Plant tissue culture detailed notes

1. CALLUS & SUSPENSION
CULTURES

2. • Types of Cultures:
CULTURES
SUSPENSION
CULTURE
BATCH
CONTINUOUS
CALLUS
CULTURE

3. • CALLUS CULTURE:
• Callus is an amorphous aggregate of loosely arranged
parenchyma cells, which proliferate from mother cells.
• Cultivation of callus usually on a solidified nutrient
medium under aseptic conditions is known as callus
culture.
• These culture need to be sub-cultured every 3-5
weeks.
• Risk free, easy to handle.

6. INITIATION OF CALLUS CULTURE:
1. Selection and preparation of explant
2. Selection of culture medium
3. Transfer of explant
4. Incubation of culture
MAINTENANCE OF CALLUS CULTURE:
1. Depletion of nutrients in medium
2. Gradual loss of water
3. Accumulation of toxic waste

7. • SUSPENSION CULTURES:
• Tissues and cells cultured in liquid medium is
known as Suspension culture.
• They often produce suspension of single cells and
clumps.
• Liquid cultures are constantly agitated, generally by a
gyratory shaker @ 100-250ppm.
• This enhances the aeration and dissociation of cell
clumps into smaller pieces.
• This suspension cultures are used for large scale
production.
• Need to be sub-cultured every week.

8. 1. INITIATION OF SUSPENSION CULTURE:
1. Isolation of cells:
a) From callus culture
b) From plant organ- by Mechanical and enzymatic
method
2. Growth Pattern of suspension culture
3.Maintenance of suspension culture:
1. Batch cultures
2. Continuous cultures

10. • Batch cultures:
• A technique used to grow microorganisms or cells.
• A limited supply of nutrients for growth is
provided; when these are used up, or some other
factor becomes limiting, the culture declines.
• Cells, or products that the organisms have made, can
then be harvested from the culture.
• The cell number or biomass of batch culture exhibits
a typical sigmoidal growth curve.

11. • LAG Phase: during this phase cell number or
biomass is unchanged.
• LOG Phase: during this phase the cell number
rapidly increases in geometrical progression.
• Stationary Phase: In this phase end phase where
cell number does not change.

12. • Batch cultures are maintained by sub-culturing.
• These cultures are mostly used in initiation of
cell suspension i.e. single cell culture.
• These are most convenient to use than
continuous cultures hence mostly used.

13. • Continuous culture:
• These cultures are maintained in a steady state for a
long period by draining out the used (or) spent
medium and adding the fresh medium.
• such subculture systems are either closed (or) open
type.
• Closed:
• The cells separated from used medium taken out for
replacement and added back to the suspension culture.
• So that the cell biomass keeps on increasing.

14. • Open:
• Both cells and the used medium are taken out from
open continuously cultures and replaced by equal
volume of fresh medium.
• The replacement volume is so adjusted that cultures
remain at sub-maximal growth indefinitely.