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European Journal of Plant Pathology

, Volume 130, Issue 2, pp 183–196 | Cite as

Molecular and serological diversity in Apple chlorotic leaf spot virus from sand pear (Pyrus pyrifolia) in China

  • Yansu Song
  • Ni Hong
  • Liping Wang
  • Hongju Hu
  • Rui Tian
  • Wenxing Xu
  • Fang Ding
  • Guoping Wang
Original Research

Abstract

Apple chlorotic leaf spot virus (ACLSV) isolates from sand pear (Pyrus pyrifolia) were characterized by analyzing the sequences of their coat protein (CP) genes and serological reactivity of recombinant coat proteins (rCPs). The sequences of CP genes from 22 sand pear isolates showed a high divergence, with 87.3–100% identities at the nucleotide (nt) level and 92.7–100% identities at the amino acid (aa) level. Phylogenetic analysis on the aa sequence of CP showed that the analyzed ACLSV isolates fell into different clusters and all isolates from sand pear were grouped into a large cluster (I) which was then divided into two sub-clusters (A and B). Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), western blot and enzyme-linked immunosorbent assay (ELISA) analyses demonstrated that rCPs of eight ACLSV isolates (PP13, PP15-2, PP24, PP43, PE, PP54, PP56 and ACLSV-C) from two sub-clusters had different mobility rates and serological reactivity. The rCPs of five isolates grouped into the sub-cluster A showed stronger reactivity with antibodies against rCPs of a sand pear isolate ACLSV-BD and virions of a Japanese apple isolate P-205 than that with the antibody against a Chinese apple isolate ACLSV-C. Three isolates grouped into the sub-cluster B showed stronger reactivity with the antibody against ACLSV-C. The antigenic determinants of CPs from these eight isolates and isolates ACLSV-BD and P-205 were predicted. These results contribute to a further understanding of molecular diversity of the virus and its implication in serological detection.

Keywords

Apple chlorotic leaf spot virus Sand pear Coat protein gene Phylogenetic tree Western blot 

Notes

Acknowledgements

This study was financially supported by Chinese Ministry of Agriculture, Industry Technology Research Project (grant no: 200903004) and the earmarked fund for Pear Modern Agro-industry Technology Research System (nycytx-29-08). The authors would like to thank Professor Yangdou Wei, University of Saskatchewan, Canada, for critical revision for the manuscript, and Professor Yanxiu Liu, Huazhong Agricultural University, China, for help with the English of the manuscript.

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Copyright information

© KNPV 2011

Authors and Affiliations

  • Yansu Song
    • 1
    • 2
  • Ni Hong
    • 1
    • 2
  • Liping Wang
    • 2
  • Hongju Hu
    • 3
  • Rui Tian
    • 3
  • Wenxing Xu
    • 2
  • Fang Ding
    • 2
  • Guoping Wang
    • 1
    • 2
  1. 1.National Key Laboratory of AgromicrobiologyHuazhong Agricultural UniversityWuhanPeople’s Republic of China
  2. 2.The Key Laboratory of Plant Pathology of Hubei ProvinceHuazhong Agricultural UniversityWuhanPeople’s Republic of China
  3. 3.Institute of Fruit and TeaHubei Academy of Agriculture SciencesWuhanPeople’s Republic of China

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