Construction of Optimal Regeneration System for Chrysanthemum ‘11-C-2’ Stem Segment with Buds
Abstract
:1. Introduction
2. Results
2.1. Optimal Medium for Adventitious Bud Differentiation of ‘11-C-2’
2.2. Optimal Rooting Medium for ‘11-C-2’
2.3. Optimal Hardening Time and Transplant Medium for ‘11-C-2’
2.4. Establishment of ‘11-C-2’ Regeneration System with Sprouted Stem Segments
3. Discussion
4. Materials and Methods
4.1. Experimental Material
4.2. Experimental Method
4.2.1. Aseptic Seedling Culture
4.2.2. Adventitious Bud Differentiation
4.2.3. Rooting Culture
4.2.4. Acclimatization and Transplantation
4.3. Data Analysis
5. Conclusions
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
Appendix A
Dependent Variable | Influencing Factor | k1 | k2 | k3 | R |
---|---|---|---|---|---|
Differentiation coefficient (Stem segment with bud) | 6-BA | 6.11 | 10.44 | 6.33 | 4.33 |
NAA | 7.33 | 6.56 | 9.00 | 2.44 | |
2,4-D | 9.33 | 7.44 | 6.11 | 3.22 | |
TDZ | 8.44 | 8.11 | 6.33 | 2.11 | |
Differentiation coefficient (Leaf) | 6-BA | 1.44 | 1.67 | 1.22 | 0.44 |
NAA | 1.78 | 1.33 | 1.22 | 0.56 | |
2,4-D | 1.56 | 1.44 | 1.33 | 0.22 | |
TDZ | 1.56 | 1.56 | 1.22 | 0.33 | |
Rooting rate | Basic medium | 94.84 | 96.43 | 96.69 | 1.85 |
NAA | 96.83 | 96.56 | 94.58 | 2.25 | |
Root number | Basic medium | 7.48 | 10.90 | 9.50 | 3.42 |
NAA | 11.99 | 8.43 | 7.46 | 4.53 | |
Root length | Basic medium | 6.00 | 7.77 | 7.02 | 1.77 |
NAA | 8.38 | 6.62 | 5.80 | 2.58 | |
Plant height (0 d) | Seedling refining time | 3.22 | 3.24 | 3.34 | 0.12 |
Substrate for transplanting | 3.25 | 3.26 | 3.29 | 0.04 | |
Plant height (5 d) | Seedling refining time | 4.12 | 4.58 | 4.98 | 0.86 |
Substrate for transplanting | 4.33 | 4.77 | 4.58 | 0.44 | |
Plant height (10 d) | Seedling refining time | 4.69 | 5.01 | 5.47 | 0.78 |
Substrate for transplanting | 4.75 | 5.35 | 5.07 | 0.60 | |
Plant height (15 d) | Seedling refining time | 5.19 | 5.48 | 5.92 | 0.73 |
Substrate for transplanting | 5.15 | 5.91 | 5.53 | 0.76 |
Dependent Variable | The Source | Sum of Squares | Mean Square | F | p |
---|---|---|---|---|---|
Differentiation coefficient (Stem segment with bud) | 6-BA × NAA | 205.63 | 25.704 | 36.526 *** | 0.000 |
Root number | Base medium × NAA | 157.076 | 19.635 | 27.369 *** | 0.000 |
Root length | 46.192 | 5.774 | 25.336 *** | 0.000 | |
Plant height (5 d) | Seedling refining time × transplanting medium | 4.38 | 0.548 | 8.167 *** | 0.000 |
Plant height (10 d) | 4.373 | 0.547 | 21.706 *** | 0.000 | |
Plant height (15 d) | 5.323 | 0.665 | 30.449 *** | 0.000 |
Dependent Variable | Univariate | Level | Sum of Squares | Mean Square | F | p |
---|---|---|---|---|---|---|
Differentiation coefficient (Stem segment with bud) | 6-BA (mg/L) | 1 | 28.222 | 14.111 | 20.053 *** | 0.000 |
