What are the sterilization techniques used in plant tissue culture?
Autoclaving, dry heat sterilization and filtration are adequate for most cell and organ culture materials. The laboratory apparatus designed to use steam under regulated pressure to achieve sterilization is called autoclave. Principle: Water boils at 100°C depending on the vapor pressure of the atmosphere.
Which is best method used for sterilization of culture media?
Sterilization of the media is most commonly achieved by applying heat and to a lesser extent by other means (physical methods, chemical treatment, and radiation). Heat sterilization: Heat is the most widely used sterilization technique.
What are the different ways in sterilizing media?
Sterilization can be achieved by a combination of heat, chemicals, irradiation, high pressure and filtration like steam under pressure, dry heat, ultraviolet radiation, gas vapor sterilants, chlorine dioxide gas etc.
Why sterilization is important in plant tissue culture?
Sterilization of explants is a key step in any plant tissue culture work, as the removal of all microorganisms, including bacteria and fungi, is essential to attain successful initiation, growth and development of the cultured tissues in vitro, which otherwise would be overwhelmed by the contaminants [2].
What is filter sterilization in tissue culture?
Filtration is used to sterilize heat-sensitive materials, such as culture media, enzymes, vaccines, and antibiotic solutions. Filtration through a membrane with 0.2 micron or smaller pore size removes biological contaminants, including bacteria, mold and yeast.
How is sterilization done in plant pathology?
Surface sterilization methods It is usually accomplished by treatment of plant tissues with oxidant or general sterilizing agent for a period, followed by a sterile rinse [28,12]. The plant samples were thoroughly washed under tap water to remove the soil debris from the leaves, stem, and roots.
Which equipment is used for sterilization of MS media?
Wet heat sterilization is done in an autoclave at 120°C at 15 lb pressure for 15 minutes. Liquid media, which are unstable at high temperature are sterilized by ultrafiltration. Chemicals, such as, alcohol is used to sterilize working area and instruments.
How do you sterilize cell culture media?
Medium sterilization we understand by fermentation media are sterilized by the use of: filtration, radiation, ultrasonic treatment, chemical treatment or heat (boiling or passing live steam through the medium, or by subjecting the medium to steam under pressure – autoclaving).
What kind of sterilization is used in tissue culture?
Different apparatus and essentials of tissue culture require different procedures for their sterilization. A list of sterilization techniques and their uses is explained below. Sterilization by heat: dry heat and moist heat are used to sterilize the equipment.
What kind of heat is used to sterilize media?
Sterilization by heat: dry heat and moist heat are used to sterilize the equipment. Autoclave works on the principle of moist heat and they can sterilize glass apparatus, solid or liquid media, distilled water, normal saline, discarded cultures, and contaminated media. Whereas, hot air ovens work on the principle of dry heat.
How is sterilization used to kill microbes?
Several sterilization techniques are designed to kill microorganisms invading your cultures. These microbes not only harm your cultures, but your hard work, time, and money you put into culturing the plants. Once the microbes enter your lab, they spread rapidly to other cultures and kill them.
What’s the best way to surface sterilize a plant?
In some cases, bromine, silver nitrate, and a mix of antibiotics are also used for surface sterilization of plant materials. Moreover for effective results, add Tween 20 or any other surfactant in the solution of sodium hypochlorite. Also, keep steering or shaken the mixture while surface sterilizing your explants.