Development of a Multiplex RT-PCR Detection for Six Viruses Infecting Strawberry

Abstract

Strawberry viruses are significant pathogenic agents in strawberry. The development and application of efficient virus detection technology can effectively reduce the economic losses incurred by virus diseases for strawberry cultivators. In order to rapidly identify strawberry virus species and prevent the spread of virus disease, a multiplex reverse transcription polymerase chain reaction system was established for the simultaneous detection and identification of strawberry mild yellow edge virus (SMYEV), strawberry vein banding virus (SVBV), strawberry mottle virus (SMoV), strawberry polerovirus 1 (SPV-1), strawberry pallidosis-associated virus (SPaV), and strawberry crinivirus 4 (SCrV-4). In this study, six pairs of specific primers were designed on the conserved genomic regions of these viruses. The primer concentration, annealing temperature, and amplification cycle number of the reaction system were optimized. Subsequent sensitivity testing and application of the optimized detection system were carried out. The results indicate the establishment of an efficient detection system for strawberry viruses. The optimal reaction can detect the six viruses at the same time, which provides technical support for the early prevention and treatment of strawberry virus diseases.