In Vitro Tetraploid Induction from Leaf and Petiole Explants of Hybrid Sweetgum (Liquidambar styraciflua × Liquidambar formosana)
Abstract
:1. Introduction
2. Materials and Methods
2.1. Plant Materials
2.2. Establishment of Aseptic Seedlings and In Vitro Multiplication
2.3. Colchicine Application
2.4. Flow Cytometric Analysis of Ploidy Level
2.5. Chromosome Counting
2.6. Statistical Analysis
3. Results
3.1. Morphologic Observations in the Initial Stage of Regeneration
3.2. Survival Rate and Regeneration of Colchicine-Treated Explants
3.3. Analysis by Flow Cytometry and Polyploid Determination
4. Discussion
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Levels | Factors | |||
---|---|---|---|---|
A | B | C | D | |
Genotype | Colchicine Concentration (mg/L) | Pre-Culture Duration (day) | Exposure Time (day) | |
1 | Z1 | 120 | 4 | 3 |
2 | Z2 | 160 | 6 | 4 |
3 | Z3 | 200 | 8 | 5 |
Treatment | Factors | Number of Shoots Examined | Survival Rate % | No. of Tetraploid | No. of Mixoploid | Tetraploid Induction % | |||
---|---|---|---|---|---|---|---|---|---|
A | B | C | D | ||||||
c1 | 1 | 1 | 1 | 1 | 50 | 95.00 | 1 | 1 | 2.00 |
c2 | 1 | 2 | 2 | 2 | 50 | 75.00 | 0 | 1 | 0.00 |
c3 | 1 | 3 | 3 | 3 | 30 | 41.67 | 1 | 2 | 3.33 |
c4 | 2 | 1 | 2 | 3 | 15 | 8.33 | 0 | 0 | 0.00 |
c5 | 2 | 2 | 3 | 1 | 50 | 81.67 | 1 | 1 | 2.00 |
c6 | 2 | 3 | 1 | 2 | 50 | 90.00 | 0 | 1 | 0.00 |
c7 | 3 | 1 | 3 | 2 | 50 | 75.00 | 0 | 1 | 0.00 |
c8 | 3 | 2 | 1 | 3 | 50 | 70.00 | 0 | 0 | 0.00 |
c9 | 3 | 3 | 2 | 1 | 50 | 85.00 | 1 | 2 | 2.00 |
Treatment | Factors | Number of Shoots Examined | Survival Rate % | No. of Tetraploid | No. of Mixoploid | Tetraploid Induction % | |||
---|---|---|---|---|---|---|---|---|---|
A | B | C | D | ||||||
c1 | 1 | 1 | 1 | 1 | 50 | 85.00 | 1 | 0 | 2.00 |
c2 | 1 | 2 | 2 | 2 | 50 | 56.67 | 2 | 1 | 4.00 |
c3 | 1 | 3 | 3 | 3 | 30 | 28.33 | 1 | 0 | 3.33 |
c4 | 2 | 1 | 2 | 3 | 15 | 10.00 | 0 | 0 | 0.00 |
c5 | 2 | 2 | 3 | 1 | 50 | 56.67 | 3 | 2 | 6.00 |
c6 | 2 | 3 | 1 | 2 | 50 | 75.00 | 0 | 0 | 0.00 |
c7 | 3 | 1 | 3 | 2 | 50 | 48.33 | 0 | 0 | 0.00 |
c8 | 3 | 2 | 1 | 3 | 50 | 51.67 | 0 | 0 | 0.00 |
c9 | 3 | 3 | 2 | 1 | 50 | 70.00 | 4 | 2 | 8.00 |
Variation Source | df | MS | F | Sig. |
---|---|---|---|---|
Genotype | 2 | 294.147 | 4.876 | 0.020 * |
Concentration | 2 | 193.362 | 3.205 | 0.064 |
Pre-culture duration | 2 | 1087.422 | 18.027 | 0.000 * |
Exposure time | 2 | 2769.584 | 45.913 | 0.000 * |
Error | 18 | 60.322 | ||
