Biotechnological Approaches on Two High CBD and CBG Cannabis sativa L. (Cannabaceae) Varieties: In Vitro Regeneration and Phytochemical Consistency Evaluation of Micropropagated Plants Using Quantitative 1H-NMR
Abstract
:1. Introduction
2. Results
2.1. Shoot Multiplication, Elongation and Regeneration
2.2. Rooting of Shoots In Vitro
2.3. Acclimatization
2.4. Chemical Analysis
3. Discussion
3.1. Shoot Multiplication, Elongation and Regeneration
3.2. Rooting of Shoots In Vitro
3.3. Acclimatization
3.4. Chemical Analysis
4. Materials and Methods
4.1. Plant Material—Explants Disinfestation—Culture Establishment
4.2. Shoot Multiplication, Elongation and Regeneration
4.3. Rooting of Shoots In Vitro
4.4. Acclimatization
4.5. Chemical Analysis
4.6. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Medium Strength | Concentration (μΜ) | Average Number of Shoots per Explant | Average Shoot Length (cm) | Average Shooting Percentage (%) |
---|---|---|---|---|
MS 1X | No PGR | 1.79 ijk | 2.42 hi | 58.33 cd |
1.0 μM BA | 2.08 ghi | 2.48 ghi | 79.17 abc | |
2.0 μM BA | 2.67 cde | 3.98 cd | 87.50 a | |
4.0 μM BA | 3.63 a | 5.66 a | 95.83 a | |
8.0 μM BA | 2.71 cd | 3.67 de | 100.00 a | |
1.0 μM TDZ | 2.21 efghi | 2.87 fgh | 87.50 a | |
2.0 μM TDZ | 2.83 cd | 3.22 defg | 100.00 a | |
4.0 μM TDZ | 3.29 ab | 5.38 a | 95.83 a | |
8.0 μM TDZ | 2.13 fghi | 3.73 de | 91.67 a | |
MS 1/2X | No PGR | 1.42 k | 2.09 i | 41.67 d |
1.0 μM BA | 1.58 jk | 2.47 ghi | 54.17 d | |
2.0 μM BA | 2.42 defgh | 4.50 bc | 87.50 a | |
4.0 μM BA | 3.13 bc | 5.09 ab | 91.67 a | |
8.0 μM BA | 2.38 defgh | 3.68 de | 87.50 a | |
1.0 μM TDZ | 1.75 ijk | 2.53 fghi | 62.50 bcd | |
2.0 μM TDZ | 2.50 defg | 3.19 efg | 83.33 ab | |
4.0 μM TDZ | 2.58 def | 4.97 ab | 91.67 a | |
8.0 μM TDZ | 1.96 hij | 3.25 def | 79.17 abc | |
MS 1X | No PGR | 0.96 g | 0.92 g | 45.83 c |
1.0 μM IBA | 1.13 fg | 1.12 efg | 54.17 bc | |
2.0 μM IBA | 3.13 a | 1.93 abc | 87.50 a | |
4.0 μM IBA | 3.17 a | 1.87 abcd | 87.50 a | |
8.0 μM IBA | 2.13 bcde | 1.54 bcdefg | 66.67 abc | |
1.0 μM NAA | 1.29 efg | 1.02 fg | 54.17 bc | |
2.0 μM NAA | 1.46 efg | 1.15 efg | 62.50 abc | |
4.0 μM NAA | 2.92 abc | 2.27 a | 83.33 ab | |
8.0 μM NAA | 2.04 cdef | 1.88 abcd | 75.00 abc | |
MS 1/2X | No PGR | 1.17 fg | 1.26 defg | 66.67 abc |
1.0 μM IBA | 1.42 efg | 1.11 efg | 54.17 bc | |
2.0 μM IBA | 2.83 abc | 1.74 abcde | 87.50 a | |
4.0 μM IBA | 2.96 ab | 1.67 abcdef | 87.50 a | |
8.0 μM IBA | 1.88 defg | 1.43 cdefg | 66.67 abc | |
1.0 μM NAA | 1.33 efg | 1.08 fg | 62.50 abc | |
2.0 μM NAA | 1.75 efg | 1.34 cdefg | 70.83 abc | |
4.0 μM NAA | 3.04 a | 2.10 ab | 83.33 ab | |
8.0 μM NAA | 2.67 abcd | 1.98 abc | 83.33 ab |
Medium Strength | Concentration (μΜ) | Average Number of Shoots per Explant | Average Shoot Length (cm) | Average Shooting Percentage (%) |
---|---|---|---|---|
MS 1X | No PGR | 1.54 hij | 2.14 i | 45.83 d |
1.0 μM BA | 1.88 fghij | 3.34 fgh | 75.00 abc | |
2.0 μM BA | 2.79 bc | 4.65 cd | 100.00 a | |
4.0 μM BA | 3.38 a | 6.23 a | 95.83 a | |
8.0 μM BA | 2.46 cde | 4.32 cde | 91.67 a | |
1.0 μM TDZ | 1.71 ghij | 2.76 hi | 54.17 cd | |
2.0 μM TDZ | 2.54 cd | 3.93 def | 95.83 a | |
4.0 μM TDZ | 3.21 a | 5.97 ab | 100.00 a | |
8.0 μM TDZ | 1.96 fghi | 4.25 def | 83.33 ab | |
MS 1/2X | No PGR | 1.46 j | 2.44 hi | 45.83 d |
1.0 μM BA | 1.67 hij | 3.00 ghi | 66.67 bcd | |
