Basics of Plant Tissue Culture
by Emirhan Turan, 29 August, 2020
Plant tissue culture is a technique to grow new plants by using a part of plants and transferring them into an artificial media where they can carry on their growth and function. Thanks to plant tissue culture, we can grow new plants outside of the original host body. In the artificial medium, we usually can use plant materials from the tips of shoots and roots as well as minuscule parts of the seed, callus, ovule, cell, pollen grain, or embryo.
There are three main stages in the tissue culture process:
The initial stage involves the establishment of the in vitro media. This stage is critical and needs to be done under sterilized conditions not to have any contamination on the plant material.
The multiplication stage aims to increase the number of shoots produced by the original plant material. Sub-culturing these new shoots on to new medium is increasing the production dramatically.
Rooting stage, roots begin to form, and plant materials are turning to plantlets. Usually, auxin is being used on micro-cutting to induce the root formation. The rooting stage is easier for the producer to control the rooting in vitro. However, ex vitro rooting has better adaption and root system then in vitro.
After these 3 steps, the plantlets are ready for acclimatization where they are adapted to ex vitro environment. This stage is very critical, and plantlets are vulnerable, so propagators can see a large number of plant losses if the stage is not cared on carefully. This step should involve gradual moving to open-air conditions where the humidity is reduced, and the light levels increase. Rooted plantlets in vitro can be acclimatized using intermittent mist in a closed polyethylene tent.
How to avoid Contamination in your Tissue Culture experiments
A human body has different bacteria species that can contaminate the plants. To avoid contamination from humans, it is always suggested to wear a lab coat and gloves while working with the samples. As jewelry, watches can be loaded with bacteria as well, it is better to remove them before starting the experiments.
Using certified, high-quality medium ingredients and sterilized water.
Lab equipment such as beakers, storage vessels, pipettes, etc. should be sterilized well before using them in the experiment. Autoclave would be a proper choice for sterilization.
Checking the laminar flow and incubators is essential before starting the experiment. Surfaces and laboratory equipment should be cleaned with 70% alcohol solution.
The experiment room and storage room should be sterilized before experimenting.
Advantages and Disadvantages of Plant Tissue Culture
There are plenty of reasons why tissue culture is being used and the demand is really high on it. These advantages include:
The speed of the process, in conventional growing, to grow plants can take a lot of time, however, in plant tissue culture the process is fast and effective. In a few weeks, thousands of plantlets can be produced from a minor amount of plant material. This includes producing, cultivating, and introducing new varieties to market at a higher speed than conventional operations.
Production space, by using plant tissue culture the required amount of space can be decreased ten times more than the traditional growing methods.
Healthy plants, if tissue culture is established in sterilized conditions, plantlets grown in vitro is generally free from diseases. Tissue culture also could help to stop spread of diseases and viruses to other plants.
The flexibility of growing, using tissue culture helps growers to produce plantlets all year round, regardless of the growing season and weather.
Beyond the advantages of Plant Tissue Culture practices, few disadvantages should be considered before starting point. These disadvantages include:
Plant Tissue Culture can be more costly and requires more skilled labor than conventional practices.
As plants are growing in vitro they might be more resilient to diseases in the acclimatization and further stages.
The plant material must be very well screened and investigated, to select already infected plants can cause big losses in the next stages.
Even the success rate is high in Tissue Culture applications, there is still a possibility due to biochemical reactions in plants that plants can die or give unexpected reactions.
Sterilization is important in tissue culture operations once the sterilization is made incorrectly during the establishment the plants can hardly survive.
How to store plants
While storing the plantlets, to be successful they should be kept in sterilized sealed containers. Tissue culture does not need light for long term storage, however, it needs a cool (between 0o and 15oC) and steady temperature. As they are growing in an artificial medium, ensure that hormones and nutrients are sufficient for the plant material, and always keep the storage room sterile. When the plants are grown enough, they should be transferred to a new medium.
There are plenty of plants that are used in Plant Tissue Culture due to the demand and yield such as banana, cassava, rice, sweet potato, tomato, etc. However, in recent years, tissue culture involves a lot of ornamental plants and other edibles depending on regional demand.
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