EVALUATION OF THE SENSITIVITY OF NITROCELLULOSE MEMBRANE LOW-COST ALTERNATIVES USED IN TISSUE BLOT IMMUNOASSAY (TBIA) FOR DETECTION OF SOME PLANT VIRUSES

Hosny Ali Younes,Maha Kawanaa

doi:10.15414/jmbfs.2020.10.1.68-72

Abstract

In an attempt of reducing the cost of tissue blot immunoassay (TBIA), some kinds of papers as alternative solid phases in place of nitrocellulose membrane (NCM), were tested for detection of three viruses: Cowpea mosaic virus (CPMV), Tomato mosaic virus (ToMV) and Zucchini yellow mosaic virus (ZYMV). Most of the examined papers developed positive reaction with all tested viruses. Canson paper (150g/m2 and 300g/m2) were sensitive for detection of the tested viruses as NCM and persist in good condition during the processing. Filter paper (Whatman No. 1), local drawing paper (60g/m2), photocopy paper (80g/m2), (56g/m2) and (70g/m2) were very weak and need to taking care in handling. High glossy photo paper (200g/m2) was not effective however, it did not show any infected or healthy tissue print. Good results were obtained when both faces of NCM and Canson paper (300g/m-2) were printed with all tested viruses. TBIA could detect CPMV in infected leaves using the unused face of NCM previously processed with 7 or 11 years ago. CPMV, ToMV and ZYMV were detected 2, 4, 6, 8 and16 days after inoculation with NCM and Canson paper (300g/m-2). Also, the tested viruses were detected in leaf, petiole, stem and root of infected plants. ToMV could be detected using NCM and Canson paper (300g/m-2) in naturally infected tomato plant samples.

Highlights

Accurate detection is an essential step to control the plant viruses (Jeong et al, 2014)
Eight different types of papers and nitrocellulose membrane (NCM) were tested for detection of Cowpea mosaic virus (CPMV), Tomato mosaic virus (ToMV) and Zucchini yellow mosaic virus (ZYMV) by Tissue blot immunoassay (TBIA)
The aforementioned results are in line with Makkouk and Kumari (2002) study results where they got the same observation with filter and photocopy paper when tested to detect Alfalfa mosaic virus (AMV), Bean yellow mosaic virus (BYMV) in faba bean and Barley yellow striate mosaic virus (BYSMV) in barley

Summary

Introduction

Accurate detection is an essential step to control the plant viruses (Jeong et al, 2014). Tissue blot immunoassay (TBIA) is widely used for detection of plant viruses in various plant tissues. It could apply even in poorly equipped laboratories. Zucchini yellow mosaic virus (ZYMV) (genus: Potyvirus; family: Potyviridae) is a major viral pathogen of cucurbits. It causes severe stunting of plants and distortion of the fruits which led to high losses of productivity (Desbiez and Lecoq, 1997). The objectives of the present study were to evaluate the sensitivity of different types of low-cost papers as solid phases alternative to the expensive NCM for detection of CPMV, ToMV and ZYMV, to assess the possibility of using both faces of the best papers, and to detect the sensitivity of the previously used NCM for detection of the tested viruses

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Conclusion