2.0 μM BA | 2.17 def | 5.15 bc | 79.17 ab | |
4.0 μM BA | 3.08 ab | 5.99 ab | 100.00 a | |
8.0 μM BA | 2.04 efgh | 4.03 def | 79.17 ab | |
1.0 μM TDZ | 1.50 j | 2.68 hi | 50.00 d | |
2.0 μM TDZ | 2.13 defg | 3.97 def | 91.67 a | |
4.0 μM TDZ | 2.42 cde | 5.67 ab | 91.67 a | |
8.0 μM TDZ | 1.75 fghij | 3.66 efg | 75.00 abc | |
MS 1X | No PGR | 0.83 g | 0.76 g | 45.83 c |
1.0 μM IBA | 0.96 fg | 0.97 fg | 54.17 bc | |
2.0 μM IBA | 2.71 a | 1.67 abc | 87.50 a | |
4.0 μM IBA | 2.88 a | 1.63 abcd | 87.50 a | |
8.0 μM IBA | 1.92 bcd | 1.35 bcdefg | 70.83 abc | |
1.0 μM NAA | 1.17 defg | 0.92 fg | 62.50 abc | |
2.0 μM NAA | 1.21 defg | 1.01 efg | 62.50 abc | |
4.0 μM NAA | 2.79 a | 1.88 a | 87.50 a | |
8.0 μM NAA | 1.75 cdef | 1.54 abcde | 75.00 abc | |
MS 1/2X | No PGR | 1.00 fg | 1.10 defg | 66.67 abc |
1.0 μM IBA | 1.25 defg | 0.90 fg | 50.00 c | |
2.0 μM IBA | 2.50 abc | 1.59 abcd | 87.50 a | |
4.0 μM IBA | 2.67 ab | 1.54 abcde | 87.50 a | |
8.0 μM IBA | 1.83 cde | 1.38 abcdef | 70.83 abc | |
1.0 μM NAA | 1.08 efg | 1.00 efg | 62.50 abc | |
2.0 μM NAA | 1.50 defg | 1.24 cdefg | 66.67 abc | |
4.0 μM NAA | 2.75 a | 1.85 ab | 83.33 ab | |
8.0 μM NAA | 2.42 abc | 1.78 abc | 83.33 ab |
CBD+CBDA (%) | CBG+CBGA (%) | ||||||
---|---|---|---|---|---|---|---|
Mother Plant | 1 | 2 | 3 | 1 | 2 | 3 | |
11.30 a | 10.40 a | 11.47 a | 10.49 b | 9.17 b | 9.87 b | ||
Clone | 1 | 10.90 a | 10.28 a | 10.57 a | 10.01 b | 9.51 b | 9.52 b |
2 | 10.22 a | 10.25 a | 11.09 a | 9.62 b | 9.75 b | 9.60 b | |
3 | 11.10 a | 10.88 a | 11.41 a | 9.41 b | 9.39 b | 9.55 b | |
Clone Mean | 10.74 a | 10.47 a | 11.02 a | 9.68 b | 9.55 b | 9.55 b | |
Overall Mother Plant Mean | 11.05 a | 9.84 b | |||||
Overall Clone Mean | 10.74 a | 9.59 b |
Cannabinoids | MW | Structure | Proton Signal | δ in ppm |
---|---|---|---|---|
CBD | 314.5 | | H-10 trans | 4.67 |
H-10 cis | 4.57 | |||
H-2 | 5.59 | |||
CBDA | 358.5 | | H-5′ | 6.27 |
H-10 trans | 4.55 | |||
H-10 cis | 4.41 | |||
H-2 | 5.56 | |||
CBG | 316.5 | | H-2/H-4 | 6.25 |
H-1′a/H-1′b | 3.43 | |||
H-2′ | 5.29 | |||
H-6′ | 5.07 | |||
CBGA | 360.5 | | H-4 | 6.27 |
H-1′a/H-1′b | 3.45 | |||
H-2′ | 5.30 | |||
H-6′ | 5.07 |
Sample Availability: Not available. |
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Ioannidis, K.; Dadiotis, E.; Mitsis, V.; Melliou, E.; Magiatis, P. Biotechnological Approaches on Two High CBD and CBG Cannabis sativa L. (Cannabaceae) Varieties: In Vitro Regeneration and Phytochemical Consistency Evaluation of Micropropagated Plants Using Quantitative 1H-NMR. Molecules 2020, 25, 5928. https://doi.org/10.3390/molecules25245928
Ioannidis K, Dadiotis E, Mitsis V, Melliou E, Magiatis P. Biotechnological Approaches on Two High CBD and CBG Cannabis sativa L. (Cannabaceae) Varieties: In Vitro Regeneration and Phytochemical Consistency Evaluation of Micropropagated Plants Using Quantitative 1H-NMR. Molecules. 2020; 25(24):5928. https://doi.org/10.3390/molecules25245928
Chicago/Turabian StyleIoannidis, Kostas, Evangelos Dadiotis, Vangelis Mitsis, Eleni Melliou, and Prokopios Magiatis. 2020. "Biotechnological Approaches on Two High CBD and CBG Cannabis sativa L. (Cannabaceae) Varieties: In Vitro Regeneration and Phytochemical Consistency Evaluation of Micropropagated Plants Using Quantitative 1H-NMR" Molecules 25, no. 24: 5928. https://doi.org/10.3390/molecules25245928