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Development of immunodiagnostics for Apple stem pitting virus and Apple mosaic virus infecting apple in India

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Abstract

Apple stem pitting virus (ASPV) and Apple mosaic virus (ApMV) are considered to be among the major viral pathogens infecting apple. These viruses have high disease incidence in the apple orchards of Jammu & Kashmir (J&K) and Himachal Pradesh (HP), so a method was required for easy virus detection. Here, we have attempted to develop a polyclonal antiserum against the coat proteins of both these viruses expressed in bacterial systems for serological based diagnostics. The purified proteins were used for rabbit immunizations and subsequently for raising antibodies. These antibodies were conjugated with alkaline phosphatase and an ELISA based protocol was standardized for detection of both the viruses. We evaluated seventy-five field samples both for ASPV and ApMV of which 25 samples for ASPV and 21 samples for ApMV were tested positive by DAS-ELISA, corroborated by RT-PCR analyses. An Immunocapture (IC)-PCR was standardized for ASPV detection using the developed antibody. This is also the first report for development of any indigenous isolate based immunodiagnostics for detection of both Apple mosaic virus and Apple stem pitting virus.

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Abbreviations

ASPV:

Apple stem pitting virus

ApMV:

Apple mosaic virus

DAS-ELISA:

Double Antibody Sandwiched-Enzyme linked immunosorbent assay

IC-RT-PCR:

Immuno-capture reverse transcriptase-polymerase chain reaction

PBS-T:

Phosphate buffered saline-Tween 20

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Acknowledgements

The authors are thankful to The Director, CSIR-Institute of Himalayan Bioresource Technology, Palampur, H.P., for providing research facilities. Academy of Scientific and Industrial Research (AcSIR) and Council of Scientific and Industrial Research (CSIR), Govt. of India are duly acknowledged for financial support. This is IHBT publication number 4612.

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  1. Sunny Dhir

    Present address: Amity Institute of Virology and Immunology, Amity University Uttar Pradesh, Noida, 201313, India

Authors and Affiliations

  1. Plant Virology Lab, Council of Scientific and Industrial Research, Institute of Himalayan Bioresource Technology, Palampur, HP, 176061, India

    Sunny Dhir, Vijay Lakshmi & Vipin Hallan

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  1. Sunny Dhir
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  2. Vijay Lakshmi
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Correspondence to Vipin Hallan.

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Dhir, S., Lakshmi, V. & Hallan, V. Development of immunodiagnostics for Apple stem pitting virus and Apple mosaic virus infecting apple in India. Indian Phytopathology 74, 189–199 (2021). https://doi.org/10.1007/s42360-020-00305-z

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