Plant Cell Culture


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1. Introduction

Authors: Victor M.

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Loyola-Vargas 2 ,. Victor M. Full text PDF Related articles. Related articles Based on techniques. References Haberlandt G Kulturversuche mit isolierten pflanzenzellen. Plant Physiol — Bot Gaz — J Am Chem Soc Nature — Nature Carlsb Res Commun — Phytochemistry — Theor Appl Genet — Plant Cell Rep — Plant Cell Tissue Org — Environ Exp Bot — Planta — J Agric Food Chem — Nat Biotechnol Front Plant Sci Curr Genomics — Hortic Environ Biotechnol — Curr Opin Biotechnol — Nat Biotechnol — These topics are all discussed in addition to applications in which cell culture has been used.


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Growth conditions for plant culture must provide it with all essential nutrients which are normally provided by the plant itself. Furthermore, these conditions must be optimized to promote the growth of the particular cell type. The medium in which the cells are grown, must contain a carbon source, vitamins, salts and other organic supplements.

The inorganic nutrients come in different concentrations of macronutrients and micronutrients. Macronutrients include nitrogen, potassium, calcium and magnesium, which are always found in a concentration greater than 0. The micronutrients include iron, copper, zinc, and cobalt and can be adjusted for maximum growth of each culture type.

In addition, medium must provide vitamins which are usually synthesized by plants.

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In addition to vitamins and nutrients, growth hormones can also influence the growth rate of plants grown in artificial environments. Many plants grown in a conventional manner produce their own growth hormones, however, in culture, artificial hormones are supplied to ensure optimal growth of the plants. Besides having the appropriate nutrients in the media, the maintenance of sterile conditions is essential for the success of the cell culture allowing it to be free of microorganisms.

This requires that all equipment used in creating a cell culture must be sterilized to ensure contamination does not occur. There are many methods of ensuring sterilization with the use of alcohol, flame, and chemicals. Furthermore, containers should be covered at all times to ensure no further airborne contamination. Many plant species can be regenerated in vitro through several approaches but all require a starting point. This can be anything from a single cell that can be reproduced, a tissue or organ part, or a cut out piece of differentiated tissue or organ known as an explant.

Once the starting point has been determined, the culture used to grow this part must be considered. There are many different plant cultures all useful for different things. Some of these culture types include the embryo culture, organ culture, callus culture, and cell culture.

Figure 1 shows schematics of the different types of culture. Figure 1: Different methods of cell culture. A Embryo culture. Cells from the embryo are grown on agar media and allowed to develop into plants. B Organ culture.

Some specialized tissues are able to regenerate the whole plant. For example, the meristem tip can be excised from a plant and used to generate a whole new plant. C Callus culture results when specific tissues are excised from the adult plant and allowed to form a structure known as a callus. The callus can then be used to generate a new tissue. D Plant cell culture involves the dissociation of cells into a single cell suspension from where new plants can be generated. Click on image to enlarge.

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The Molecular Biology of Plant Cells

The main culture of interest is the cell culture, which allows for the culture of isolated cells from a very small cluster dispersed in a liquid media. This is the culture type that we will be focusing on throughout the remainder of this article. Cell suspension cultures are rapidly dividing homogenous suspensions of cells grown in liquid nutrient media from which samples can be taken [2]. In a cell suspension, a mass of cells, called a callus, must first be collected. The callus can then be suspended in a liquid callus induction media containing all the required nutrients and elements to allow for optimal growth which acts to turn all cells into undifferentiated cells.

The cell suspension is then placed on a shaker to allow the cell aggregates to disperse to form smaller clumps and single cells that are equally distributed throughout the liquid media. The cells will continuously grow until one of the factors becomes limiting causing cell growth to slow. Before this happens, cells are collected and grown on a media containing hormones to activate specific cell growth. For example if the media contains auxin, it could activate formation of adventitious roots, whereas cytokinin would stimulate axillary and adventitious shoot proliferation and regulate differentiation.


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Once the plants have developed, they can be transplanted into soil where they can continue to grow. Tissue culture techniques are becoming increasingly popular as an alternative means of plant vegetative propagation, mass production of chemicals, and genetic engineering [3]. The primary goal of plant tissue culture is crop management.

YOUR GUIDE TO PLANT CELL CULTURE

This involves asexual methods of propagation to generate whole plants from single cells [1]. In order for successful selection and genetic manipulation to occur, one must have successful in vitro plant regeneration, making this topic very important. In vitro clonal propagation is referred to as micropropagation [1]. In this context clonal means the production of genetically identical plants grown from parts of plants and reproduced by asexual reproduction.

This procedure allows for many thousands of plants to be derived from a single cell or tissue in a relatively short amount of time [1]. This appears to be advantageous over conventional plant reproduction in that only a small amount of plant tissue is required as the initial explant for regeneration of millions of clonal plants in one year [1]. Therefore, this in vitro technique allows for speedy international exchange of plant materials. It also eliminates the danger of introducing disease to the crop if performed under sterile conditions.

Plant Cell Culture Plant Cell Culture
Plant Cell Culture Plant Cell Culture
Plant Cell Culture Plant Cell Culture
Plant Cell Culture Plant Cell Culture
Plant Cell Culture Plant Cell Culture
Plant Cell Culture Plant Cell Culture
Plant Cell Culture Plant Cell Culture

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