1 Cycle Sequencing Kits (��Applied Biosystems,�� USA) and DNA-analyser 3130 Genetic Analyser (��Applied Biosystems,�� USA). Abiraterone price Analysis of the nucleotide sequences was done in Sequencing Analysis 5.3.1 software (��Applied Biosystems,�� USA) and NCBI (National Center for Biotechnology Information, http://blast.ncbi.nlm.nih.gov/Blast.cgi) database.3. Results and DiscussionThe presence of nucleotide sequences of some mycoplasma genes within EMVs of A. laidlawii PG8 allowed a possibility to use PCR to perform differential detection of the corresponding structures of the bacterium in samples under study. Despite growth conditions, we detected sequences of genes for pdhC, rpoB, pnp, and tufB genes in EMVs, but not for ftsZ and 16S�C23S rRNA genes.

This allowed us to use a combination of primers for differential detection of cells and membrane vesicles in the tested samples.Previously [2], we reported about the ability of A. laidlawii PG8 cells to enter through the root system of O. sativa L. into overground parts of plants and induce there changes in ultrastructural organization. This conclusion was based on data of TEM and PCR obtained 9 days after rice cultivation with mycoplasma cells.

As a result of use of primers for amplification of the nucleotide sequences for the following genes��pdhC (codes dihydrolipoamide acetyltransferase), rpoB (codes ��-subunit of RNA-polymerase), pnp (codes polyribonucleotide-nucleotidyl transferase, a global regulator of virulence in phytopathogenic bacteria [14]), and tufB (codes elongation factor Tu) permitting to detect cells as well vesicles of the mycoplasma, and primers for amplification of the nucleotide sequences of ftsZ gene (codes FtsZ, a protein for cell division), and 16S�C23S rRNA gene (codes a spacer zone and flankings) permitting to detect cells but not bacterial vesicles, we found that PCR signals were presented for all indicated genes in tissues of plant leaves grown in medium with mycoplasma cells for 1�C9 days (Figure 1(c)). Also, amplicon specific to pnp gene of the mycoplasma was found 2 hours since the beginning of plant incubation with mycoplasma cells (Figure 1(b)). Sequencing results confirmed the belonging of a sequence of this amplicon to A. laidlawii PG8 pnp-gene (Figure 2). The obtained data allow to suggest that (1) EMVs are able to display virulent features Dacomitinib linked with infectivity and invasivity in plants; (2) penetration of EMVs to plant tissues precedes to plant infection with mycoplasma cells; (3) EMVs are heterogeneous on the content and functions; (4) virulent features connected with infectivity and invasivity are different in vesicles varying on the content.