Methods for the Diagnosis of Grapevine Viral Infections: A Review

Abstract

The yielding capacity of grapevine growth and the quality of the resulting product heavily depend on the health of the cultivated plants. The phytopathogens affecting the vineyards can cause a significant reduction in the yield and quality of the product. For this reason, it is extremely important to use diagnostic methods that make it possible to identify the pathogens, and to choose the correct method of plant protection. This review considers the main viral grapevine pathogens, and the existing methods of their diagnosis. The limitations of conventional diagnostic methods that are based either on the visual assessment of symptoms, or on bio-testing, are analyzed. A major focus is placed on two intensively developed approaches of diagnosis, molecular genetic and immunochemical methods. Applications of amplification techniques and DNA chips are presented, as well as opportunities for next-generation sequencing. A reduction of assay duration and labor intensity in combination with the assay shifts from specialized laboratories toward the places of sampling are considered as the main factors influencing the development of immunodiagnostic techniques. The potential place of diagnostic tests in vine-growing practices, and the requirements for their most efficient applications for early disease diagnosis is also discussed.

Determination of the grapevine virome by high-throughput sequencing and grapevine viruses detection in Serra Gaúcha, Brazil

“…Parameters such yield and grape quality are heavily affected by viral pathogens. As a consequence, adequate diagnosis and identification of viruses is of fundamental relevance for efficient plant protection (Zherdev et al, 2018).…”

Grapevine Virology in the Third-Generation Sequencing Era: From Virus Detection to Viral Epitranscriptomics

“…Grapevine fleck virus (GFkV), Grapevine Red Blotch-associated Virus (GRBV), GLRaV-1-5, -7, and -9, Grapevine virus A (GVA), Grapevine virus B (GVB) and Grapevine virus D (GVD), have been detected through PCR amplification-based methods in the laboratory or on-site [19]. Some techniques, in particular LAMP, have improved the nucleic acid amplification-based detection of grapevine viruses.…”

“…Some techniques, in particular LAMP, have improved the nucleic acid amplification-based detection of grapevine viruses. Indeed, high speed and specificity, resistance to inhibitors, and colorimetric detection features have made LAMP a popular diagnostic method [19]. LAMP uses a set of four or six different primers from various regions of a viral genome.…”

“…These methods include virion-associated nucleic acids (VANAs) and doublestranded RNA (dsRNA) extraction, small interfering RNA (siRNA), and rRNA depleted and polyadenylated RNA isolation. Many known and novel grapevine viruses have been detected and identified with these methods, such as Grapevine Syrah virus 1 (GSyV-1), through total RNA and dsRNA sequencing, Grapevine vein-clearing virus (GVCV) and Grapevine Pinot gris virus (GPGV), through small RNA sequencing, Grapevine virus F (GVF) and Grapevine red blotch associated virus (GRBaV), through dsRNA sequencing, and Grapevine Roditis leaf discolorationassociated virus (GRLDaV), through siRNA sequencing [19,[25][26][27][28]. Although second-generation sequencing has been broadly used in virus identification and detection, several limitations remain, such as read lengths, GC content and amplification biases, laborious and costly library preparation methods, data management, and requirement of sophisticated technical expertise for data analysis [29,30].…”

Analysis of Virome by High-Throughput Sequencing Revealed Multiple Infection and Intra-Virus Diversity in a Single Grapevine Plant

“…A rapid, specific and effective diagnose integrated to the certification schemes is one of the most important tools to control grapevine viruses (Zherdev et al, 2018). However, the effectiveness of such measures can be negatively affected by a high virus genetic variability and occurrence of divergent variants escaping the detection (Glasa et al, 2015).…”