Wheat Virus Identification Within Infected Tissue Using Nanopore Sequencing Technology

• DOI: 10.1094/pdis-09-18-1700-re
• Publication Date: 2019-02-07T19:29:51Z
• Authors: John P. Fellers; Christian Webb; Madison C. Fellers; Jessica Shoup Rupp; Erick De Wolf
• Keywords: Sanger sequencing; Barley yellow dwarf

Viral diseases are a limiting factor to wheat production. Viruses difficult diagnose in the early stages of disease development and often confused with nutrient deficiencies or other abiotic problems. Immunological methods useful identify viruses, but specific antibodies may not be available require high virus titer for detection. In 2015 2017, plants containing Wheat streak mosaic (WSMV) resistance gene, Wsm2, were found have symptoms characteristic WSMV. Serologically, WSMV was detected all four samples. Additionally, High Plains (HPWMoV) also one Barley yellow dwarf (BYDV) detected, detection kit readily Triticum (TriMV). Initially, cDNA cloning Sanger sequencing used determine presence WSMV; however, process time-consuming expensive. Subsequently, from infected tissue sequenced single-strand, Oxford Nanopore technology (ONT). ONT able confirm TriMV samples BYDV three Deep coverage full-length, single-strand revealed variation compared Sidney-81 reference strain represent new variants which overcome Wsm2. These results demonstrate that can more accurately causal agents has sufficient resolution provide evidence variants.