Abstract
Wheat dwarf virus (WDV) is considered as one of the most common viruses on cereal crops. Recently, severe outbreaks of WDV have been observed especially on winter wheat in southwestern part of Poland. Moreover, the presence of genetically different WDV-barley-specific and WDV-wheat-specific forms (WDV-B and WDV-W, respectively) was confirmed. In this study, a loop-mediated isothermal amplification assay (LAMP) was developed for the first time for efficient and rapid detection of WDV-B and WDV-W in infected plants. The reaction was performed using a set of three primer pairs: WDVF3/WDVB3, WDVFIB/WDVBIP and WDVLoopF/WDVLoopB specific for coat protein coding sequence. The amplified products were analyzed by direct staining of DNA, gel electrophoresis and real-time monitoring of the amplification curves. The sensitivity of optimized reaction was tenfold higher in comparison with conventional PCR. LAMP assay developed here is a useful and practical method for the rapid detection of different WDV isolates and can be implemented by phytosanitary services.
Highlights
Wheat dwarf virus (WDV) is a member of the genus Mastrevirus in the family Geminiviridae
Taking into account the genetic diversity of the WDV population, the primers for the loop-mediated isothermal amplification assay (LAMP) assay were designed based on the alignment of coat protein (CP) sequences
LAMP technique developed here was capable of the detection of both WDV-B and WDV-W forms isolated from different hosts in less than hour
Summary
Wheat dwarf virus (WDV) is a member of the genus Mastrevirus in the family Geminiviridae. The virus is transmitted by leafhoppers Psammotettix alienus (Dahlb.) [1] and P. provincialis [2] and it infects numerous species within the family Poaceae including important cereals (mainly wheat and barley) [3]. The virus was first reported in Czechoslovakia [5] and subsequently in many countries in Europe, Asia and North Africa [6]. Severe outbreaks of WDV have been reported in Sweden [7], Czech Republic [8], Austria [6] and recently in Poland [9]. The virus has monopartite single-stranded circular (ss) DNA genome of ~2.7 kb which encodes four proteins: the movement protein (MP), the coat protein (CP) and two others
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