17 Aug 2021

For Beginners: Requirements to Prepare Tissue Culture Media

Anjali Singh, MS

As a content and community manager, I leverage my expertise in plant biotechnology, passion for tissue culture, and writing skills to create compelling articles, simplifying intricate scientific concepts, and address your inquiries. As a dedicated science communicator, I strive to spark curiosity and foster a love for science in my audience

Table of Contents

What is Tissue Culture and its Advantages?

One of the most commonly asked questions by our beginner tissue culture enthusiasts is “What are the requirements to prepare the growth media and how to do it?”

But first, what is growth media, what does it do, and why is it required?

Tissue culture is a technique to grow plants under a laboratory environment, where we provide the cultured plants the same environment that they would have required in nature. The technique has been popular among scientists and growers alike because it allows them to address some major issues with the cultivation of plants.

Some advantages of tissue culture:

Tissue culture can rapidly produce plants in a large number irrespective of their growing season.
Only from a few tissues of plants, hundreds of plants can be obtained using the tissue culture technique.
It’s used to conserve and protect endangered plant species.
It’s effective to grow plants that are difficult to grow using conventional techniques.
It’s used to produce haploid or hybrid plants.
It’s able to produce disease-free plants.

To culture plants in a lab environment, some specific requirements need to be fulfilled, which include:

Aseptic environment (that is free from any microorganisms or a completely clean environment).
Sterilized equipment and chemicals.
Aseptic culture area
Equipment like burner, forceps, sharp blades, etc.
Materials like culture vessels/containers, flasks, beakers, Petri dishes, etc.
Machines like autoclave, hot air oven, sterilizer, and laminar flow hood.
The culture room is equipped with machines that help in regulating the room condition favorable to plants.
Chemicals like surfactants, plants growth regulators, agar, and MS media.

Each of these is essential to properly grow plants under in vitro conditions. Agar and MS media together constitute the growth media for plants.

What are tissue culture growth medias and its requirements?

The growth media, also known as culture media, is a recipe that contains all the nutrients required for the growth and development of plants. It consists of basic media containing macronutrients and micronutrients, agar, and plant growth regulators.

Basic Growth Media

It’s a mixture of macronutrients and micronutrients. These nutrients are involved in essential metabolic functions of the plants, required for their living.

Based on the concentrations of different elements present, the basic media are of different types. It includes MS media, White’s media, Gamborg (B5) media, Nitsch and Nitsch (NN) media, and Linsmaier and Skoog (LS) media.

The most common of all these media is MS media or Murashige & Skoog media. This medium is used for organogenesis, callus culture, micropropagation, and cell suspension. Gamborg medium is best suited for protoplast culture, the NN media is fit for anther culture, LS media works great for callus culture and cell suspension, and White’s medium is used for shoot culture.

You can find the best MS medium here!

Agar or Gellan gum

Agar is a solidifying agent in the tissue culture media. It’s a jelly-like substance that chemically consists of a mixture of agarose and agaropectin. These are obtained from the outer layer of red algae (Rhodophytes) seaweeds like Gelidium and Gracilaria.

You need to mix the basic growth media with agar by continuous stirring. This will lead to an even distribution of MS nutrients. After autoclaving, the prepared mixture of media and agar, pour into the culture vessel. After the mixture is cooled, it will be solidified, and ready to culture.

Similar to agar, gellan gum is also a solidifying agent that is more transparent than agar and is impurity-free. Based on your protocol, you can choose either of the gelling agents for your experiment.

You can find the best quality agar and gellan gum from here.

Plant Growth Regulators (PGRs)

PGRs induce the growth and development of plants. In natural conditions, plants synthesize these hormones themselves. However, under in vitro conditions, where a few tissues are used to culture the plants, the plants can’t produce these hormones on their own. And, that’s why the hormone requirements are fulfilled by supplying them from outside, mixed with culture media.

The PGRs are generally added to the media at multiplication and rooting stages. The most common hormones used in the tissue culture are auxin and cytokinin. The ratio of these hormones determines the proper development of shoots and roots of the in vitro plants.

Cytokinin induces cell division leading to the formation of shoots, whereas, auxins induce both, cell division and elongation in plants, leading to the formation of adventitious roots.

With plant cell technology you can get different cytokinins including metatopolin, kinetin, and Benzylaminopurine (BAP). It also has auxins like Indole-3-Acetic Acid (IAA) and Indole-3-Butyric Acid (IBA).

Buy the best quality PGRs from the PCT store now!

Plant Preservative Mixtures (PPM)—Optional

Contamination is one of the major problems in tissue culture experiments. The microorganisms can attack and spoil your cultures leading to loss of time, effort, and money.

To avoid these kinds of problems, some culturists use PPM or antibiotics in their media. These chemicals prevent the contaminant from attacking your cultures.

PPM is most preferred over antibiotics because of their efficacy. Antibiotics change the genetics of the plants, negatively affect their growth, and are not 100% effective in eliminating contamination.

On the other hand, PPM has the potential to prevent the growth of any microbe, including endophytes. It can be autoclaved with the media as its heat stable and doesn’t alter the genetics and affect the growth and development of plants at any stage.

Shop for PPM here.

Let us know at our given email ID if you need any help with your tissue culture processes. Plant Cell Technology is doing its best to provide tissue culturists the best of their requirements and services. You can also share your experience with our products. We will be delighted to feature the stories on our social media platforms.

Happy culturing!