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Biotechnological Problems Associated with Plants - From a biotechnological point of view, if genes are to be modified and correct genes returned to plants, we have to have a clear understanding of the various problems specific to plant material.

Plant tissue can present a number of practical problems for the isolation of nucleic acids.

Tough cell walls can make effective homogenization of the tissue difficult and this fact coupled with the large amount of nucleases (in particular ribonuclease) present in many plant tissues, makes for very low yields of nucleic acid per gram of tissue.

This is especially true if mRNA must be extracted from a difficult tissue such as mature leaves or certain seeds. RNA extraction is much easier if young seedlings can be used. Purity of RNA preparations can also be a problem.

Plant cells contain large quantities of polysaccharide which, being a negatively charged polymer can sometimes copurify with RNA. Persistent polysaccharide contamination is a difficult problem to solve.

Isolation of DNA from the nucleus requires removal of both chloroplast and mitochondrial DNA, although chloroplast DNA is quantitatively the larger problem.

This problem, coupled with that of low yield of DNA, can be circumvented to some extent by careful choice of tissue.

Nuclear DNA is often isolated from embryonic axes which have a high proportion of nuclear to organelle DNA; collecting the embryonic axes can, however, be a tedious process.

None of these problems are insoluble but it is important that they be considered when planning plant genetic engineering experiments.

 
 

 
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