Development of a Real-Time PCR Assay for the Early Detection of the Eucalyptus Pathogen <i>Quambalaria eucalypti</i>
<i>Quambalaria eucalypti</i> is a fungal pathogen that causes leaf spot, shoot blight, and stem canker on <i>Eucalyptus</i> spp. Early diagnosis of the disease is difficult, although the symptoms are clear in its advanced phase. To enable a rapid and sensitive screening of as...
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-02-01
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| Series: | Forests |
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| Online Access: | https://www.mdpi.com/1999-4907/15/2/375 |
| _version_ | 1797298197472739328 |
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| author | Roberto Faedda Gabriela B. Silva |
| author_facet | Roberto Faedda Gabriela B. Silva |
| author_sort | Roberto Faedda |
| collection | DOAJ |
| description | <i>Quambalaria eucalypti</i> is a fungal pathogen that causes leaf spot, shoot blight, and stem canker on <i>Eucalyptus</i> spp. Early diagnosis of the disease is difficult, although the symptoms are clear in its advanced phase. To enable a rapid and sensitive screening of asymptomatic or latently infected plant material for <i>Q. eucalypti</i>, a SYBR green-based real-time PCR assay targeting the partial histone-H3 region was developed. The assay demonstrated specificity for <i>Q. eucalypti</i>, not showing cross-reactivity with other <i>Quambalaria</i> species or the other eucalyptus fungal pathogens tested. The primers developed in this study ensured high analytical sensitivity, allowing the detection of <i>Q. eucalypti</i> DNA concentrations as low as 10 fg DNA from asymptomatic plants. The robustness and efficacy of the assay was demonstrated by interlaboratory comparisons with similar results. This newly developed quantitative PCR assay can be used for more comprehensive epidemiological investigations, testing the plant material in known <i>Q. eucalypti</i> distribution areas for early management strategies, or collecting data for resistance breeding programs. |
| first_indexed | 2024-03-07T22:31:34Z |
| format | Article |
| id | doaj.art-4ef5f4fc8852448c954741379874c1ae |
| institution | Directory Open Access Journal |
| issn | 1999-4907 |
| language | English |
| last_indexed | 2024-03-07T22:31:34Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Forests |
| spelling | doaj.art-4ef5f4fc8852448c954741379874c1ae2024-02-23T15:17:05ZengMDPI AGForests1999-49072024-02-0115237510.3390/f15020375Development of a Real-Time PCR Assay for the Early Detection of the Eucalyptus Pathogen <i>Quambalaria eucalypti</i>Roberto Faedda0Gabriela B. Silva1Department of Agriculture, Food and Environment, University of Catania, 95123 Catania, ItalyAmarelo Agroflorestal Ltda., Porto Alegre 90020180, Brazil<i>Quambalaria eucalypti</i> is a fungal pathogen that causes leaf spot, shoot blight, and stem canker on <i>Eucalyptus</i> spp. Early diagnosis of the disease is difficult, although the symptoms are clear in its advanced phase. To enable a rapid and sensitive screening of asymptomatic or latently infected plant material for <i>Q. eucalypti</i>, a SYBR green-based real-time PCR assay targeting the partial histone-H3 region was developed. The assay demonstrated specificity for <i>Q. eucalypti</i>, not showing cross-reactivity with other <i>Quambalaria</i> species or the other eucalyptus fungal pathogens tested. The primers developed in this study ensured high analytical sensitivity, allowing the detection of <i>Q. eucalypti</i> DNA concentrations as low as 10 fg DNA from asymptomatic plants. The robustness and efficacy of the assay was demonstrated by interlaboratory comparisons with similar results. This newly developed quantitative PCR assay can be used for more comprehensive epidemiological investigations, testing the plant material in known <i>Q. eucalypti</i> distribution areas for early management strategies, or collecting data for resistance breeding programs.https://www.mdpi.com/1999-4907/15/2/375<i>Quambalaria eucalypti</i><i>Quambalaria</i> shoot blightqPCRdiagnosticsmolecular detection<i>Eucalyptus</i> |
| spellingShingle | Roberto Faedda Gabriela B. Silva Development of a Real-Time PCR Assay for the Early Detection of the Eucalyptus Pathogen <i>Quambalaria eucalypti</i> Forests <i>Quambalaria eucalypti</i> <i>Quambalaria</i> shoot blight qPCR diagnostics molecular detection <i>Eucalyptus</i> |
| title | Development of a Real-Time PCR Assay for the Early Detection of the Eucalyptus Pathogen <i>Quambalaria eucalypti</i> |
| title_full | Development of a Real-Time PCR Assay for the Early Detection of the Eucalyptus Pathogen <i>Quambalaria eucalypti</i> |
| title_fullStr | Development of a Real-Time PCR Assay for the Early Detection of the Eucalyptus Pathogen <i>Quambalaria eucalypti</i> |
| title_full_unstemmed | Development of a Real-Time PCR Assay for the Early Detection of the Eucalyptus Pathogen <i>Quambalaria eucalypti</i> |
| title_short | Development of a Real-Time PCR Assay for the Early Detection of the Eucalyptus Pathogen <i>Quambalaria eucalypti</i> |
| title_sort | development of a real time pcr assay for the early detection of the eucalyptus pathogen i quambalaria eucalypti i |
| topic | <i>Quambalaria eucalypti</i> <i>Quambalaria</i> shoot blight qPCR diagnostics molecular detection <i>Eucalyptus</i> |
| url | https://www.mdpi.com/1999-4907/15/2/375 |
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