Ntification of those microorganisms [24]. The 18 selected strains represented, altogether, the six rep-PCR clusters. All strains yielded single amplification goods from the anticipated size (about 1,500 bp) for the 16S rRNA genes and showed identical restriction RsaI profiles (information not shown), characteristic on the genus Azotobacter [2, 24]. When ARDRA was performed applying HhaI, six different profiles were obtained. Cluster evaluation of HhaI restriction profiles revealed 4 distinct clusters at 80 similarity level (Figure two). Due to the fact all strains grouped incluster I showed profiles distinctive of A. chroococcum, as reported by Aquilanti et al. 2004 [2], and identical to these of A. chroococcum reference strain BNM 272, they have been assigned to this species. Cluster II included only strain AT33, which showed a characteristic banding profile in the species A. armeniacus [2], whereas cluster III contained only the 3 A. vinelandii strains used as reference. The ARDRA profiles of strains in cluster IV, obtained experimentally, had been comparable to these of A. salinestris reference strains ATCC 49674T and I-A accomplished in silico. In line with these final results, the strains of heterogeneous cluster IV (Figure 2) had been assigned to A. salinestris. To confirm species identification of isolates, partial sequencing from the 16S rRNA gene was performed for seven strains representing ARDRA clusters. Determined by the similarity observed amongst these sequences, strains AT25 and AT31 in cluster I (Figure two) were associated to A. chroococcum LMG 8756T (99 identity), strain AT33 in cluster II was relatedTable 1: Geographical origin and land use of soil samples from which Azotobacter isolates have been obtained. Summary of fingerprinting and identification benefits of isolates and soil chemical qualities. Isolate OM ( ) three.38 five.72 1.86 1.05 0.98 8.00 8.45 eight.20 five.80 1.21 0.43 1.45 0.48 7.30 0.48 AT25 AT22 AT30 AT31 AT4 AT5 AT9 AT24 AT28 AT43 1 I 1 I 1 nd A. chroococcum A. chroococcum A. chroococcum 1 I A. chroococcum 1 nd A. chroococcum 5.72 two.74 3.15 1.02 0.19 0.17 A. chroococcum A. armeniacus HQ623182 four.47 3.12 7.83 six.40 8.30 six.60 eight.77 eight.60 7.00 6.13 0.80 0.49 0.66 1.58 0.28 0.20 0.93 0.69 eight.50 40.40 45.80 8.ten 4.80 4.50 13.50 10.10 1 I A. chroococcum 1 I A. chroococcum HQ623181 1 I A. chroococcum 1 nd A. chroococcum 1 I A. chroococcum HQ623180 rep-PCR group ARDRA cluster Species assignments Partial 16S rDNA sequence (accession quantity) Soil chemical parameters pH EC (mS cm-1 ) P (ppm) 7.40 51.00 1.90 7.70 eight.Sampling siteMaize stubbleThe Scientific World JournalAgricultural bareLagoon bankLagoon bankGeographical origin Buenos Aires (Azul) Buenos Aires (Balcarce) Buenos Aires (Mar IL-10 Activator medchemexpress Chiquita) Buenos Aires (Mar Chiquita) Buenos Aires (Santa Clara del Mar)Urban landSide of roadNatural pastureNatural pastureRiver bankBuenos Aires (Santa Clara del Mar) Chubut (Esquel) Chubut (Gaiman) Chubut (Trevel ) i Jujuy (CYP2 Activator supplier Tilcara) AT11 AT13 AT26 AT27 AT39 AT32 II 2 nd 1 nd 1 1 1 1 nd nd nd I A. chroococcum A. chroococcum A. chroococcum A. chroococcumSide of routeaJujuy (Tilcara)Natural pastureWheat cropWheat cropWheat crop two nd IV IV ndATA. armeniacus3.six.0.11.Wheat crop 3 AT19 AT14 AT1 3ATA. armeniacus HQ591467 A. salinestris A. salinestris A. salinestris3.15 1.78 1.64 3.six.06 six.40 7.80 7.0.52 0.21 2.24 nd11.10 48.80 3.30 127.Soybean cropNatural pastureEntre R s i (Paran? a C?rdoba o (Corral de Bustos) C?rdoba o (Corral de Bustos) C?rdoba o (Corral de Bustos) Salta (Embarcaci?n) o Salta (Joaqu V. i Gonz e.
