植物学报 ›› 2019, Vol. 54 ›› Issue (4): 539-546.DOI: 10.11983/CBB18161
• 技术方法 • 上一篇
收稿日期:
2018-07-18
接受日期:
2018-12-10
出版日期:
2019-07-10
发布日期:
2020-01-08
通讯作者:
于贤昌
基金资助:
Jia Guo,Yansu Li,Chaoxing He,Yan Yan,Xianchang Yu()
Received:
2018-07-18
Accepted:
2018-12-10
Online:
2019-07-10
Published:
2020-01-08
Contact:
Xianchang Yu
摘要:
南瓜(Cucurbita moschata)再生率较低, 为建立高效的南瓜再生体系, 以南瓜子叶为外植体, 进行35组不同激素浓度的不定芽诱导研究。结果表明, 南瓜再生受培养基中激素浓度和配比的影响, 适宜浓度6-苄氨基腺嘌呤(6-BA)能有效促进不定芽形成; 单独使用脱落酸(ABA)诱导使南瓜子叶发黄, 但与6-BA组合使用可显著提高外植体的再生能力, 1.0 mg∙L -16-BA与0.5 mg∙L -1ABA组合南瓜芽再生率高达90.26%。将不定芽置于MS培养基中进行生根培养, 再生苗移栽易成活。从子叶接种到苗再生约需70天。
郭佳,李衍素,贺超兴,闫妍,于贤昌. 南瓜高效再生体系的建立. 植物学报, 2019, 54(4): 539-546.
Jia Guo,Yansu Li,Chaoxing He,Yan Yan,Xianchang Yu. Establishing a High-efficiency Regeneration System in Pumpkin (Cucurbita moschata). Chinese Bulletin of Botany, 2019, 54(4): 539-546.
Source | Type III Sum of squares | df | Mean square | F | Sig. |
---|---|---|---|---|---|
Corrected model | 1.194a | 10 | 0.119 | 19.893 | 0.000 |
Intercept | 11.738 | 1 | 11.738 | 1955.511 | 0.000 |
6-BA concentration | 0.932 | 5 | 0.186 | 31.040 | 0.000 |
ABA concentration | 0.291 | 5 | 0.058 | 9.699 | 0.000 |
Error | 0.144 | 24 | 0.006 | ||
Total | 13.664 | 35 | |||
Corrected total | 1.338 | 34 |
表1 南瓜再生率方差分析
Table 1 Variance analysis of regeneration rate of pumpkin
Source | Type III Sum of squares | df | Mean square | F | Sig. |
---|---|---|---|---|---|
Corrected model | 1.194a | 10 | 0.119 | 19.893 | 0.000 |
Intercept | 11.738 | 1 | 11.738 | 1955.511 | 0.000 |
6-BA concentration | 0.932 | 5 | 0.186 | 31.040 | 0.000 |
ABA concentration | 0.291 | 5 | 0.058 | 9.699 | 0.000 |
Error | 0.144 | 24 | 0.006 | ||
Total | 13.664 | 35 | |||
Corrected total | 1.338 | 34 |
Source | Type III Sum of squares | df | Mean square | F | Sig. |
---|---|---|---|---|---|
Corrected model | 20.734a | 10 | 2.073 | 6.731 | 0.000 |
Intercept | 116.473 | 1 | 116.473 | 378.098 | 0.000 |
6-BA concentration | 11.488 | 5 | 2.298 | 7.458 | 0.000 |
ABA concentration | 10.032 | 5 | 2.006 | 6.513 | 0.001 |
Error | 7.393 | 24 | 0.308 | ||
Total | 151.118 | 35 | |||
Corrected total | 28.127 | 34 |
表2 南瓜平均再生芽数方差分析
Table 2 Variance analysis of average number of induced shoots of pumpkin
Source | Type III Sum of squares | df | Mean square | F | Sig. |
---|---|---|---|---|---|
Corrected model | 20.734a | 10 | 2.073 | 6.731 | 0.000 |
Intercept | 116.473 | 1 | 116.473 | 378.098 | 0.000 |
6-BA concentration | 11.488 | 5 | 2.298 | 7.458 | 0.000 |
ABA concentration | 10.032 | 5 | 2.006 | 6.513 | 0.001 |
Error | 7.393 | 24 | 0.308 | ||
Total | 151.118 | 35 | |||
Corrected total | 28.127 | 34 |
Treatments | 6-BA (mg∙L-1) | Number of explants | Number of explants with adventitious shoots | Number of shoots | Average number of induced shoots | Rate of plant regeneration (%) |
---|---|---|---|---|---|---|
M1 | 0.5 | 60 | 39.78 | 73.60 | 1.85 | 66.31 |
M2 | 1.0 | 60 | 41.51 | 72.22 | 1.74 | 69.18 |
M3 | 1.5 | 60 | 34.01 | 50.00 | 1.47 | 56.69 |
