捕虫堇叶片高效再生体系建立

doi:10.11983/CBB23118

植物学报 ›› 2024, Vol. 59 ›› Issue (4): 626-634.DOI: 10.11983/CBB23118 cstr: 32102.14.CBB23118

捕虫堇叶片高效再生体系建立

杨佳丽¹^,², 饶羽菲¹^,², 张润花³, 周国林³, 林处发³, 何燕红¹^,²^,^*(), 宁国贵¹^,²^,^*()

¹华中农业大学园艺林学学院, 果蔬园艺作物种质创新与利用全国重点实验室, 武汉 430070
²华中农业大学花卉研究所, 武汉 430070
³武汉市农业科学院蔬菜研究所, 武汉 430035

收稿日期:2023-08-29 接受日期:2023-11-02 出版日期:2024-07-10 发布日期:2024-07-10
通讯作者: 何燕红,宁国贵
基金资助:
现代农业产业技术体系(CARS-23)

Establishment of an Efficient Leaf Regeneration System for Pinguicula

Jiali Yang¹^,², Yufei Rao¹^,², Runhua Zhang³, Guolin Zhou³, Chufa Lin³, Yanhong He¹^,²^,^*(), Guogui Ning¹^,²^,^*()

¹National Key Laboratory for Germplasm Innovation & Utilization of Horticultural Crops, College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan 430070, China
²The Institute of Flowers, Huazhong Agricultural University, Wuhan 430070, China
³Institute of Vegetable Research, Wuhan Academy of Agricultural Sciences, Wuhan 430035, China

Received:2023-08-29 Accepted:2023-11-02 Online:2024-07-10 Published:2024-07-10
Contact: Yanhong He, Guogui Ning

摘要/Abstract

摘要： 以2个不同基因型的捕虫堇圆切捕虫堇(Pinguicula cyclosecta)和塞提捕虫堇(P. ‘Sethos’)叶片为外植体, 通过探究不定芽再生的影响因素, 成功建立了捕虫堇高效再生体系。结果表明, 2种捕虫堇对消毒方式和基本培养基种类要求相同, 但再生能力有显著差异。圆切捕虫堇叶片在MS+1.0 mg∙L^-1 6-BA+0.2 mg∙L^-1 NAA培养基上再生能力最强, 再生率为92.22%, 再生系数达4.84, 在MS+0.6 mg∙L^-1 6-BA+0.1 mg∙L^-1NAA培养基上, 不定芽增殖系数达4.98。塞提捕虫堇叶片在MS+2.0 mg∙L^-16-BA+0.2 mg∙L^-1NAA培养基中再生率最高, 为77.78%, 再生系数为6.12, 在增殖培养基MS+0.3 mg∙L^-1 6-BA+0.1 mg∙L^-1NAA上, 增殖系数可达4.84。2种捕虫堇在1/2MS+0.1 mg∙L^-1 IBA培养基上进行生根培养, 根系生长状态最佳。该研究解决了捕虫堇繁殖系数低及工厂化育苗难等问题, 为其规模化生产和育种改良提供了技术支持。

关键词: 食虫植物, 捕虫堇属, 叶片再生, 不定芽

Abstract: Using two different genotypes of Pinguicula cyclosecta and P. ‘Sethos’ as explants, an efficient system for adventitious bud regeneration of Pinguicula was established by studying the factors affecting regeneration. The results showed that the two kinds of Pinguicula had the same requirements for the disinfection scheme and basic medium, but the regeneration ability differed. The best medium for P. cyclosecta leaf regeneration was MS+1.0 mg∙L^-1 6-BA+0.2 mg∙L^-1NAA, with a regeneration rate of 92.22% and a regeneration coefficient was 4.84. On MS+0.6 mg∙L^-16-BA+0.1 mg∙L^-1NAA media, the proliferation coefficient of adventitious buds reached 4.98. The best medium for the leaf regeneration of P. ‘Sethos’ was MS+2.0 mg∙L^-1 6-BA+0.2 mg∙L^-1 NAA, with a regeneration rate of 77.78% and a regeneration coefficient of 6.12. On the MS+0.3 mg∙L^-1 6-BA+0.1 mg∙L^-1NAA media, the proliferation coefficient reached 4.84. After root culture (1/2MS+0.1 mg∙L^-1IBA), the root state of the two kinds of Pinguicula was the best. This study solved the problems of a low reproduction coefficient and difficulty in factory breeding of Pinguicula and provided technical support for its large-scale production and breeding improvement.

Key words: carnivorous plant, Pinguicula, leaf regeneration, adventitious bud

杨佳丽, 饶羽菲, 张润花, 周国林, 林处发, 何燕红, 宁国贵. 捕虫堇叶片高效再生体系建立. 植物学报, 2024, 59(4): 626-634.

