Introduction:
- A
Plant Tissue Culture Lab is a specialized facility for the in
vitro cultivation of plant cells, tissues, or organs.
- Establishing
a well-equipped lab ensures sterility, efficiency, and the
successful propagation of plant materials for research or commercial
purposes.
Key
Components of a Plant Tissue Culture Lab:
1. Laboratory Setup:
A typical plant tissue
culture lab includes the following functional areas:
·
Media
Preparation Room:
This is for preparing culture media and sterilization.
·
Inoculation/Transfer
Room: For
aseptic handling and inoculation.
·
Culture/Incubation
Room: For
maintaining cultures under controlled conditions.
·
Growth
Room: For
plantlet growth and hardening.
·
Washing/Sterilization
Area: This
area is for cleaning and sterilizing equipment.
2. Essential Equipment:
1.
Laminar
Air Flow (LAF) Cabinet:
o Function: Provides a sterile
environment for tissue transfer.
o Types: Vertical and Horizontal
flow.
2.
Autoclave:
o Function: Sterilizes media,
glassware, and instruments using high-pressure steam.
o Typical Conditions: 121°C at 15 psi for 20
minutes.
3.
pH
Meter:
o Function: Measures and adjusts the pH
of the culture media.
4.
Incubators/Growth
Chambers:
o Function: Provides controlled
temperature, humidity, and light conditions for cultures.
o Temperature Range: Typically, 25-28°C.
5.
Refrigerator/Cold
Storage:
o Function: Stores media, chemicals,
and plant materials.
6.
Hot
Air Oven:
o Function: Sterilizes glassware
through dry heat.
7.
Microscope:
o Function: Observes plant tissues and
detects contamination.
o Types: Simple light microscopes
or stereo microscopes.
8.
Analytical
Balance:
o Function: Accurately measures
chemicals for media preparation.
9.
Shaker/Orbital
Shaker:
o Function: For liquid cultures and
suspension cultures, providing continuous agitation.
10. Distillation Unit/RO
System:
·
Function: Provides pure,
deionized water for media preparation.
11. UV Sterilizer:
·
Function: Sterilizes the working
area in LAF cabinets.
3. Culture Vessels:
1.
Petri
Dishes:
o Material: Glass or disposable
plastic.
o Uses: For culturing small plant
segments or callus induction.
2.
Test
Tubes:
o Material: Glass or plastic with
cotton plugs or screw caps.
o Uses: For culturing small
explants or germinating seeds.
3.
Conical
Flasks/Erlenmeyer Flasks:
o Sizes: 50 ml to 1 L.
o Uses: For suspension cultures
and liquid media.
4.
Magenta
Boxes/Phyto culture Boxes:
o Material: Plastic.
o Uses: For rooting and shooting
stages of micropropagation.
5.
Culture
Bottles/Jars:
o Material: Glass or plastic with a vented
cap.
o Uses: For growing larger
explants or multiple shoots.
6.
Beakers:
o Uses: For media preparation and
transfer of solutions.
7.
Watch
Glasses:
o Uses: For weighing small
quantities of chemicals.
4. Basic Consumables:
·
Media
Components:
Agar, sucrose, Murashige and Skoog (MS) medium, plant growth regulators.
·
Sterile
Tools:
Forceps, scalpels, inoculation loops.
·
Glassware: Measuring cylinders,
pipettes, graduated flasks.
·
Filter
Papers:
For filtration and aseptic transfers.
5. Sterilization and
Aseptic Techniques:
·
Media
Sterilization:
Done using an autoclave.
·
Surface
Sterilization of Explants:
Uses disinfectants like sodium hypochlorite or ethanol.
·
Working
in LAF:
Ensures no microbial contamination during culture transfers.
Conclusion:
- Establishing
a plant tissue culture lab requires careful planning of equipment, functional
areas, and culture vessels to maintain sterility and
ensure successful plant growth.
- A
well-equipped lab facilitates research and large-scale production of
plantlets for agriculture, horticulture, and pharmaceutical
applications