M4 | 2.0 | 60 | 29.88 | 33.46 | 1.12 | 49.79 |
M5 | 2.5 | 60 | 27.16 | 31.50 | 1.16 | 45.26 |
表3 不同浓度6-BA对南瓜芽再生的影响
Table 3 Effects of different concentrations of 6-BA on the bud induction of pumpkin
Treatments | 6-BA (mg∙L-1) | Number of explants | Number of explants with adventitious shoots | Number of shoots | Average number of induced shoots | Rate of plant regeneration (%) |
---|---|---|---|---|---|---|
M1 | 0.5 | 60 | 39.78 | 73.60 | 1.85 | 66.31 |
M2 | 1.0 | 60 | 41.51 | 72.22 | 1.74 | 69.18 |
M3 | 1.5 | 60 | 34.01 | 50.00 | 1.47 | 56.69 |
M4 | 2.0 | 60 | 29.88 | 33.46 | 1.12 | 49.79 |
M5 | 2.5 | 60 | 27.16 | 31.50 | 1.16 | 45.26 |
Treatments | ABA (mg∙L-1) | Number of explants | Number of explants with adventitious shoots | Number of shoots | Average number of induced shoots | Rate of plant regeneration (%) |
---|---|---|---|---|---|---|
M6 | 0.5 | 60 | 15.35 | 16.27 | 1.06 | 25.58 |
M7 | 1.0 | 60 | 18.63 | 20.31 | 1.09 | 31.05 |
M8 | 1.5 | 60 | 21.52 | 24.75 | 1.15 | 35.87 |
M9 | 2.0 | 60 | 13.12 | 13.26 | 1.01 | 21.87 |
M10 | 2.5 | 60 | 14.85 | 15.74 | 1.06 | 24.74 |
表4 不同浓度ABA对南瓜芽再生的影响
Table 4 Effects of different concentrations of ABA on the bud induction of pumpkin
Treatments | ABA (mg∙L-1) | Number of explants | Number of explants with adventitious shoots | Number of shoots | Average number of induced shoots | Rate of plant regeneration (%) |
---|---|---|---|---|---|---|
M6 | 0.5 | 60 | 15.35 | 16.27 | 1.06 | 25.58 |
M7 | 1.0 | 60 | 18.63 | 20.31 | 1.09 | 31.05 |
M8 | 1.5 | 60 | 21.52 | 24.75 | 1.15 | 35.87 |
M9 | 2.0 | 60 | 13.12 | 13.26 | 1.01 | 21.87 |
M10 | 2.5 | 60 | 14.85 | 15.74 | 1.06 | 24.74 |
Treat- ments | 6-BA (mg∙L-1) | ABA (mg∙L-1) | Number of explants | Number of explants with adventitious shoots | Number of shoots | Average number of induced shoots | Rate of plant regeneration (%) |
---|---|---|---|---|---|---|---|
T1 | 0.5 | 0.5 | 60 | 48.64 | 148.36 | 3.05 | 81.07 |
T2 | 0.5 | 1.0 | 60 | 44.53 | 95.74 | 2.15 | 74.22 |
T3 | 0.5 | 1.5 | 60 | 46.31 | 135.24 | 2.92 | 77.19 |
T4 | 0.5 | 2.0 | 60 | 36.35 | 68.70 | 1.89 | 60.58 |
T5 | 0.5 | 2.5 | 60 | 34.88 | 60.00 | 1.72 | 58.14 |
T6 | 1.0 | 0.5 | 60 | 53.87 | 228.00 | 4.21 | 90.26 |
T7 | 1.0 | 1.0 | 60 | 51.52 | 170.53 | 3.31 | 85.86 |
T8 | 1.0 | 1.5 | 60 | 52.39 | 193.33 | 3.69 | 87.32 |
T9 | 1.0 | 2.0 | 60 | 50.55 | 92.00 | 1.82 | 84.25 |
T10 | 1.0 | 2.5 | 60 | 37.36 | 54.17 | 1.45 | 62.26 |
T11 | 1.5 | 0.5 | 60 | 51.50 | 183.33 | 3.56 | 85.83 |
T12 | 1.5 | 1.0 | 60 | 52.80 | 192.20 | 3.64 | 88.01 |
T13 | 1.5 | 1.5 | 60 | 36.28 | 46.07 | 1.27 | 60.46 |
T14 | 1.5 | 2.0 | 60 | 41.36 | 64.53 | 1.56 | 68.94 |
T15 | 1.5 | 2.5 | 60 | 26.59 | 32.17 | 1.21 | 44.32 |
T16 | 2.0 | 0.5 | 60 | 43.36 | 114.47 | 2.64 | 72.27 |
T17 | 2.0 | 1.0 | 60 | 40.25 | 80.91 | 2.01 | 67.09 |
T18 | 2.0 | 1.5 | 60 | 35.53 | 57.21 | 1.61 | 59.22 |
T19 | 2.0 | 2.0 | 60 | 31.78 | 39.73 | 1.25 | 52.97 |
T20 | 2.0 | 2.5 | 60 | 14.29 | 15.00 | 1.05 | 23.81 |
