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LABORATORY PROCEDURES FOR PLANT CELL VIRUSES

REFERENCE NO.: PVC/1998/3.01


TITLE: PRESERVATION OF VIRUS INFECTED PLANT MATERIAL BY FREEZING AND LOW TEMPERATURE STORAGE IN LIQUID NITROGEN


INTRODUCTION

Original virus infected samples arriving in the plant virus collection will be stored in liquid nitrogen for the duration of the authentication process (PVC/1998/2.01). This sample will be destroyed after an ordinary database entry has been assigned and the authentication process is completed. Material preserved in liquid nitrogen is not meant for any distribution but as nuclear seed stock to serve as back up and inoculum to infect virus propagation plants which are subsequently used to replenish stocks preserved by freeze-drying (PVC/1998/3.02) which are for distribution. Virus preservation in liquid nitrogen is also indicated for viruses which loose their infectivity upon prolonged storage or which are sensitive to other preservation and storage conditions.

 

PROCEDURE

A small sample of symptomatic plant tissue infected with virus is excised and placed directly in a 2 ml cryo-vial and subjected to liquid nitrogen without any prior manipulation to the material. The infected tissue is stored in liquid nitrogen and removed to be used for inoculum when required. Infectivity tests are performed after a maximum of 3 years storage in liquid nitrogen. Nuclear virus seed stocks are replenished only after renewed authentication (PVC/1998/2.04).


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