Ofiles had been analyzed with GelCompar II and compared making use of the Dice similarity coefficient to construct the similarity matrix. The dendrogram was obtained by UPGMA. In silico ARDRA was carried out with HhaI making use of the restriction mapper software program (http://www.restrictionmapper .org/) and 16S rRNA gene sequences AB175656 (A. salinestris ATCC 49674T ) and FJ032010 (A. salinestris IA), both obtained from GenBank. 2.4. 16S rRNA Gene Sequencing. The partial 16S rRNA gene sequence was amplified utilizing primers Y1 and Y2 [15]. Then, amplicons (290 bp) were purified working with the QIAquick PCR purification kit (Qiagen, GmbH) and sequenced by Unidad de Genmica (Instituto de Biotecnolog , INTA, Buenos o i Aires, Argentina) in each directions applying the same primers. The obtained sequences had been compared with these from GenBank applying BLASTN 2.2.16 [16]. two.five. Nucleotide Sequence Accession Numbers. The obtained 16S rRNA gene sequences had been deposited at the GenBank/EMBL/DDBJ database under the following accession numbers: HQ541448, HQ591467, HQ623180, HQ623181, HQ623182, HQ623178, and HQ623179. 2.6. Determination of Prospective Plant GrowthPromoting Traits. Eighteen selected strains have been assessed for siderophore production in line with the OCAS method [17]. Phosphatesolubilizing activity was tested on Pikovskaya medium [18], NBRIP medium [19] and modified Burk’s agar medium [1], adding 0.five of Ca3 (PO4 )2 to every single medium as insoluble P supply. In each assays, Pseudomonas fluorescens2. Components and Methods2.1. Soil Sampling, Bacterial Isolation, and Azotobacter Reference Strains. In total, 74 bulk soil samples (00 cm) have been collected from agricultural (53 samples) and nonagricultural websites (21 samples) in the course of spring 2006. Samples belonged to 38 distinctive places of Northwest, Pampas, and Patagonia regions of Argentina (see Supplementary Material available on-line at http://dx.doi.org/10.1155/2013/519603). Soil aggregates (2 mm) had been spread onto the surface of Petri dishes containing Nfree Burk’s agar medium with mannitol as Csource [1]. Soon after 5 days at 28 C, slimy and glistening Azotobacterlike colonies expanding around soil particles have been chosen and further purified in Nfree LG with bromothymol blue agar medium [1]. Motility, pigment production, and encystment had been determined as previously described [1].The Scientific Globe Journal BNM233 (Banco Nacional de Microorganismos, Buenos Aires, Argentina) was used as a good control. Auxin production was determined using a colorimetric assay [20], with measurements following 1, 2, three, and five days of development in modified LG (LGSP) liquid medium containing 1 sucrose and 0.five soymeal peptone. At each and every time interval, the number of cells (cfu mL1 ) was determined by plate counting on LG agar.Price of Aminoethyl-SS-propionic acid Nitrogenase activity was estimated by the acetylene reduction assay.Methyl 5-bromo-3-hydroxypicolinate Chemscene Bacterial cultures have been grown in Nfree Burk’s agar medium at 28 C for 24 h and ethylene production was measured by gas chromatography [21], utilizing a Hewlett Packard Series II 5890 equipped with a flame ionization detector (FID) along with a stainlesssteel Porapak N column (three.PMID:33752488 two mm two m; 80/100 mesh). The injector, oven, and detector temperatures have been 110 C, 90 C, and 250 C, respectively. N2 was used as carrier gas (4.five cm s1 linear gas velocity). Total protein concentration of bacterial cells was determined by the Lowry approach together with the DC Protein Assay kit (BioRad, USA). Nitrogenase activity was expressed as mmol ethylene produced per mg of protein in 24 h. Indole3acetic acid (IAA),.