Cloning of the Coat Protein Gene of Little Cherry Virus 1 and the Preparation of a Polyclonal Antibody

Little cherry disease (LChD) is a serious viral disease that significantly reduces the yield and quality of sweet cherries. And Little cherry virus 1 (LChV-1) is a major pathogen. To facilitate the rapid and efficient detection of LChV-1, we successfully generated polyclonal antibodies targeting its coat protein (CP). The LChV-1 CP gene was cloned and submitted to the GenBank database. Sequence analysis of 26 reference isolates revealed nucleotide and amino acid sequence identities ranging from 72.59% to 99.84% and 74.26% to 100%, respectively. Bioinformatics analysis identified the 27th to 180th amino acid residues as the optimal immunogenic region for antibody production. This antigenic region was expressed and used to immunize rabbits, yielding polyclonal antibodies with an antiserum titer of 1:81,000 as determined by indirect ELISA. Western blot analysis demonstrated high specificity, with the antibody diluted at 1:3,000 detecting CP both in recombinant protein expression and LChV-1-infected sweet cherry leaf samples. These findings establish a foundation for developing reliable diagnostic tools and advancing research on LChV-1 pathogenesis.