Abstract
The citrus systemic diseases, including citrus Huanglongbing (caused by Candidatus Liberibacter asiaticus (CLas)), citrus tristeza (caused by citrus tristeza virus (CTV)), citrus tatter leaf (caused by citrus tatter leaf virus (CTLV)), and citrus exocortis (caused by citrus exocortis viroid (CEVd)), are threats to citrus production in Taiwan. Reliable diagnostic methods are important for the management of these systemic diseases. In this study, we developed a multiplex reverse transcription–polymerase chain reaction (RT-PCR) assay to detect four pathogens simultaneously. Herein, the specific amplicons from each pathogen (295 bp for CLas, 468 bp for CTV, 120 bp for CTLV, and 196 bp for CEVd) were successfully produced using the optimized multiplex RT-PCR described here. The sensitivity evaluation showed that low titers of pathogens could be detected using this multiplex RT-PCR. Compared with the published simplex assays, the detection of field samples using the multiplex RT-PCR developed in this study showed a better performance. The detections using multiplex RT-PCR revealed that these four citrus systemic pathogens were commonly found in fields, and 30.0% of field samples were mix-infected. To our knowledge, this is the first study of a survey of the four important citrus systemic diseases in Taiwan, and it provides insights for improving disease management. Therefore, the multiplex RT-PCR assay provides a useful method for routine disease surveying and the production of pathogen-free citrus plants.
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Claims 8
Claim · model-written summary
The multiplex assay produced a 295 bp amplicon for CLas.
CitrusOrganisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“295 bp for CLas”
e1“specific amplicons”
t0“Citrus”
Claim · model-written summary
The multiplex assay produced a 468 bp amplicon for CTV.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“468 bp for CTV”
e1“specific amplicons”
t0“Citrus”
Claim · model-written summary
The multiplex assay produced a 120 bp amplicon for CTLV.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“120 bp for CTLV”
e1“specific amplicons”
t0“Citrus”
Claim · model-written summary
The multiplex assay produced a 196 bp amplicon for CEVd.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“196 bp for CEVd”
e1“specific amplicons”
t0“Citrus”
Claim · model-written summary
The multiplex assay detected low pathogen titers.
- Outcome
- low titers of pathogense0
Not reportedComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“low titers of pathogens could be detected using this multiplex RT-PCR”
t0“Citrus”
Claim · model-written summary
The multiplex assay performed better than published simplex assays on field samples.
- Outcome
- detection of field samplese0
- Direction
- better performancee0
- Comparator
- published simplex assayse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Compared with the published simplex assays, the detection of field samples using the multiplex RT-PCR developed in this study showed a better performance”
t0“Citrus”
Claim · model-written summary
All four citrus systemic pathogens were commonly found in fields.
- Outcome
- commonly found in fieldse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“these four citrus systemic pathogens were commonly found in fields”
Claim · model-written summary
Thirty percent of field samples had mixed infection.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“30.0% of field samples were mix-infected”
t0“Citrus”