Rapid Detection of Pine Pathogens Lecanosticta acicola, Dothistroma pini and D. septosporum on Needles by Probe-Based LAMP Assays
| Autor: | Irene Barnes, Colton D. Meinecke, Luisa Ghelardini, Ariska van der Nest, Chiara Aglietti, Caterina Villari |
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| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
Diagnostic methods forest health protection Loop-mediated isothermal amplification red band needle blight Early detection Biology 01 natural sciences Rapid detection phytosanitary surveillance 030308 mycology & parasitology Microbiology 03 medical and health sciences field-portable fungal diagnostics diagnostica molecolare malattia delle bande rosse del pino loop-mediated isothermal amplification brown spot needle blight red band needle blight Pinus Mycosphaerella dearnessii field-portable fungal diagnostics forest health protection phytosanitary surveillance Mycosphaerella dearnessii Blight Phytosanitary certification 0303 health sciences Lecanosticta acicola Dothistroma pini Forestry lcsh:QK900-989 15. Life on land lcsh:Plant ecology brown spot needle blight Pinus loop-mediated isothermal amplification 010606 plant biology & botany |
| Zdroj: | Università degli studi di Firenze-IRIS Forests Volume 12 Issue 4 Forests, Vol 12, Iss 479, p 479 (2021) |
| ISSN: | 1999-4907 |
| DOI: | 10.3390/f12040479 |
| Popis: | Needle blights are serious needle fungal diseases affecting pines both in natural and productive forests. Among needle blight agents, the ascomycetes Lecanosticta acicola, Dothistroma pini and D. septosporum are of particular concern. These pathogens need specific, fast and accurate diagnostics since they are regulated species in many countries and may require differential management measures. Due to the similarities in fungal morphology and the symptoms they elicit, these species are hard to distinguish using morphological characteristics. The symptoms can also be confused with those caused by insects or abiotic agents. DNA-based detection is therefore recommended. However, the specific PCR assays that have been produced to date for the differential diagnosis of these pathogens can be applied only in a well-furnished laboratory and the procedure takes a relatively long execution time. Surveillance and forest protection would benefit from a faster diagnostic method, such as a loop-mediated isothermal amplification (LAMP) assay, which requires less sophisticated equipment and can also be deployed directly on-site using portable devices. LAMP assays for the rapid and early detection of L. acicola, D. pini and D. septosporum were developed in this work. Species-specific LAMP primers and fluorescent assimilating probes were designed for each assay, targeting the beta tubulin (β-tub2) gene for the two Dothistroma species and the elongation factor (EF-1α) region for L. acicola. Each reaction detected its respective pathogen rapidly and with high specificity and sensitivity in DNA extracts from both pure fungal cultures and directly from infected pine needles. These qualities and the compatibility with inexpensive portable instrumentation position these LAMP assays as an effective method for routine phytosanitary control of plant material in real time, and they could profitably assist the management of L. acicola, D. pini and D. septosporum. |
| Databáze: | OpenAIRE |
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