Biopolym. Cell. 2007; 23(6):489-494.
Molecular and Cell Biotechnologies
Construction of vectors for plant transformation in order to confer resistance to potyviruses
1Korniichuk I. V., 1Polischuk V. P., 2Yeh S. D.
  1. Taras Shevchenko National University of Kyiv
    64, Volodymyrska Str., Kyiv, Ukraine, 01033
  2. National Chung Hsing University
    250, Kuo Kuang Rd., Taichung 402, Taiwan

Abstract

Genes HC-Pro, CIP, NIb, and CP are known to be the most conservative in genome of the potyviruses. The comparison of nucleotide sequences of these genes for 32 viruses of Potyvirus genus was carried out. The fragments with the highest percent of identity were selected, amplified from full size cDNA of Turnip mosaic virus (TuMV), and inserted into a high-copy TOPO-vector, used for the transformation of competent Escherichia coli cells. The construction of vectors for plant transformation on the basis of commercial vector pBI121 was performed. cDNA corresponding to one separate (HC-Pro, CIP, NIb or CP), two (HC-Pro and CIP), and three (HC-Pro, CIP, and NIb) conservative regions was inserted into the vector. The transformed cells of Agrobacterium tumefaciens carrying plasmids pBI121 were obtained.
Keywords: potyviruses, resistance, cDNA, PCR, plant transformation

References

[1] Baulcombe DC. Mechanisms of Pathogen-Derived Resistance to Viruses in Transgenic Plants. Plant Cell. 1996;8(10):1833-1844.
[2] Burch-Smith TM, Schiff M, Liu Y, Dinesh-Kumar SP. Efficient virus-induced gene silencing in Arabidopsis. Plant Physiol. 2006;142(1):21-7.
[3] Chumakov MI. Transfer of genetic information from agrobacteria to bacterial and plant cells: membrane and supramembrane structures involved in transfer. Membr Cell Biol. 2000;14(3):309-31.
[4] Di Nicola-Negri E, Brunetti A, Tavazza M, Ilardi V. Hairpin RNA-mediated silencing of Plum pox virus P1 and HC-Pro genes for efficient and predictable resistance to the virus. Transgenic Res. 2005;14(6):989-94.
[5] Fuchs M, Gonsalves D. Resistance of Transgenic Hybrid Squash ZW-20 Expressing the Coat Protein Genes of Zucchini Yellow Mosaic Virus and Watermelon Mosaic Virus 2 to Mixed Infections by Both Potyviruses. Bio/Technology. 1995;13(12):1466–73.
[6] Germundsson A, Valkonen JP. P1- and VPg-transgenic plants show similar resistance to Potato virus A and may compromise long distance movement of the virus in plant sections expressing RNA silencing-based resistance. Virus Res. 2006;116(1-2):208-13.
[7] Hily JM, Scorza R, Malinowski T, Zawadzka B, Ravelonandro M. Stability of gene silencing-based resistance to Plum pox virus in transgenic plum (Prunus domestica L.) under field conditions. Transgenic Res. 2004;13(5):427-36.
[8] Jan FJ, Fagoaga C, Pang SZ, Gonsalves D. A single chimeric transgene derived from two distinct viruses confers multi-virus resistance in transgenic plants through homology-dependent gene silencing. J Gen Virol. 2000;81(Pt 8):2103-9.
[9] Komari T, Takakura Y, Ueki J, Kato N, Ishida Y, Hiei Y. Binary vectors and super-binary vectors. Methods Mol Biol. 2006;343:15-41.
[10] Koonin EV. The phylogeny of RNA-dependent RNA polymerases of positive-strand RNA viruses. J Gen Virol. 1991;72 ( Pt 9):2197-206.
[11] Mahajan S, Dolja VV, Carrington JC. Roles of the sequence encoding tobacco etch virus capsid protein in genome amplification: requirements for the translation process and a cis-active element. J Virol. 1996;70(7):4370-9.
[12] Meynard JM, Dor? T, Lucas P. Agronomic approach: cropping systems and plant diseases. C R Biol. 2003;326(1):37-46.
[13] Ratcliff F, Harrison BD, Baulcombe DC. A similarity between viral defense and gene silencing in plants. Science. 1997;276(5318):1558-60.
[14] Ravelonandro M, Scorza R, Callahan A, Levy L, Jacquet C, Monsion M, Damsteegt V. The use of transgenic fruit trees as a resistance strategy for virus epidemics: the plum pox (sharka) model. Virus Res. 2000;71(1-2):63-9.
[15] Rychahou PG, Jackson LN, Farrow BJ, Evers BM. RNA interference: mechanisms of action and therapeutic consideration. Surgery. 2006;140(5):719-25.
[16] Sanford JC, Johnson SA. The concept of pathogen derived resistance: Deriving resistabce genes from the parasite's own genome. J Theor Biol. 1985; 113(2): 395-405.
[17] Scorza R, Callahan A, Levy L, Damsteegt V, Webb K, Ravelonandro M. Post-transcriptional gene silencing in plum pox virus resistant transgenic European plum containing the plum pox potyvirus coat protein gene. Transgenic Res. 2001;10(3):201-9.
[18] Shukla DD, Ward CW, Brunt AA. The Potyviridae. Ed. D. D. Shukla. Wallingford: CAB Int., 1994: 1-26.
[19] Stram Y, Kuzntzova L. Inhibition of viruses by RNA interference. Virus Genes. 2006;32(3):299-306.
[20] Timmons L. Construction of plasmids for RNA interference and in vitro transcription of double-stranded RNA. Methods Mol Biol. 2006;351:109-17.
[21] Waterhouse PM, Fusaro AF. Plant science. Viruses face a double defense by plant small RNAs. Science. 2006;313(5783):54-5.