Simultaneous detection of Apple mosaic virus in cultivated hazelnuts by one-tube RT-PCR

Abstract

The most economically damaging ilarvirus affecting hazelnut on a worldwide scale is the related apple mosaic virus (ApMV). Attempts were made to isolate the virus RNA from hazelnut tissues using different extraction methods. The most suitable extraction method that could detect the virus occurring naturally in hazelnut by reverse-transcription polymerase chain reaction (RT-PCR) methodology was selected. RT-PCR was applied successfully using flower, husk and leaf tissues. The most suitable extraction method and hazelnut tissues determined were sensitive, simple, rapid and reliable for simultaneous detection of ApMV in hazelnut tissues. To our knowledge, this is the first report of the simultaneous detection of the virus by RT-PCR, an alternative detection of ApMV in hazelnut hosts.

Is It Possible to Produce Certified Hazelnut Plant Material in Sicily? Identification and Recovery of Nebrodi Genetic Resources, in vitro Establishment, and Innovative Sanitation Technique From Apple Mosaic Virus

“…The obtained results confirmed the detection of the most economically damaging Ilarvirus affecting hazelnut by one step RT-PCR applied on a set of local Sicilian cultivars using dormant tissues, as described by Ertunc et al (2014) , overcoming the problem of having high amount of phenolic compounds, tannins in particular. The latter are considered a problem for the extraction of good quality RNA by several researchers that tried the herbaceous host inoculation ( Sokmen, 2003 ) and flower tissues extraction ( Akbas and Degirmenci, 2010 ). Moreover, this study provided further information on the sanitary status and ApMV incidence within eight Italian hazelnut genotypes showing the presence of the virus in all of them.…”

Tissue and Time Optimization for Real-Time Detection of Apple Mosaic Virus and Apple Necrotic Mosaic Virus Associated with Mosaic Disease of Apple (Malus domestica)

“…Due to the non-availability of chemical viricides, it is difficult to control viral pathogens. Hence, the only effective way to prevent viral spread in perennial plants is through the use of virus-free planting material via indexing of scion wood and rootstocks using robust, timely and precise detection methods [ 19 , 20 ]. Diagnostic assays play an important role in the detection of plant viruses through their spatial and temporal distribution or selected expression in symptomatic, as well as asymptomatic, host plants [ 21 , 22 ].…”

“…Although DAS-ELISA and symptomatic visualization can help in the detection of viruses, a low titer of virus combined with a limitation to certain time period in the growing season can give false results. To avoid such ambiguity, reverse transcription-polymerase chain reaction (RT-PCR) can provide an alternate route [ 19 ]. The technique is sensitive even during seasons of low virus titer and reproducible with limited risk of contamination compared to end-point RT-PCR [ 23 , 24 ].…”