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Different target genes and chimeric-gene hairpin structures affect virus resistance mediated by RNA silencing in transgenic tobacco

  • Original Papers
  • Published:
Biologia Plantarum

Abstract

Two types of hairpin RNA (hpRNA) constructions were designed using a chimeric gene formed from two genes, the coat protein (CP) gene or the silencing suppressor gene, from the Cucumber mosaic virus (CMV) and the Potato virus Y (PVYN), respectively; one type generated a single hairpin structure, whereas the other formed a two-hairpin structure. Four constructs, pDCPSH (double CP gene fragments, single hairpin), pDCPDH (double CP gene fragments, double hairpins), pHC2bSH (two silencing suppressor gene fragments, single hairpin), and pHC2bDH (two silencing suppressor gene fragments, double hairpins), were individually introduced into tobacco plants. A transcript analysis demonstrates that the small interference RNA (siRNA) processing efficiency was greater with the double-hairpin construct than with the single-hairpin construct, although the expression of their target genes were similar. A viral resistance assay shows that the transgenic tobacco plants effectively resisted a mixed infection of CMV and Potato virus Y (PVYN) and that pDCPDH exhibited the highest silencing efficiency. The accumulation of siRNA in the inoculated transgenic plants expressing different hairpin structures was similar. A genetic analysis reveals that viral resistance in the transgenic plants was stably inherited from the T0 to T1 generation. A transcript analysis and a viral resistance assay indicate that the double-hairpin structure of the same target sequences tended to produce more siRNA before the virus inoculation and thus strengthened RNA-mediated viral resistance.

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Abbreviations

CMV:

Cucumber mosaic virus

CP:

coat protein

hpRNA:

hairpin RNA

PVY:

Potato virus Y

siRNA:

small interference RNA

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Authors and Affiliations

  1. State Key Laboratory of Crop Biology, Shandong Key Laboratory of Crop Biology, Shandong Agricultural University, Tai’an, Shandong, 271018, P.R. China

    X. Xie, Y. Song, X. Liu, S. Wang, C. Zhu & F. Wen

Authors
  1. X. Xie
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  2. Y. Song
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  3. X. Liu
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  4. S. Wang
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  5. C. Zhu
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  6. F. Wen
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Corresponding authors

Correspondence to C. Zhu or F. Wen.

Additional information

Acknowledgments: This work was financially supported by the National Natural Science Foundation of China (No. 31272113) and the National Natural Science Foundation of Shandong Province (ZR2012CM001).

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Xie, X., Song, Y., Liu, X. et al. Different target genes and chimeric-gene hairpin structures affect virus resistance mediated by RNA silencing in transgenic tobacco. Biol Plant 58, 575–581 (2014). https://doi.org/10.1007/s10535-014-0422-9

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  • DOI: https://doi.org/10.1007/s10535-014-0422-9

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