Next Generation Sequencing, and Development of a Pipeline as a Tool for the Detection and Discovery of Citrus Pathogens to Facilitate Safer Germplasm Exchange

Citrus is affected by many diseases, and hence, the movement of citrus propagative materials is highly regulated in the USA. Currently used regulatory pathogen detection methods include biological and laboratory-based technologies, which are time-consuming, expensive, and have many limitations. Ther...

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Bibliographic Details
Published in:Plants (Basel) 2024-01, Vol.13 (3), p.411
Main Authors: Keremane, Manjunath, Singh, Khushwant, Ramadugu, Chandrika, Krueger, Robert R, Skaggs, Todd H
Format: Article
Language:English
Subjects:
Bioinformatics
biological indexing
budwood certification
Causes of
Certification
citrus germplasm
Citrus greening
Disease
DNA sequencing
Fruits
Genetic aspects
Genomes
Germplasm
Germplasm resources, Plant
Hybrids
Identification and classification
Laboratories
Next-Generation Sequencing
Nucleotide sequencing
pathogen testing
Pathogenic microorganisms
Pathogens
Plant bacterial diseases
Plant viruses
Quarantine
real time PCR
Serology
Success
Transcriptomes
Viruses
Workflow
Yellows
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Summary:Citrus is affected by many diseases, and hence, the movement of citrus propagative materials is highly regulated in the USA. Currently used regulatory pathogen detection methods include biological and laboratory-based technologies, which are time-consuming, expensive, and have many limitations. There is an urgent need to develop alternate, rapid, economical, and reliable testing methods for safe germplasm exchange. Citrus huanglongbing (HLB) has devastated citrus industries leading to an increased need for germplasm exchanges between citrus growing regions for evaluating many potentially valuable hybrids for both HLB resistance and multilocational performance. In the present study, Next-Generation Sequencing (NGS) methods were used to sequence the transcriptomes of 21 test samples, including 15 well-characterized pathogen-positive plants. A workflow was designed in the CLC Genomics Workbench software, v 21.0.5 for bioinformatics analysis of the sequence data for the detection of pathogens. NGS was rapid and found to be a valuable technique for the detection of viral and bacterial pathogens, and for the discovery of new citrus viruses, complementary to the existing array of biological and laboratory assays. Using NGS methods, we detected beet western yellows virus, a newly reported citrus virus, and a variant of the citrus yellow vein-associated virus associated with the "fatal yellows" disease.
ISSN:2223-7747
2223-7747
DOI:10.3390/plants13030411