Skip to main content

CTV Vectors and Interactions with the Virus and Host Plants

  • Protocol
  • First Online:
Book cover Citrus Tristeza Virus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2015))

Abstract

Citrus is a graft-propagated perennial crop, and Citrus tristeza virus (CTV) is readily graft-transmissible. CTV is comprised of a complex of strains and isolates and, in nature, is spread semi-persistently by aphid vectors. Therefore, citrus trees become infected with multiple CTV strains over time. An important step in characterizing a CTV field isolate is to use aphid vectors to “clean” up the CTV population of a source tree to separate strains and eliminate other graft-transmissible agents. Use of Toxoptera citricida or Aphis gossypii will expedite efficient CTV transmission. CTV vector studies require critical coordination of abundant robust and virus-free vector-competent aphid colonies and an insect-proof, climate-controlled greenhouse or growth chamber. CTV donor and healthy receptor plants with young flush growth must be available for virus acquisition and inoculation. Vector optimums for virus acquisition and inoculation are 24 h for each. CTV infection is readily determined by serology using a polyclonal antiserum or a monoclonal antiserum cocktail; whereas, molecular genotyping is conducted with reverse transcription polymerase chain (RT-PCR) or real time quantitavtive RT-PCR (RT-qPCR) with strain-specific primers and probes. However, the phenotype of the aphid-transmitted isolate still requires virus indexing by graft inoculation to a citrus host range and evaluating symptoms such as stem pitting, vein clearing, stunting, and chlorosis.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Raccah B, Loebenstein G, Bar-Joseph M (1976) Transmission of Citrus tristeza virus by the melon aphid. Phytopathology 66:1102–1104

    Article  Google Scholar 

  2. Yokomi RK, Lastra R, Stoetzel MB et al (1994) Establishment of the brown citrus aphid (Homoptera: Aphididae) in Central America and the Caribbean Basin and transmission of Citrus tristeza virus. J Econ Entomol 87:1078–1085

    Article  Google Scholar 

  3. Rocha-Peña MA, Lee RF, Lastra R et al (1995) Citrus tristeza virus and its vector Toxoptera citricida. Plant Dis 79:437–445

    Article  Google Scholar 

  4. Dickson RC, Johnson MM, Flock RA et al (1956) Flying aphid populations in southern California citrus groves and their relation to the transmission of tristeza virus. Phytopathology 46:204–210

    Google Scholar 

  5. Roistacher CN, Bar-Joseph M (1984) Transmission of tristeza and seedling yellows tristeza by small populations of Aphis gossypii. Plant Dis 68:494–496

    Article  Google Scholar 

  6. Yokomi RK, Polek M, Gumpf DJ (2010) Transmission and spread of Citrus tristeza virus in Central California. In: Karasev AV, Hilf ME (eds) Citrus tristeza virus complex and tristeza diseases. APS Press, St. Paul, MN, pp 151–166

    Google Scholar 

  7. Raccah B, Loebenstein G, Singer S (1980) Aphid transmissibility variants of Citrus tristeza virus in infected citrus trees. Phytopathology 70:89–93

    Article  Google Scholar 

  8. Yokomi RK, DeBorde RL (2005) Incidence, transmissibility and genotype analysis of Citrus tristeza virus (CTV) from CTV eradicative and non-eradicative districts in central California. Plant Dis 89:859–866

    Article  CAS  Google Scholar 

  9. Yokomi RK, Garnsey SM (1988) Host effects on natural spread of Citrus tristeza virus in Florida. In: Timmer LW, Garnsey SM, Navarro L (eds) Proceeding 10th Conference of the International Organization of Citrus Virologists, Valencia, November 1986. IOCV, Riverside, pp 77–81

    Google Scholar 

  10. Yokomi RK, Garnsey SM (1987) Transmission of Citrus tristeza virus by Aphis gossypii and Aphis citricola in Florida. Phytophylactica 19:169–172

    Google Scholar 

  11. Gottwald TR, Gibson GJ, Garnsey SM et al (1999) Examination of the effect of aphid vector population composition on the spatial dynamics of Citrus tristeza virus spread by stochastic modelling. Phytopathology 89:603–608

    Article  CAS  Google Scholar 

  12. Harper SJ, Yokomi RK, Dawson WO (2016) Citrus tristeza virus-aphid interactions. In: Brown JK (ed) Vector-mediated transmission of plant pathogens. APS Press, St. Paul, MN, pp 121–130

    Chapter  Google Scholar 

  13. Herron CM, Mirkov TE, da Graça JV et al (2006) Citrus tristeza virus transmission by the Toxoptera citricida vector: in vitro acquisition and transmission and infectivity immunoneutralization experiments. J Virol Methods 134:205–211

    Article  CAS  Google Scholar 

  14. Killiny N, Harper SJ, Alfaress S et al (2016) Minor coat and heat shock proteins are involved in the binding of citrus tristeza virus to the foregut of its aphid vector, Toxoptera citricida. Appl Environ Microbiol 82:6294–6302

