Production of policlonal antisera specific to plant viruses by rabbit oral immunization

Abstract

Serological techniques are of great importance for plant virus identification and characterization. The major limiting factor for using these techniques for plant virus identification is the requirement of a good virus purified preparation to be used in immunizing animals for antiserum production. In the present study, two New Zealand rabbits were orally immunized with extracts from cowpea (Vigna unguiculata) plants systemically infected with Cowpea severe mosaic virus (CPSMV) and with extracts from papaya (Carica papaya) infected with Papaya lethal yellowing virus (PLYV). The leaf extracts were prepared in saline solution 0.15 M in the rate of 1:1 (w/v) and clarified by a centrifugation of 10,000 g for 10 min. The clarified extracts containing the viruses were orally administered to the New Zealand rabbits in two series of five daily doses of 1.0 ml each. The obtained policlonal antisera were shown to be very specific to their respective viruses in double immunodiffusion and indirect ELISA. These seem to be the first antisera specific for plant virus obtained by rabbit oral immunization. The results open up some possibilities for producing antisera to plant viruses of difficult purification. It is a simple, fast and inexpensive method for production of antisera for plant viruses when compared to the traditional techniques that involve rabbit injections with purified virus preparations.