Highly specific qPCR and amplicon sequencing method for detection of quarantine citrus pathogen Phyllosticta citricarpa applicable for air samples

Zajc, Janja (avtor), Kogej Zwitter, Zala (avtor), Fišer, Sara (avtor), Gostinčar, Cene (avtor), Vicent, Antonio (avtor), Galvañ Domenech, Anaïs (avtor), Riccioni, Luca (avtor), Boonham, Neil (avtor), Ravnikar, Maja (avtor), Kogovšek, Polona (avtor)

The fungus Phyllosticta citricarpa is a quarantine pathogen in the EU and is of high economic importance in many parts of the world where favourable climate conditions drive the development of citrus black spot (CBS) disease. Disease symptoms include necrotic lesions on leaves and fruits. Low disease pressure can reduce crop market-ability, while higher disease pressure can cause premature fruit drop, significantly increasing crop losses. The wind-dispersed spores of P. citricarpa are especially prob-lematic for rapid pathogen dispersal, but also provide an opportunity for early detec-tion of the disease spreading into a new area. In this study we have developed and validated a quantitative PCR (qPCR) assay based on the TEF1-α sequence. Specificity testing demonstrated that it is currently the only qPCR assay that does not cross- react with closely related Phyllosticta species. The assay is sensitive and can detect a single copy of the TEF1 gene in a reaction, it is highly repeatable and reproducible and can be used for testing of the sticky tapes from spore traps as well as citrus fruit sam-ples. High-throughput sequencing (HTS) of the DNA barcodes ITS1 and TEF1 was also explored for the detection and discrimination of P. citricarpa. The limit of detection of the HTS was 1000 spores on a daily spore trap tape. This study makes an important improvement to the diagnostics of the CBS and the methods developed can also be applied to improve the surveillance and early detection of the pathogen when linked to spore samplers in the field.

• Jezik: Angleški jezik
• Ključne besede: detection, fungal spore sampling, internal transcribed region, ITS, translation elongation factor 1-α, TEF1
• Vrsta gradiva: Članek v reviji
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: BF - Biotehniška fakulteta
• Status publikacije: Objavljeno
• Različica publikacije: Objavljena publikacija
• Leto izida: 2023
• Št. strani: Str. 548-563
• Številčenje: Vol. 72, iss. 3
• PID: 20.500.12556/RUL-145002
• UDK: 632
• ISSN pri članku: 1365-3059
• DOI: 10.1111/ppa.13679
• COBISS.SI-ID: 136443139
• Datum objave v RUL: 29.03.2023

Gradivo je del revije

• Naslov: Plant pathology
• Skrajšan naslov: Plant pathol.
• Založnik: British Society for Plant Pathology, Wiley
• ISSN: 1365-3059
• COBISS.SI-ID: 517809945

Licence

• Licenca: CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by/4.0/deed.sl
• Opis: To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

• Jezik: Slovenski jezik
• Ključne besede: Phyllosticta citricarpa, detekcija, citrusi, zrak