1.5 | 67.556 | 33.778 | 48.000 *** | 0.000 | ||
2 | 2.667 | 1.333 | 1.895 | 0.179 | ||
NAA (mg/L) | 0.1 | 6.000 | 3.000 | 4.263 * | 0.031 | |
0.2 | 57.556 | 28.778 | 40.895 *** | 0.000 | ||
0.3 | 114.000 | 57.000 | 81.000 *** | 0.000 | ||
Root number | Base medium | 1/4 MS | 29.609 | 14.804 | 20.636 *** | 0.000 |
1/2 MS | 42.007 | 21.003 | 29.277 *** | 0.000 | ||
MS | 32.54 | 16.27 | 22.679 *** | 0.000 | ||
NAA (mg/L) | 0.1 | 24.949 | 12.474 | 17.388 *** | 0.000 | |
0.2 | 9.887 | 4.943 | 6.891 ** | 0.006 | ||
0.3 | 20.16 | 10.08 | 14.051 *** | 0.000 | ||
Root length | Base medium | 1/4 MS | 11.752 | 5.876 | 25.784 *** | 0.000 |
1/2 MS | 11.674 | 5.837 | 25.613 *** | 0.000 | ||
MS | 8.502 | 4.251 | 18.653 *** | 0.000 | ||
NAA (mg/L) | 0.1 | 5.431 | 2.716 | 11.916 *** | 0.001 | |
0.2 | 2.835 | 1.417 | 6.22 ** | 0.009 | ||
0.3 | 6.579 | 3.289 | 14.433 *** | 0.000 | ||
Plant height (5 d) | Seedling refining time | 1 d | 0.136 | 0.068 | 1.011 | 0.384 |
3 d | 0.242 | 0.121 | 1.807 | 0.193 | ||
5 d | 0.616 | 0.308 | 4.591 * | 0.024 | ||
Substrate for transplanting | 1:1 | 0.676 | 0.338 | 5.039 * | 0.018 | |
1:2 | 1.5 | 0.75 | 11.188 *** | 0.001 | ||
1:3 | 1.309 | 0.654 | 9.762 *** | 0.001 | ||
Plant height (10 d) | Seedling refining time | 1 d | 0.429 | 0.214 | 8.515 ** | 0.002 |
3 d | 0.389 | 0.194 | 7.721 ** | 0.004 | ||
5 d | 0.807 | 0.403 | 16.015 *** | 0.000 | ||
Substrate for transplanting | 1:1 | 0.669 | 0.334 | 13.279 *** | 0.000 | |
1:2 | 1.182 | 0.591 | 23.471 *** | 0.000 | ||
1:3 | 0.96 | 0.48 | 19.059 *** | 0.000 | ||
Plant height (15 d) | Seedling refining time | 1 d | 0.329 | 0.164 | 7.525 ** | 0.004 |
3 d | 0.909 | 0.454 | 20.797 *** | 0.000 | ||
5 d | 1.629 | 0.814 | 37.271 *** | 0.000 | ||
Substrate for transplanting | 1:1 | 0.242 | 0.121 | 5.542 ** | 0.013 | |
1:2 | 1.402 | 0.701 | 32.085 *** | 0.000 | ||
1:3 | 1.109 | 0.554 | 25.373 *** | 0.000 |
Ingredient | 1/4 MS (mg/L) | 1/2 MS (mg/L) | MS (mg/L) |
---|---|---|---|
KNO3 | 475 | 950 | 1900 |
NH4NO3 | 412.5 | 825 | 1650 |
KH2PO4 | 42.5 | 85 | 170 |
MgSO4·7H2O | 92.5 | 185 | 370 |
CaCl2·2H2O | 110 | 220 | 440 |
KI | 0.83 | ||
H3BO3 | 6.2 | ||
MnSO4·H2O | 22.30 | ||
ZnSO4·7H2O | 8.6 | ||
Na2MoO4·2H2O | 0.25 | ||
CuSO4·5H2O | 0.025 | ||
CoCl2·6H2O | 0.025 | ||
Na2–EDTA | 37.3 | ||
FeSO4·4H2O | 27.8 | ||
C6H12O6 | 100 | ||
C2H5NO2 | 2 | ||
C12H17ClN4OS·HCl | 0.1 | ||
C8H12ClNO3 | 0.5 | ||
C6H5NO2 | 0.5 | ||
Sucrose | 30,000 | ||
Agar | 7000 |
Treatment | 6-BA (mg/L) | NAA (mg/L) | 2,4-D (mg/L) | TDZ (mg/L) |
---|---|---|---|---|
A-1 | 1 | 0.1 | 0 | 0 |
A-2 | 1 | 0.3 | 0.3 | 0.1 |
A-3 | 1 | 0.5 | 0.1 | 0.3 |
A-4 | 1.5 | 0.1 | 0.3 | 0.3 |
A-5 | 1.5 | 0.3 | 0.1 | 0 |
A-6 | 1.5 | 0.5 | 0 | 0.1 |
