1351. https://doi.org/10.3390/plants9101351
Chen C-C, Lee M-R, Wu C-R, Ke H-J, Xie H-M, Tsay H-S, Agrawal DC, Chang H-C. LED Lights Affecting Morphogenesis and Isosteroidal Alkaloid Contents in Fritillaria cirrhosa D. Don—An Important Chinese Medicinal Herb. Plants. 2020; 9(10):1351. https://doi.org/10.3390/plants9101351
Chicago/Turabian StyleChen, Chia-Chen, Maw-Rong Lee, Chi-Rei Wu, Hsin-Ju Ke, Hui-Min Xie, Hsin-Sheng Tsay, Dinesh Chandra Agrawal, and Hung-Chi Chang. 2020. "LED Lights Affecting Morphogenesis and Isosteroidal Alkaloid Contents in Fritillaria cirrhosa D. Don—An Important Chinese Medicinal Herb" Plants 9, no. 10: 1351. https://doi.org/10.3390/plants9101351