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RNAi-Mediated Transgenic Tospovirus Resistance Broken by IntraspeciesSilencing Suppressor Protein Complementation

October 2009 , Volume 22 , Number  10
Pages  1,250 - 1,257

Afshin Hassani-Mehraban,1 A. B. Brenkman,2 N. J. F. van den Broek,2 Rob Goldbach,1 and Richard Kormelink1

1Laboratory of Virology, Wageningen University, Binnenhaven 11, 6709 PD, Wageningen, The Netherlands; 2Department of Metabolic and Endocrine Diseases and Netherlands Metabolomics Centre, University Medical Centre, 3508 AB, Utrecht, The Netherlands

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Accepted 26 May 2009.

Extension of an inverted repeat transgene cassette, containing partial nucleoprotein (N) gene sequences from four different tomato-infecting Tospovirus spp. with a partial N gene sequence from the tomato strain of Tomato yellow ring virus (TYRV-t), renders transgenic Nicotiana benthamiana plants additionally resistant to this strain but not to the soybean strain of this Tospovirus sp. (TYRV-s), both strains exhibiting 14.4% nucleotide sequence divergence in their N genes. Surprisingly, coinoculation of the TYRV-t-resistant transgenic lines with both TYRV-t and TYRV-s resulted in rescue of the former. Mass-spectrometric analysis of the viral ribonucleocapsids accumulating in the transgenic plants showed the presence of the N proteins of both strains excluding hetero-encapsidation as rescue mechanism and indicating suppression of TYRV-t N gene transcript breakdown by RNA interference. Prior (Potato virus X [PVX]-vector-mediated) expression of the TYRV-s silencing suppressor (NSs) gene also allowed TYRV-t to break the resistance. This phenomenon was also observed when the homologous (TYRV-t) NSs gene was provided from a PVX replicon, demonstrating that TYRV can break RNA-mediated host resistance upon a priori expression of its NSs protein. Remarkably, mixed inoculation of TYRV-t with other Tospovirus spp. or nonrelated viruses did not result in resistance breaking, indicating that the rescuing activity of NSs—though based on suppressing RNA silencing—is species-dependent.

© 2009 The American Phytopathological Society