T21 | 2.5 | 0.5 | 60 | 35.98 | 63.33 | 1.76 | 59.97 |
T22 | 2.5 | 1.0 | 60 | 34.11 | 50.82 | 1.49 | 56.85 |
T23 | 2.5 | 1.5 | 60 | 29.30 | 35.45 | 1.21 | 48.84 |
T24 | 2.5 | 2.0 | 60 | 30.97 | 38.40 | 1.24 | 51.61 |
T25 | 2.5 | 2.5 | 60 | 28.36 | 33.75 | 1.19 | 47.27 |
表5 不同浓度6-BA与ABA配比对南瓜芽再生的影响
Table 5 Effects of different concentrations of 6-BA and ABA on the bud induction of pumpkin
Treat- ments | 6-BA (mg∙L-1) | ABA (mg∙L-1) | Number of explants | Number of explants with adventitious shoots | Number of shoots | Average number of induced shoots | Rate of plant regeneration (%) |
---|---|---|---|---|---|---|---|
T1 | 0.5 | 0.5 | 60 | 48.64 | 148.36 | 3.05 | 81.07 |
T2 | 0.5 | 1.0 | 60 | 44.53 | 95.74 | 2.15 | 74.22 |
T3 | 0.5 | 1.5 | 60 | 46.31 | 135.24 | 2.92 | 77.19 |
T4 | 0.5 | 2.0 | 60 | 36.35 | 68.70 | 1.89 | 60.58 |
T5 | 0.5 | 2.5 | 60 | 34.88 | 60.00 | 1.72 | 58.14 |
T6 | 1.0 | 0.5 | 60 | 53.87 | 228.00 | 4.21 | 90.26 |
T7 | 1.0 | 1.0 | 60 | 51.52 | 170.53 | 3.31 | 85.86 |
T8 | 1.0 | 1.5 | 60 | 52.39 | 193.33 | 3.69 | 87.32 |
T9 | 1.0 | 2.0 | 60 | 50.55 | 92.00 | 1.82 | 84.25 |
T10 | 1.0 | 2.5 | 60 | 37.36 | 54.17 | 1.45 | 62.26 |
T11 | 1.5 | 0.5 | 60 | 51.50 | 183.33 | 3.56 | 85.83 |
T12 | 1.5 | 1.0 | 60 | 52.80 | 192.20 | 3.64 | 88.01 |
T13 | 1.5 | 1.5 | 60 | 36.28 | 46.07 | 1.27 | 60.46 |
T14 | 1.5 | 2.0 | 60 | 41.36 | 64.53 | 1.56 | 68.94 |
T15 | 1.5 | 2.5 | 60 | 26.59 | 32.17 | 1.21 | 44.32 |
T16 | 2.0 | 0.5 | 60 | 43.36 | 114.47 | 2.64 | 72.27 |
T17 | 2.0 | 1.0 | 60 | 40.25 | 80.91 | 2.01 | 67.09 |
T18 | 2.0 | 1.5 | 60 | 35.53 | 57.21 | 1.61 | 59.22 |
T19 | 2.0 | 2.0 | 60 | 31.78 | 39.73 | 1.25 | 52.97 |
T20 | 2.0 | 2.5 | 60 | 14.29 | 15.00 | 1.05 | 23.81 |
T21 | 2.5 | 0.5 | 60 | 35.98 | 63.33 | 1.76 | 59.97 |
T22 | 2.5 | 1.0 | 60 | 34.11 | 50.82 | 1.49 | 56.85 |
T23 | 2.5 | 1.5 | 60 | 29.30 | 35.45 | 1.21 | 48.84 |
T24 | 2.5 | 2.0 | 60 | 30.97 | 38.40 | 1.24 | 51.61 |
T25 | 2.5 | 2.5 | 60 | 28.36 | 33.75 | 1.19 | 47.27 |
图1 南瓜子叶外植体的离体再生体系建立 (A) MS培养基中萌发3天的种子(Bar=1 cm); (B)-(E) 在不定芽诱导培养基(含激素的MS培养基)中分别培养3天、10天、20天和4周时的外植体, 其中, (D) 在切口处已见愈伤组织产生; (E) 已见芽原基((B)-(C) Bars=2 cm; (D) Bar=22.5 mm; (E) Bar=15 mm); (F) 外植体接种7周产生的不定丛生芽(Bar=15 mm); (G) 伸长芽(Bar=1 cm); (H) 再生植株(Bar=2 cm); (I), (J) 炼苗移栽((I) Bar=35 mm; (J) Bar=40 mm)。
Figure 1 Establishment of plant regeneration of cotyledons of pumpkin (A) Cultivation for 3 days: seed germination in MS (Bar=1 cm); (B)-(E) Cultivation of explants in shoot regeneration medium (SRM, MS with hormone) for 3 days, 10 days, 20 days and 4 weeks, respectively, (D) Callus induction from explants; (E) Differentiation of bud primordium from callus (Bars=2 cm in (B) and (C), Bar=22.5 mm in (D), 15 mm in (E)); (F) Cultivation for 7 weeks: shoots induction in SRM (Bar=15 mm); (G) Elongating of shoots (Bar=1 cm); (H) Regenerated plant (Bar=2 cm); (I), (J) Exercising and transplanting((I) Bar=35 mm; (J) Bar=40 mm).
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