Improvements in the detection of pea seed-borne mosaic virus by ELISA

The detection by ELISA of pea seed-borne mosaic virus (PSbMV) in pea leaves and seeds was improved by the addition of cellulase or Triton X-100 to the extraction fluid, probably because the additives aided the release of virus particles from host materials. With leaf extracts the additives were most effective at 0.1%. In initial tests cellulase was used with macerozyme, but the latter enzyme was then shown to decrease the effectiveness of cellulase. Triton X-100 was as effective as cellulase and the absorbance values obtained in ELISA of infected leaf extracts, diluted to 1/10 in extraction fluid containing the additive, were about six times greater than those of infected extracts diluted in normal extraction fluid. Five named isolates of PSbMV, in addition to the homologous isolate, were readily detected in infected leaves extracted in fluid containing Triton X-100. In tests on seeds and seedlings of seven infected seed lots of pea cv. Waverex, using Triton X-100 in the extraction fluid, PSbMV was detected in five times as many seeds as seedlings, probably mainly because in many infected seeds the virus was in the testa and not in the embryo. About 9% of infected seedlings were without recognisable symptoms 4 wk after emergence.