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Publikation

Carbonell, A.; Flores, R.; Gago, S. Trans -cleaving hammerhead ribozymes with tertiary stabilizing motifs: in vitro and in vivo activity against a structured viroid RNA Nucleic Acids Research 39, 2432-2444, (2011) DOI: 10.1093/nar/gkq1051

Trans -cleaving hammerheads with discontinuous or extended stem I and with tertiary stabilizing motifs (TSMs) have been tested previously against short RNA substrates in vitro at low Mg 2+ concentration. However, the potential of these ribozymes for targeting longer and structured RNAs in vitro and in vivo has not been examined. Here, we report the in vitro cleavage of short RNAs and of a 464-nt highly structured RNA from potato spindle tuber viroid (PSTVd) by hammerheads with discontinuous and extended formats at submillimolar Mg 2+ . Under these conditions, hammerheads derived from eggplant latent viroid and peach latent mosaic viroid (PLMVd) with discontinuous and extended formats, respectively, where the most active. Furthermore, a PLMVd-derived hammerhead with natural TSMs showed activity in vivo against the same long substrate and interfered with systemic PSTVd infection, thus reinforcing the idea that this class of ribozymes has potential to control pathogenic RNA replicons.
Publikation

Renovell, Á.; Gago, S.; Ruiz-Ruiz, S.; Velázquez, K.; Navarro, L.; Moreno, P.; Vives, M. C.; Guerri, J. Mapping the subgenomic RNA promoter of the Citrus leaf blotch virus coat protein gene by Agrobacterium-mediated inoculation Virology 406, 360-369, (2010) DOI: 10.1016/j.virol.2010.07.034

Citrus leaf blotch virus has a single-stranded positive-sense genomic RNA (gRNA) of 8747 nt organized in three open reading frames (ORFs). The ORF1, encoding a polyprotein involved in replication, is translated directly from the gRNA, whereas ORFs encoding the movement (MP) and coat (CP) proteins are expressed via 3' coterminal subgenomic RNAs (sgRNAs). We characterized the minimal promoter region critical for the CP-sgRNA expression in infected cells by deletion analyses using Agrobacterium-mediated infection of Nicotiana benthamiana plants. The minimal CP-sgRNA promoter was mapped between nucleotides −67 and + 50 nt around the transcription start site. Surprisingly, larger deletions in the region between the CP-sgRNA transcription start site and the CP translation initiation codon resulted in increased CP-sgRNA accumulation, suggesting that this sequence could modulate the CP-sgRNA transcription. Site-specific mutational analysis of the transcription start site revealed that the + 1 guanylate and the + 2 adenylate are important for CP-sgRNA synthesis.
Publikation

Carbonell, A.; Martínez de Alba, A.E.; Flores, R.; Gago, S. Double-stranded RNA interferes in a sequence-specific manner with the infection of representative members of the two viroid families Virology 371, 44-53, (2008) DOI: 10.1016/j.virol.2007.09.031

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Publikation

Carbonell, A.; de la Peña, M.; Flores, R.; Gago, S. Effects of the trinucleotide preceding the self-cleavage site on eggplant latent viroid hammerheads: difference in co- and post-transcriptional self-cleavage may explain the lack trinucleotide AUC in most natural hammerheads Nucleic Acids Research 34, 5613-5622, (2006) DOI: 10.1093/nar/gkl717

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Publikation

Fortes, A.M.; Miersch, O.; Lange, P.R.; Malho, R.; Testillano, P.S.; del Risueno, M.C.; Wasternack, C.; Pais, M.S. Expression of allene oxide cyclase and accumulation of jasmonates during organogenic nodule formation from hop (<EM>Humulus lupulus</EM> var. Nugget) internodes Plant Cell Physiol. 46, 1713-23, (2005)

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Publikation

Schüler, G.; Mithöfer, A.; Baldwin, I.T.; Berger, S.; Ebel, S.; Santos, J.G.; Herrmann, G.; Hölscher, D.; Kramell, R.; Kutchan, T.M.; Maucher, H.; Schneider, B.; Stenzel, I.; Wasternack, C.; Boland, W. Coronalon: a powerful tool in plant stress physiology FEBS Lett. 563, 17-22, (2004) DOI: 10.1016/S0014-5793(04)00239-X

Coronalon, a synthetic 6-ethyl indanoyl isoleucine conjugate, has been designed as a highly active mimic of octadecanoid phytohormones that are involved in insect and disease resistance. The spectrum of biological activities that is affected by coronalon was investigated in nine different plant systems specifically responding to jasmonates and/or 12-oxo-phytodienoic acid. In all bioassays analyzed, coronalon demonstrated a general strong activity at low micromolar concentrations. The results obtained showed the induction of (i) defense-related secondary metabolite accumulation in both cell cultures and plant tissues, (ii) specific abiotic and biotic stress-related gene expression, and (iii) root growth retardation. The general activity of coronalon in the induction of plant stress responses together with its simple and efficient synthesis suggests that this compound might serve as a valuable tool in the examination of various aspects in plant stress physiology. Moreover, coronalon might become employed in agriculture to elicit plant resistance against various aggressors.
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