Publicação: Colletotrichum abscissum: Detection on symptomless citrus leaves and symptomatic citrus flowers using high-resolution melting analysis
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Data
2022
Orientador
Coorientador
Pós-graduação
Curso de graduação
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Editor
Wiley
Tipo
Artigo
Direito de acesso
Acesso aberto
Resumo
High-resolution melting (HRM) analysis has been recently applied for the diagnosis of plant diseases. This
cost-effective method is advantageous over standard PCR as it does not require DNA band visualization or the use of probes necessary for quantitative PCR. Colletotrichum
abscissum causes postbloom fruit drop of citrus (PFD) and survives asymptomatically on
vegetative tissue. Quiescent appressoria on citrus leaves are the primary source of inoculum
between flowering periods. Early PFD symptoms may be misidentified as physical damage on
citrus flowers. Our objectives were to validate an HRM-based method to rapidly identify C.
abscissum on citrus leaves and flowers. We screened seven previously published primer pairs
and concluded that the most effective set of primers was CaITS-F and CaITS-R815. We
evaluated three different DNA extraction methods, two with a purification step (DNeasy Plant
Mini and PowerSoil kits) and a faster method without a purification step (buffer GEB2). We
inoculated citrus leaves with C. abscissum conidial suspensions of varying concentrations and
HRM detected as few as 200 C. abscissum spores using DNeasy Plant Mini or DNeasy
PowerSoil kits for DNA extraction. As expected, samples extracted with GEB2 buffer reduced
HRM sensitivity. We further collected field samples from areas with high and low C. abscissum
inoculum levels. The HRM method could detect the pathogen in high inoculum areas from both
leaves and flowers regardless of the extraction method used. In summary, we describe a new
molecular diagnostic tool to detect C. abscissum on citrus during its asymptomatic and
symptomatic stages.
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