Category: Biology Second Paper

After placing the explants in the culture medium, the container is kept in a controlled environment. Room temperature is kept at 17-20°C, light intensity 3,000-5,000 lux or 1000-3000 lux and relative humidity 70-75%. Within a few days, the cell divides repeatedly to form a multicellular nucleus. This multicellular body is called a callus. After 5-7 days numerous buds or microflora are produced in the callus. Buds grow due to totipotency of cells. New seedlings are then produced from the buds.

Another important step in tissue culture technology is the preparation of culture media. Usually this culture media is made up of essential plant nutrients. The culture media is prepared with the main and secondary plant nutrients, vitamins, phytohormones, agar agar, glucose, sucrose (2-4%), fructose, maltose etc. in a container. This culture medium prepared with basic nutrients is called basal medium. The pH of the growing medium is kept between 5.5-6.0. The most commonly used culture media are MS medium (Murashige & Skoog, 1962) and B5 (Gamborg et al, 1968) medium.

The first and most important step in tissue culture technology is explant selection and isolation. First the explants are selected. While selecting the explant, it should be kept in mind that it should be healthy, vigorous, disease free and of improved variety. Besides, care is taken to ensure that the desired features are present in it. Later, the required parts such as apical bud, lateral bud or leaf part, dividing cells, meristem, pollen, ovule, embryo, nucellus, protoplast etc. are separated from the explant.

1. Explant selection (Ex-plant selection): The first and most important step in tissue culture technology is explant selection and isolation. First the explants are selected. While selecting the explant, it should be kept in mind that it should be healthy, vigorous, disease free and of improved variety. Besides, care is taken to ensure that the desired features are present in it. Later, the necessary parts such as apical bud, side bud or leaf part, dividing cells, meristem, pollen, ovule, embryo, nucellus, protoplast etc. are separated from the explant.
2. Preparation of culture media: Another important step in tissue culture technology is the preparation of culture media. Usually this culture media is made up of essential plant nutrients. The culture media is prepared with the main and secondary plant nutrients, vitamins, phytohormones, agar agar, glucose, sucrose (2-4%), fructose, maltose etc. in a container. This culture medium prepared with basic nutrients is called basal medium. The pH of the growing medium is kept between 5.5 and 6.0. The most commonly used culture media are MS medium (Murashige & Skoog, 1962) and B5 (Gamborg et al, 1968) medium.
3. Sterilization: Both culture media and explants used in tissue culture technology must be sterilized. The culture media is taken in a conical flask or large test tube. Then, both the culture medium and the explants were placed in an autoclave at 121°C for 20 minutes under 15 pounds of pressure. As a result it becomes germ free.
4. Placing explants in culture medium: Sterile explants are placed in sterile culture medium. The knives, tongs and hands used during this time are disinfected with ethyl alcohol. This work is done under a laminar air flow machine in a sterile environment.
5. Callus creation and increase in number (Callus create): After placing the explants in the culture medium, the container is kept in a controlled environment. The room temperature is kept at 17-20°C, light intensity is 3,000-5,000 lux or 1000-3000 lux and relative humidity is kept between 70-75%. Within a few days, the cell divides repeatedly to form a multicellular nucleus. This multicellular body is called a callus. After 5-7 days numerous buds or microflora are produced in the callus. Buds grow due to totipotency of cells. New seedlings are then produced from the buds.
6. Plantlets production: Buds are cut from the callus with a sterile knife. The buds are then placed in seedling producing culture medium. Auxin is applied in the culture medium for root formation. As a result, seedlings are produced within a few days. The method of making seedlings from explants through artificial cultivation is in-vitro culture.
7. Plant transfer on Tub: When the roots of the seedlings are well formed, the seedlings are carefully placed on the agar soil of the tub. The tub is kept in a light and airy place. The tub is watered occasionally. Within a few days, the seedlings grow and become suitable for planting in the field.
8. Plant transfer in field: Tubs with seedlings are sometimes kept in natural environment. In this the plants can adapt to the natural environment. Later, healthy, vigorous and disease-free seedlings are planted in the field.

The environment for tissue culture must be sterile. Culture medium containers and other glassware are sterilized by placing them in an oven at 160-180°C for 1-2 hours. Forceps, needle, scalpel etc. are sterilized by immersion in 95% alcohol.

1. Meristem Culture: Plants are produced by meristem culture of plants.
2. Pollination culture: Complete plants are produced by culturing the pollinators and anthers of plants.
3. Room Bud Culture: Plants are created from plant buds.
4. Callus Culture: Numerous seedlings are produced from the multicellular peduncle or callus of the plant.
5. Micropropagation: Using small plant parts to produce whole plants.
6. Somatic culture: New seedlings are created by culturing the dividing somatic cells of plants.

In 1901, the American scientist Morgan (Morgan) was the first to think that every living plant cell has the ability to transform into a complete plant and this ability is called totipotency. In 1920, Gottlieb Haberlandt first invented tissue culture technology. That is why he is called the father of tissue culture. In 1939, Gautheret and Nabercourt were the first to do plant tissue culture.

The process in which a plant’s dividing cells or organs are grown in complete sterile culture medium to produce new seedlings or plants is called tissue culture. Complete plants are formed by culturing plant dividing cells, meristem, pollen, ovule, embryo, nucellus, protoplast etc. This process is called micropropagation as a whole plant is created using small parts of the plant. It is also called cloning technology as identical clones of plants are made. At present, virus vaccines, hormones, insulin, alkaloids etc. are being made using this technology. Vitamins, enzymes, hormones, sugars, mineral salts etc. are used in tissue culture.

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Solving and analyzing various problems in biology by computer is called bioinformatics.