New Antisense RNA Systems Targeted Against Plant Pathogens

  • Matousek, J. (IPMB AS CR) ;
  • Vrba, L. (IPMB AS CR, South Bohemian University) ;
  • Kuchar, M. (South Bohemian University) ;
  • Pavingerova, D. (IPMB AS CR) ;
  • Orctova, L. (IPMB AS CR) ;
  • Ptacek, J. (Institute for Potato Research) ;
  • Schubert, J. (BAZ, Institute for Resistance Research and Pathogen Diagnostics) ;
  • Steger, G. (Heinrich -Heine Universitat Dusseldorf Universityatstrasse) ;
  • Beier, H. (Institute of Biochemistry, Biozentrum, Am Hubland) ;
  • Riesner, D. (Heinrich -Heine Universitat Dusseldorf Universityatstrasse)
  • Published : 2000.09.01

Abstract

tRNA and 7SL RNA based antisense vehicles were prepared by inserting conserved anti-viral and anti-viroid domains. Anti-PVS coat protein leader sequence (ACPL) and antistructural antihairpin domain of PSTVd (AHII) were inserted in tRNA cassette; anti- zing finger domain of PVS, AHII and anti hop latent viroid ribozyme were inserted in 7SL RNA gene isolated from A. thaliana. These constructs were shown to be transcribed both, in in vitro and in in vivo conditions. However, it followed from our work that closely linked position of PoIII reference genes and PoIIII antisense genes within T-DNA lead to the impairment of RNA expression in transgenic plants. To assay in vivo transcription of antisense genes, hairy root potato cultures were established using h. tumefaciens A4-24 bearing both, Ri plasmid and PoIII-promoterless plant expression vectors with antisense RNA genes. Expression of antisense RNA in transgenic potato tissues was proven by specific RT-PCR reactions.

