Abstract
Rice black-streaked dwarf virus (RBSDV), classified under the Reoviridae, Fijivirus genus, caused an epidemic in the eastern provinces of China and other East Asian countries and resulted in severe yield loss in rice and wheat production. RBSDV is transmitted by the small brown planthopper (SBPH, Laodelphax striatellus Fallén) in a persistent manner. In order to provide a stable and cost-effective detection probe, in this study we selected three DNA aptamers (R3, R5 and R11) by an optimized, standardized and time saving emulsion PCR-based SELEX, for the detection of RBSDV outer-shell P10 protein for in situ localization studies in the midgut of SBPH. The specificity of these three DNA aptamers was tested through detection of the P10 protein using an enzyme-linked oligonucleotide assay (ELONA) and aptamer-based dot-blot ELISA. All three DNA aptamers can be used to detect RBSDV P10 protein by immunofluorescent labeling in the midgut of RBSDV-infected SBPH. These data show that the selected aptamers can be used for the detection of RBSDV P10 protein in vitro and in vivo. This is the first report of aptamers being selected for detection of a rice virus capsid protein.
Highlights
Rice black-streaked dwarf virus (RBSDV) belongs to the Reoviridae Family, Spinareovirinae, Fijivirus genus [1,2], causing rice black-streaked dwarf disease and maize rough dwarf disease [3,4]
A variety of fusion tags to the RBSDV P10 protein were screened for expression (Figure S1A,B)
A plasmid construct of the RBSDV His-P10 protein expressed in E. coli BL21-CodonPlus (DE3)-RIPL
Summary
Rice black-streaked dwarf virus (RBSDV) belongs to the Reoviridae Family, Spinareovirinae, Fijivirus genus [1,2], causing rice black-streaked dwarf disease and maize rough dwarf disease [3,4]. RBSDV is transmitted by the small brown plant hopper (SBPH, Laodelphax striatellus Fallén) in a persistent propagative manner and infects many plant species in Gramineae including rice, maize, wheat and barley [1,2]. Typical symptoms of infected rice plants include stunting, darkening of leaf color, and tumors on leaves and stems. When infection occurs at the tillering stage, the panicles are usually closed, causing severe loss of crop production [5,6]. In 2013, a field survey confirmed that 333 out of plants from five provinces surveyed in China were infected with RBSDV [7]
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