Plant tissue culture and genetic engineering are the most widely used methods for crop improvement in plant breeding. Plant tissue culture is a technique of in vitro twining and growing plant cells, tissues aseptically on an artificial medium in suitable containers under controlled environmental conditions. It is based on the ‘totipotent’ nature of plant cell or the phenomenon of totipotency which indicate that each plant cell has inherent capacity to develop into a complete plant. This concept was introduced by Haberlandt in 1902, also known as “Father of Plant Tissue Culture”.
What Is the Technique of Plant Tissue Culture?
There are 3 fundamental techniques to successfully develop a plant tissue culture.
Pre - experimental phase
Conduct you experiment in biological safety cabinet or laminar flow cabinet. You don’t want all your tissue culture to be contaminated with all the fungus and ruin your culture. Make sure to apply the aseptic technique throughout this process and fumigate the working area with alcohol or bleach. In addition to this, always wash, and sterile all the glassware and apparatus required for this process including the beaker, test tubes, forceps, and cutting knife.
Media for plant tissue culture: selecting the right culture media is important as tissue culture needs special components to support its growth. The main component required by tissue culture in order to grow the explant successfully are nutrients, carbohydrate source, solidifying agents (agar), water, growth regulators, and special additives including auxin, cytokinin, and gibberellins. However, depending on the type of tissue culture you want to grow, it might require the proper combination of culture media components. Our HiMedia plant tissue culture media are the perfect solution for you to achieve successful totipotency as it offers a wide selection of media and reagents, suitable for your need. it comes with tested formula for every tissue culture you wish to grow.
In this phase, tissue culture media with the addition of hormones need to be prepared according to the manufacturer procedure and always ensure the pH level is around 5.5 to 5.8. The prepared media then need to undergo sterilization to kill all the unwanted microorganisms within the media that could potentially interrupt the callus growth.
The selection of explant also plays a major role in determining the success of this process. The plant shoot is selected either from the tip of the new shoot, the stem, the auxiliary bud of the leaf, or at the root. All the collected leave needs to undergo a sterilization process using Tween20 and bleach before putting into the solidified agar. Allow the explant to incubate under 16 hours proper light condition and 8 hours in dark condition with temperature around 25ᵒ to 30ᵒC and observe them 1 week and afterward with a proper subculture.
Post- experimental phase
After complete plant development, the culture plant needs to be acclimatized in the greenhouse, allowing it to develop into a stronger plant. This acclimatization process allows the plant to adjust to changes in its environment, hence enabling it to survive in harsh environments such as temperature change, water, and food availability.
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