Chinese Bulletin of Botany ›› 2024, Vol. 59 ›› Issue (3): 433-440.DOI: 10.11983/CBB23094 cstr: 32102.14.CBB23094
• TECHNIQUES AND METHODS • Previous Articles Next Articles
Hao Zeng1,2, Peifang Li1,2, Zhihui Guo1,2, Chunlin Liu1,3, Ying Ruan1,2,*()
Received:
2023-07-18
Accepted:
2023-12-19
Online:
2024-05-10
Published:
2024-05-10
Contact:
E-mail: Hao Zeng, Peifang Li, Zhihui Guo, Chunlin Liu, Ying Ruan. Establishment of a Regeneration System for Lunaria annua[J]. Chinese Bulletin of Botany, 2024, 59(3): 433-440.
Number | 75% ethanol (s) | 0.1% HgCl2 (min) | Number of explants | Contamination rate (%) | Survival rate (%) |
---|---|---|---|---|---|
1 | 45 | 8 | 69 | 0.90±0.79 c | 3.01±0.44 c |
2 | 45 | 6 | 96 | 13.10±0.79 b | 86.70±1.29 b |
3 | 45 | 4 | 75 | 47.33±1.15 a | 98.88±0.96 a |
Table 1 Effect of the duration of HgCl2 sterilization on the disinfection of explants
Number | 75% ethanol (s) | 0.1% HgCl2 (min) | Number of explants | Contamination rate (%) | Survival rate (%) |
---|---|---|---|---|---|
1 | 45 | 8 | 69 | 0.90±0.79 c | 3.01±0.44 c |
2 | 45 | 6 | 96 | 13.10±0.79 b | 86.70±1.29 b |
3 | 45 | 4 | 75 | 47.33±1.15 a | 98.88±0.96 a |
Number | 6-BA (mg∙L-1) | NAA (mg∙L-1) | Number of explants | Callus induction rate (%) |
---|---|---|---|---|
1 | 0.5 | 0 | 90 | 0.67±0.67 g |
2 | 1.0 | 0 | 90 | 1.22±1.17 g |
3 | 2.0 | 0 | 90 | 2.22±0.51 g |
4 | 0.5 | 1.0 | 90 | 40.89±2.14 d |
5 | 1.0 | 1.0 | 90 | 75.60±1.06 b |
6 | 2.0 | 1.0 | 90 | 52.00±0.78 c |
7 | 0.5 | 2.0 | 90 | 32.74±0.56 f |
8 | 1.0 | 2.0 | 90 | 78.04±1.20 a |
9 | 2.0 | 2.0 | 90 | 35.52±0.83 e |
Table 2 Effects of 6-BA and NAA combination on callus induction of Lunaria annua leaf explants
Number | 6-BA (mg∙L-1) | NAA (mg∙L-1) | Number of explants | Callus induction rate (%) |
---|---|---|---|---|
1 | 0.5 | 0 | 90 | 0.67±0.67 g |
2 | 1.0 | 0 | 90 | 1.22±1.17 g |
3 | 2.0 | 0 | 90 | 2.22±0.51 g |
4 | 0.5 | 1.0 | 90 | 40.89±2.14 d |
5 | 1.0 | 1.0 | 90 | 75.60±1.06 b |
6 | 2.0 | 1.0 | 90 | 52.00±0.78 c |
7 | 0.5 | 2.0 | 90 | 32.74±0.56 f |
8 | 1.0 | 2.0 | 90 | 78.04±1.20 a |
9 | 2.0 | 2.0 | 90 | 35.52±0.83 e |
Number | 6-BA (mg∙L-1) | 2,4-D (mg∙L-1) | Number of explants | Callus induction rate (%) |
---|---|---|---|---|
1 | 0.5 | 0 | 90 | 1.22±0.51 g |
2 | 1.0 | 0 | 90 | 2.26±0.51 fg |
3 | 2.0 | 0 | 90 | 3.00±0.62 f |
4 | 0.5 | 1.0 | 90 | 82.48±1.08 b |
5 | 1.0 | 1.0 | 90 | 69.08±0.63 c |
6 | 2.0 | 1.0 | 90 | 25.19±1.19 e |
7 | 0.5 | 2.0 | 90 | 93.37±1.07 a |
8 | 1.0 | 2.0 | 90 | 81.26±1.14 b |
9 | 2.0 | 2.0 | 90 | 31.63±0.56 d |
Table 3 Effects of 6-BA and 2,4-D combination on callus induction of Lunaria annua leaf explants
Number | 6-BA (mg∙L-1) | 2,4-D (mg∙L-1) | Number of explants | Callus induction rate (%) |
---|---|---|---|---|
1 | 0.5 | 0 | 90 | 1.22±0.51 g |
2 | 1.0 | 0 | 90 | 2.26±0.51 fg |
3 | 2.0 | 0 | 90 | 3.00±0.62 f |
4 | 0.5 | 1.0 | 90 | 82.48±1.08 b |
5 | 1.0 | 1.0 | 90 | 69.08±0.63 c |
6 | 2.0 | 1.0 | 90 | 25.19±1.19 e |
