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  Section: Biotechnology Methods » Tissue Culture Techniques
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Preparation of Plant Tissue Culture Media

Tissue Culture Techniques
  Tissue Culture Methods
  Plant Tissue Culture
  Plant Tissue Culture (Cont.)
  Many Dimensions of Plant Tissue Culture Research
  What is Plant Tissue Culture?
  Uses of Plant Tissue Culture
  Plant Tissue Culture demonstration by Using Somaclonal Variation to Select for Disease Resistance
  Demonstration of Tissue Culture for Teaching
  Preparation of Plant Tissue Culture Media
  Plant Tissue Culture Media
  Preparation of Protoplasts
  Protoplast Isolation, Culture, and Fusion
  Agrobacterium Culture and Agrobacterium — Mediated transformation
  Isolation of Chloroplasts from Spinach Leaves
  Preparation of Plant DNA using
  Suspension Culture and Production of Secondary Metabolites
  Protocols for Plant Tissue Culture
  Sterile Methods in Plant Tissue Culture
  Media for Plant Tissue Culture
  Safety in Plant Tissue Culture
  Preparation of Media for Animal Cell Culture
  Aseptic Technique
  Culture and Maintenance of Cell Lines
  Trypsinizing and Subculturing Cells from a Monolayer
  Cellular Biology Techniques
  In Vitro Methods
  Human Cell Culture Methods

Since the pioneer plant tissue culture studies, one of the main objectives was to design a proper medium that supports sufficient growth of the explants in a totally artificial environment. The first medium formulations used for plant culture work were based on experience of microorganisms. These media contained several distinct classes of compounds:
  • Carbon source
  • Organic supplements
  • Inorganic salts
  • Trace elements.
Composition of commonly used tissue culture media
Table 1 Composition of commonly used tissue culture media
in molar concentration
As works on plant cell cultures progressed, attention was paid in order to specifically define media for an optimal growth of the variable plant cells, tissue, and organs. Today, the most commonly used culture media are based on following components:
  • Macroelements—N, P, K, Ca, Mg, and S
  • Microelements—Fe, Cu, Mn, Co, Mo, B, I, Zn, Cl and sometimes Al, Ni, and Si
  • Carbon source—Sucrose, glucose, fructose, or sorbitol
  • Organic compounds (Vitamins)—thiamine, niacin, pyridoxine, biotin, folic acid, ascorbic acid, tocopherol
  • Myo-inositol or casein hydrolysate
  • Complex organics—Coconut milk and water, yeast extract, fruit juices, or pulps
  • Plant growth regulators—Auxins, cytokinins, Gibberellins, abscisic acid
  • Gelling agents—Agar, agarose, gellan gum
  • Other components—MES, activated charcoal, antibiotics, fungicides.
The number of plant species that have been cultured and the different media used are extensive. Once the plant species to be cultured is decided upon, a suitable medium formulation should be selected. One of the most successful media, is the MS medium, was devised by Murashige and Skoog (1962), based on the constitution of tobacco plants. The choice or formulation of the media basically depends on the requirements of the cultivated explant and species.

Media Formulation
Table 2 Composition of commonly used tissue culture media in
As mentioned above, MS medium was developed for culture of tobacco explants and the formulation was based on the analysis of the mineral compounds present in the tobacco tissue itself (Table 1). It is noted for its high salt levels, particularly K and N salts. LM medium (Linsmaier and Skoog, 1965) is a revision of the MS medium, in which the organic constituents have been modified and only thiamine and myo-inositol retained. White’s medium (White, 1963) was devised for the culture of tomato roots and has lower concentrations of salts than MS medium. Gamborg’s B5 medium (Gamborg et al., 1968) was devised for soybean callus culture and contains a much greater proportion of nitrate compared to the ammonium. SH medium (Schenk and Hildebrandt, 1972) was developed for callus culture of both monocotyledons and dicotyledons. Nitsch’s medium (Nitsch and Nitsch, 1969) was developed for another culture. It contains lower salt concentrations than that of MS, but not as low as that of White’s medium. KM medium (Kao and Michayluck, 1975) was designed to grow cells and protoplasts of Vicia at a very low cell density in liquid media. The medium of Chu (N6) is defined to improve the formation, growth, and differentiation of pollen in rice. The concentration of ammonium proved to be crucial for the development of callus. Recently other media have been developed for specific explants and species.

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