Jiali Yang, Yufei Rao, Runhua Zhang, Guolin Zhou, Chufa Lin, Yanhong He, Guogui Ning. Establishment of an Efficient Leaf Regeneration System for Pinguicula. Chinese Bulletin of Botany, 2024, 59(4): 626-634.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB23118

https://www.chinbullbotany.com/CN/Y2024/V59/I4/626

图/表 7

表1 不同消毒方式对2种捕虫堇污染率和存活率的影响

Table 1 Effect of disinfection scheme on pollution rate and survival rate of two Pinguicula species

Disinfection scheme	Pollution rate (%)		Survival rate (%)
Disinfection scheme	P. cyclosecta	P. ‘Sethos’	P. cyclosecta	P. ‘Sethos’
0.1% HgCl₂2 min	40.00±3.33 a	37.78±5.09 a	97.78±3.85 a	89.25±1.29 a
0.1% HgCl₂5 min	31.11±3.85 b	18.89±1.92 b	94.44±5.09 a	84.44±1.93 b
0.1% HgCl₂8 min	13.33±3.34 d	5.56±1.93 d	81.11±1.92 bc	74.44±1.93 c
75% alcohol 10 s+0.1% HgCl₂2 min	17.78±1.92 c	12.22±1.92 c	95.55±3.85 a	81.11±1.92 b
75% alcohol 10 s+0.1% HgCl₂5 min	7.78±1.92 e	4.44±1.93 de	86.67±3.34 b	71.11±1.92 c
75% alcohol 10 s+0.1% HgCl₂8 min	1.11±1.92 f	0.00±0.00 f	75.56±1.93 cd	64.45±3.85 d
75% alcohol 20 s+0.1% HgCl₂2 min	4.44±1.93 ef	1.11±1.92 ef	74.45±3.85 d	66.67±3.34 d
75% alcohol 20 s+0.1% HgCl₂5 min	0.00±0.00 f	0.00±0.00 f	64.78±2.51 e	57.78±1.92 e
75% alcohol 20 s+0.1% HgCl₂8 min	0.00±0.00 f	0.00±0.00 f	61.11±1.92 e	47.78±1.92 f

图1 不同基本培养基对圆切捕虫堇和塞提捕虫堇叶片再生的影响 (A) 圆切捕虫堇再生率; (B) 塞提捕虫堇再生率。不同小写字母表示各处理间差异显著(P<0.05)。

Figure 1 Effects of different basic media on the leaf regeneration of Pinguicula cyclosecta and P. ‘Sethos’ (A) Regeneration rate of P. cyclosecta; (B) Regeneration rate of P. ‘Sethos’. Different lowercase letters indicate significant differences among different treatments (P<0.05).

表2 植物生长调节剂对2种捕虫堇叶片再生的影响

Table 2 Effect of plant growth regulators on leaf regeneration of two Pinguicula species

6-BA (mg·L^-1)	NAA (mg·L^-1)		Regeneration rate (%)		Regeneration coefficient
6-BA (mg·L^-1)	NAA (mg·L^-1)		P. cyclosecta	P. ‘Sethos’	P. cyclosecta	P. ‘Sethos’
0.5		0	87.78±3.85 bc	72.22±5.09 c	3.60±0.98 cd	3.52±0.99 d
0.5		0.2	92.22±3.85 b	81.11±1.93 b	3.96±1.28 bc	4.14±1.03 bc
0.5		0.5	83.33±3.33 c	75.56±3.85 bc	3.61±0.82 cd	4.02±0.96 c
1.0		0	98.89±1.93 a	91.11±1.93 a	3.60±0.96 cd	3.94±0.87 c
1.0		0.2	92.22±1.93 b	91.11±3.85 a	4.84±0.95 a	4.04±0.96 c
1.0		0.5	82.22±1.93 c	77.78±1.93 bc	4.32±0.86 b	3.84±0.79 cd
2.0		0	85.56±1.93 c	76.67±3.33 ab	4.04±0.85 b	4.38±0.81 b
2.0		0.2	83.33±6.67 c	77.78±3.85 bc	3.36±0.79 d	6.12±0.97 a
2.0		0.5	84.44±5.09 c	72.22±3.85 c	2.54±0.88 ef	5.96±0.82 a
3.0		0	84.44±3.85 c	71.11±5.09 c	2.82±0.95 e	1.90±0.71 e
3.0		0.2	82.22±1.93 c	75.56±5.09 bc	2.24±0.70 fg	2.08±0.60 e
3.0		0.5	72.22±1.93 d	72.22±3.85 c	2.02±0.68 g	2.11±0.72 e