Keywords

References

  1. Gene v.44 The human $tRNA^{Val}$ gene family: organization, nucleotide sequences and homologous transcription of three single-copy genes Amold, J.G.;Schmutzler, C.;Thomann, U.;van Tol, H.;Gross, H.J.
  2. Plant Mol Biol v.19 Suppression of gene expression in plant cells utilizing antisense sequences transcribed by RNA polymerase Ⅲ Bourque, J.E.;Folk, R.W.
  3. EMBO J v.14 A plant basal in vitro system supporting accurate transcription of both RNA polymerase Ⅱ- and Ⅲ-dependent genes: supplement of green leaf component(s) drives accurate transcription of a light-responsive rbcS gene Fan, H.;Sugiura, M.
  4. Plant Mol Biol v.20 The $tRNA^{Tyr}$ multigene family of Nicotiana rustica genome organization, sequence analyses and expression in vitro Fuchs, T.;Beier, D.;Beier, H.
  5. Trends of General Virology v.68 Zinc fingers : a novel protein motif for nucleic acid recognition Klug, A.;Rhodes, D.
  6. EMBO J v.10 formation of a thermodynamically metastable structure containing hairpin Ⅱ is critical for infectivity of potato spindle tuber viroid RNA Loss, P.;Schmitz, M.;Steger, G.;Riesner, D.
  7. Nucleic Acids Res. v.21 Sequence of the Arabidopsis thaliana 7SL RNA gene Marques, J.P.;Gualberto, J. M.;Palme, K.
  8. Biol Chem Hoppe-Seyler v.375 Inhibition of viroid infection by antisense RNA expression in transgenic plants Matou$\ˇ{s}$ek, J.;Schr$\"{o}$der A.R.W.;Trn$\ˇ{s}$n$\'{a}$ L.;Reimers, M.;Baumstark, T.;D$\ˇ{e}$di$\ˇ{c}$, P.;Vlas$\'{a}$k, J.;Becker, I.;Kreuzaler, F.;Fladung, M.;Riesner, D.
  9. Brichte aus der BAZ-Zeuchtungsforschung v.Heft 1 The design and analysis of anti-HLVd ribozymes Matou$\ˇ{s}$ek, J.;Oriniakov$\'{a}$, P.;Trn$\ˇ{e}$n$\'{a}$, L.;Steger, G.;Riesner, D
  10. V$\ˇ{e}$deck$\'{e}$ pr$\'{a}$ce, V$\'{y}$zkum$\'{y}$ $\'{u}$stav brambor$\'{a}ˇ{r}$sk$\'{y}$, Havl$\'{i}\ˇ{r}$k$\{uu}$v Brod v.13 The design of antisense RNA genes targeted against putative potato viroid promoter element Matou$\ˇ{s}$ek, J.;Vrba, L.;Junker, V.;Schr$\"{o}$der, A.;Steger, G.;Riesner, D
  11. Gene v.239 Molecular characterization and genome organization of 7SL RNA genes from hop (Humulus lupulus L.) Matou$\ˇ{s}$ek, J.;Junker, V.;Vrba, L.;Schubert, J.;Patzak, J.;Steger, G.
  12. Can J Plant Pathol v.22 A broad variability of potato virus S (PVS) revealed by analysis of virus sequences amplified by reverse transcriptase-polymerase chain reaction Matou$\ˇ{s}$ek, J.;Schubert, J.;D$\ˇ{e}$di$\ˇ{c}$, P.;Pt$\'{a}\ˇ{c}$ek, J.
  13. Acta virologica v.44 Analysis of variability of $PVY^{N}$ P1 region using TGGE and electrophoresis of heteroduplexes Matou$\ˇ{s}$ek, J.;Pt$\'{a}\ˇ{c}$ek, J.;D$\ˇ{e}$di$\ˇ{c}$, P.;Schubert, J.
  14. Proceedings from the $7^{th}$ Aschersleben Symposium: New Aspects of Resistance Research on Cultivated Plants The sequence variation of of hop latent viroid increases upon the conditions of thermo-therapy. Beitraege zur Zuechtungsforschung, 6. 3. 2000 Matou$\ˇ{s}$ek, J.;Patzak, J.;Orctov$\'{a}$, L.;Svoboda, P.;Schubert, J.;Riesner, D.
  15. Arch Phytopathol Pflanzenschutz The design and partial analysis of RNAse Ⅲ-anti-PVS antisense complex system to induce plant resistance Matou$\ˇ{s}$ek, J.;Schubert, J.;Kucha$\ˇ{r}$, M.;D$\ˇ{e}$di$\ˇ{c}$, P.;Pt$\'{a}\ˇ{c}$ek, J.;Vrba, L.;Lichtenstein, C.P.
  16. Transgenic Research v.5 Strategies for the suppression of peroxidase gene expression in tobacco. Ⅱ. In vivo suppression of peroxidase activity in transgenic tobacco using ribozyme and antisense constructs McIntyre, C.L.;Bettenay, H.M.;Manners, J.M.
  17. Physiol Plant v.15 A revised medium for rapid growth and bioassays with tobacco tissue cultures Murashige, T.;Skoog, F.
  18. Genetics v.147 Dissection of a synthesized quantitative trait to characterize transgene interactions Nap, J-P;Conner, A.J.;MlyN$\'{a}$rov$\'{a}$, L.;Stiekema, W.J.;Jansen, R.C.
  19. Proc Natl Acad Sci USA v.92 Effective ribozyme delivery in plant cells Perriman, R.;Bruening, G.;Dennis, E.S.;Peacock, W.J.
  20. EMBO J 12 v.1993 Multiple pathways of reversion in viroids for conservation of structural elements Qu, F.;Heinrich, C.;Loss, P.;Steger, G.;Tien, P.;Riesner, D.
  21. Viroids and viroid-like RNAs v.1 Landoldt -bornstein, Numerical Data and Functional Relationships in Science and Technology. Group Ⅶ, Biophysics Vol.1, Nucleic Acids; Subvol. d, Physical Data Ⅱ, Teoretical Investigations Riesner, D.;Steger, G.;Saenger, W.(ed.)
  22. Meth Enzymol v.118 Gene transfer in plants: production of transformed plants using Ti plasmid vectors Rogers, S.G.;Horsch, R.B.;Fraley, R.T.
  23. Annals of Botany v.79 The silence of genes in transgenic plants Stam, M.;Mol, J.N.M.;Kooter, J.M.
  24. EMBO J v.6 A cell-free plant extract for accurate pre-tRNA processing, splicing and modification Stange, N.;Beier, H.
  25. Arch Virol v.142 Deletion analysis of a translational enhancer upstream from the coat protein open reading frame of potato virus S Turner, R.;Foster, G.D.
  26. Antisense Research and Applications Modulation of gene expression in plants by antisense genes Van Blokland, R.;De Lange, P.;Mol, J.N.M.;Kooter, J.M.;Crooke, S.T.(ed.);Lebleu, B.(ed.)
  27. Cell v.76 RNA-directed 'de novo' methylation of genomic sequence in plants Wassenegger, M.;Heimes, S.;Riedel, L.;S$\"{a}$nger, H.L.
  28. Proc Natl Acad Sci USA v.90 Strategies to protect plants against viruses: Pathogen-derived resistance blossoms Wilson, T.M.A.