TaqMan qPCR Detection and Quantification of Phytophthora cinnamomi in Soil and Plant Tissues for Walnut Disease Management

Anita Haegi (), Laura Luongo, Salvatore Vitale, Lorenza Tizzani and Alessandra Belisario
Additional contact information
Anita Haegi: Council for Agricultural Research and Economics, (CREA)—Plant Protection and Certification (DC), Via C. G. Bertero 22, 00156 Roma, Italy
Laura Luongo: Council for Agricultural Research and Economics, (CREA)—Plant Protection and Certification (DC), Via C. G. Bertero 22, 00156 Roma, Italy
Salvatore Vitale: Council for Agricultural Research and Economics, (CREA)—Plant Protection and Certification (DC), Via C. G. Bertero 22, 00156 Roma, Italy
Lorenza Tizzani: Council for Agricultural Research and Economics, (CREA)—Plant Protection and Certification (DC), Via C. G. Bertero 22, 00156 Roma, Italy
Alessandra Belisario: Council for Agricultural Research and Economics, (CREA)—Plant Protection and Certification (DC), Via C. G. Bertero 22, 00156 Roma, Italy

Agriculture, 2024, vol. 14, issue 7, 1-17

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. 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.

Keywords: real-time PCR; tree decline; diagnostic; early detection; soil detection (search for similar items in EconPapers)
JEL-codes: Q1 Q10 Q11 Q12 Q13 Q14 Q15 Q16 Q17 Q18 (search for similar items in EconPapers)
Date: 2024
References: View complete reference list from CitEc
Citations:

Downloads: (external link)
https://www.mdpi.com/2077-0472/14/7/999/pdf (application/pdf)
https://www.mdpi.com/2077-0472/14/7/999/ (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:gam:jagris:v:14:y:2024:i:7:p:999-:d:1422189

Access Statistics for this article

Agriculture is currently edited by Ms. Leda Xuan

More articles in Agriculture from MDPI
Bibliographic data for series maintained by MDPI Indexing Manager ().