Widespread Occurrence of Wheat spindle streak mosaic virus in Belgium

Abstract

In order to assess the occurrence of Wheat spindle streak mosaic virus (WSSMV) in Belgium, a reverse-transcription polymerase chain reaction (RT-PCR) was developed, targeting WSSMV isolates from Canada, France, Germany, Italy, and the United States. The primers also were designed for virus quantification by real-time RT-PCR with SYBR-Green. No cross-reaction with soilborne cereal viruses such as Barley mild mosaic virus, Barley yellow mosaic virus, Soilborne cereal mosaic virus, and Soil-borne wheat mosaic virus was observed. The RT-PCR and real-time quantitative RT-PCR allowed a more sensitive detection of WSSMV than enzymelinked immunosorbent assay. The incidence of WSSMV in Belgium was evaluated using a bioassay with wheat cvs. Cezanne and Savannah and rye cv. Halo, grown in 104 Belgian soils. The presence of WSSMV was detected from plants grown in 32% of the soils. The RT-PCR methods developed here, combined with large sampling, allowed WSSMV to be detected for the first time in Belgium. The real-time quantitative RT-PCR was developed as a tool for evaluating the resistance to WSSMV by quantifying the virus concentration in wheat cultivars.

Polymyxa graminis Isolates from Australia: Identification in Wheat Roots and Soil, Molecular Characterization, and Wide Genetic Diversity

“…Leaves and roots were tested for the presence of soil-bome viruses by RT-PCR, using a OneStep RT-PCR kit (Qiagen), as per manufacturers' instructions, with specific primer pairs WSSMVc2F/WSSMVc2R for WSSMV (35), SBWMV-EF/SBWMV-UNIR for SBCMV (8), SBMV L2/SBMV R2 for SBWMV (9), and universal primers Furol-F/Furo2-F for furoviruses (28). Root samples (80 mg/sample) ground under liquid nitrogen as described above for DNA extraction were also used for RNA extraction.…”

Response of UK winter wheat cultivars to Soil-borne cereal mosaic and Wheat spindle streak mosaic viruses across Europe.

“…SBCMV has been found subsequently at 12 further sites in the UK, including the Isle of Wight, Kent and Northampton. In Europe, WSSMV was first detected in France in 1977 (Signoret et al 1977), and is now known to occur throughout Belgium, France, Germany and Italy (Gitton et al 1999;Huth 2000;Rubies Autonell and Vallega 1987;Vaianopoulos et al 2006). WSSMV is often found infecting wheat in combination with SBCMV, but this virus has not yet been reported in the UK.…”

Epistatic determinism of durum wheat resistance to the wheat spindle streak mosaic virus

“…20) of randomly drawn leaves, while an expert could spot one individual plant with symptoms among 30–40 plants to score SS; and (3) symptomless carriers may exist if some plants/genotypes have WSSMV tolerance (Carroll et al 1997). Although qPCR is described as a very sensitive (and expensive) method (Vaïanopoulos et al 2006), it could even be more dependent on the sampling procedure than ELISA since only 0.1 g of leaves are sampled to represent the two rows of an experimental unit. Indeed, qPCR yielded lower heritability in our study than SS and ELISA.…”

“…Visual scoring is an efficient phenotyping method, but WSSMV infected fields are frequently infected by other related viruses, such as SBCMV, which may lead to some confusion (Carroll et al 1997; Vallega et al 2003). Thus, the use of more discriminant viral content assessment methods such as DAS-ELISA (double antibody sandwich-enzyme-linked immunosorbent assay) based on specific serum (Clark and Adams 1977), and PCR, based on specific primers (Vaïanopoulos et al 2006) is necessary. Although ELISA and PCR are more precise, they are more prone to sampling bias when the viral infection is heterogeneous within the plot.…”