Abstract
For the detection of wheat yellow mosaic virus (WYMV), we established a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method. Using Primer Explorer software, four sets of primers were designed and RT-LAMP assay reaction conditions were optimized. The RT-LAMP was performed at different times by four primer sets. Agarose gel analysis showed that WYMV could be detected after 30 min with the primer set III and after 45 min with the other three primer sets, both under the 80-min reaction time. RT-LAMP had the same results with the four primer sets, thus primer set III and 65°C for 80 min reaction were selected for virus detection. There was no significant different when avian myeloblastosis virus (AMV) and moloney murine leukemia virus (M-MLV) RT-LAMP with the four primer sets and M-MLV was chosen due to its relatively cheap price. The result on specificity showed that the assay could amplify WYMV specifically, and the sensitivity comparison showed that the RT-LAMP was 100 times more sensitive than conventional reverse-transcriptase-polymerase chain reaction (RT-PCR). Overall, RT-LAMP was found to be a simple, specific, sensitive, convenient and time-saving method for WYMV detection.
Highlights
Wheat yellow mosaic is one of the most devastating soil-borne diseases of winter wheat (Triticum aestivum L.)
The Chinese wheat mosaic virus (CWMV)-infected samples were collected from Yantai, Shandong Province; the barley stripe mosaic virus (BSMV)-infected samples were fresh wheat leaves inoculated with BSMV in our laboratory in May 2011
Primer sets I and II were located at 665-875 nt (3’ terminal) and 106-332 nt of the coat protein (CP) gene; and primer sets III and IV were located at 715916 nt and 1335-1557 nt of the 72 kDa gene (Figure 1)
Summary
Wheat yellow mosaic is one of the most devastating soil-borne diseases of winter wheat (Triticum aestivum L.). It was first reported in Japan in the 1920s and China in the 1960s [1,2], and spread continually in Japan and China [3,4]. Wheat yellow mosaic virus (WYMV), the causal agent of wheat yellow mosaic, belongs to the genus Bymovirus within the family Potyviridae. It is a soil-borne pathogen and is transmitted by the fungus-like organism Polymyxa graminis [5]. WYMV can serologically cross-react with wheat spindle streak mosaic virus [17], and RT-PCR is not perfect either
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
