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Plant expression vector pBI 121 containing the gene encoding coat protein of Plum Pox Virus of the Skierniewice isolate (CP PPV-S) was prepared (clone pCMl). The construct was used for transformation of Nicotiana tabacum plants using an Agrobacterium tumefaciens based system. About 82% of kanamycin resistant plant lines contained a transgene (the sequence of CP PPV-S) but only 81% of them actively expressed the PPV-S coat protein gene as measured by RT-PCR.
Several methods of transformation are currently available for delivering exogenous DNA to plant cells. Agrobacterium-mediated transformation, microprojectile bombardment and direct protoplast transformation are routinely used today. However, each of them has certain disadvantages, which led to research into the development of novel alternative systems such as infiltration, electroporation of cells and tissues, electrophoresis of embryos, microinjection, pollen-tube pathway, silicon carbide- and liposome-mediated transformation. The low efficiency of transformation is considered to be the main reason for the limited popularity of the alternative transformation methods, other than infiltration and silicon carbide-mediated transformation, which seem to be the most promising ones for practice.
Matrix attachment regions (MARs) are thought to participate in the organization and segregation of independent chromosomal loop domains. Although there are sev­eral reports on the action of natural MARs in the context of heterologous genes in transgenic plants, in our study we tested a synthetic MAR (sMAR) with the special property of unpairing when under superhelical strain, for its effect on reporter gene expression in tobacco plants. The synthetic MAR was a multimer of a short sequence from the MAR 3 end of the immunoglobulin heavy chain (IgH) enhancer. This sMAR sequence was used to flank the β-glucuronidase (GUS) reporter gene within the T-DNA of the binary vector pBI121. Vectors with or without the sMARs were then used to transform tobacco plants by Agrobacterium tumefaciens. Transgenic plants containing the sMAR sequences flanking the GUS gene exhibited higher levels of transgene expression compared with transgenic plants which lacked the sMARs. This effect was observed independently of the position of the sMAR at the 5 side of the re­porter gene. However, variation of the detected transgene expression was significant in all transformed plant populations, irrespective of the construct used.
Standard program of plant protection against fire blight consists of use of :management practices and chemical control method. Recently a new, non-conventional possibilities based on application of biocontrol agents (two biopreparations have been already introduced into practice: Bliteban A506 Pseudomonas fluorescens) and BIossomBless (Pantoea agglomerans), plant active against Erwinia amylovora (AkseBio containing extracts from spicata and Biomit Plussz with extracts from various plant species and ents) and resistance inducers (Regalis, Bion and plant extracts) are of great interest. Also plant transformation with resistance genes such as: hrpN (harpin), dpo (EPS depolymerase) and lytic protein genes (attacin E, cecropin 'SB-37, T4 lysozyme) is a promising perspective.
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