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ISHS Acta Horticulturae 550: XVIII International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops - Top Fruit Diseases

THE USE OF A SIMPLIFIED DAS-ELISA PROCEDURE FOR LARGE-SCALE DETECTION OF APPLE CHLOROTIC LEAF SPOT AND APPLE STEM GROOVING VIRUSES IN APPLE TREES OVER EXTENDED SAMPLING PERIODS

Authors:   L.M. Corvo, M.T.F. Barros
Abstract:
ACLSV is the most frequently encountered virus of apple orchards. As antisera of high quality are commercially available this should be a virus suited for routine detection by ELISA. However, different studies using DAS-ELISA have shown that, to be reliable, ACLSV detection should be done during a limited period in the growing season. In an attempt to overcome this limitation, we have conducted a series of experiments using different apple tissues including winter budsticks, flowers, leaves and growing shoots. Procedures adapted from the modified Flegg and Clark (1979) version of DAS-ELISA were compared with standard DAS-ELISA procedures. The former were clearly advantageous due to their simplicity, increased rapidity and adequacy to detect ACLSV throughout extended sampling periods, including dormancy. The experiments herein reported aimed at further simplification and cost reduction in the detection of ACLSV and ASGV. The overall work enabled us to develop a much faster and convenient procedure to detect ACLSV in apple tree tissues, as compared to standard procedures in current use. The fact that the optimised procedure is highly suitable for screening ACLSV in winter budsticks has obvious advantages for use in the control of circulating propagative material. Moreover, this procedure was found adequate for detection of ASGV using the same extraction conditions, allowing the simultaneous screening of both viruses without any additional effort in sample preparation.
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