银扇草再生体系的建立

doi:10.11983/CBB23094

植物学报 ›› 2024, Vol. 59 ›› Issue (3): 433-440.DOI: 10.11983/CBB23094

银扇草再生体系的建立

曾浩¹^,², 李佩芳¹^,², 郭至辉¹^,², 刘春林¹^,³, 阮颖¹^,²^,^*()

¹湖南农业大学, 作物表观遗传调控与发育重点实验室, 长沙 410128
²湖南农业大学生物科学技术学院,长沙 410128
³湖南农业大学农学院, 长沙 410128

收稿日期:2023-07-18 接受日期:2023-12-19 出版日期:2024-05-01 发布日期:2023-12-29
通讯作者: label>*阮颖, 湖南农业大学生物科学技术学院二级教授, 博士生导师, 作物表观遗传调控与发育湖南省重点实验室主任, 植物遗传与分子生物学湖南省高等学校重点实验室主任; 兼任国家自然科学基金外围评审专家, 国家科技奖励专家库专家, 教育部博士后基金评审专家, 湖南省植物学会常务理事, 担任Frontier in Plant Science、Current Genomics等杂志的审稿专家。近5年, 共发表研究论文35篇, 其中SCI收录16篇, 申请授权发明专利4项, 编写教材3部。目前研究团队主要以油料作物油菜、蓖麻、银扇草等为材料, 重点研究植物开花及品质性状的表观遗传调控机制, 为提高作物产量和改良作物品质提供理论依据。E-mail: yingruan@hotmail.com
基金资助:
国家自然科学基金委联合重点项目(U20A2028)

Establishment of a Regeneration System for Lunaria annua

Hao Zeng¹^,², Peifang Li¹^,², Zhihui Guo¹^,², Chunlin Liu¹^,³, Ying Ruan¹^,²^,^*()

¹Key Laboratory of Crop Epigenetic Regulation and Development, Hunan Agricultural University, Changsha 410128, China
²College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, China
³College of Agronomy, Hunan Agricultural University, Changsha 410128, China

Received:2023-07-18 Accepted:2023-12-19 Online:2024-05-01 Published:2023-12-29
Contact: *E-mail: yingruan@hotmail.com

摘要/Abstract

摘要： 为建立银扇草(Lunaria annua)体外再生体系, 以其真叶为外植体, 探讨消毒条件、植物生长调节剂组合及浓度对愈伤组织诱导、不定芽和不定根分化的影响; 进一步分析了生根方式对成苗和幼苗生长的影响。结果表明, 用75%乙醇消毒45秒配合0.1%升汞溶液消毒6分钟为银扇草叶片外植体最佳消毒处理; 真叶愈伤组织诱导及不定芽分化的最适培养基为MS+0.5 mg∙L^-16-BA+2.0 mg∙L^-1 2,4-D, 愈伤组织诱导率达93.37%, 不定芽分化率达84.08%; 最佳生根培养基为MS+ 0.1 mg∙L^-1NAA, 从接种叶片外植体到获得再生植株约90天。该研究建立了银扇草稳定的再生体系, 为开发利用银扇草资源及挖掘功能基因奠定了基础。

关键词: 银扇草, 愈伤组织, 组织培养, 再生体系

Abstract: In order to establish an in vitro regeneration system for Lunaria annua, its true leaves were used as explants to study the effects of sterilization conditions, combinations and concentrations of plant growth regulator on the induction of callus and on the differentiation of adventitious buds and roots; The effect of rooting methods on the growth of seedlings and young plants was further explored. The results showed that the best disinfection treatment for leaf explants was a combination of 75% alcohol for 45 seconds and 0.1% HgCl₂ solution for 6 minutes. The most suitable medium for induction of callus and differentiation of adventitious buds was MS+0.5 mg∙L^-16-BA+2.0 mg∙L^-12,4-D. The induction rate of callus reached 93.37% and the differentiation rate of adventitious buds reached 84.08%. The best rooting medium was MS+0.1 mg∙L^-1NAA, and the time from inoculating leaf explants to obtaining regenerated plants was about 90 days. In this study, a stable regeneration system was established, which laid a foundation for the development and utilization of L. annua resources and digging for functional genes.

Key words: Lunaria annua, callus, tissue culture, regeneration system

曾浩, 李佩芳, 郭至辉, 刘春林, 阮颖. 银扇草再生体系的建立. 植物学报, 2024, 59(3): 433-440.

Hao Zeng, Peifang Li, Zhihui Guo, Chunlin Liu, Ying Ruan. Establishment of a Regeneration System for Lunaria annua. Chinese Bulletin of Botany, 2024, 59(3): 433-440.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB23094

https://www.chinbullbotany.com/CN/Y2024/V59/I3/433

图/表 8

表1 氯化汞消毒时间对外植体消毒效果的影响

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

表2 6-BA和NAA组合对银扇草叶片外植体愈伤组织诱导的影响

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

表3 6-BA和2,4-D组合对银扇草叶片外植体愈伤组织诱导的影响

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

图1 银扇草再生体系的建立 (A) 刚接种的外植体; (B) 愈伤组织诱导; (C) 不定芽分化; (D) 不定芽; (E) 不定芽增殖; (F) 组培苗; (G) 移栽。Bars=1 cm

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

表4 不同植物生长调节剂组合对银扇草愈伤组织不定芽诱导的影响

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

表5 不同培养基配方对银扇草不定根诱导的影响

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^-1NAA	15	91.63±1.67 a	8.14±0.26 a	8.67±1.54 a
2	MS+0.5 mg∙L^-1NAA+0.5 mg∙L^-1IBA	15	81.78±1.56 b	5.73±0.71 b	6.33±0.88 b

表6 预培养时间对IAA处理后银扇草不定芽生根的影响

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

表7 银扇草再生植株炼苗与移栽

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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银扇草再生体系的建立

Establishment of a Regeneration System for Lunaria annua

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