Skip to Content
Merck
HomePlant Tissue CultureGrowth Regulators – Plant Tissue Culture Protocol

Growth Regulators – Plant Tissue Culture Protocol

The importance of plant growth regulators in plant tissue culture is well documented. We offer a broad range of plant growth regulators specifically tested for plant cell culture. Each product is assayed for physical and chemical characteristics, then is biologically tested following the criteria established for powdered media. Each auxin is tested for enhancement of callus growth and/or root initiation in vitro. Each cytokinin is tested for stimulation of shoot production.

The plant growth regulator solutions are sterile filtered through a double 0.2 µm filter unit into sterile bottles. Each lot of plant growth regulator solution is tested for sterility according to specifications established by U.S. Pharmacopeia Vol. XXI, as well as biological testing in plant cell culture using criteria identical to those established for our plant tissue culture media.

FOR LABORATORY USE, PLANT TISSUE CULTURE MEDIA PREPARATION AND PLANT RESEARCH PURPOSES ONLY. NOT FOR USE AS A PLANT GROWTH REGULATOR ON DEVELOPED PLANTS. NOT FOR DRUG OR HOUSEHOLD USES.

Preparation and Use

To prepare a 1 mg/mL stock solution: Add 100 mg of the plant growth regulator to a 100 mL volumetric flask or other glass container. Add 2-5 mL of solvent to dissolve the powder. Once completely dissolved, bring to volume with double processed water (Product No. W3500). Stirring the solution while adding water may be required to keep the material in solution. Store the stock solution as recommended in the tables. Add 1.0 mL of the stock solution to 1 liter of medium to obtain a final concentration of 1.0 mg/L of the plant growth regulator in the culture medium (Tables 1-3).

Volume of Stock Solution = (Desired Hormone Conc. X Medium Volume) / Stock Solution Conc.

Plant Cell Culture Tested Auxins are generally used in plant cell culture at a concentration range of 0.01-10.0 mg/L. When added in appropriate concentrations, they may regulate cell elongation, tissue swelling, cell division, formation of adventitious roots, inhibition of adventitious and axillary shoot formation, callus initiation and growth, and induction of embryogenesis.

Plant Cell Culture Tested Cytokinins are generally used in plant cell culture at a concentration range of 0.1-10.0 mg/L. When added in appropriate concentrations, they may regulate cell division, stimulate axillary and adventitious shoot proliferation, regulate differentiation, inhibit root formation, activate RNA synthesis and stimulate protein and enzyme activity.

Table for Molar Equivalence and Solution Preparation

Auxins

Molar EquivalenceSolution Preparation
Product NameProduct No.Mol. Wt.µM for 1 mg/LSolventDiluentPowder StorageLiquid StorageSteriliz-
ation*
Working Conc. (mg/L)
p-Chlorophenoxyacetic acid (4-CPA)C0413186.65.36EtOHRT2-8 °CCA0.1-10.0
2,4-Dichlorophenoxyacetic acidD72992214.53RT2-8 °CCA0.01-6.0
2,4-Dichlorophenoxyacetic acid Sodium saltD66792434.12WaterRT2-8 °CCA0.01-6.0
Indole-3-acetic acid Free acid (IAA)I2886175.25.71EtOH/1N NaOHWater-0 °C-0 °CCA/F0.01-3.0
Indole-3-acetic acid Sodium saltI5148197.25.07WaterWater2-8 °C-0 °CCA/F0.01-3.0
Indole-3-acetic acid methyl esterI9770189.25.292-8 °C2-8 °C
Indole-3-acetyl-L-aspartic acidI9387290.33.450.5N NaOHWater-0 °C-0 °CF0.01-5.0
Indole-3-butyric acid (IBA)I5386203.24.90EtOH/1N NaOHWater2-8 °C-0 °CCA/F0.1-10.0
Indole-3-butyric acid Potassium salt (K-IBA)I7512241.34.14Water2-8 °C-0 °CCA/F0.1-10.0
alpha-Naphthaleneacetic acid Free acid (NAA)N0640186.25.371N NaOHWaterRT2-8 °CCA0.1-10.0
beta-Naphthoxyacetic acid Free acid (NOA)N3019202.24.951N NaOHWaterRT2-8 °CCA0.1-10.0
Phenylacetic acid (PAA)P6061136.27.34EtOHRT2-8 °CCA/F0.1-50.0
PicloramP5575241.54.14DMSORT2-8 °CCA0.01-10.0
2,4,5-Trichlorophenoxyacetic acid (2,4,5-T)T5785255.53.91EtOHRT2-8 °CCA0.01-5.0
2,3,5-Triiodobenzoic acid Free acid (TIBA)T5910499.82.001N NaOHWater-0 °C-0 °CF0.05-5.0

*CA = coautoclavable with other media components. F = filter sterlize. CA/F = coautoclavable with other media components, however, some loss of activity may occur. This can be compensated for by increasing component concentration. Component may be filter sterilized.