表3 植物生长调节剂对2种捕虫堇不定芽增殖的影响

Table 3 Effects of plant growth regulators on the adventitious bud proliferation of two Pinguicula species

6-BA (mg·L^-1)	NAA (mg·L^-1)	Proliferation coefficient		Growth situation
6-BA (mg·L^-1)	NAA (mg·L^-1)	P. cyclosecta	P. ‘Sethos’	P. cyclosecta	P. ‘Sethos’
0.3	0.05	3.24±0.87 d	2.66±0.87 c	+	++
0.3	0.1	3.50±0.95 d	4.84±0.93 a	++	++++
0.6	0.05	4.36±0.92 b	3.52±0.85 b	+++	+
0.6	0.1	4.98±0.94 a	3.96±0.84 b	++++	++
1.0	0.05	4.04±1.03 c	2.90±0.93 bc	++	++
1.0	0.1	4.18±0.77 bc	2.80±0.85 bc	++	++

表4 植物生长调节剂对2种捕虫堇不定芽生根的影响

Table 4 Effects of plant growth regulators on the adventitious bud rooting of two Pinguicula species

No.	NAA (mg·L^-1)	IBA (mg·L^-1)	Average rooting number		Root length (cm)
No.	NAA (mg·L^-1)	IBA (mg·L^-1)	P. cyclosecta	P. ‘Sethos’	P. cyclosecta	P. ‘Sethos’
1	0.1	0	7.13±1.33 d	9.95±1.08 d	0.89±0.96 d	3.97±0.80 c
2	0.2	0	6.97±1.11 d	12.03±1.70 b	0.76±1.40 e	4.86±0.84 b
3	0.3	0	5.30±0.74 e	9.90±0.90 d	0.63±0.74 f	3.02±0.86 d
4	0	0.1	11.00±1.15 a	13.25±1.28 a	1.69±0.75 a	5.58±0.68 a
5	0	0.2	9.70±0.78 b	11.73±0.93 b	1.55±0.84 b	4.23±1.03 c
6	0	0.3	8.07±0.89 c	10.88±1.26 c	1.31±0.81 c	4.08±0.89 c

图2 圆切捕虫堇叶片再生过程 (A) 圆切捕虫堇; (B) 外植体; (C), (D) 叶片在MS+1.0 mg∙L-1 6-BA+0.2 mg∙L-1 NAA培养基上培养30天; (E), (F) 不定芽在MS+ 0.6 mg∙L-1 6-BA+0.1 mg∙L-1 NAA培养基上增殖培养30天; (G), (H) 增殖芽在1/2MS+0.1 mg∙L-1 IBA培养基上生根培养30天; (I) 移栽30天后成活的再生植株。Bars=1 cm

Figure 2 Leaf regeneration process of Pinguicula cyclosecta (A) P. cyclosecta; (B) Explants; (C), (D) Leaves regenerated on MS+1.0 mg∙L-1 6-BA+0.2 mg∙L-1 NAA media for 30 days; (E), (F) Buds grew on MS+0.6 mg∙L-1 6-BA+0.1 mg∙L-1 NAA media for 30 days; (G), (H) The fascicular buds rooted on 1/2MS+ 0.1 mg∙L-1 IBA media for 30 days; (I) Regenerated plants that are viable after 30 days of transplanting. Bars=1 cm

图3 塞提捕虫堇叶片再生过程 (A) 塞提捕虫堇; (B) 外植体; (C), (D) 叶片在MS+2.0 mg∙L-1 6-BA+0.2 mg∙L-1 NAA培养基上培养30天; (E), (F) 不定芽在MS+ 0.3 mg∙L-1 6-BA+0.1 mg∙L-1 NAA培养基上增殖培养30天; (G), (H) 增殖芽在1/2MS+0.1 mg∙L-1 IBA培养基上生根培养30天; (I) 移栽30天后成活的再生植株。Bars=1 cm

Figure 3 Leaf regeneration process of Pinguicula ‘Sethos’ (A) P. ‘Sethos’; (B) Explants; (C), (D) Leaves regenerated on MS+2.0 mg∙L-1 6-BA+0.2 mg∙L-1 NAA media for 30 days; (E), (F) Buds grew on MS+0.3 mg∙L-1 6-BA+0.1 mg∙L-1 NAA media for 30 days; (G), (H) The fascicular buds rooted on 1/2MS+ 0.1 mg∙L-1 IBA media for 30 days; (I) Regenerated plants that are viable after 30 days of transplanting. Bars=1 cm

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捕虫堇叶片高效再生体系建立

Establishment of an Efficient Leaf Regeneration System for Pinguicula

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