A-7 | 2 | 0.1 | 0.1 | 0.1 |
A-8 | 2 | 0.3 | 0 | 0.3 |
A-9 | 2 | 0.5 | 0.3 | 0 |
Treatment | Base Medium | NAA (mg/L) |
---|---|---|
B-1 | 1/4 MS | 0.1 |
B-2 | 0.2 | |
B-3 | 0.3 | |
B-4 | 1/2 MS | 0.1 |
B-5 | 0.2 | |
B-6 | 0.3 | |
B-7 | MS | 0.1 |
B-8 | 0.2 | |
B-9 | 0.3 |
Treatment | Seedling Refining Time (d) | Substrate for Transplanting |
---|---|---|
C-1 | 1 | 1:1 |
C-2 | 1:2 | |
C-3 | 1:3 | |
C-4 | 3 | 1:1 |
C-5 | 1:2 | |
C-6 | 1:3 | |
C-7 | 5 | 1:1 |
C-8 | 1:2 | |
C-9 | 1:3 |
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Treatment | Factor A | Factor B | Factor C | Factor D | Differentiation Coefficient | |
---|---|---|---|---|---|---|
6-BA (mg/L) | NAA (mg/L) | 2,4-D (mg/L) | TDZ (mg/L) | Stem Segment with Bud | Leaf | |
A-1 | 1.0 | 0.1 | 0 | 0 | 8.33 ± 0.58 c | 2.00 ± 1.00 a |
A-2 | 1.0 | 0.3 | 0.3 | 0.1 | 4.00 ± 1.00 f | 1.33 ± 0.58 a |
A-3 | 1.0 | 0.5 | 0.1 | 0.3 | 6.00 ± 1.00 de | 1.00 ± 0 a |
A-4 | 1.5 | 0.1 | 0.3 | 0.3 | 7.33 ± 0.58 cd | 1.67 ± 0.58 a |
A-5 | 1.5 | 0.3 | 0.1 | 0 | 10.00 ± 1.00 b | 1.67 ± 0.58 a |
A-6 | 1.5 | 0.5 | 0 | 0.1 | 14.00 ± 1.00 a | 1.67 ± 0.58 a |
A-7 | 2.0 | 0.1 | 0.1 | 0.1 | 6.33 ± 0.58 de | 1.67 ± 0.58 a |
A-8 | 2.0 | 0.3 | 0 | 0.3 | 5.67 ± 0.58 e | 1.00 ± 1.00 a |
A-9 | 2.0 | 0.5 | 0.3 | 0 | 7.00 ± 1.00 cde | 1.00 ± 0 a |
Dependent Variable | Source of Variance | Sum of Squares | Mean Square | F | p |
---|---|---|---|---|---|
Differentiation coefficient (Stem segment with bud) | 6-BA | 107.185 | 53.593 | 76.158 *** | 0.000 |
NAA | 28.074 | 14.037 | 19.947 *** | 0.000 | |
2,4-D | 47.185 | 23.593 | 33.526 *** | 0.000 | |
TDZ | 23.185 | 11.593 | 16.474 *** | 0.000 | |
Differentiation coefficient (Leaf) | 6-BA | 0.889 | 0.444 | 1.091 | 0.357 |
NAA | 1.556 | 0.778 | 1.909 | 0.177 | |
2,4-D | 0.222 | 0.111 | 0.273 | 0.764 | |
TDZ | 0.667 | 0.333 | 0.818 | 0.457 |
Treatment | Factor A | Factor B | Rooting Rate (%) | Root Number | Root Length (cm) |
---|---|---|---|---|---|
Basic Medium | NAA (mg/L) | ||||
B-1 | 1/4 MS | 0.1 | 95.24 ± 4.12 a | 9.87 ± 1.47 c | 7.46 ± 0.90 c |
B-2 | 0.2 | 94.84 ± 4.51 a | 7.12 ± 0.40 e | 5.87 ± 0.35 e | |
B-3 | 0.3 | 94.44 ± 4.81 a | 5.45 ± 0.59 f | 4.67 ± 0.11 f | |
B-4 | 1/2 MS | 0.1 | 97.62 ± 4.12 a | 13.92 ± 0.70 a | 9.36 ± 0.71 a |
B-5 | 0.2 | 97.22 ± 4.81 a | 9.71 ± 1.15 c | 7.22 ± 0.40 c | |
B-6 | 0.3 | 94.44 ± 4.81 a | 9.08 ± 0.43 cd | 6.74 ± 0.32 cd | |
B-7 | MS | 0.1 | 97.62 ± 4.12 a | 12.18 ± 0.69 b | 8.32 ± 0.20 b |
B-8 | 0.2 | 97.62 ± 4.12 a | 8.45 ± 1.07 | 6.77 ± 0.38 cd | |
B-9 | 0.3 | 94.84 ± 4.51 a | 7.86 ± 0.45 de | 5.98 ± 0.40 de |
Character | Source of Variance | Sum of Squares | Mean Square | F | p |
---|---|---|---|---|---|
Rooting rate | Basic medium | 18.042 | 9.021 | 0.547 | 0.587 |
