M. G. Lu, R. Gao, R. R. Chen, B. Wu, Z. X. Zhang, and S. F. Li,

Abstract

Sweet cherry (Prunus avium) is one of the important crops in China. So far, eleven viruses have been reported in cherry in China, including Apple mosaic virus (ApMV), Prunus necrotic ringspot virus (PNRSV), Prune dwarf virus (PDV), Apple chlorotic leaf spot virus (ACLSV), Cherry rasp leaf virus (CRLV), Cherry virus A (CVA), Cucumber mosaic virus (CMV), Cherry green ring mottle virus (CGRMV), Plum bark necrosis stem pitting-associated virus (PBNSPaV), Cherry necrotic rusty mottle virus (CNRMV), and Little cherry virus 2 (LChV-2). Two members of the family Closteroviridae, Little cherry virus 1 (LChV-1) and LChV-2, have been associated with little cherry disease (LChD), one of the major viral diseases of sweet cherry worldwide (Jelkmann 2011; Candresse et al. 2013). From April to August 2014, virus diseases of sweet cherry were investigated in 8 cherry orchards of Shandong, Liaoning provinces and Beijing City of China. Leaf symptoms of shrivel, deformity, leaf roll, and yellow and green mottle were observed in these orchards. In total, 28 symptomatic or asymptomatic leaf and bark samples were collected. RT-PCR was used for the detection of eleven viruses listed above and LChV-1. Total nucleic acids were extracted using RNAprep Pure Plant Kit (TIANGEN Biotech (Beijing) Co., Ltd) and subjected to RT-PCR using virus-specific primers. Seven viruses, CVA, PNRSV, PDV, CGRMV, PBNSPaV, and ACLSV, were detected with LChV-1 also detected using LChV-1 primer pair reported byBajet et al. (2008). LChV-1-specific PCR product of 300 bp (nucleotides 7,090 to 7,389) spanned the open reading frame ORF1b encoding the putative replicase. Out of 28 samples, 1 symptomatic leaf sample of Prunus avium cv. Napoleon from Beijing (ChBJ28) and 1 asymptomatic bark sample of Prunus avium cv. Tieton from Daliang of Liaoning (ChDL2) were positive for LChV-1. To further confirm this result, LChV-1 primer pair IN1 for/rev reported by Matic et al. (2009) was used to amplify a 1,774-bp fragment (nucleotides 1 to 1,774), spanning ORF1a and 5′UTR, and encoding papain-like protease from sample ChDL2. PCR products of the expected size, 1775 bp, were detected. Both 1,775-bp and 300-bp amplicons were cloned and sequenced. BLAST analysis revealed that the 1,775-bp and 300-bp fragments shared 73% and 98% nucleotide identity with isolate V2356 deposited in GenBank (Accession No. JX669615.1), respectively, and 71% and 99% deduced amino acid identity with isolate V2356 deposited in GenBank (Accession No. AGB06245.1), respectively, and two obtained sequences was submitted to GenBank with Accession Nos. KP781913 and KM921658. To our knowledge, this is the first report of LChV-1 in cherries in China.

Plant Disease, 2015, Volume 99, Number 8, Page 1191
Go to the article: http://dx.doi.org/10.1094/PDIS-11-14-1200-PDN