Archivio Journal of Plant Pathology

Abstract:

Cherry rasp leaf virus (CRLV, genus Cheravirus, family Secoviridae) may cause symptoms on cherry such as leaf distortion and enations on the underside of the leaves along the midribs, whereas limbs of infected trees might become bare near the base (Adaskaveg and Caprile, 2014). In 2002, CRLV was first reported in Liaoning Province based on the result of field survey, virus purification and electron microscope observations (Tan et al., 2002). Shandong province is one of the most important cherry production areas in China. During May and June in 2013, a survey was conducted for the occurrence of CRLV in a sweet cherry orchard in the countryside of Zoucheng city (Shandong). Although no characteristic CRLV symptoms such as leaf distortion and enations were observed, leaf samples were anyhow randomly collected from 20 cherry trees, each sample consisting of 10 leaves. These samples were subjected to RNA was extracted and subjected to RT-PCR assay for amplifying the vp24 gene of CRL V RNA2 using primer pairs vp24F (5’-GGCCCTGACCCTTTTTCCTTTCATTTG-3’) and vp24R (5’-GGTGTACTCAGCTTTGAGGGCTC-3’). DNA fragments of ca. 580 bp were amplified from 14 out of the 20 cherry leaf samples. PCR products of two randomly selected samples were cloned into pMD18-T vector (TaKaRa, Japan) and sequenced in both directions. Sequence alignment and BLAST analysis showed that the nucleotide sequences of both fragments were 100% identical to vp24 gene of a flat apple isolate of CRLV (GenBank Accession No. AY122330). To the best of out knowledge, this is the first report of the occurrence of CRLV in Shandong province based on molecular assays.