Chinese Bulletin of Botany ›› 2020, Vol. 55 ›› Issue (5): 605-612.DOI: 10.11983/CBB20030
• TECHNIQUES AND METHODS • Previous Articles Next Articles
Jianfei Liu1, Yan Liu1, Kejian Liu2, Yang Chi3, Zhifa Huo3, Yonghong Huo3, Xiangling You1,*()
Received:
2020-02-26
Accepted:
2020-06-05
Online:
2020-09-01
Published:
2020-09-03
Contact:
Xiangling You
Jianfei Liu, Yan Liu, Kejian Liu, Yang Chi, Zhifa Huo, Yonghong Huo, Xiangling You. Optimization of the Regeneration System from Somatic Embryogenesis in Larix olgensis[J]. Chinese Bulletin of Botany, 2020, 55(5): 605-612.
Figure 1 Callus and its microscopic images of Larix olgensis (A) Callus; (B) Embryogenic callus; (C) Non-embryogenic callus; (D) The image of embryogenic callus; (E) The image of non-embryogenic callus. (A)-(C) Bars=3 mm; (D) Bar= 200 μm; (E) Bar=500 μm
Excellent trees | Callus induction rate (%) | Embryogenic callus formation rate (%) |
---|---|---|
061-1009 | 89.43±48.59 a | 0 |
091-1008 | 81.56±47.91 c | 0 |
058-842 | 91.25±53.17 a | 4.53±2.73 a |
024-922 | 85.91±37.29 b | 0 |
Table 1 Callus and embryogenic callus induction in different Larix olgensis lines (means±SD)
Excellent trees | Callus induction rate (%) | Embryogenic callus formation rate (%) |
---|---|---|
061-1009 | 89.43±48.59 a | 0 |
091-1008 | 81.56±47.91 c | 0 |
058-842 | 91.25±53.17 a | 4.53±2.73 a |
024-922 | 85.91±37.29 b | 0 |
The combination of plant growth regulators in the medium | Somatic embryogenesis (%) | Somatic embryo amount (numbers·g-1) |
---|---|---|
0.1 mg?L-1 2,4-D+0.04 mg·L-1 BA+0.02 mg·L-1 KT | 45.27±23.31 | 16.47±1.67 |
0.5 mg?L-1 NAA+0.5 mg·L-1 BA+0.5 mg·L-1 KT | 52.65±31.83 b | 25.31±9.85 c |
0.4 mg?L-1 NAA+0.5 mg·L-1 BA+0.5 mg·L-1 KT | 57.38±15.41 ab | 31.53±5.37 b |
0.3 mg?L-1 NAA+0.5 mg·L-1 BA+0.5 mg·L-1 KT | 64.41±11.79 a | 39.74±4.29 ab |
0.2 mg?L-1 NAA+0.5 mg·L-1 BA+0.5 mg·L-1 KT | 72.36±10.57 a | 48.56±3.77 a |
0.1 mg?L-1 NAA+0.5 mg·L-1 BA+0.5 mg·L-1 KT | 63.70±43.86 a | 41.92±7.22 a |
Table 2 Effects of plant growth regulators at different concentration ratios on somatic embryogenesis and somatic embryo amount in Larix olgensis (means±SD)
The combination of plant growth regulators in the medium | Somatic embryogenesis (%) | Somatic embryo amount (numbers·g-1) |
---|---|---|
0.1 mg?L-1 2,4-D+0.04 mg·L-1 BA+0.02 mg·L-1 KT | 45.27±23.31 | 16.47±1.67 |
0.5 mg?L-1 NAA+0.5 mg·L-1 BA+0.5 mg·L-1 KT | 52.65±31.83 b | 25.31±9.85 c |
0.4 mg?L-1 NAA+0.5 mg·L-1 BA+0.5 mg·L-1 KT | 57.38±15.41 ab | 31.53±5.37 b |
0.3 mg?L-1 NAA+0.5 mg·L-1 BA+0.5 mg·L-1 KT | 64.41±11.79 a | 39.74±4.29 ab |
0.2 mg?L-1 NAA+0.5 mg·L-1 BA+0.5 mg·L-1 KT | 72.36±10.57 a | 48.56±3.77 a |
0.1 mg?L-1 NAA+0.5 mg·L-1 BA+0.5 mg·L-1 KT | 63.70±43.86 a | 41.92±7.22 a |
