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Questions and Answers
MS medium is widely used for many types of ______ systems
culture
B5 medium was originally designed for cell suspension and ______ cultures
callus
N6 medium is used for cereal anther ______
culture
MS medium is most frequently used in plant tissue ______ work
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Synthetic media have almost replaced the ______ media for tissue culture
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White's medium is one of the earliest plant tissue culture media developed for root ______
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When a medium contains chemically undefined compounds, it is regarded as a ______ medium
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The concentrations of inorganic and organic constituents in culture media are usually expressed as mass values (mg/l or ppm or mg ______)
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Nitsch's medium is frequently used for ______ cultures
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Synthetic medium is composed of chemically defined components, hence referred to as a ______ medium
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Study Notes
Plant Growth Regulators (PGRs)
- Gibbrellins (e.g. GA3) are used for internode elongation and meristem growth.
- Abscissic acid (ABA) is used for somatic embryogenesis and culturing woody species.
Solidifying Agents
- Agar is a high molecular weight polysaccharide obtained from seaweeds and is used to prepare semisolid tissue culture media.
- Agar is added to the medium in a concentration ranging from 0.5% to 1% (w/v) and is inert, non-reactive with media constituents, and not digested by plant enzymes.
- Alternative gelling agents like Gelrite form clear gels, making it easier to detect contamination.
pH
- pH affects the absorption of ions and solidification of gelling agents.
- The optimum pH for culture media is 5.8 before sterilization, and it can drop by 0.3 to 0.5 units after autoclaving.
- pH values lower than 4.5 or higher than 7.0 can greatly inhibit growth and development in vitro.
Preparation of Media
- Media preparation is a crucial step in the experiment.
- Stock solutions are prepared separately for major salts, minor salts, iron, and organic nutrients.
- The required quantities of agar, sucrose, and organic supplements are added separately.
Selection of Suitable Media
- A suitable medium can be devised by trying different combinations of PGRs, salts, sucrose, and other components.
- The exact conditions required for plant cells in culture or to regenerate intact plants from cultured cells differ for each plant species.
Micronutrients
- Manganese is essential for photosynthesis and enzymatic activities, and is supplied as Manganese Sulphate.
- Copper plays an important role in photosynthesis and is supplied as Copper Sulphate.
- Molybdenum is essential for converting nitrate to ammonium and is supplied as Sodium molybdate.
- Boron is involved in different enzymatic activities and is supplied as Boric acid.
- Iron is an important enzyme co-factor, and its deficiency can have severe effects on plant growth and development.
Setting Up a Tissue Culture Lab
- A tissue culture lab requires a general washing area, media preparation area, aseptic transfer area, environmentally controlled incubators, and an observation/data collection area.
Nutrient Media
- The type and composition of culture media strongly govern the growth and morphogenesis of plant tissues.
- The choice of tissue culture medium depends on the species to be cultured.
Components of Tissue Culture Medium
- Inorganic nutrients include macro and micronutrients like nitrogen, potassium, phosphorus, calcium, magnesium, and sulphur.
- Nitrogen is a major component of all plant tissue culture media and is supplied in the form of ammonium and nitrate ions.
Examples of Tissue Culture Media
- White's medium was developed for root culture.
- MS medium is widely used for many types of culture systems.
- B5 medium was developed for cell suspension and callus cultures.
- Nitsch's medium is frequently used for anther cultures.
- MS medium is the most frequently used in plant tissue culture work due to its success with several plant species and culture systems.
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Description
Test your knowledge on plant growth regulators like gibberellins and abscisic acid, and the use of solidifying agents in tissue culture media preparation. Learn about the specific functions of different PGRs and the importance of solidifying agents in tissue culture techniques.
