Introduction:
Cell suspension culture is a type
of plant tissue culture where single cells or small groups of cells are grown
in a liquid medium under controlled conditions. It is widely used for producing
secondary metabolites, studying cell behaviour, and large-scale propagation of
plants.
Principle:
Cells derived from a callus, or
an explant are suspended in a liquid nutrient medium and kept under continuous
agitation. This process ensures that the cells receive adequate nutrients and
oxygen, promoting their growth and division.
Procedure:
- Selection of Explant:
- A healthy leaf or other plant tissue is chosen.
- The tissue is sterilized to prevent contamination.
- Callus Formation:
- The explant is placed on a solid agar medium
containing cytokinin and other nutrients.
- This promotes callus (undifferentiated cell mass)
formation.
- Preparation of Cell Suspension:
- The callus is transferred to a liquid medium or
agar plate.
- The medium contains essential nutrients, growth
regulators, and vitamins.
- Continuous shaking ensures the callus breaks down
into individual cells.
- Growth in Liquid Medium:
- Cells are grown in a flask under controlled
conditions (temperature, pH, light).
- Agitation provides proper aeration and prevents
clumping.
- Subculturing:
- After a certain period, cells are transferred to a
fresh medium to maintain their growth and viability.
Support
Systems for Plant Tissue Culture
1. Liquid
vs Solid Medium:
·
Liquid Medium: Used for the production of
secondary metabolites.
·
Solid Medium: Not used in bioreactors.
2. Nutrient
Requirements:
·
Macronutrients: N, P, K, S.
·
Micronutrients: Fe, Zn, B, Cu, Mn.
Plant
Tissue Culture Process
Steps:
1.
Callus Formation:
o Shaking
the medium and callus vigorously.
o Result:
Mass of individual cells.
2.
Isolation of Individual Cells:
o Centrifugation
to settle cells at the bottom.
o Transfer
individual cells to a test tube.
3.
Agar Plate Cultivation:
o Individual
cells are placed on an agar plate.
o Incubation
for growth.
4.
Cell Colonies:
o Selection
of more productive clones.
o Choose
highly reactive, functional clones with more pigmentation.
5.
Metabolite Production:
o Primary
Metabolite: The selection process ensures optimal clones.
o Secondary
Metabolite: Transfer to a bioreactor for secondary metabolite production.
§ Examples:
Plant compounds like phenols, alkaloids, etc., used for medicine production.
Functions:
- Production of Secondary Metabolites: Useful
for producing plant-derived compounds like alkaloids, phenols, and other
pharmaceuticals.
- Plant Propagation: Helps in large-scale
production of plants.
- Genetic Studies: Facilitates research on
plant genetics and metabolic pathways.
Uses:
- Pharmaceutical Industry: Production of
valuable compounds like medicines and vaccines.
- Agriculture: Propagation of
disease-resistant plant varieties.
- Research: Studying cellular processes and
responses to environmental changes
Conclusion:
Cell suspension culture is a
powerful tool in plant biotechnology, offering a scalable method for producing
important plant compounds and facilitating advanced research. It plays a
significant role in industries like pharmaceuticals, agriculture, and research
laboratories.