Plant tissue culture techniques involve two main methods: static culture using solid agar medium, which is easier for initial cell line maintenance and investigation of organ formation; and suspension cultures using liquid medium, which allows for higher growth rates and superior biomass control as cells are fully surrounded by nutrients. Key tools and techniques for plant tissue culture include sterilization using autoclaves, pH meters to monitor medium pH, laminar airflow benches for aseptic work, and various analytical tools like colorimeters and chromatography for biochemical analysis. Plant growth chambers are also used to control environmental factors like light, temperature, and humidity.

1. Tools and Techniques Used for Plant
Tissue Culture

2. Plant Tissue Culture Techniques
There are mainly two major techniques in plant
tissue culture.
1-Static culture (Solid-agar Medium):
• It can also be called as callus plant tissue
culture. In this procedure, the plant-tissue is
grown on solid agar medium and always gives
rise to tissue mass called a callus.

3. • This callus culture technique is easier as it is
easier and even convenient for initial
maintenance of cell-lines, and also for carrying
out the investigation studies related to
organogenesis i.e organ formation.

4. 2- Suspension cultures (Liquid media):
• In this cell aggregates, or single cells are grown in
liquid culture. The cells are kept suspended by
using shakers/ impellers. The actual growth rate
is higher in comparison to grown solid-agar
medium.
• Besides, this technique provide much superior
control over the growth of biomass as the cells
are always surrounded by the nutrient medium
completely

5. Tools and Steps
1-Methods of Sterilization:
Asepsis:
• Plant tissue culture requires contamination
free environment, tools and cultures or strict
maintenance of germ free system in all the
operations, known as asepsis.

7. 2-pH and pH Meter:
• Applications:
• 1. To adjust pH of different solutions, preparation of
buffers and culture media.
• 2. Determination of pH of cells (cell sap) and in
analytical techniques.
• ADVERTISEMENTS:
To monitor pH of the medium in a
bioreactor.

9. 3- Autoclave:
• Autoclave is used to sterilize medium,
glassware and tools for the purpose of plant
tissue culture. The same equipment is used in
hospitals to sterilize gauge, cotton, tools and
linen, etc.

10. • Sterilization of material is carried out by
increasing moist heat (121 °C) due to
increased pressure inside the vessel (15-22
psi, pounds per square inch or 1.02 to 1.5
kg/cm2) for 15 minutes for routine
sterilization. Moist heat kills the
microorganism and makes the material free
from microbes

12. 4- Plant Growth Chamber:
• Plant growth chambers can be constructed in
a suitable sized room or can be purchased as
commercially available equipment. Thermal
insulation of walls increases the efficiency of
the cooling system.

13. • Essentially plant growth chamber has three
environmental control systems:
• 1. Light-intensity and duration cycle control.
• 2. Temperature control and regulation.
• 3. Humidity control and regulation.

14. 5- Laminar Air Flow Bench:
• Laminar air flow (LAF) bench is the main working
table for aseptic manipulations related to plant
tissue culture.
• This is equipment fitted with High Efficiency
Particulate Air (HEPA) Filters, which allow air to
pass but retain all the particles and micro-
organisms. These HEPA Filters have a very small
pore size (0.3 µm) with 99.97-99.99% efficiency.

16. 6- Colorimeter:
• The most commonly used method for
determining the concentration of biochemical
compounds is colorimetry.
• It uses the property of light such that when
white light passes through a coloured
solution, some wavelengths are absorbed
more than others.

17. • Hyaline solution can be made coloured by
specific reactions with suitable reagents.
These reactions are generally very sensitive to
determine quantities of material in the region
of millimole per litre concentration.

18. 7- Chromatography:
• Chromatography (meaning ‘colored writing’) is
a technique to separate molecules on the
basis of differences in size, shape, mass,
charge and adsorption properties.
• The term chromatography was used by the
Russian botanist Tswett to describe the
separation of plant pigments on a column of
alumina.

19. • There are different types of chromatography
but they all involve interactions between
these components: the mixture to be
separated, a solid phase, and a solvent.