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Lecture notes – The plant tissue culture
Biology (C100)
University of Salford
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Lecture notes – The plant tissue culture
The Plant Tissue culture is the in vitro aseptic culture of cells, tissues, organs, or whole plant under controlled nutritional and environmental conditions often to produce the clones of plants.
It refers to a collection of techniques used to maintain or grow plant cells, tissues, or organs under sterile conditions on a nutrient culture medium of the known composition.
The resultant clones are true-to-type of the selected genotype.
Plant Tissue Culture
The Principle of Plant Tissue Culture
The Plant tissue culture relies on the fact that many plant cells have the ability to regenerate a whole plant (totipotency).
The Single cells, plant cells without cell walls (protoplasts), pieces of leaves, stems or roots can often be used to generate a new plant on culture media given the required nutrients and plant hormones.
The controlled conditions provide the culture an environment conducive for growth and multiplication.
These conditions include the proper supply of nutrients, pH medium, adequate temperature, and proper gaseous and liquid environment.
The Plant Tissue Culture Technique
Preparation of plant tissue for tissue culture is performed under aseptic conditions under HEPA filtered air provided by a laminar flow cabinet.
The tissue is grown in sterile containers, such as Petri dishes or flasks in a growth room with controlled temperature and light intensity.
The Living plant materials from the environment are naturally contaminated on their surfaces (and sometimes interiors) with microorganisms, so their surfaces are sterilized in chemical solutions (usually alcohol and sodium or calcium hypochlorite) before suitable samples (known as explants) are taken.
The sterile explants are then usually placed on the surface of a sterile solid culture medium but are sometimes placed directly into a sterile liquid medium, particularly when cell suspension cultures are desired.
The Solid and liquid media are generally composed of inorganic salts plus a few organic nutrients, vitamins, and plant hormones. Solid media are prepared from liquid media with the addition of a gelling agent, usually purified agar.
There are composition of the medium, particularly the plant hormones and the nitrogen source (nitrate versus ammonium salts or amino acids) has profound effects on the morphology of the tissues that grow from the initial explant.
This For example, an excess of auxin will often result in a proliferation of roots, while an excess of cytokinin may yield shoots.
A balance of both auxin and cytokinin will often produce an unorganized growth of cells or callus, but the morphology of the outgrowth will depend on the plant species as well as the medium composition.
The cultures grow, pieces are typically sliced off and subcultured onto new media to allow for growth or to alter the morphology of the culture. The skill and experience of the tissue culturist are important in judging which pieces to culture and which to discard.
This shoots emerge from culture, they may be sliced off and rooted with auxin to produce plantlets which, when mature, can be transferred to potting soil for further growth in the greenhouse as normal plants.
Lecture notes – The plant tissue culture
Module: Biology (C100)
University: University of Salford
