Introduction:

Definition

Plant tissue culture is the technique of growing plant cells, tissues, or organs on a nutrient medium under sterile laboratory conditions.

The principle of plant tissue culture is based on totipotency — the ability of a single plant cell to develop into a complete plant under suitable conditions.

The concept of plant tissue culture was discovered by Gottlieb Haberlandt in 1902. He is known as the Father of Plant Tissue Culture.

Aim

To learn and perform plant tissue culture techniques for growing plants under sterile laboratory conditions.

Objectives

  • To understand the basic concept of Totipotency
  • To learn sterilization and aseptic techniques
  • To prepare nutrient culture media
  • To inoculate and grow plant explants in culture bottles
  • To observe callus formation, shoot growth, and rooting
  • To gain hands-on practical skills in Plant Biotechnology and plant tissue culture methods

I started my one month internship at Vigyan Ashram on 8 May to gain hands-on training and practical knowledge in plant tissue culture.

The theory related to plant tissue culture was covered from 8th May to 10th May 2026.9

Plant Tissue Culture

Plant tissue culture is the technique of growing plant cells, tissues, or organs under sterile laboratory conditions. It is carried out on a nutrient medium. Aseptic conditions are necessary to avoid contamination.

Important Terms

  • Explant → A small part of a plant used to start the culture.
    Examples: shoot tip, leaf, node, root, meristem.
  • In vitro → “In glass,” meaning growth in a test tube, flask, or culture bottle.

Historical Background

Gottlieb Haberlandt introduced the concept of plant tissue culture in 1902. He is known as the Father of Plant Tissue Culture.

Principle – Totipotency

A single plant cell has the ability to develop into a complete plant. With suitable nutrients, hormones, temperature, and light, cell division occurs and forms:

  • Callus
  • Shoot
  • Root
  • Complete plantlet

Applications

  • Micropropagation – production of a large number of plants
  • Production of disease-free plants
  • Conservation of rare plants
  • Year-round production
  • Genetic improvement

Advantages

  • Rapid multiplication
  • Disease-free plants
  • Uniform plants
  • Space saving
  • Germplasm conservation

Limitations

  • Expensive setup
  • Skilled labour required
  • Risk of contamination

Laboratory Safety in Plant Tissue Culture

  • Laboratory safety in plant tissue culture is important to prevent contamination, protect workers, and ensure successful plant growth under sterile conditions.
  • Important Safety Rules
  • Wear gloves, face mask, lab coat, and clean footwear.
  • Tie long hair properly.
  • Clean the laminar airflow surface with 70% ethanol before work.
  • Sterilize instruments like forceps and scalpels before use.
  • Avoid talking, coughing, or sneezing near culture materials.
  • Handle culture bottles carefully near the flame.
  • Follow fire safety rules while using spirit lamps and ethanol.
  • Never leave flames unattended.
  • Dispose of contaminated materials properly.
  • Maintain a clean and aseptic working environment.
  • These safety practices help maintain contamination-free tissue culture conditions.

Aseptic Practices in Plant Tissue Culture

A) Laminar Air Flow

  • Clean the surface with 70% ethanol before and after work.
  • Switch on UV light for 15–20 minutes before use.
  • Work close to the flame or sterile area.
  • Avoid unnecessary movement and talking during work.

B) Tool Sterilization

  • Sterilize forceps and scalpels by flame sterilization.
  • Use clean and dry instruments.
  • Re-sterilize tools before handling another sample.
  • Avoid touching sterile tools with bare hands.

C) Chemical Safety

  • Handle ethanol and chemicals carefully.
  • Wear gloves and mask while preparing solutions.
  • Label all chemical bottles properly.
  • Keep flammable chemicals away from flame.

D) Autoclave Safety

  • Do not overfill the autoclave.
  • Check water level before operation.
  • Open the lid only after pressure becomes normal.
  • Wear heat-resistant gloves while removing materials.

E) Hot Air Oven Safety

  • Use only dry glassware and metal instruments.
  • Do not touch hot equipment directly.
  • Maintain proper temperature and timing.
  • Allow materials to cool before handling.

F) Glassware Safety

  • Use clean and crack-free glassware.
  • Handle culture bottles carefully.
  • Wash and sterilize glassware properly before use.
  • Store glassware in a clean place.

G) Waste Disposal Safety

  • Dispose contaminated cultures carefully.
  • Separate glass, chemical, and biological waste.
  • Sterilize biological waste before disposal.
  • Keep the laboratory clean and hygienic.

Cleaning and sterilization of glassware and instruments

Instruments

Autoclave:An autoclave is a sterilization machine used in plant tissue culture for sterilizing glassware, media, instruments, and other laboratory materials using steam under pressure.

Principle of Autoclave

The autoclave works on the principle of steam under pressure (moist heat sterilization).

When water boils inside the autoclave, steam is produced. The pressure increases, which raises the temperature above 100°C. The hot steam penetrates materials and kills microorganisms, bacteria, fungi, viruses, and spores by denaturing their proteins.

Standard Conditions

Time: 15–30 minutes

Temperature: 121°C

Pressure: 15 psi

Hot Air Oven:Hot Air Oven

A hot air oven is used for sterilizing dry glassware and metal instruments in plant tissue culture laboratories.

Principle of Hot Air Oven

The hot air oven works on the principle of dry heat sterilization.
Hot air circulates inside the oven at high temperature and kills microorganisms by oxidation and protein denaturation.

Standard Conditions

Time: 1–2 hours

Temperature: 160–180°C