دورية أكاديمية
High-resolution melting analysis : a new molecular approach for the early detection of Diplodia pinea in Austrian pine
| العنوان: | High-resolution melting analysis : a new molecular approach for the early detection of Diplodia pinea in Austrian pine |
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| المؤلفون: | Luchi, Nicola, Pratesi, Nicola, Simi, Lisa, Pazzagli, Mario, Capretti, Paolo, Scala, Aniello, Slippers, Bernard, Pinzani, Pamela |
| بيانات النشر: | Elsevier |
| سنة النشر: | 2011 |
| المجموعة: | University of Pretoria: UPSpace |
| مصطلحات موضوعية: | High-resolution melting analysis (HRMA), mt SSU rDNA, Pinus nigra, Botryosphaeriaceae, Austrian pine -- Diseases and pests -- Identification, Diplodia |
| الوصف: | The differentiation of Diplodia pinea from closely related species, such as Diplodia scrobiculata and Diplodia seriata, and its detection in plant tissue, represented a critical issue for a long time. Molecular screening tools have recently been developed to address this topic. In this study we applied one of the most sensitive and rapid diagnostic screening method so far developed, called High-Resolution Melting Analysis (HRMA), to detect D. pinea in Austrian pine (Pinus nigra). HRMA exploits differences in the melting behaviour of PCR products to rapidly identify DNA sequence variants without the need for cumbersome post-PCR methods. We developed a HRMA method to detect specific fungal sequences in the mitochondrial small subunit ribosome gene (mt SSU rDNA). The reliability of this technique was firstly assessed on DNA extracted from pure cultures of D. pinea and closely related species. Amplicon differences were screened by HRMA and the results confirmed by direct DNA sequencing. Subsequently, HRMA was tested on DNA from symptomatic and symptomless pine shoots, and the presence of the fungus was also confirmed by both conventional and molecular quantitative approaches. The HRMA allowed the distinction of D. pinea from closely related species, showing specific melting profiles for the each pathogen. This new molecular technique, here tested in a plantefungus pathosystem for the first time, was very reliable in both symptomatic and symptomless shoots. HRMA is therefore a highly effective and accurate technique that permits the rapid screening of pathogens in the host. ; This work was supported by grant to Nicola Luchi from the Consorzio Interuniversitario Biotecnologie (CIB, Italy). ; http://www.elsevier.com/locate/funbioTest ; nf2012 |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| العلاقة: | http://hdl.handle.net/2263/18979Test; Nicola Luchi, Nicola Pratesi, Lisa Simi, Mario Pazzagli, Paolo Capretti, Aniello Scala, Bernard Slippers & Pamela Pinzani, High-resolution melting analysis : a new molecular approach for the early detection of Diplodia pinea in Austrian pine, Fungal Biology, vol. 115, no. 8, pp. 715-723 (2011), doi:10.1016/j.funbio.2011.05.005; 1878-6146 (print); 1878-6162 (online) |
| DOI: | 10.1016/j.funbio.2011.05.005 |
| الإتاحة: | https://doi.org/10.1016/j.funbio.2011.05.005Test http://hdl.handle.net/2263/18979Test |
| حقوق: | © 2011 The British Mycological Society. Published by Elsevier Ltd. All rights reserved. Notice : this is the author’s version of a work that was accepted for publication in Fungal Biology. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Fungal Biology, vol. 115, issue 8, August 2011, doi:10.1016/j.funbio.2011.05.005. |
| رقم الانضمام: | edsbas.3E0A57F5 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1016/j.funbio.2011.05.005 |
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