Total | 27 |
Variation Source | df | MS | F | Sig. |
---|---|---|---|---|
Genotype | 2 | 138.765 | 6.238 | 0.009 * |
Concentration | 2 | 97.631 | 4.389 | 0.028 * |
Pre-culture duration | 2 | 811.981 | 36.502 | 0.000 * |
Exposure time | 2 | 1612.270 | 72.478 | 0.000 * |
Error | 18 | 22.245 | ||
Total | 27 |
Explant Type | A | B | C | D | |
---|---|---|---|---|---|
Leaf | K1 | 211.67 | 178.33 | 255.00 | 261.67 |
K2 | 180.00 | 226.67 | 168.33 | 240.00 | |
K3 | 230.00 | 216.67 | 198.33 | 120.00 | |
k1 | 70.56 | 59.44 | 85.00 | 87.22 | |
k2 | 60.00 | 75.56 | 56.11 | 80.00 | |
k3 | 76.67 | 72.22 | 66.11 | 40.00 | |
R | 16.67 | 16.11 | 28.89 | 47.22 | |
Petiole | K1 | 170.00 | 143.33 | 211.67 | 211.67 |
K2 | 141.67 | 165.00 | 136.67 | 180.00 | |
K3 | 170.00 | 173.33 | 133.33 | 90.00 | |
k1 | 56.67 | 47.78 | 70.56 | 70.56 | |
k2 | 47.22 | 55.00 | 45.56 | 60.00 | |
k3 | 56.67 | 57.78 | 44.44 | 30.00 | |
R | 9.44 | 10.00 | 26.11 | 40.56 |
Explant Type | A | B | C | D | |
---|---|---|---|---|---|
Leaf | K1 | 5.33 | 2.00 | 2.00 | 6.00 |
K2 | 2.00 | 2.00 | 2.00 | 0.00 | |
K3 | 2.00 | 5.33 | 5.33 | 3.33 | |
k1 | 1.78 | 0.67 | 0.67 | 2.00 | |
k2 | 0.67 | 0.67 | 0.67 | 0.00 | |
k3 | 0.67 | 1.78 | 1.78 | 1.11 | |
R | 1.11 | 1.11 | 1.11 | 2.00 | |
Petiole | K1 | 9.33 | 2.00 | 2.00 | 16.00 |
K2 | 6.00 | 10.00 | 12.00 | 4.00 | |
K3 | 8.00 | 11.33 | 9.33 | 3.33 | |
k1 | 3.11 | 0.67 | 0.67 | 5.33 | |
k2 | 2.00 | 3.33 | 4.00 | 1.33 | |
k3 | 2.67 | 3.78 | 3.11 | 1.11 | |
R | 1.11 | 3.11 | 3.33 | 4.22 |
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Zhang, Y.; Wang, Z.; Qi, S.; Wang, X.; Zhao, J.; Zhang, J.; Li, B.; Zhang, Y.; Liu, X.; Yuan, W. In Vitro Tetraploid Induction from Leaf and Petiole Explants of Hybrid Sweetgum (Liquidambar styraciflua × Liquidambar formosana). Forests 2017, 8, 264. https://doi.org/10.3390/f8080264
Zhang Y, Wang Z, Qi S, Wang X, Zhao J, Zhang J, Li B, Zhang Y, Liu X, Yuan W. In Vitro Tetraploid Induction from Leaf and Petiole Explants of Hybrid Sweetgum (Liquidambar styraciflua × Liquidambar formosana). Forests. 2017; 8(8):264. https://doi.org/10.3390/f8080264
Chicago/Turabian StyleZhang, Yan, Zewei Wang, Shuaizheng Qi, Xiaoqi Wang, Jian Zhao, Jinfeng Zhang, Bailian Li, Yadong Zhang, Xuezeng Liu, and Wei Yuan. 2017. "In Vitro Tetraploid Induction from Leaf and Petiole Explants of Hybrid Sweetgum (Liquidambar styraciflua × Liquidambar formosana)" Forests 8, no. 8: 264. https://doi.org/10.3390/f8080264