    Article  CAS  Google Scholar 

  15. Harper SJ, Killiny N, Tatineni S et al (2016) Sequence variation in two genes determines the efficacy of transmission of citrus tristeza virus by the brown citrus aphid. Arch Virol 161:3555–3559

    Article  CAS  Google Scholar 

  16. Yokomi RK, Oldfield GN (1991) Seasonal fluctuations of alate aphid activity in California citrus groves. In: Brlansky RH, Lee RF, Timmer LW (eds) Proceeding of the 11th Conference of the International Organization of Citrus Virologists, Orlando, November 1989. IOCV, Riverside, pp 71–76

    Google Scholar 

  17. Roistacher CN (1991) The plant laboratory. In: Graft-transmissible disease of citrus: Handbook for detection and diagnosis. International Organization of Citrus Virologists and Food and Agriculture Organization of the United Nations, Rome, pp 159–189

    Google Scholar 

  18. Bar-Joseph M, Garnsey SM, Gonsalves D et al (1979) The use of enzyme-linked immunosorbent assay for detection of citrus tristeza virus. Phytopathology 69:190–194

    Article  Google Scholar 

  19. Garnsey SM, Cambra M (1991) Enzyme-linked immunosorbent assay (ELISA) for citrus pathogens. In: Roistacher CN (ed) Graft-transmissible diseases of citrus handbook for detection and diagnosis. FAO, Rome, pp 193–209

    Google Scholar 

  20. Hilf ME, Mavrodieva VA, Garnsey SM (2005) Genetic marker analysis of a global collection of isolates of Citrus tristeza virus: Characterization and distribution of CTV genotypes and association with symptoms. Phytopathology 95:909–917

    Article  CAS  Google Scholar 

  21. Saponari M, Keremane M, Yokomi RK (2008) Quantitative detection of Citrus tristeza virus in citrus and aphids by real-time reverse transcription-PCR (TaqMan®). J Virol Methods 147:43–53

    Article  CAS  Google Scholar 

  22. Saponari M, Yokomi RK (2010) Use of the coat protein (CP) and minor CP intergene sequence to discriminate severe strains of Citrus tristeza virus in three U.S. CTV isolate collections. In: Hilf ME, Milne RG, Timmer LW et al (eds) Proceedings of the 17th Conference of the International Organization of Citrus Virologists, Adana, October 2007. IOCV, Riverside, pp 43–57

    Google Scholar 

  23. Yokomi RK, Saponari M, Sieburth PJ (2010) Rapid differentiation and identification of potential severe strains of Citrus tristeza virus by real-time reverse transcription-polymerase chain reaction assays. Phytopathology 100:319–327

    Article  CAS  Google Scholar 

  24. Harper SJ, Cowell SJ, Dawson WO (2015) With a little help from my friends: complementation as a survival strategy for viruses in a long-lived host system. Virology 478:123–128

    Article  CAS  Google Scholar 

  25. Yokomi R, Selvaraj V, Maheshwari Y et al (2018) Molecular and biological characterization of a novel mild strain of citrus tristeza virus in California. Arch Virol 163:1795–1804

    Article  CAS  Google Scholar 

  26. Garnsey SM, Civerolo EL Gumpf DJ et al (2005) Biocharacterization of an international collection of citrus tristeza virus (CTV) isolates. In: Hilf ME, Durán-Villa N, Rocha-Peña MA (eds) Proceedings of the 16th Conference of the International Organization of Citrus Virologists, Monterrey, November 2004. IOCV, Riverside, pp 75–93

    Google Scholar 

  27. Blackman RL, Eastop VF (1984) Aphid on the world’s crops: an identification guide. Wiley, Chichester

    Google Scholar 

  28. Kono T, Papp CS (1977) Aphids. In: Handbook of agricultural pests: aphids, thrips, mites, snails and slugs. CDFA, DPI, Laboratory Services – Entomology, Sacramento, pp 13–86

    Google Scholar 

  29. Garnsey SM, Gumpf DJ, Roistacher CN et al (1987) Toward a standardized evaluation of the biological properties of citrus tristeza virus (CTV) isolates. Phytophylactica 19:151–157

    Google Scholar 

  30. Irwin ME, Ruesink WG (1986) Vector intensity: a product of propensity and activity. In: McLean GD, Garrett RG, Ruesink WG (eds) Plant virus epidemics. Academic Press, Sydney, pp 13–33

    Google Scholar 

  31. Marroquín C, Olmos A, Gorris MT et al (2004) Estimation of the number of aphids carrying Citrus tristeza virus that visit adult citrus trees. Virus Res 100:101–108

    Article  Google Scholar 

Download references

Acknowledgments

This work was supported by the USDA Agricultural Research Service, In-House Appropriated Project Number 2034-22000-013-00-D.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Raymond Yokomi .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

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

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Yokomi, R. (2019). CTV Vectors and Interactions with the Virus and Host Plants. In: Catara, A., Bar-Joseph, M., Licciardello, G. (eds) Citrus Tristeza Virus. Methods in Molecular Biology, vol 2015. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9558-5_4

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9558-5_4

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9557-8

  • Online ISBN: 978-1-4939-9558-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics