1; 003.4; 003.5; 003.32; 003.34; 006.1; 006.2; 006.4; 006.7; 006.8; 006.10; 104.24; 105.1; 105.28 and 003.10; 003.12; 003.23; 003.24; 006.13; 006.16; 006.17; 006.18; 006.51; 104.10; 105.6; 105.12, respectively. To determine the susceptibility of the bacterial isolates to the essential oil obtained from L. sidoides genotypes LSID006 and LSID104 containing contrasting amounts of LY2874455 price thymol and carvacrol (Table 1), MICs were determined by a doubling dilution technique using the two essential oils at eight concentrations (from 4 to 0.03 mg ml-1). From the MIC
determination (Figure 5), 85.7% and 74.6% of the strains tested presented a MIC ≥ 0.25 mg ml-1 for the essential oil from genotypes LSID006 and LSID104, respectively, suggesting an intermediate susceptibility learn more of the isolates to the presence of both essential oils. When a paired two-sample t-test was used, the strain susceptibility pattern against each of the essential oils was considered statistically significant (P = 0.05). Figure 5 Minimum inhibitory concentration (MIC) determination of the isolated strains for the essential oil from genotypes LSID006 and LSID104. The bacterial
community in the stems and leaves of four L. sidoides genotypes as determined by a cultivation-independent approach In a cultivation-independent approach (PCR-DGGE), the endophytic bacterial, actinobacterial and fungal communities were STA-9090 research buy evaluated with respect to their structures in the stems
and leaves of L. sidoides genotypes. Highly reproducible PCR-DGGE profiles were obtained from triplicate samples (stems and leaves from the four genotypes) from all communities evaluated in Farnesyltransferase our experiment, indicating the robustness of the PCR-DGGE analyses (data not shown). To facilitate the comparison and further extraction of bands, two replicates per sample were loaded onto each gel. The total bacterial community was first evaluated using the 16S rRNA primer pairs described by Nübel et al. . The DGGE profiles were found to be very similar when DNA samples (stems or leaves) obtained from the four genotypes were compared. However, the same was not observed when the stem-derived samples were compared to leaf-derived samples (Figure 1a). Although certain common bands were detected in all of the samples, it appears that the colonization of the interior of the stems of L. sidoides is dominated by strains that are different from those found in the leaves. Cluster analysis corroborated the visual interpretation of the DGGE profiles, as stem-derived samples were separated from leaf-derived samples at approximately 50% (Figure 1a). Some bands (marked with the letter A, followed by a number) were retrieved from the gel, reamplified and sequenced. Phylogenetic comparison of 14 bands revealed seven sequences affiliated with Enterobacter sp. (A2-A4, A7-A10), one with Pantoea sp. (A5) and six with chloroplast DNA (A1, A6, A11-A14).