engleski

Stability of plant LTR-retrotransposons in Agrobacterium tumefaciens

LTR-retrotransposons are the most common class of eukaryotic mobile elements and have been recently shown to occupy a large proportion of all plant genomes (e.g. more than 70% of maize genome). However, only few transpositionally active plant LTR-retrotransposons have been isolated so far. An impediment to investigation of retrotransposon activity in native host is the considerable background of copies. The aim of our study was to transform A. thaliana with hybrid common bean Tpv2 retrotransposon to investigate its activity. The hybrid Tpv2 element was made out of the fragments of two non-functional copies isolated from the genome of Phaseolus vulgaris. In addition, the intron of glutamine synthetase gene of Lotus japonicus has been inserted into one Tpv2 construct named Tpv2i. Roots of A. thaliana WS and Col-O ecotypes were transformed with either Tpv2 or Tpv2i element via Agrobacterium-mediated transformation. Four types of transgenic Arabidopsis plants were obtained this way. Upon the isolation of plant genomic DNA, the activity of retroelements was analysed by PCR, using primers flanking intron, and by Southern blotting. The activity of neither Tpv2 nor Tpv2i has been detected in any transgenic plant studied.

Agrobacterium tumefaciens

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