7 | 0.5 | 2.0 | 90 | 93.37±1.07 a |
8 | 1.0 | 2.0 | 90 | 81.26±1.14 b |
9 | 2.0 | 2.0 | 90 | 31.63±0.56 d |
Figure 1 Establishment of regeneration system for Lunaria annua (A) Freshly inoculated explants; (B) Induction of callus; (C) Differentiation of adventitious buds; (D) Adventitious buds; (E) Adventitious buds proliferation; (F) Seedlings from tissue culture; (G) Transplanting. Bars=1 cm
Number | 6-BA (mg∙L-1) | 2,4-D (mg∙L-1) | Number of explants | Adventitious bud induction rate (%) | Proliferation coefficient |
---|---|---|---|---|---|
1 | 0.25 | 2.0 | 40 | 34.08±1.42 d | 1.83±0.15 d |
2 | 0.5 | 2.0 | 40 | 84.08±1.38 a | 3.63±0.35 a |
3 | 0.75 | 2.0 | 40 | 65.91±2.04 b | 2.93±0.15 b |
4 | 1.0 | 2.0 | 40 | 43.41±0.88 c | 2.30±0.26 c |
Table 4 Effects of different combinations of plant growth regulator on the induction of adventitious bud in callus of Lunaria annua
Number | 6-BA (mg∙L-1) | 2,4-D (mg∙L-1) | Number of explants | Adventitious bud induction rate (%) | Proliferation coefficient |
---|---|---|---|---|---|
1 | 0.25 | 2.0 | 40 | 34.08±1.42 d | 1.83±0.15 d |
2 | 0.5 | 2.0 | 40 | 84.08±1.38 a | 3.63±0.35 a |
3 | 0.75 | 2.0 | 40 | 65.91±2.04 b | 2.93±0.15 b |
4 | 1.0 | 2.0 | 40 | 43.41±0.88 c | 2.30±0.26 c |
Number | Medium formulation | Number of explants | Rooting rate (%) | Average root length (cm) | Average root number |
---|---|---|---|---|---|
1 | MS+0.1 mg∙L-1 NAA | 15 | 91.63±1.67 a | 8.14±0.26 a | 8.67±1.54 a |
2 | MS+0.5 mg∙L-1 NAA+0.5 mg∙L-1 IBA | 15 | 81.78±1.56 b | 5.73±0.71 b | 6.33±0.88 b |
Table 5 Effects of different formulation of culture media on adventitious root induction of Lunaria annua
Number | Medium formulation | Number of explants | Rooting rate (%) | Average root length (cm) | Average root number |
---|---|---|---|---|---|
1 | MS+0.1 mg∙L-1 NAA | 15 | 91.63±1.67 a | 8.14±0.26 a | 8.67±1.54 a |
2 | MS+0.5 mg∙L-1 NAA+0.5 mg∙L-1 IBA | 15 | 81.78±1.56 b | 5.73±0.71 b | 6.33±0.88 b |
Number | Pre-incubation time (d) | Number of explants | Rooting rate (%) |
---|---|---|---|
1 | 0 | 8 | 0 |
2 | 5 | 8 | 20.83±7.22 b |
3 | 10 | 8 | 95.83±7.22 a |
Table 6 Effect of pre-incubation time on rooting of adventitious buds of Lunaria annua after IAA treatment
Number | Pre-incubation time (d) | Number of explants | Rooting rate (%) |
---|---|---|---|
1 | 0 | 8 | 0 |
2 | 5 | 8 | 20.83±7.22 b |
3 | 10 | 8 | 95.83±7.22 a |
Rooting method | Number of explants | Survival rate (%) | Growth situation |
---|---|---|---|
Medium induced rooting | 25 | 93.33±0.58 a | ++ |
Rooting in soil after pre-incubation | 25 | 98.67±0.58 a | + |
Table 7 Hardening-seedling and transplanting of the regenerated plantlet of Lunaria annua
Rooting method | Number of explants | Survival rate (%) | Growth situation |
---|---|---|---|
Medium induced rooting | 25 | 93.33±0.58 a | ++ |
Rooting in soil after pre-incubation | 25 | 98.67±0.58 a | + |
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