Cytokinins

Molar EquivalenceSolution Preparation
Product NameProduct No.Mol. Wt.µM for 1 mg/LSolventDiluentPowder StorageLiquid StorageSteriliz-
ation*
Working Conc. (mg/L)
Adenine Free baseA5665135.17.401.0 HClWaterRT2-8 °CCA50-250
Adenine hemisulfate Hemisulfate saltA2545184.25.43WaterRT2-8 °CCA50-250
6-Benzylaminopurine (BA)B3408225.34.441N NaOHWaterRT2-8 °CCA/F0.1-5.0
6-Benzylaminopurine HydrochlorideB5920261.73.82WaterRT2-8 °CCA/F0.1-5.0
6-Benzylaminopurine (BA)B3274225.34.441N NaOHWaterRT2-8 °CCA/F0.1-5.0
N-Benzyl-9-(2-tetrahydropyranyl)adenine (BPA)B2275309.43.23EtOH-0 °C-0 °CCA/F0.1-5.0
N-(2-Chloro-4-pyridyl)-N'-phenylurea (4-CPPU)C2791247.74.04DMSO2-8 °C2-8 °CF0.001-1.0
6-(gamma,gamma-Dimethylallylamino)purine (2iP)D7674203.24.921N NaOHWater-0 °C-0 °CCA/F1.0-30.0
6-(gamma,gamma-Dimethylallylamino)purine (2iP)D5912203.24.921N NaOHWater-0 °C-0 °CCA/F1.0-30.0
1,3-Diphenylurea (DPU)D7535212.34.71DMSORT2-8 °CF0.1-1.0
KinetinK0753215.24.651N NaOHWater-0 °C-0 °CCA/F0.1-5.0
KinetinK3378215.24.651N NaOHWater-0 °C-0 °CCA/F0.1-5.0
KinetinK3253215.24.651N NaOHWater-0 °C-0 °CCA/F0.1-5.0
Kinetin HydrochlorideK1885251.73.97Water-0 °C-0 °CCA/F0.1-5.0
1-Phenyl-3-(1,2,3-thiadiazol-5-yl)ureaP6186220.24.54DMSORT2-8 °CCA/F0.001-0.05
trans-Zeatin Free baseZ0876219.24.561N NaOHWater-0 °C-0 °CCA/F0.01-5.0
ZeatinZ0164219.24.561N NaOHWater-0 °C-0 °CCA/F0.01-5.0
trans-Zeatin HydrochlorideZ2753255.73.91Water-0 °C-0 °CCA/F0.01-5.0
trans-Zeatin ribosideZ3541351.42.851N NaOHWater-0 °C-0 °CF0.01-5.0

*CA = coautoclavable with other media components. F = filter sterlize. CA/F = coautoclavable with other media components, however, some loss of activity may occur. This can be compensated for by increasing component concentration. Component may be filter sterilized.

Miscellaneous Plant Growth Regulators

Molar EquivalenceSolution Preparation
Product NameProduct No.Mol. Wt.µM for 1 mg/LSolventDiluentPowder StorageLiquid StorageSteriliz-
ation*
Working Conc. (mg/L)
(±)-cis,trans-Abscisic acid (ABA)A1049264.33.781N NaOHWater-0 °C-0 °CCA/F0.1-10.0
AncymidolA9431256.33.90DMSO2-8 °C-0 °CCA/F1.0-10.0
Chlorocholine chloride (CCC)C4049158.16.33WaterRT2-8 °CFup to 500
3,6-Dichloro-o-anisic acid (Dicamba)D5417221.04.52EtOH/Water2-8 °C2-8 °CF0.01-10.0
Gibberellic acid (GA3)G7645346.42.89EtOHRT2-8 °CCA/F0.01-5.0
Gibberellic acid Potassium salt (K-GA3)G1025384.52.60Water2-8 °C-0 °CCA/F0.01-5.0
Gibberellin A4 Free acid (GA4)G7276332.43.01EtOH-0 °C-0 °CF0.01-5.0
(±)-Jasmonic acidJ2500210.34.76EtOH2-8 °C-0 °CF0.01-100.0
PhloroglucinolP1178126.17.93WaterRT2-8 °CCA/Fup to 162
N-(Phosphonomethyl)glycine (Glyphosate)P9556169.15.911N NaOHWaterRT2-8 °CF
Succinic acid 2,2-dimethylhydrazideS2022160.26.24Water2-8 °C2-8 °CCA/F0.1-10.0

*CA = coautoclavable with other media components. F = filter sterlize. CA/F = coautoclavable with other media components, however, some loss of activity may occur. This can be compensated for by increasing component concentration. Component may be filter sterilized.

Sign In To Continue

To continue reading please sign in or create an account.

Don't Have An Account?