Transformation of Mexican lime with an intron-hairpin construct expressing untranslatable versions of the genes coding for the three silencing suppressors of Citrus tristeza virus confers complete resistance to the virus
Article first published online: 8 MAR 2012
© 2012 The Authors. Plant Biotechnology Journal © 2012 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd
Plant Biotechnology Journal
Volume 10, Issue 5, pages 597–608, June 2012
How to Cite
Soler, N., Plomer, M., Fagoaga, C., Moreno, P., Navarro, L., Flores, R. and Peña, L. (2012), Transformation of Mexican lime with an intron-hairpin construct expressing untranslatable versions of the genes coding for the three silencing suppressors of Citrus tristeza virus confers complete resistance to the virus. Plant Biotechnology Journal, 10: 597–608. doi: 10.1111/j.1467-7652.2012.00691.x
- Issue published online: 1 MAY 2012
- Article first published online: 8 MAR 2012
- Received 23 December 2011; revised 31 January 2012; accepted 1 February 2012.
Figure S1 (a) Diagram of the T-DNA from the binary vector pCambia-SIAS with the SIAS cassette controlled by the CaMV 35S-pro and the nos-ter and flanked by the cassettes 35S-pro/nptII/35S-ter and 35S-pro/uidA/nos-ter. Transcription orientation for each cassette is indicated by arrows, and restriction sites HindIII and EcoRI by vertical broken lines. (b,c) Southern blot hybridization of nucleic acid preparations from Mexican lime transformed with the SIAS construct (lines 46, 22, 41, 1, 2, 27, 26, 28, 43 and 39) and with the empty vector (EV). DNA was digested with EcoRI (b), which cuts once the T-DNA, or with EcoRI and HindIII (c), which excise the SIAS expression cassette. Size of DNA markers is indicated at the right. Hybridization was with a DIG-labelled DNA probe for detecting the coding region of p23. The higher intensity of the two bands in line SIAS-26 suggests multiple integrations as concatamers at the two loci.
Figure S2 Symptoms of Mexican lime propagations from a susceptible (SIAS-39) (a) and a resistant (SIAS-2) (b) SIAS transgenic plants expressed in the third flush after graft inoculating the CTV-T36 clonal strain in the Carrizo citrange rootstock in comparison with a noninoculated EV control (c). Susceptible transgenic SIAS scions showed vein clearing (upper), young leaf epinasty (middle) and adult leaf cupping (lower).
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