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Example text

There are three main steps in the purification of a plant virus; these are: first, the extraction of the sap; secondly, clarification of the crude extract; and thirdly, the isolation of the virus from this. Unless the quantity of infected material is very small, it is better to put the whole plant through an ordinary domestic meat grinder fitted with a worm which compresses the material before it reaches the cutters. Extraction of the sap is helped if the leaves are first frozen at about -10°C and stored in polythene bags but not allowed to dry.

Ample, the virus constitutes 25 per cent of the dry weight of the whole larva). easily attainable. In the virus diseases of the higher animals there is usualJy n9 more ti:}an one part of virus per 108 _109 parts of wet host tissue (Cramer, 1964). )90f propagatin it in the chorioallantoic fluid of 10-13 day-old chick embryos is often used. Jes. One procedure for the preliminary pur1ficatiqn and cQn~entration of influenza virus not now in general use is to adsorb it on to chick,en red blood cells in the cold.

There are several points of practical importance to be considered before the purification of the virus can be accomplished. A high virus content is essential, the minimum quantity of virus necessary is of the order 5 to 10 mg of dry virus per kilogram me of fresh leaf tissue. The concentration of virus can be tested fairly easily by a series of simple dilution tests, provided, of course, that the virus is transmissible by mechanical means. If it is not, but is dependent on an arthropod or other type of vector, then the situation becomes much more difficult.

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