Adjustment of RT-PCR-based molecular techniques for virus detection in strawberry plants

Abstract

Strawberry (Fragaria × ananassa Duch.), a globally consumed berry, holds major socio-economic significance in producing countries. Viral diseases significantly impact strawberry yield and quality. In Argentina, the predominant viruses affecting strawberry crops are: Strawberry Crinkle Virus (SCV), Strawberry Mild Yellow Edge Virus (SMYEV), Strawberry Mottle Virus (SMoV), and Strawberry Polerovirus 1 (SPV1). Tucumán, a key strawberry-producing province in Argentina, faces high viral disease incidence. Implementing highly sensitive diagnostic methods is therefore essential for effective phytosanitary monitoring and crop management. This study adapted molecular techniques for the concurrent detection of these four viruses in Argentina strawberries. We detail a reliable method for high-quality mRNA from strawberry leaves. Furthermore, novel specific primers were designed and RT-PCR conditions optimized for SCV, SMYEV, SMoV, and SPV1 detection. While SPV1 was detected solely in the positive control (submitted by IPAVE), SCV, SMYEV, and SMoV were identified in plants from a commercial field. None of the viruses were detected in plants from in vitro strawberry tissue culture. The established protocol provides a valuable tool for enhancing strawberry plant health surveillance programs.