Establishment of an Efficient Leaf Regeneration System for Pinguicula

doi:10.11983/CBB23118

Abstract

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

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

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB23118

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

Figures/Tables 7

References 26

[1]	陈英转, 孟新亚, 宋希强, 田代科, 周韬 (2023). 盾叶秋海棠叶片高频再生体系的建立. 植物生理学报 59, 165-171.
[2]	李春华, 李柯澄 (2020). 捕虫堇温室生产与繁殖. 中国花卉园艺 (14), 24-28.
[3]	梁展钊, 梁达新, 赖一航, 叶燕 (2020). 一种捕虫堇组培快繁的方法. 中国专利, CN109220802B. 2020-02-14.
[4]	廖敏凌, 蒲娅, 武晓云, 马朝峰, 王文奎, 戴思兰 (2023). 平潭野菊混合瓣型株系再生体系的建立. 植物学报 58, 449- 460. DOI
[5]	逯锦春, 曹丽娜, 佟冠杰, 王鑫颖, 张利英, 喻锌, 李荟芳, 李彦慧 (2022). 大花银莲花愈伤组织诱导及再生体系的建立. 植物学报 57, 217-226. DOI
[6]	彭爱红, 何永睿, 邹修平, 许兰珍, 邹哨兵 (2002). 观赏植物组织培养与基因工程研究进展(综述). 亚热带植物科学 31(2), 58-63.
[7]	曲晓慧 (2020). 几种菊属野生植物再生体系的建立. 硕士论文. 南京: 南京农业大学. pp. 1-43.
[8]	任毅, 刘明时, 田联会, 田先华, 李智军 (2006). 太白山自然保护区生物多样性研究与管理. 北京: 中国林业出版社. pp. 256.
[9]	宋倩, 卢家仕, 周锦业, 周主贵, 李春牛, 卜朝阳 (2015). 食虫植物保护现状及其研究进展. 北方园艺 (3), 171-175.
[10]	王春夏, 张梦迪, 王锦霞, 王志平, 孙红梅 (2020). 朱顶红无菌苗叶片高效再生体系. 园艺学报 47, 301-309. DOI
[11]	王江英, 汤雪燕, 葛金涛, 邵小斌, 孙明伟 (2023). 四季茶花夏咏国色叶片愈伤诱导与植株再生研究. 浙江农业科学 64, 390-395. DOI
[12]	王亚琴, 韦陆丹, 王文静, 刘宝骏, 张春玲, 张俊卫, 何燕红 (2020). 万寿菊再生体系的建立及优化. 植物学报 55, 749- 759.
[13]	吴之坤, 张长芹, 程治英 (2005). 不同培养基质及条件对三种报春花种子萌发及幼苗生长的影响. 种子 24(4), 1-5.
[14]	咸宏康, 支秋娟, 李卉, 王长泉 (2019). 金樱子(Rosa laevigata Michx.)叶片直接再生不定芽体系的建立. 北方园艺 (13), 93-100.
[15]	张清凤, 赵洁, 普春, 李啟菊, 马梅见 (2022). 蝴蝶兰组织培养再生体系的建立. 现代农业科技 (22), 116-118.
[16]	张彦文, 王海洋 (2000). 食虫植物研究的现状和趋势. 广西植物 20, 88-93.
[17]	Corredoira E, Costa RL (2021). Application of tissue culture in plant reproduction. Forests 12, 342.
[18]	Grogg D, Rohner M, Yates S, Manzanares C, Bull SE, Dalton S, Bosch M, Studer B, Broggini GAL (2022). Callus induction from diverse explants and genotypes enables robust transformation of perennial ryegrass (Lolium perenne L.). Plants 11, 2054.
[19]	Ikeuchi M, Sugimoto K, Iwase A (2013). Plant callus: mechanisms of induction and repression. Plant Cell 25, 3159-3173.
[20]	Karlsson PS, Nordell KO, Eirefelt S, Svensson A (1987). Trapping efficiency of three carnivorous Pinguicula species. Oecologia 73, 518-521. DOI PMID
[21]	Krowiak A, Herren CM, Webert KC, Einarsson Á, Hoekman D, Jackson RD, Ives AR (2017). Resource gradients and the distribution and flowering of butterwort, a carnivorous plant. Ann Zool Fenn 54, 163-173.
[22]	Mehbub H, Akter A, Akter MA, Mandal MSH, Hoque MA, Tuleja M, Mehraj H (2022). Tissue culture in ornamentals: cultivation factors, propagation techniques, and its application. Plants 11, 3208.
[23]	Phillips GC, Garda M (2019). Plant tissue culture media and practices: an overview. In Vitro Cell Dev Biol-Plant 55, 242-257.
[24]	Villegas SG, Alcalá RE (2018). Reproductive ecology of the carnivorous plant Pinguicula moranensis (Lentibulariaceae). Plant Biol 20, 205-212.
[25]	Wójciak M, Feldo M, Stolarczyk P, Płachno BJ (2023). Carnivorous plants from Nepenthaceae and Droseraceae as a source of secondary metabolites. Molecules 28, 2155.
[26]	Zhai N, Xu L (2021). Pluripotency acquisition in the middle cell layer of callus is required for organ regeneration. Nat Plants 7, 1453-1460. DOI PMID

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

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

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

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

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	++	++