S with a wide range of values for organic matter content material (0.19.72 ), pH (5.eight.7), electrical conductivity (0.22.two mS cm1 ), and extractable phosphorus (1.927.8 ppm) (Table 1). We obtained 31 bacterial isolates that had been preliminary characterized on the basis of pigment production and cell morphology. All of them made nondiffusible brown pigments in agar medium, showed motile cells, formed cysts in butanolcontaining medium, and showed no fluorescent pigments under UV light (data not shown). 3.2. Genomic Fingerprinting by repPCR. The intraspecific diversity amongst 31 isolates was assessed by means of repPCR. Most isolates showed distinctive banding profiles, reflecting the genetic diversity among them. The cluster analysis of fingerprints revealed six significant groups among all isolates at 55 similarity level (Figure 1). Isolates showing hugely comparable fingerprints (similarity 90 ) have been deemed clonemates. As a result, 23 distinct strains have been obtained. No clear connection may be established amongst repPCR clustering along with the geographical origin of isolates. As an example, group 1 integrated strains which had been isolated from four provinces (Buenos Aires, Chubut, Entre R s, and Jujuy) of your 3 i regions (Pampas, Northwest, and Patagonia).27194-74-7 Chemscene Nonetheless, some tendencies among clustering and the origin of soil samples had been observed. Group 2 clustered all isolates from Crdoba o province (Pampas area), group three integrated strains isolated from Salta and Santiago del Estero provinces (NorthwestSimilarity ( ) 40 60 80 one hundred AT4 AT27 ATBNM 272 A.chroococcummThe Scientific Planet JournalAT13 AT28 AT25 AT39 AT30 AT43 AT31 AT11 AT24 AT5 AT9 AT22 AT32 AT33 AT36 AT1 AT2 AT16 AT17 AT18 AT14 AT19 AT29 AT42 AT37 AT38 AT12 AT4 5Figure 1: Genetic diversity of azotobacteria isolated from agricultural and nonagricultural soils from diverse regions of Argentina revealed by repPCR genomic fingerprinting evaluation. The dendrogram was constructed by using the Pearson correlation coefficient () and also the UPGMA approach working with GelCompar II version 6.5 software. The groups indicated by 1 to six numbers were defined in the 55 similarity level (vertical dashed line).2-Bromo-4-fluoro-5-methylpyridine Purity The cophenetic correlation value for this dendrogram was 0.PMID:24187611 92.area), and group 4 integrated two strains obtained from Chubut province (Patagonia area) (Figure 1 and Table 1). We chose representative strains of each group to classify them using ARDRA. three.3. ARDRA and 16S rRNA Gene Sequence Evaluation. ARDRA with RsaI and HhaI restriction enzymes was utilized to recognize Azotobacter strains to genus and species level, as previously suggested for the molecular identification of those microorganisms [24]. The 18 chosen strains represented, altogether, the six repPCR clusters. All strains yielded single amplification items with the anticipated size (about 1,500 bp) for the 16S rRNA genes and showed identical restriction RsaI profiles (information not shown), characteristic in the genus Azotobacter [2, 24]. When ARDRA was performed using HhaI, six unique profiles have been obtained. Cluster analysis of HhaI restriction profiles revealed 4 distinct clusters at 80 similarity level (Figure 2). Given that all strains grouped incluster I showed profiles distinctive of A. chroococcum, as reported by Aquilanti et al. 2004 [2], and identical to those of A. chroococcum reference strain BNM 272, they had been assigned to this species. Cluster II incorporated only strain AT33, which showed a characteristic banding profile from the species A. armeniacus.
