Biologia plantarum 58:575-581, 2014 | DOI: 10.1007/s10535-014-0422-9

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

X. Xie1, Y. Song1, X. Liu1, S. Wang1, C. Zhu1,*, F. Wen1,*
1 State Key Laboratory of Crop Biology, Shandong Key Laboratory of Crop Biology, Shandong Agricultural University, Tai'an, Shandong, P.R. China

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.

Keywords: coat protein; Cucumber mosaic virus; dual-virus resistance; Nicotiana tabacum; Potato virus Y; silencing suppressor
Subjects: hairpin; coat protein; dual-virus resistance; gene silencing; transgenic plants; tobacco
Species: Nicotiana tabacum; Cucumber mosaic virus; Potato virus Y

Received: June 4, 2013; Revised: September 25, 2013; Accepted: October 25, 2013; Published: September 1, 2014Show citation

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Xie, X., Song, Y., Liu, X., Wang, S., Zhu, C., & Wen, F. (2014). Different target genes and chimeric-gene hairpin structures affect virus resistance mediated by RNA silencing in transgenic tobacco. Biologia plantarum, 58(3), 575-581. doi: 10.1007/s10535-014-0422-9.
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