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The six Tomato yellow leaf curl virus genes expressed individually in tomato induce different levels of plant stress response attenuation

Abstract

Tomato yellow leaf curl virus (TYLCV) is a begomovirus infecting tomato plants worldwide. TYLCV needs a healthy host environment to ensure a successful infection cycle for long periods. Hence, TYLCV restrains its destructive effect and induces neither a hypersensitive response nor cell death in infected tomatoes. On the contrary, TYLCV counteracts cell death induced by other factors, such as inactivation of HSP90 functionality. Suppression of plant death is associated with the inhibition of the ubiquitin 26S proteasome degradation and with a deactivation of the heat shock transcription factor HSFA2 pathways (including decreased HSP17 levels). The goal of the current study was to find if the individual TYLCV genes were capable of suppressing HSP90-dependent death and HSFA2 deactivation. The expression of C2 (C3 and CP to a lesser extent) caused a decrease in the severity of death phenotypes, while the expression of V2 (C1 and C4 to a lesser extent) strengthened cell death. However, C2 or V2 markedly affected stress response under conditions of viral infection. The downregulation of HSFA2 signaling, initiated by the expression of C1 and V2, was detected in the absence of virus infection, but was enhanced in infected plants, while CP and C4 mitigated HSFA2 levels only in the infected tomatoes. The dependence of analyzed plant stress response suppression on the interaction of the expressed genes with the environment created by the whole virus infection was more pronounced than on the expression of individual TYLCV genes.

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Acknowledgements

We thank Prof. D. Baulcombe for providing us with the pGR106 (PVX) vector, Prof. E. Bejarano for the recombinant constructs PVX-C2, -V2, Prof. A. Grover for providing us with anti-HSP17 antibodies, Dr. K.D. Scharf for the anti-HSFA2 antibody. This research was supported by a grant from the Israel Science Foundation award (1037/13) and by a grant from the U.S. Agency for International Development, Middle East Research and Cooperation (MERC) program (GEG-G-00-02-00003-00).

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Correspondence to Rena Gorovits.

Electronic supplementary material

Supplementary Figure 1

HR response of N. benthamiana to the presence of expressed TYLCV genes. A. N. benthamiana 14 days after infiltration with the PVX-V2 demonstrated an obvious decline in growth and symptoms of leaf necrosis, while PVX-V2 presence in tomatoes did not affect plant growth and did not cause HR phenotype on leaf and stems. B. Hsp90-silenced N. benthamiana plants (blank) were agroinfiltrated with PVX, PVX-C2, −CP, −C1, −V2 constructs. Fifteen days after agroinfiltration, changes in growth and CD were detected. C. Comparative growth of HSP90-silenced tomatoes infiltrated with the PVX vector, and with the PVX-C2 and PVX-V2 constructs during TYLCV infection (14, 21, 28 dpi). (GIF 789 kb)

High Resolution Image (TIFF 1462 kb)

(GIF 537 kb)

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Supplementary Figure 2

qRT-PCR analysis of Hsp90 gene expression in tomatoes agro-inoculated with TRV- Hsp90 silencing construct (TRV- Hsp90 ). 24 h after TRV-Hsp90 inoculation, half of the plants were inoculated with the PVX- derived vector (TRV-Hsp90 + PVX-0). Plants were either infected (i) or uninfected (o) with TYLCV 5 days after TRV-Hsp90 inoculation. qRT-PCR was performed immediately before TRV inoculation (0), and 5, 19 days after. The relative expression of each gene was calculated in relation to TRV-Hsp90 plants before TRV treatment. The results were normalized using β-actin as an internal marker. Bars represent the average and standard deviation of the relative expression from three independent biological repeats. (GIF 110 kb)

High Resolution Image (TIFF 89 kb)

Supplementary Table 1

Primers used for the amplification of the TYLCV genes C1, C3, C4 and CP. (PDF 171 kb)

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Gorovits, R., Moshe, A., Amrani, L. et al. The six Tomato yellow leaf curl virus genes expressed individually in tomato induce different levels of plant stress response attenuation. Cell Stress and Chaperones 22, 345–355 (2017). https://doi.org/10.1007/s12192-017-0766-0

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Keywords

  • Geminivirus
  • Viral proteins
  • Stress response
  • Cell death