This document discusses plant tissue culture media. It begins by defining synthetic and natural media and noting that synthetic media have largely replaced natural media for tissue culture. It then discusses important media such as White's, MS, B5, N6 and Nitsch's media. The document outlines the macro and micronutrients used in media, including their roles and common salts. It also discusses organic compounds, growth regulators like auxins and cytokinins, and natural complexes sometimes used in media. The overall purpose is to provide an overview of the composition and components of plant tissue culture media.

1. Dr. B. KARUNAI SELVI
ASSISTANT PROFESSOR OF BOTANY
V.V.VANNIAPERUMAL COLLEGE FOR WOMEN
VIRUDHUNAGAR- 626 001,
TAMIL NADU, INDIA
V.V.VANNIAPERUMAL COLLEGE FOR WOMEN
(Belonging to Virudhunagar Hindu Nadars)
An Autonomous Institution Affiliated to Madurai Kamaraj University
Re–accredited with ‘A’ Grade (3rd cycle) by NAAC
VIRUDHUNAGAR
PLANT TISSUE CULTURE MEDIA

2. Plant Tissue Culture Media
SYNTHETIC AND NATURAL MEDIA:
• Medium is composed of chemically defined components, it is
referred to as a synthetic medium.
• Medium contains chemically undefined compounds (e.g.,
vegetable extract, fruit juice, plant extract), it is regarded as a
natural medium.
• Synthetic media have almost replaced the natural media for tissue
culture.
• Expression of concentrations in media:
– The concentrations of inorganic and organic constituents in culture media
are usually expressed as mass values (mg/l or ppm or mg I-1). However, as
per the recommendations of the International Association of Plant
Physiology, the concentrations of macronutrients should be expressed as
mmol/l– and micronutrients as µmol/l–.

3. Plant Tissue Culture Media
• White’s medium:
– root culture.
• MS medium:
– organogenesis,
– regeneration of plants in cultured tissues.
• B5 medium:
– cell suspension and callus cultures
– protoplast culture.
• N6 medium: anther cultures
• Nitsch’s medium: anther cultures

4. Plant Tissue Culture Media
MACRO-NUTRIENT SALTS
• NH4NO3 Ammonium nitrate
• KNO3 Potassium nitrate
• CaCl2 -2 H2O Calcium chloride (Anhydrous)
• MgSO4 -7 H2O Magnesium sulfide (Epsom Salts)
• KH2PO4 Potassium hypophosphate
• FeNaEDTA Fe/Na ethylene-diamine-tetra acetate
• H3BO3 Boric Acid
• MnSO4 - 4 H2O Manganese sulfate
• ZnSO4 - 7 H2O Zinc sulfate
• KI Potassium iodide
• Na2MoO4 - 2 H2O Sodium molybdate
• CuSO4 - 5 H2O Cupric sulfate

5. Plant Tissue Culture Media
MACRONUTRIENT SALTS
NITROGEN
• Influences plant growth rate
• Essential in plant nucleic acids (DNA), proteins, chlorophyll, amino
acids and hormones.
PHOSPHORUS
• Abundant in meristematic and fast growing tissue
• Essential for photosynthesis and respiration.
POTASSIUM
• Necessary for cell division, meristematic tissue
• Helps in the pathways for carbohydrate, protein and chlorophyll
synthesis.

6. Plant Tissue Culture Media
MACRONUTRIENT SALTS
CALCIUM
• Involved in formation of cell walls and root and leaf development.
• Participates in translocation of sugars, amino acids, and ties up
oxalic acid (toxin).
IRON
• Involved in respiration , chlorophyll synthesis and photosynthesis.
FeNaEDTA =
• Sodium salt of EDTA sequesters iron, making it available to plants.

7. Plant Tissue Culture Media
MICRONUTRIENT SALTS
MAGNESIUM
• Involved in photosynthetic and respiration systems.
• Active in uptake of phosphate and translocation of phosphate and
starches.
SULFUR
• Involved in formation of nodules and chlorophyll synthesis, structural
component of amino acids and enzymes.
Manganese
• Involved in regulation of enzymes and growth hormones.
Molybdenum
• Involved in enzymatic reduction of nitrates to ammonia.
• Assists in conversion of inorganic phosphate to organic form.

8. Plant Tissue Culture Media
MICRONUTRIENT SALTS
ZINC
• Involved in production of growth hormones and chlorophyll.
• Active in respiration and carbohydrate synthesis.
BORON
• Involved in production of growth hormones and chlorophyll.
• Active in respiration and carbohydrate synthesis.
COPPER
• Involved in photosynthetic and respiration systems.
• Assists chlorophyll synthesis

9. Plant Tissue Culture Media
ORGANIC COMPOUNDS ,ORGANIC ACIDS AND PHENOLIC
COMPOUNDS
CARBON SOURCES
• Sucrose, sometimes Glucose or Fructose
VITAMINS
• Adenine – part of RNA and DNA
• Inositol – part of the B complex, in phosphate form is part of cell membranes
THIAMINE
• essential as a coenzyme in the citric acid cycle.
ORGANIC ACIDS
• Citric acid used with ascorbic acid as an antioxidant.
PHENOLIC COMPOUNDS
• Phloroglucinol - Stimulates rooting of shoot sections.

10. Plant Tissue Culture Media
NATURAL COMPLEXES
• Coconut endosperm
• Protein hydrolysates
• Tomato juice
• Yeast extracts
• Malt extract
• Potato agar

11. Plant Tissue Culture Media
GROWTH REGULATORS: AUXIN - ROOTS & CYTOKININ - SHOOTS
• Plant Cell Growth regulators (e.g. Auxins, Cytokinins and
Gibberellins) - Plant hormones play an important role in growth
and differentiation of cultured cells and tissues.
• There are many classes of plant growth regulators used in culture
media
• Auxin – Roots
• Cytokinin – Shoots

12. Plant Tissue Culture Media
Auxins
• The Auxins facilitate cell division and root differentiation.
• Auxins induce cell division, cell elongation, and formation of callus
in cultures.
• Indole-3-acetic acid =IAA
• Naphthalene acetic acid NAA
• Indole-3-butyric acid
• 2,4-D
• 2,4,5-T
• 2,4-dichlorophenoxy acetic acid is one of the most commonly
added auxins in plant cell cultures.

13. Plant Tissue Culture Media
CYTOKININS
• The Cytokinins induce cell division and differentiation.
• Cytokinins promote RNA synthesis and stimulate protein and
enzyme activities in tissues.
• Enhances adventitious shoot formation
• Benzyl adenine
• 2iP
• Kinetin
• Kinetin and benzyl-aminopurine are the most frequently used
cytokinins in plant cell cultures

14. Dr.B. Karunai Selvi, Assistant Professor of Botany, VVV College for Women, VNR
Plant Tissue Culture Media