Plant tissue culture is a collection of techniques used to maintain or grow plant cells, tissues, or organs under sterile conditions. The document discusses the basic requirements for a plant tissue culture media including a complex mixture of salts (macroelements and microelements), organic supplements like vitamins and amino acids, a carbon source like sucrose, gelling agents, plant growth regulators, and antibiotics. It then provides details on the composition and use of Gamborg's media and White's media, which are commonly used plant tissue culture media developed in the 1960s.

1. v.Kanthasamy
M.Sc.,Life Sciences
Bharahidasan university
Tiruchyrappalli-24
PLANT TISSUE CULTURE PRESENTATION

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

4. 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

5. 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

6. 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

7. 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

8. 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.

9. 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.

10. 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

11. WHITES AND GAMBORG’S
MEDIA
Use
Medium is used for callus culture and cell
suspension culture
Summary
• 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.

12. • 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

13. 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.

14. • 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

15. 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.

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

17. 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.

18. 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.

19. 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

20. 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 8000.00
TOTAL: 23.23 gm/litre

21. 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

22. • 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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