当归愈伤组织诱导、继代培养条件及生长动力学的优化

Optimization of induction, subculture conditions, and growth kinetics of Angelica sinensis (Oliv.) Diels callus.

作者信息

Huang Bing, Han Lijuan, Li Shaomei, Yan Chunyan

机构信息

Department of Natural Medicinal Chemistry, College of Pharmacy, Guangdong Pharmaceutical University, Guangzhou, China.

出版信息

Pharmacogn Mag. 2015 Jul-Sep;11(43):574-8. doi: 10.4103/0973-1296.160443.

DOI:10.4103/0973-1296.160443

PMID:26246734

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4522845/

Abstract

BACKGROUND

Angelica sinensis (Oliv.) Diels is an important traditional Chinese medicine, and the medicinal position is its root. This perennial herb grows vigorously only in specific areas and the environment. Tissue culture induction of callus and plant regeneration is an important and effective way to obtain large scale cultures of A. sinensis.

OBJECTIVE

The objective was to optimize the inductive, subculture conditions, and growth kinetics of A. sinensis.

MATERIALS AND METHODS

Tissue culture conditions for A. sinensis were optimized using leaves and petioles (types I and II) as explants source. Murashige and Skoog (MS) and H media supplemented with 30 g/L sucrose, 7.5 g/L agar, and varying concentrations of plant growth regulators were used for callus induction. In addition, four different basal media supplemented with 1.0 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D), 0.2 mg/L 6-benzyladenine (BA) and 30 g/L sucrose were optimized for callus subculture. Finally, growth kinetics of A. sinensis cultured on different subculture media was investigated based on callus properties, including fresh weight, dry weight, medium pH, callus relative fresh weight growth, callus relative growth rate (CRGR), and sucrose content.

RESULTS

MS medium supplemented with 5 mg/L α-naphthaleneacetic acid, 0.5 mg/L BA, 0.7 mg/L 2,4-D, 30 g/L sucrose and 7.5 g/L agar resulted in optimal callus induction in A. sinensis while petiole I was found as the best plant organ for callus induction. The B5 medium supplemented with 1.0 mg/L 2,4-D, 0.2 mg/L BA and 30 g/L sucrose displayed the best results in A. sinensis callus subculture assays.

CONCLUSION

The optimized conditions could be one of the most potent methods for large-scale tissue culture of A. sinensis.

摘要

背景

当归是一种重要的传统中药，药用部位为其根部。这种多年生草本植物仅在特定区域和环境中生长旺盛。通过组织培养诱导愈伤组织和植株再生是获得大规模当归培养物的重要且有效途径。

目的

优化当归的诱导、继代培养条件及生长动力学。

材料与方法

以叶片和叶柄（I型和II型）作为外植体来源，优化当归的组织培养条件。使用添加30 g/L蔗糖、7.5 g/L琼脂及不同浓度植物生长调节剂的Murashige和Skoog（MS）培养基及H培养基进行愈伤组织诱导。此外，优化了添加1.0 mg/L 2,4-二氯苯氧乙酸（2,4-D）、0.2 mg/L 6-苄基腺嘌呤（BA）和30 g/L蔗糖的四种不同基本培养基用于愈伤组织继代培养。最后，基于愈伤组织特性，包括鲜重、干重、培养基pH、愈伤组织相对鲜重生长、愈伤组织相对生长速率（CRGR）和蔗糖含量，研究了在不同继代培养基上培养的当归的生长动力学。

结果

添加5 mg/Lα-萘乙酸、0.5 mg/L BA、0.7 mg/L 2,4-D、30 g/L蔗糖和7.5 g/L琼脂的MS培养基能实现当归愈伤组织的最佳诱导，而I型叶柄是愈伤组织诱导的最佳植物器官。添加1.0 mg/L 2,4-D、0.2 mg/L BA和30 g/L蔗糖的B5培养基在当归愈伤组织继代培养试验中效果最佳。