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A Short Note on Reflections and Publications on Citrus tristeza virus (CTV) Methodologies

  • Moshe Bar-Joseph
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2015)

Abstract

My PhD thesis work of Citrus tristeza virus (CTV) purification was aimed to develop a rapid serological assay to replace biological indexing. The task turned difficult and was achieved after a lengthy struggle, rewarded by allowing (1) the rapid diagnosis of the first incidences of natural spread of a severe CTV-VT strain in our region and (2) finding that the CTV particle isolation protocol, with some modifications, was also useful for Beet yellows virus (BYV) particles, leading to their assignment in the Closterovirus group, the first group of elongated plant viruses with different modal lengths. Later, following the introduction of ELISA for large-scale diagnosis of tristeza-infected citrus trees, the CTV infection rates through the coastal citrus production areas were continually increasing, with many ELISA-positive samples appearing symptomless, prompting the need to develop strain-specific assays. Using CTV-VT cDNA fragments, as hybridization probes, the genetic diversity among local CTV isolates was demonstrated. With the emergence of the PCR technology, we developed a CTV-dsRNA cloning method based on the ligation of known oligonucleotide molecules to dsRNA ends and the use of complementary oligonucleotides for cDNA synthesis and PCR amplification. The method allowed the cloning of a cDNA molecule complementary to a defective dsRNA of 2.4 kb with intact 5 and 3 ends of the CTV-VT genome. A list of publications, resulting from continuous collaborative work with local and foreign associates and students on the development and adaptation of novel CTV methodologies, is present.

Key words

DsRNA Defective RNA Oligonucleotide dsRNA ligation ELISA cDNA cloning Hybridization Diagnosis Virus strain differentiation Symptomless virus isolate 

Notes

Acknowledgment and Dedication

My thanks and gratitude goes to my family and fellow friends with whom I was fortunate to collaborate through the years. This short biographical note was edited by Dr. W.O. Dawson and is dedicated to the memory of our dear friend and collaborator, Dr. Stephen M. Garnsey.

References

  1. 1.
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Selected Additional Personal Publication List

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Reviews and Chapters

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Moshe Bar-Joseph
    • 1
  1. 1.The S. Talkowski Laboratory, Department of Plant Pathology and Weed Research, The Volcani CenterAgricultural Research OrganizationBet DaganIsrael

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