ISHS Acta Horticulturae 236: V International Symposium on Small Fruit Virus Diseases
CONTROLLING THE PH OF BLUEBERRY SAP FOR EFFICIENT DETECTION OF VIRUSES IN BLUEBERRY LEAVES BY ELISA |
| Authors: | S.G. MacDonald, R.R. Martin, J.M. Gillett, D.C. Ramsdell |
| Abstract:
Detection of tomato ringspot virus (TomRSV), blueberry scorch virus (BBScV) and blueberry red ringspot virus (BBRRSV) from blueberry leaves by ELISA was not possible with standard ELISA grinding buffer. We compared standard PBS-Tween-Ov-PVP (0.015 M phosphate), 0.15 M PBS-Tween-Ov-PVP, pH 7.5, and 0.1 M borate-Ov-PVP, pH 8.0, each with and without 0.5% nicotine, alkaloid, as buffers to control the pH of blueberry leaf sap and to improve the detection of viruses in blueberry by ELISA. Detection by ELISA of TomRSV, BBScV and BBRRSV in blueberry was improved by the addition of nicotine to each of the buffers. With BBRRSV the use of 0.5% nicotine added to standard PBS-Tween-Ov-PVP gave differences between infected and healthy tissues similar to those obtained with partially purified samples previously used to detect this virus in leaf tissues. With TomRSV and BBScV the use of nicotine increased ELISA A405 values to 10–15 times greater than those obtained with the standard ELISA buffer for infected samples without any increase in the A405 values of healthy samples. The use of 0.15 M phosphate also improved the detection of each virus but not as much as the addition of nicotine, for all the viruses tested. Blueberry shoestring virus was detected successfully with or without the addition of nicotine to any of the buffers tested. | |
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