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Detection of European mountain ash ringspot-associated virus-specific RNA and protein P3 in the pear leaf blister mite Phytoptus pyri (Eriophyidae)

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Abstract

The means by which European mountain ash ringspot-associated virus (EMARaV), a minus-strand ssRNA virus and the type member of the genus Emaravirus, is naturally spread, is unknown. In attempts to identify an EMARaV vector, galls induced by the eriophyid mite Phytoptus pyri were frequently found on infected leaves. By immunofluorescence microscopy, the presence of EMARaV nucleocapsid protein P3 was demonstrated in P. pyri individuals collected from diseased plants. Furthermore, RT-PCR analysis of entire P. pyri individuals revealed the presence of both viral genomic ss(−)RNAs and antigenomic ss(+)RNAs, suggesting that P. pyri might be a candidate vector of EMARaV.

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Acknowledgments

We thank Dr. Joachim Schliesske for extensive information about the family Eriophyidae and for help in identifying P. pyri. Thanks are also due to Dr. Dietmar Keyser (University of Hamburg, Biocentre Grindel) for his help in SEM studies. The work was funded by the Deutsche Forschungsgemeinschaft.

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Authors and Affiliations

  1. Department of Molecular Phytopathology and Genetics, Biocentre Klein Flottbek, University of Hamburg, Ohnhorststrasse 18, 22609, Hamburg, Germany

    Nicole Mielke-Ehret, Joscha Thoma, Nanette Schlatermund & Hans-Peter Mühlbach

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  1. Nicole Mielke-Ehret
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  2. Joscha Thoma
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  3. Nanette Schlatermund
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  4. Hans-Peter Mühlbach
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Correspondence to Hans-Peter Mühlbach.

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705_2010_667_MOESM1_ESM.tif

Supplementary Fig. 1 Detection of EMARaV-specific RNAs and 18S rRNA in entire P. pyri (Eriophyidae) from EMARaV-infected S. aucuparia by standard RT-PCR. (A): Amplification of 18S rRNA from total RNA extracted from five mites and using 30 and 35 PCR cycles. (B): Amplification of vRNA 3 from different numbers of mites. (C): Amplification of vRNAs 1-4 from five mites collected from infected and non-infected plants. (D): Amplification of genomic vRNA 3 and antigenomic vcRNA 3 from five entire mites. M = GeneRulerTM DNA Ladder Low Range (Fermentas, St. Leon-Rot, Germany), S.a. = Purified total RNA from EMARaV-infected S. aucuparia. (TIFF 11100 kb).

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Mielke-Ehret, N., Thoma, J., Schlatermund, N. et al. Detection of European mountain ash ringspot-associated virus-specific RNA and protein P3 in the pear leaf blister mite Phytoptus pyri (Eriophyidae). Arch Virol 155, 987–991 (2010). https://doi.org/10.1007/s00705-010-0667-3

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