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Authors = Anita Haegi ORCID = 0000-0003-0214-7623

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17 pages, 1572 KiB  
Article
TaqMan qPCR Detection and Quantification of Phytophthora cinnamomi in Soil and Plant Tissues for Walnut Disease Management
by Anita Haegi, Laura Luongo, Salvatore Vitale, Lorenza Tizzani and Alessandra Belisario
Agriculture 2024, 14(7), 999; https://doi.org/10.3390/agriculture14070999 - 26 Jun 2024
Cited by 4 | Viewed by 2216
Abstract
Phytophthora cinnamomi is a devastating soil-borne plant pathogen. The primary source of P. cinnamomi infection is the soil, where the pathogen can persist for long periods. Effective prevention and management of this pathogen in tree crops requires an early and reliable detection method. [...] Read more.
Phytophthora cinnamomi is a devastating soil-borne plant pathogen. The primary source of P. cinnamomi infection is the soil, where the pathogen can persist for long periods. Effective prevention and management of this pathogen in tree crops requires an early and reliable detection method. In this study, we developed a simple, fast, reliable, and sensitive method based on real-time quantitative PCR (qPCR) for P. cinnamomi detection and quantification directly in plant or soil samples. Primers were developed targeting the nuclear single-copy ras-related protein gene Ypt1, suitable for Phytophthora-specific PCR. The specificity of the assay was confirmed by testing it against genomic DNA from 50 isolates across eight different Phytophthora clades, including the very similar P. parvispora. The efficiency and reliability of the qPCR protocol were evaluated in challenging environmental samples, such as plant tissue of different host trees (walnut, chestnut, oak) and naturally infected soils in walnut orchards. The main outcome was the development of a qPCR method for the specific identification and quantification of P. cinnamomi in natural soil samples. Additionally, this study established a systematic and repeatable soil sampling method and developed an efficient soil DNA extraction technique to apply the developed qPCR in naturally infested soils of walnut orchards. Full article
(This article belongs to the Section Crop Protection, Diseases, Pests and Weeds)
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21 pages, 1920 KiB  
Review
New-Generation Sequencing Technology in Diagnosis of Fungal Plant Pathogens: A Dream Comes True?
by Maria Aragona, Anita Haegi, Maria Teresa Valente, Luca Riccioni, Laura Orzali, Salvatore Vitale, Laura Luongo and Alessandro Infantino
J. Fungi 2022, 8(7), 737; https://doi.org/10.3390/jof8070737 - 16 Jul 2022
Cited by 36 | Viewed by 6541
Abstract
The fast and continued progress of high-throughput sequencing (HTS) and the drastic reduction of its costs have boosted new and unpredictable developments in the field of plant pathology. The cost of whole-genome sequencing, which, until few years ago, was prohibitive for many projects, [...] Read more.
The fast and continued progress of high-throughput sequencing (HTS) and the drastic reduction of its costs have boosted new and unpredictable developments in the field of plant pathology. The cost of whole-genome sequencing, which, until few years ago, was prohibitive for many projects, is now so affordable that a new branch, phylogenomics, is being developed. Fungal taxonomy is being deeply influenced by genome comparison, too. It is now easier to discover new genes as potential targets for an accurate diagnosis of new or emerging pathogens, notably those of quarantine concern. Similarly, with the development of metabarcoding and metagenomics techniques, it is now possible to unravel complex diseases or answer crucial questions, such as “What’s in my soil?”, to a good approximation, including fungi, bacteria, nematodes, etc. The new technologies allow to redraw the approach for disease control strategies considering the pathogens within their environment and deciphering the complex interactions between microorganisms and the cultivated crops. This kind of analysis usually generates big data that need sophisticated bioinformatic tools (machine learning, artificial intelligence) for their management. Herein, examples of the use of new technologies for research in fungal diversity and diagnosis of some fungal pathogens are reported. Full article
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