| In order to understand the genetic evolution of native sweet potato feather mottle virus（SPFMV）population in Hainan,the sweet potato samples collected from Hainan were identified by small RNA sequencing,then 85 contigs were obtained with having high similarity to sequences of SPFMV and NC001841.First,small fragment primers were designed,Primer combinations used as SPFMV-1F/1R,SPFMV-11F/1R,SPFMV-2F/2R,SPFMV-3F/3R and SPFMV-3F/33 R,target fragments of 3 987 bp,1 996 bp and 4 730 bp were amplified by PCR,respectively.The primers were designed in sections to perform full-length PCR amplification of the SPFMV-HN genome.The complete sequence of the SPFMV-HN genome was obtained with sequence splicing,including the complete coding reading frame.The results of sequence analysis showed that the genome length of the SPFMV-HN was 10820 nucleotides（Gen Bank number: MN852852）,5’ and3’ UTR contain 117 and 221 nucleotides,respectively.The 3’ end of poly（A）tail sequence contains 30 nucleotides,which belongs to a typical Potyvirus genus virus genome structure with only a large open reading coding frame consisting of 10446 nucleotides that translate a polyprotein consisting of 3494 amino acids.It is the first full report of the genome of SPFMV isolates from Hainan in China.And another frame element（periodic input same period output,PISPO）was predicted in the P1 region,and the polymerase slip mechanism could generate transcript variants with additional nucleotides that could be translated into P1N-PISPO and P3N-PIPO.This results should provide a basis for further understanding the pathogenic molecular mechanism of SPFMV-HN and the cultivation of disease-resistant sweet potato variety.Restructuring polymerase enzyme amplification（Recombinase polymerase amplification,RPA）is a simple,rapid,high sensitivity and specificity.In this study,the fluorescence probe RPA detection method and the side flow chromatography strip RPA detection method for the rapid detection of sweet potato pink-mottled virus SPFMV were established.Among them,each reaction sensitivity of fluorescence RPA detection method was 102 copies,and the detection rate in field sample detection was 94% compared with the real-time fluorescence quantitative PCR method.The sensitivity of the flow-testing strip RPA method was 101 copies/reaction.Compared with the real-time quantitative PCR method,the detection rate in the field samples was 97%. |
