4-Callus Culture.pptx

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1. Callus Culture M.PhooL Badshah

2. Callus culture Formed by the proliferation of parent tissue. Cells are parenchymatous, amorphous and unorganized. Callus is formed as a result of injury at the cut ends of stem or a root. When tissue on culture produce unorganized mass of callus with no regular form called callus culture. Rechinger , 1893 was firstly observed. Production and maintenance of callus on a solidified nutrient medium under aseptic conditions.

3. Steps of Callus Culture: Selection and Collection of explant material. Selection and preparation of culture media. Surface sterilization of explant material. Preparation of explant. Inoculation (transfer) of explant in culture medium. Incubation of culture. Sub-culturing of culture. Regeneration of plant from callus (Organogenesis and embryogenesis method).

4. Explant Material: Callus may initiate from explant s from any multicellular plants. Explant from stem, root, leaf, flower, fruit or seed etc. may be taken for culture. Examples of callus formation are: Storage parenchyma, Pericyclic cells of root, Cambial cells of vascular bundles, Perivascular cells, Secondary phloem, Pith cells, mesophyll cells and Cotyledons.

5. Reason of sub-culturing cells: Callus culture Nutrient may be exhausted Callus culture Agar may be desiccated. Callus culture Cell metabolites may accumulate and cause toxicity.

6. Callus Culture medium: Murashige and Skoog medium (MS). Kinetin used for initiation and maintaining callus. Usually required an exogenous supply of hormone for Initiation of callus. According to hormone requirement callus culture are: Auxin Cytokinins. Culture required both Auxin and Cytokinins Gibberellins.

7. Stages of Callus Culture development: Callus formation from an explants occurs in three stages: Induction stage: Metabolism is stimulated and the cells prepare to divide. Cell size remains unchanged. Cell division stage: Cells divide actively and the cell size decreases. Cell division is mainly periclinal and occurs towards the periphery giving rise to wound cambial cells.

8. adsdcf Differentiation stage: Cells differentiate by expansion and maturation. Rapidly growing calluses are more or less alike but as the growth rate decrease the calluses show their characteristic structures and forms. But all calluses have some similarities. Contain nodular or sheet meristems in group or scattered through the tissue.

9. Significance of Callus Culture: Determine the total amount of nucleic acid (DNA or RNA) rapidly from callus culture. By using tracer elements in callus culture the pith of various metabolic processes has been determined. Friable callus is suitable for cell suspension culture.