WHITES and

GAMBORG’S MEDIA

composition …v.KanthasamyM.Sc.,Life SciencesBharahidasan universityTiruchyrappalli-24

PLANT TISSUE CULTURE PRESENTATION

Reference:Gamborg O.L., Miler R.A. And Ojima K., 1968. page no:151-158.

Plant tissue culture

Plant tissue culture

• Plant tissue culture is a collection of techniques

used to maintain or grow plant cells, tissues or

organs under sterile conditions on a

nutrient culture medium of known composition.

• Plant tissue culture is widely used to produce

clones of a plant in a method known as

micropropagation

BASIC REQUIREMENT OF A PLANT TISSUE CULTURE MEDIA

1. Complex mixture of salts: Essential elements, or

mineral ions.

2. Organic supplement: vitamins and/or amino acids.

3. Carbon source: usually sugar sucrose.

4. Gelling agents

5. Plant Growth Regulators

6. Antibiotics

1. Complex mixture of salts: Essential elements, or mineral ions.

The essential elements can further be divided

into the following categories:

a. Macroelements (or macronutrients)

b. Microelements (or micronutrients)

c. Iron Source

Macroelements:• These elements are required in large amounts for

plant growth and development.Ex : Nitrogen, phosphorus, potassium, magnesium, calcium etc..Microelements:These elements are required in trace amounts for plant growth and development. Ex : Manganese, iodine, copper, cobalt, boron, molybdenum, iron

Organic supplements:

• These include vitamins and amino acids. Two

vitamins, i.e., thiamine (vitamin B1) and

myoinositol (a vitamin B) are essential for the

culture of plant cells in vitro.

• The most commonly used amino acid is glycine.Carbon source:• The most commonly used carbon source is

sucrose.

Gelling agents:

• Plant tissue culture media can be used in either liquid or

'solid' forms, depending on the type of culture being

grown.

• Agar, produced from seaweed, is the most common type of

gelling agent .

Plant growth regulators:

• Specific media manipulations can be used to direct the

development of plant cells in culture due to plasticity and

totipotency.

Plant growth regulators:

• There are five main classes of plant growth regulator used

in plant cell culture, namely:

a. Auxins

b. Cytokinins

c. Gibberellins

d. Abscisic acid

e. Ethylene

WHITES AND GAMBORG’S MEDIA

Use Medium is used for callus culture and cell suspension cultureSummary• established by Gamborg O.L. (1968) for callus and

cellsuspension culture of Glycine max (Family- Fabaceae).

• This medium is widely used for in vitro plant cell, tissue and organ culture.

• Principle

• Gamborg Medium provides all essential

Macroelements, Microelements, &Vitamins for the

growth of plant cell, tissue and organ culture

invitro.

• The medium generally contains greater proportion

of nitrate and potassium but low concentration of

ammonia

Media composition (gamborgs)

Macroelements mg/L

• Potassium nitrate 2500.00• Ammonium sulphate 134.00• Calcium chloride anhydrous 113.24• Magnesium sulphate 122..09• Sodium phosphate monobasic 130.42 These elements are required in large amounts for plant growth and development.

• Need of macro nutrients is higher.

• It is present in milli molar (mM) quantities (more than 30 ppm/1 or mg/1)

• Macro nutrients provide both anions and cations for the plant cells

Phosphorus• Very important for energy metabolism.

• Essential element for DNA & RNA.

• Deficiency may cause delayed growth and dark green

coloure of leaves.

• Supplied instead of sodium hydrogen phosphate or

potassium hydrogen phosphate.

Magnesium• Essential for enzymatic reactions, energy metabolism(ATP synthesis).

• Supplied as magnesium sulphate.

sulphur

• Important substance.• It increases the cholophyll in leaves.• Supplied as magnesium Sulphate and

Potassium Sulphate.

Microelements mg/L• Manganese sulphate. 10.00• Boric Acid 3.00• Potassium iodide 0.75• Molybdic acid (sodium salt). 0.21• Zinc sulphate. 2.00• Copper sulphate. 0.025• Cobalt chloride. 0.025• Ferrous sulphate. 27.80• Na .EDTA 37.26 These elements are required in trace amounts for plant growth and development.

Zink• Zn plays an active role in protein synthesis .• Supplied as Zinc Sulphate

Manganese• Plays an important role in the Hill reaction of

photasynthesis.• Required in many enzymatic activities.• Supplied as Manganese Sulphate.

Copper• Copper plays important role in photosynthesis.• Supplied as Copper Sulphate.

Hexitols

• Most tissue culture media have this compound.

• Essential for seed germination, sugar transport,

carbohydrate metabolism, membrane structure

and cell wall formation

Vitamins mg/L• Myo-Inositol 100.00• Thiamine HCL 10.00• Pyridoxine HCL 1.00• Nicotinic acid (Free acid) 1.00 Thiamine content had been increased in the medium which supported the growth of cell suspension culture. The medium lacks glycine.Gelling Agent• Agar 8000.00TOTAL: 23.23 gm/litre

Whites media

• White medium was established by white (1963) for

root culture of tomato

• This is one of the earliest media with low salt

formulation

• Nitrate concentration is 19 % less than MS media

• Prominence was given to MgSO4

• White Medium Composition :• Ingredients milligrams/litre• Potassium nitrate 80.00• Calcium nitrate 221.96• Magnesium sulphate 360.00• Sodium phosphate monobasic 18.98• Potassium chloride 65.00• Sodium sulphate 200.00• Manganese sulphate.H2O 5.04• Boric acid 1.50• Potassium iodide 0.75• Molybdenum trioxide 0.001• Zinc sulphate.7H2O 2.67• Copper sulphate.5H2O 0.01• Ferrous sulphate.7H2O 2.50• myo - Inositol 100.00• Thiamine hydrochloride 0.10• Pyridoxine hydrochloride 0.10• Nicotinic acid (Free acid) 0.50• Glycine (Free base) 3.00• Sucrose 20000.00• TOTAL gm/litre 21.06

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