Figure 3 Effects of different factors on the rate of somatic embryogenesis in Larix olgensis (means±SE) (A) ABA; (B) PEG4000; (C) Sucrose and maltose; (D) Cultured with sucrose for 6 weeks; (E) Cultured with maltose for 6 weeks. (D), (E) Bars=2.5 mm. Different lowercase letters indicate significant differences (P<0.05). * P<0.05; ** P<0.01
Figure 4 Somatic embryo seedlings at different developmental states of Larix olgensis and their transplanting (A) Cotyledonary embryo; (B) Cotyledons germinate; (C) Control of somatic embryo seedling; (D) Optimization somatic embryo seedling; (E) Rooting somatic embryo seedlings; (F) Transplanting somatic embryo seedling. (A) Bar=1.5 mm; (B) Bar=6.5 mm; (C), (D) Bars=1.0 cm; (E), (F) Bars=1.5 cm
Growth regulator combination | Somatic embryo seedlings | |||
---|---|---|---|---|
NAA (mg·L-1) | IBA (mg·L-1) | Seedling rate (%) | Stem length (mm) | Main root length (mm) |
0.4 | 0.5 | 38.83±29.38 ab | 1.59±1.44 c | 16.38±12.74 b |
0.4 | 1.0 | 46.21±41.29 a | 1.75±0.77 ab | 19.17±14.52 a |
0.4 | 1.5 | 35.37±33.34 b | 1.67±1.75 b | 12.60±10.34 c |
0.8 | 0.5 | 22.57±25.43 c | 1.76±0.91 ab | 12.31±9.54 c |
0.8 | 1.0 | 22.65±15.83 c | 1.81±0.88 a | 15.18±13.37 bc |
0.8 | 1.5 | 21.43±19.68 c | 1.78±0.67 ab | 11.40±8.53 c |
Table 3 Effects of growth regulator combination on the growth of somatic embryo seedling in Larix olgensis (means±SD)
Growth regulator combination | Somatic embryo seedlings | |||
---|---|---|---|---|
NAA (mg·L-1) | IBA (mg·L-1) | Seedling rate (%) | Stem length (mm) | Main root length (mm) |
0.4 | 0.5 | 38.83±29.38 ab | 1.59±1.44 c | 16.38±12.74 b |
0.4 | 1.0 | 46.21±41.29 a | 1.75±0.77 ab | 19.17±14.52 a |
0.4 | 1.5 | 35.37±33.34 b | 1.67±1.75 b | 12.60±10.34 c |
0.8 | 0.5 | 22.57±25.43 c | 1.76±0.91 ab | 12.31±9.54 c |
0.8 | 1.0 | 22.65±15.83 c | 1.81±0.88 a | 15.18±13.37 bc |
0.8 | 1.5 | 21.43±19.68 c | 1.78±0.67 ab | 11.40±8.53 c |
Plant regeneration | GA3 concentration (mg·L-1) | ||
---|---|---|---|
0 | 0.5 | 1.0 | |
Seedling rate (%) | 45.61±52.27 b | 55.44±37.91 a | 45.91±35.81 b |
Stem length (mm) | 1.63±1.33 b | 2.65±2.61 a | 0.91±1.11 c |
Main root length (mm) | 17.98±14.22 a | 12.41±16.55 b | 8.65±5.49 c |
Table 4 Effects of different concentration of GA3 on the growth of somatic embryo seedling in Larix olgensis (means± SD)
Plant regeneration | GA3 concentration (mg·L-1) | ||
---|---|---|---|
0 | 0.5 | 1.0 | |
Seedling rate (%) | 45.61±52.27 b | 55.44±37.91 a | 45.91±35.81 b |
Stem length (mm) | 1.63±1.33 b | 2.65±2.61 a | 0.91±1.11 c |
Main root length (mm) | 17.98±14.22 a | 12.41±16.55 b | 8.65±5.49 c |
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