Discrimination and genetic diversity of Wheat dwarf virus in the Czech Republic

Abstract

Cereal-infecting Mastreviruses are one of the most ubiquitous of viruses, having caused huge yield losses during the last decade in the Czech Republic. The presence of two strains of Wheat dwarf virus (WDV), one of which is wheat adapted and one barley adapted, have been confirmed from among field samples of wheat and barley plants. The virus typing was conducted by both PCR-RFLP and sequencing-based methods. The Czech WDV isolates of the barley strain are more variable than the isolates of the wheat strain, separating them into two clades, one representing a pool of divergent isolates. The RFLP analysis confirmed the separation of the Czech isolates into two strains and highlights a powerful method for the rapid diagnosis of both the wheat and barley strains. Additionally, both RFLP and sequence analysis have shown that the barley strain is restricted to the barley host, while the wheat strain is present in both wheat and barley plants.

Real-time PCR assay for the discrimination and quantification of wheat and barley strains of Wheat dwarf virus

“…Our results demonstrate that several techniques, using TaqMan Ò and SYBR Ò Green chemistry, are available to identify, discriminate and quantify both WDV strains commonly found in cereal crops in the Czech Republic [8]. These qPCR methods have potential for monitoring the virus in the field, forecasting virus epidemic development and studying virus-plant-vector interactions.…”

“…Fifty-two field samples of wheat and barley isolates were sequenced by Kundu et al [8], and others collected in 2009. Confirmation of the virus was performed by PCR-RFLP [8].…”

“…Confirmation of the virus was performed by PCR-RFLP [8]. Samples that discriminated for the presence of wheat-or barley-adapted WDV strains were chosen as positive controls for further development of real-time qPCR.…”

“…An oat-adapted isolate, distinct from wheat and barley strains, was reported from Germany [7]. Several well-established methods for identification of wheat and barley strains have been reported including PCR, Rolling-circle amplification (RCA)-Restriction fragment length polymorphism (RFLP) [7], and PCR-RFLP [8]. These methods discriminate both strains based on viral DNA amplification and identification of one and/or several restriction endonuclease cleavage sites.…”

Widespread occurrence and molecular characterization of Wheat dwarf virus in Iran

“…While barley yellow dwarf viruses (BYDVs, family Luteoviridae) are regarded as the most common viruses associated with this disease (Miller and Rasochová 1997), considerable yield losses of cereal crops are also caused by Wheat dwarf virus (WDV, family Geminiviridae) in certain regions including China, Europe, and North Africa (Bendahmane et al 1995;Bisztray et al 1989;Kundu et al 2009;Kvarnheden et al 2002;Köklu et al 2007;Najar et al 2000;Schubert et al 2007;Xie et al 2007). …”