Development of Multiplex RT-PCR Assay for the Simultaneous Detection of Four Systemic Diseases Infecting Citrus

Shun-Min Yao, Meng-Ling Wu and Ting-Hsuan Hung ()
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Shun-Min Yao: Department of Plant Pathology and Microbiology, National Taiwan University, Taipei 10617, Taiwan
Meng-Ling Wu: Division of Forest Protection, Taiwan Forestry Research Institute, Taipei 10066, Taiwan
Ting-Hsuan Hung: Department of Plant Pathology and Microbiology, National Taiwan University, Taipei 10617, Taiwan

Agriculture, 2023, vol. 13, issue 6, 1-14

Abstract: The citrus systemic diseases, including citrus Huanglongbing (caused by Candidatus Liberibacter asiaticus ( C Las)), 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 C Las, 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.

Keywords: citrus Huanglongbing; citrus tristeza; citrus tatter leaf; citrus exocortis; multiplex RT-PCR (search for similar items in EconPapers)
JEL-codes: Q1 Q10 Q11 Q12 Q13 Q14 Q15 Q16 Q17 Q18 (search for similar items in EconPapers)
Date: 2023
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