• MDMV is a major threat for maize crops worldwide.
  • RT-LAMP assay was developed for the first time for MDMV detection.
  • Results can be obtained within 1 hr in isothermal conditions.
  • Low-cost detection assay can be routinely used in diagnostics of maize crops.
Maize dwarf mosaic virus (MDMV) is a serious and widespread virus pathogen of maize plants. This +ssRNA virus belongs to the Potyvirus genus in the Potyviridae family. Together with sugarcane mosaic virus (SCMV) it causes one of the most important viral diseases on maize crops in the world – maize dwarf mosaic. Both viruses are transmitted in the same non-persistent manner by several aphid species. They induce similar symptoms of leaf mosaic or mottling, stunting and a reduction in plant weight and grain yield. Available MDMV diagnostics include primarily commercialized enzyme-linked immunosorbent assays (ELISA) and reverse transcription-polymerase chain reactions (RT-PCR). Here, laborsaving reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay was optimized for identification of genetically different MDMV isolates. For this purpose, primer sets, MDMVF3/MDMVB3 and MDMVFIP/MDMVBIP amplifying fragments of coat protein coding sequence of MDMV, were used. The specificity of the reaction was verified using three MDMV (-P1, -Sp, -PV0802-DSMZ) and three SCMV (-P1, -PV0368- -DSMZ, -PV1207-DSMZ) isolates. Obtained products were visualised by DNA staining, electrophoretic separation as well as by real-time monitoring of the reaction. The sensitivity of RT-LAMP and conventional RT-PCR reactions was comparable. Both methods could detect virus as low as 550 fg · μl–1 of total RNA. This technique has application value for screening MDMV by phytosanitary services.
The authors would like to thank Dr Maria Angeles Achon from the University of Lleida for providing the MDMV-Sp isolate used in this study.
This study was financed by the Polish Ministry of Science and Higher Education, by Statutory activity project: WIB03.
Natasza Borodynko-Filas
The authors have declared that no conflict of interests exist.
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