The document discusses plant tissue culture media, detailing its essential components such as inorganic nutrients, carbon sources, organic supplements, and growth regulators. It elaborates on specific media types like Murashige and Skoog, Gamborg's B5, Nitsch and Nitsch, and White's medium, highlighting their unique compositions and purposes in tissue culture. Key nutrients are also listed, emphasizing their roles in plant growth and metabolic processes.

1. PLANT TISSUE CULTURE
MEDIA
Dr. Deepa M.A
Assistant Professor of Botany
Government Arts College (Autonomous)
Coimbatore – 641 018
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2. • Plant tissue culture media contains macronutrients,
micronutrients, vitamins, amino acids or nitrogen
supplements, source(s) of carbon, undefined organic
supplements, growth regulators and solidifying agents

3. Composition
1. Inorganic nutrients
2. Carbon and energy sources
3. Organic supplements
4. Growth regulators
5. Solidifying agents
6. pH of medium

4. Inorganic nutrients
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Biology/Book%3A_General_Biology_(Boundless)/31%3A_Soil_a
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5. Nitrogen: It is required for the division of cells. It is the major constituent of DNA,
RNA, proteins, vitamins, hormones etc.
Phosphorus: It is a constituent of cell membranes, proteins, and nucleic acids and ATP
Potassium: It is also required for the opening and closing of stomata. It helps in the
activation of enzymes and helps in maintaining the turgidity of the cell.
Calcium: It is required during cell division and in the normal functioning of the cell
membrane. Metabolic activities are also regulated by this mineral.
Magnesium: It is involved in the activation of enzymes in respiration and
photosynthesis – chlorophyll pigments and helps in the synthesis of DNA and RNA. It
is also an important constituent of ribosomes and helps in maintaining the structure of
the ribosome.

6. Sulfur: Formation of chlorophyll pigments. It is an important constituent of
amino acids, coenzymes, and vitamins. It is crucial in nodule development and
efficient nitrogen fixation in legumes. Sulphur is also important in
photosynthesis and contributes to crop winter hardiness
Iron: it plays critical role in metabolic processes such as DNA synthesis,
respiration, and photosynthesis. It is an important constituent of protein and
helps with the activation of enzymes.
Manganese: It is an essential cofactor for the oxygen-evolving complex (OEC)
of the photosynthetic machinery, catalyzing the water-splitting reaction in
photosystem II (PSII).

7. Boron: Along with Calcium boron is important for cell wall synthesis; cell
division. Requires for reproduction growth, helps in pollination and fruit
development
Chlorine: It plays some important roles in plants, including in
photosynthesis, osmotic adjustment and suppression of plant disease
Copper (cu): It acts as a catalyst in photosynthesis and respiration and plays
an important role in the formation of lignin in the cell wall
Cobalt: It is a transition element, is an essential component of several
enzymes and co-enzymes

8. Molybdenum: Molybdenum is an essential component in two enzymes
that convert nitrate into nitrite (a toxic form of nitrogen) and then into
ammonia before it is used to synthesize amino acids within the plant.
Plants also use molybdenum to convert inorganic phosphorus into organic
forms in the plant
Zinc: It is used in the formation of chlorophyll and some carbohydrates,
conversion of starches to sugars and its presence in plant tissue helps
the plant to withstand cold temperatures. Zinc is essential in the formation
of auxins, which help with growth regulation and stem elongation.

9. Carbon and energy sources
• Sucrose – Source of Energy
• Why Sucrose?
• Why not Glucose or Fructose?

10. Organic supplements
• The organic supplements include vitamins (thiamine,
riboflavin, niacin, pyridoxine, folic acid, pantothenic acid,
biotin, ascorbic acid, myoinositol, Para amino benzoic acid
and vitamin E.), amino acids
• Organic nitrogen (in the form of amino acids such as L-
glutamine, L-asparagine, L- arginine, L-cysteine) is more
readily taken up than inorganic nitrogen by the plant cells.).

11. • Organic acids - citrate, malate, succinate or fumarate. –
Biostimulants and importnat intermediates in krebs cycle –
leads to active metabolism
• Organic extracts- yeast, casein hydrolysate, coconut milk,
orange juice, tomato juice and potato extract.
• Activated charcoal and antibiotics.

12. Murashige and Skoog (MS) medium
• MS medium was invented by two scientists named Toshio
Murashige and Folke K. Skoog in 1962 while the two
scientists were working on the discovery of plant growth
regulators. It is the most commonly used medium in the tissue
culture lab.
•

14. Gamborg’s (B5) Medium
• This medium was developed by O. L. Gamborg in 1968. He
used the media for the callus and cell suspension culture of
Glycine max belonging to the family of Fabaceae. This
medium is a blend of nutrients like inorganic salts, vitamins,
and carbohydrates.
• The medium has a higher concentration of nitrate and
potassium and a lower concentration of ammonia. Potassium
nitrate is useful in inducing the soybean root callus formation
and ammonium sulfate plays an essential role in cell growth.

16. Nitsch and Nitsch (NN) medium
• The medium was developed by J. P. Nitsch in 1969 for the
establishment of the in vitro anther culture of Nicotiana, from
family Solanaceae.
• It contains a high concentration of thiamine, biotin, and folic
acid that supports anther callus.
• Designed to establish the in vitro anther culture.

18. White’s Medium
• The medium was developed by P. R. White in 1963 for the establishment
of the root culture of tomato. This was the earliest plant tissue culture
media developed for root culture. It has a lower salt concentration and a
higher concentration of MgSO4. The concentration of nitrate is 19% lesser
than the MS media.
• Purpose: White’s medium can be used for the purpose of shoot culture and
callus culture. It is suitable for culture Musa and Daucus species.

19. It is a very crucial step in tissue culture
It cannot be choose in one step
Require complete understanding of the plants requirement
Or use two or three different media that contains low, medium and high
salt concentrations and based on the response, it can be finalised