TAILIEUCHUNG - Plant tissue culture

SPB2 2/27/2003 4:06 PM Page 35 2 Plant tissue culture Introduction Most methods of plant transformation applied to GM crops require that a whole plant is regenerated from isolated plant cells or tissue which have been genetically transformed. This regeneration is conducted in vitro so that the environment and growth medium can be manipulated to ensure a high frequency of regeneration. In addition to a high frequency of regeneration, the regenerable cells must be accessible to gene transfer by whatever technique is chosen (gene transfer methods are described in Chapter 3). The primary aim is therefore to produce, as easily and as quickly as possible, a large. | SPB2 2 27 2003 4 06 PM Page 35 2 Plant tissue culture Introduction Most methods of plant transformation applied to GM crops require that a whole plant is regenerated from isolated plant cells or tissue which have been genetically transformed. This regeneration is conducted in vitro so that the environment and growth medium can be manipulated to ensure a high frequency of regeneration. In addition to a high frequency of regeneration the regenerable cells must be accessible to gene transfer by whatever technique is chosen gene transfer methods are described in Chapter 3 . The primary aim is therefore to produce as easily and as quickly as possible a large number of regenerable cells that are accessible to gene transfer. The subsequent regeneration step is often the most difficult step in plant transformation studies. However it is important to remember that a high frequency of regeneration does not necessarily correlate with high transformation efficiency. This chapter will consider some basic issues concerned with plant tissue culture in vitro particularly as applied to plant transformation. It will also look at the basic culture types used for plant transformation and cover some of the techniques that can be used to regenerate whole transformed plants from transformed cells or tissue. Plant tissue culture Practically any plant transformation experiment relies at some point on tissue culture. There are some exceptions to this generalisation Chapter 3 will look at some but the ability to regenerate plants from isolated cells or tissues in vitro underpins most plant transformation systems. Plasticity and totipotency Two concepts plasticity and totipotency are central to understanding plant cell culture and regeneration. SPB2 2 27 2003 4 06 PM Page 36 36 2 Plant tissue culture Plants due to their sessile nature and long life span have developed a greater ability to endure extreme conditions and predation than have animals. Many of the processes involved in plant growth

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