Fast and Sensitive Detection of Soil-Borne Cereal Mosaic Virus in Leaf Crude Extract of Durum Wheat

Monica Marra; Chiara D'Errico; Cinzia Montemurro; Claudio Ratti; Elena Baldoni; Slavica Matic; Gian Paolo Accotto

doi:10.3390/v15010140

Fast and Sensitive Detection of Soil-Borne Cereal Mosaic Virus in Leaf Crude Extract of Durum Wheat

Viruses. 2022 Dec 31;15(1):140. doi: 10.3390/v15010140.

Authors

Monica Marra^{1

2}, Chiara D'Errico^{1

3}, Cinzia Montemurro^{2

4}, Claudio Ratti⁵, Elena Baldoni⁶, Slavica Matic¹, Gian Paolo Accotto¹

Affiliations

¹ Institute for Sustainable Plant Protection, National Research Council, 10135 Turin, Italy.
² Department of Soil, Plant and Food Science, University of Bari Aldo Moro, 70126 Bari, Italy.
³ European Laboratory for Non-Linear Spectroscopy, LENS, 50019 Sesto Fiorentino, Italy.
⁴ Institute for Sustainable Plant Protection, National Research Council, 70126 Bari, Italy.
⁵ Department of Agricultural and Food Sciences, University of Bologna, 40127 Bologna, Italy.
⁶ Institute of Agricultural Biology and Biotechnology, National Research Council, 20133 Milan, Italy.

Abstract

Soil-borne cereal mosaic virus (SBCMV) is a furovirus with rigid rod-shaped particles containing an ssRNA genome, transmitted by Polymyxa graminis Led., a plasmodiophorid that can persist in soil for up to 20 years. SBCMV was reported on common and durum wheat and it can cause yield losses of up to 70%. Detection protocols currently available are costly and time-consuming (real-time PCR) or have limited sensitivity (ELISA). To facilitate an efficient investigation of the real dispersal of SBCMV, it is necessary to develop a new detection tool with the following characteristics: no extraction steps, very fast results, and high sensitivity to allow pooling of a large number of samples. In the present work, we have developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) protocol with such characteristics, and we have compared it with real-time PCR. Our results show that the sensitivity of LAMP and real-time PCR on cDNA and RT-LAMP on crude extracts are comparable, with the obvious advantage that RT-LAMP produces results in minutes rather than hours. This paves the way for extensive field surveys, leading to a better knowledge of the impact of this virus on wheat health and yield.

Keywords: Furovirus; RT-LAMP; SBCMV; durum wheat; real-time PCR.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Complex Mixtures
Nucleic Acid Amplification Techniques / methods
Plant Leaves
Plant Viruses* / genetics
Reverse Transcription
Sensitivity and Specificity
Triticum*

Substances

Complex Mixtures

Supplementary concepts

Soil-borne cereal mosaic virus

Grants and funding

This research was funded by Regione Lombardia, dds n. 4403 28/03/2018, grant n° 42—“SURF” and supported through the Euphresco network for phytosanitary research coordination and funding, 2021-A-374: “Diagnosis and epidemiology of viruses infecting cereal crops”. This publication was also supported by the European Virus Archive GLOBAL (EVA-GLOBAL) project that has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No. 871029. M.M. was supported by the “SURF” Project.