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CTV Vectors and Interactions with the Virus and Host Plants
1San Joaquin Valley Agricultural Sciences Center, USDA-ARS, Parlier, CA, USA. Ray.Yokomi@ARS.USDA.GOV.
Methods in Molecular Biology (Clifton, N.J.)
|June 22, 2019
Summary
Characterizing Citrus tristeza virus (CTV) isolates requires aphid vectors to separate strains. This method, using Toxoptera citricida or Aphis gossypii, aids in understanding CTV diversity and disease management.
Area of Science:
- Plant Pathology
- Entomology
- Horticultural Science
Background:
- Citrus tristeza virus (CTV) is a graft-transmissible pathogen affecting perennial citrus crops.
- CTV exists as a complex of strains and isolates, often transmitted semi-persistently by aphid vectors.
- Citrus trees frequently harbor multiple CTV strains due to natural spread.
Purpose of the Study:
- To outline the methodology for using aphid vectors to purify and characterize Citrus tristeza virus (CTV) field isolates.
- To detail the requirements for establishing and conducting CTV transmission studies using aphid vectors.
- To emphasize the importance of virus indexing for confirming the phenotype of aphid-transmitted CTV isolates.
Main Methods:
- Utilizing specific aphid vectors, Toxoptera citricida or Aphis gossypii, for efficient CTV transmission.
- Maintaining virus-free aphid colonies and insect-proof, climate-controlled facilities for vector studies.
- Employing a 24-hour acquisition and inoculation feeding period for optimal virus transmission by vectors.
- Determining CTV infection via serological assays (polyclonal or monoclonal antiserum) and molecular genotyping (RT-PCR, RT-qPCR).
- Conducting virus indexing through graft inoculation to citrus hosts to evaluate CTV-induced symptoms.
Main Results:
- Aphid vectors effectively 'clean up' CTV populations from source trees, facilitating strain separation.
- Established protocols enable efficient transmission and characterization of CTV isolates.
- Serological and molecular methods confirm CTV presence, but graft indexing is crucial for phenotypic analysis.
Conclusions:
- Aphid-mediated transmission is a critical technique for isolating and characterizing individual CTV strains from complex field populations.
- Robust experimental conditions, including aphid colonies and controlled environments, are essential for successful CTV vector studies.
- Phenotypic characterization of CTV isolates, post-vector transmission, necessitates host range evaluation and symptom assessment.
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