NAA | 27.168 | 13.584 | 0.823 | 0.452 | |
Root number | Basic medium | 53.295 | 26.647 | 39.101 *** | 0.000 |
NAA | 102.348 | 51.174 | 75.091 *** | 0.000 | |
Root length | Basic medium | 14.263 | 7.132 | 33.498 *** | 0.000 |
NAA | 31.347 | 15.674 | 73.621 *** | 0.000 |
Treatment | Factor A | Factor B | Plant Height (0 d) | Plant Height (5 d) | Plant Height (10 d) | Plant Height (15 d) |
---|---|---|---|---|---|---|
Seedling Refining Time (d) | Substrate for Transplanting | |||||
C-1 | 1 | 1:1 | 3.21 ± 0.10 ab | 3.98 ± 0.31 e | 4.43 ± 0.23 f | 4.97 ± 0.15 e |
C-2 | 1:2 | 3.26 ± 0.06 ab | 4.26 ± 0.32 de | 4.97 ± 0.12 cd | 5.43 ± 0.15 c | |
C-3 | 1:3 | 3.18 ± 0.12 b | 4.11 ± 0.20 e | 4.67 ± 0.12 ef | 5.17 ± 0.06 cde | |
C-4 | 3 | 1:1 | 3.12 ± 0.06 b | 4.38 ± 0.21 cde | 4.72 ± 0.15 de | 5.12 ± 0.21 de |
C-5 | 1:2 | 3.28 ± 0.15 ab | 4.78 ± 0.15 bc | 5.23 ± 0.06 bc | 5.90 ± 0.10 b | |
C-6 | 1:3 | 3.31 ± 0.20 ab | 4.59 ± 0.20 bcd | 5.07 ± 0.21 c | 5.41 ± 0.10 cd | |
C-7 | 5 | 1:1 | 3.42 ± 0.15 a | 4.64 ± 0.35 bcd | 5.10 ± 0.10 c | 5.37 ± 0.06 cd |
C-8 | 1:2 | 3.23 ± 0.15 ab | 5.27 ± 0.21 a | 5.84 ± 0.21 a | 6.40 ± 0.10 a | |
C-9 | 1:3 | 3.37 ± 0.12 ab | 5.03 ± 0.31 ab | 5.47 ± 0.15 b | 6.00 ± 0.26 b |
Character | Source of Variance | Sum of Squares | Mean Square | F | p |
---|---|---|---|---|---|
Plant height (0 d) | Seedling refining time | 0.092 | 0.046 | 2.36 | 0.118 |
Substrate for transplanting | 0.003 | 0.001 | 0.076 | 0.927 | |
Plant height (5 d) | Seedling refining time | 3.387 | 1.693 | 28.559 *** | 0.000 |
Substrate for transplanting | 0.896 | 0.448 | 7.552 ** | 0.003 | |
Plant height (10 d) | Seedling refining time | 2.749 | 1.374 | 58.651 *** | 0.000 |
Substrate for transplanting | 1.562 | 0.781 | 33.332 *** | 0.000 | |
Plant height (15 d) | Seedling refining time | 2.456 | 1.228 | 39.137 *** | 0.000 |
Substrate for transplanting | 2.570 | 1.285 | 40.943 *** | 0.000 |
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Chen, Q.; Gao, K.; Pan, B.; Wang, Y.; Chen, L.; Yu, J.; Wang, L.; Fan, Y.; Li, H.; Huang, C. Construction of Optimal Regeneration System for Chrysanthemum ‘11-C-2’ Stem Segment with Buds. Plants 2024, 13, 2403. https://doi.org/10.3390/plants13172403
Chen Q, Gao K, Pan B, Wang Y, Chen L, Yu J, Wang L, Fan Y, Li H, Huang C. Construction of Optimal Regeneration System for Chrysanthemum ‘11-C-2’ Stem Segment with Buds. Plants. 2024; 13(17):2403. https://doi.org/10.3390/plants13172403
Chicago/Turabian StyleChen, Qingbing, Kang Gao, Bo Pan, Yaoyao Wang, Lijie Chen, Junjun Yu, Lili Wang, Yongming Fan, Haiying Li, and Conglin Huang. 2024. "Construction of Optimal Regeneration System for Chrysanthemum ‘11-C-2’ Stem Segment with Buds" Plants 13, no. 17: 2403. https://doi.org/10.3390/plants13172403