Plant transformation techniques

Prior to tissue culture and transformation, plant material must be surface sterilized to remove contaminating micro-organisms. 

Remove leaves from tobacco plants, wash briefly under running cold tap-water. 

Wash leaves in three changes of sterile distilled water. 

Preparation of culture medium for adventitious shoot regeneration on leaf discs of tobacco (Nicotlana tabacum) 

For 1 litre of medium, dissolve the appropriate mass of Murashige and Skoog basal medium in 800 ml of distilled water. Add 30 g of sucrose, 1 mg of BAP, 0.1 mg of NAA, and stir until sucrose is fully dissolved. 

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Describes the theory and practice of modern plant transformation techniques with respect to nuclear and plastic genomes... 

Plant transformation techniques allow the delivery of the transforming DNA through the cell wall and plasma and nuclear membranes, without compromising the viability of the cell. Gene delivery can be performed either via a biological vector (plant viruses or bacteria) or by non-biological vector-free procedures (chemical methods, microinjection, particle bombardment, etc.) (Birch 1997). 

High sensitivity of this system to genotoxic damage allows selection of chemicals that have no or a negligible impact on plant genome integrity (Kovalchuk et al. 2001). Therefore, this test system makes it possible to develop clean transformation techniques in contrast to those that usually result in extensive DNA damage (Kohler et al. 1989, Leskov et al. 2001). 

To date, most examples of plant produced antibodies have been in tobacco. This is simply because it is eaq to transform and rapid to regenerate, and is one of the best studied and mderstood of the plant species. Historically, plants of the dicotyledonous group have been the easiest to transform, using Agrobacterium, consequently most of the early work has been concentrated in these plants. Other examples are Arabidopsis (another model plant), potato, tomato, and soy. More recently, with the development of ballistic bombardment techniques for plant transformation, many other plant species have become amenable to transformation, including important crop plants such as maize, rice, and wheat. The range of plants that could be used as vehicles for antibody production is now close to including all the commercially important crop species. 

Methods and iastmments that are used to monitor phosgene content ia air are well developed and have been reviewed (46—48). One detection iastmment is a porous tape that measures the concentration of phosgene ia air ia quantities as small as 6 ppb (49). Fourier transform ir spectrometry techniques have been developed to permit line and area monitoring ia the area around phosgene plants (50). 

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