Shoot Regenerative Capacity Assays in Arabidopsis and Tobacco
拟南芥和烟草中嫩芽再生能力的测试
发布: 2016年03月05日第6卷第5期 DOI: 10.21769/BioProtoc.1753 浏览次数: 10474
评审: Marisa RosaYuan ChenAnonymous reviewer(s)
下载 PDF 
Q&A
参见作者原研究论文
The authors used this protocol in:
Feb 2015
Abstract
Plant regeneration refers to a process through which an explant is differentiated to a whole plant under certain conditions. It has been shown that two plant hormones, auxin and cytokinin, play critical roles within this process (Duclercq et al., 2011). Cytokinin induces shoot regeneration, whereas auxin promotes root production. In addition to hormones, recent study has revealed that age cue serves as a common element behind plant cell totipotency (Toledano et al., 2012). Here we present an easy protocol to assess the shoot regenerative capacity of Arabidopsis and tobacco leaves of different ages.
Keywords: Shoot regeneration (再生)
Cytokinin (细胞分裂素)
Auxin (生长素)
Plant age (植物的年龄)
Materials and Reagents
- Surgical tape (3M, catalog number: 15300 )
- Sterile petri dishes
- Seeds: Arabidopsis thaliana (Col-0 ecotype) and tobacco (Nicotiana tabacum cv SR1)
- Bacterial strain: Agrobacterium tumefaciens EHA105
- Sodium hypochlorite (Sigma-Aldrich, catalog number: L099100 )
Note: The Pricing and availability are not currently available. - Kanamycin sulfate Streptomyces kanamyceticus (Sigma-Aldrich, catalog number: K0879 )
- Timentin (Yeasen, catalog number: 60230ES07 )
- Agar (Sangon Biotech, catalog number: A100637 )
- Methylester sulfonate (Sangon Biotech, catalog number: MB0341 )
- Sucrose (Sigma-Aldrich, catalog number: E001888 )
- Glucose (Sigma-Aldrich, catalog number: G8270 )
- Ethyl alcohol, Pure (Sigma-Aldrich, catalog number: 459844 )
- Deionized water
- Soil
- Murashige and Skoog (MS) basal medium with vitamin powder (PhytoTechnology, Laboratories®, catalog number: 15B0519117B ) (1 L) (see Recipes)
- 1/2 MS medium (1 L) (see Recipes)
- MS1 medium (1 L) (see Recipes)
- MS2 medium (1 L) (see Recipes)
- MS3 medium (1 L) (see Recipes)
- Callus-inducing medium (1 L) (see Recipes)
- Shoot-inducing medium (1 L) (see Recipes)
- 22 mM 2, 4-dichlorophenoxy (2, 4-D) stock solution (Sigma-Aldrich, catalog number: 31518 ) (see Recipes)
- 9 mM 3-indoleacetic acid (IAA) (Sigma-Aldrich, catalog number: I2886 ) stock solution (see Recipes)
- 20 mM Kinetin (Sigma-Aldrich, catalog number: K0753 ) stock solution (see Recipes)
- 50 mM 6-(γ, γ-Dimethylallylamino)purine (2-IP) (Sigma-Aldrich, catalog number: D5912 ) stock solution (see Recipes)
- 2 mg/ml 6-benzylaminopurine (6-BA) (Sigma-Aldrich, catalog number: B3408 ) stock solution (see Recipes)
- 1 M Potassium hydroxide (KOH) (Sigma-Aldrich, catalog number: P5958 ) (see Recipes)
- 15% bleach (see Recipes)
- 10% bleach (see Recipes)
- Infection buffer (see Recipes)
Equipment
- Clean bench (horizontal laminar flow)
- Autoclave (ZEALWAY)
- Incubator (25 °C/80 UML) (Percival-scientific, model: CU-41L4 )
- Growth chamber
- Glass culture bottles (130 mm x 100 mm x 62 mm)
- Sterile 5 mm-hole punch
- Scalpel (10 cm in length)
- Forceps (20 cm in length)
- Scissor (15 cm in length)
Procedure
文章信息
版权信息
© 2016 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Zhang, T. and Wang, J. (2016). Shoot Regenerative Capacity Assays in Arabidopsis and Tobacco. Bio-protocol 6(5): e1753. DOI: 10.21769/BioProtoc.1753.
Download Citation in RIS Format
分类
植物科学 > 植物发育生物学 > 综合
植物科学 > 植物生理学 > 植物生长
您对这篇实验方法有问题吗？
在此处发布您的问题，我们将邀请本文作者来回答。同时，我们会将您的问题发布到Bio-protocol Exchange，以便寻求社区成员的帮助。
提问指南
+ 问题描述
写下详细的问题描述，包括所有有助于他人回答您问题的信息（例如实验过程、条件和相关图像等）。
发布问题
