Whites and gamborg’s media in plant tissue culture

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1. WHITES and GAMBORG’S MEDIA composition … 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

3. 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

4. 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

5. 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

6. 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

7. 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.

8. 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 .

9. 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

10. 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.

11. 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

12. 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.

13. 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

14. 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 .

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

16. 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.

17. 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 .

18. 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

19. 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

20. 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 i s 19 % less than MS media Prominence was given to MgSO 4

21. 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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