Growth Regulators – Plant Tissue Culture Protocol

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
  
• Cytokinins
  
• Miscellaneous Plant Growth Regulators

Auxins

*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

*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

*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.