What are the application of protoplast culture?

What are the application of protoplast culture?

Organelle Isolation: Protoplasts are very convenient material for the isolation of chloroplasts, mitochondria, nuclei and even chromosomes. It has been demonstrated that chloroplasts particularly isolated from cereal protoplast have higher capacity for CO2 fixation than those obtained by mechanical grinding.

What are protoplasts and what are their uses?

Protoplasts can be used to study membrane biology, including the uptake of macromolecules and viruses . These are also used in somaclonal variation. Protoplasts are widely used for DNA transformation (for making genetically modified organisms), since the cell wall would otherwise block the passage of DNA into the cell.

What is the function of protoplasts?

functions include: (1) providing the protoplast, or living cell, with mechanical protection and a chemically buffered environment, (2) providing a porous medium for the circulation and distribution of water, minerals, and other small nutrient molecules, (3) providing rigid building blocks from which stable structures …

Which of the following is used for protoplast fusion in plant tissue culture?

Glucose is the preferred carbon source for protoplast culture. A high Auxin/kinetin ratio is preferred to induce cell division. A high kinetin/Auxin ratio is preferred for regeneration.

What are the possible outcome of protoplast fusion?

By protoplast fusion it is possible to transfer some useful genes such as diesese resistance,nitrogen fixation ,rapid growth rate ,more product formation rate,protein quality,frost hardiness,drought resistance,herbicide resistance ,heat and cold resistance from one species to another.

What are the advantages of protoplast fusion?

By protoplast fusion, it is possible to transfer some useful genes such as disease resistance, nitrogen fixation, rapid growth rate, protein quality, frost hardiness, drought resistance etc. from one species to another and thereby widen the genetic base for plant breeding.

What are the advantages of protoplast culture?

Advantages of protoplast transformation include: (i) delivery of multiple plasmids with high levels of cotransformation, (ii) no binary vector required, (iii) high frequency transformation, and (iv) most plant species are amenable to protoplast isolation and transformation.

What is protoplast fusion technology?

Somatic fusion, also called protoplast fusion, is a type of genetic modification in plants by which two distinct species of plants are fused together to form a new hybrid plant with the characteristics of both, a somatic hybrid.

How does protoplast fusion work?

Protoplast fusion permits the production of the somatic hybrids. One cell can be induced to fuse with another cell to form the heterokaryon which consists of cytoplasms containing chloroplasts, mitochondria and nuclei from both parent cells.

Which is the most effective chemical fusion used in protoplast fusion?

polyethylene glycol (PEG)
Several chemicals have been used successfully to induce fusion but the most successful and the one most widely used at the present time is polyethylene glycol (PEG). The effects of PEG are not specific and it will promote the aggregation and fusion of protoplasts from the same or different species.

What is protoplast technology?

Protoplast technology includes the isolation, culture and fusion of higher plant protoplasts leading to the production of entire plants. You will be studying the method of isolation and fusion of protoplast in this chapter. A hybrid produced by fusion of somatic cells of two varieties (or) species is called somatic hybrid.

What is the best method for fusion between protoplasts?

Subsequently, Michel (1937) demonstrated fusion between protoplasts using potassium nitrate as plasmolyticum. Power etal (1970) reported sodium nitrate induced fusion of cereal root proto­plasts. By this method, equal densities of proto­plasts from two different sources are mixed and then centrifuged at 100 g for 5 minutes to get a dense pellet.

How to prepare suspended protoplast in centrifugation?

The suspended protoplasts are kept in water-bath at 35° C for 5 minutes and again centrifuged at 200 g for 5 minutes. The pellet is once again kept in water-bath at 30°C for 30 minutes. Fusion of protoplast takes place at the time of incubation. The, pellet is again suspended by 0.1% sodium nitrate for 5-10 minutes.

What are chemical fusogens and how do they affect protoplasts?

Chemical fusogens cause the isolated protoplasts to adhere to one another and leads to tight agglutination followed by fusion of protoplast (Fig. 6.14). The adhesion of isolated protoplast takes place either due to reduction of negative charges of protoplast or due to attraction of protoplast by electrostatic forces caused by chemical fusogens.

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