*2.2. Molecular Identification and Characterization of the Isolate ADR1*

Molecular identification of the strain ADR1 was based on 16S rRNA gene sequence analysis. The genomic DNA of the strain ADR1 was isolated using the method developed for Gram-positive bacteria [20] with a few modifications. Briefly, amplification of 16S rRNA gene was carried out using universal primers: V1f (50-AGAGTTTGATCMTGGCTCAG-30), V9r (50-AAGGAGGTGATCCANCCRCA-30), V3f (50-CCAGACTCCTACGGGAGGCAG-30) and V6r (50-ACGAGCTGACGACARCCATG-30) in a PCR machine (Mastercycler® nexus, Eppendorf International, Germany) by using the programme described elsewhere [25]. The amplified product was sequenced by Sanger's method using a 3130XL sequencer (Applied Biosystems, California, USA) for the 16S rRNA gene using universal primers as described above. The sequences were aligned in MEGA 6.0 to generate single consensus sequence. Homology search was performed using the standard Basic Local Alignment Search Tool (BLAST) sequence similarity search tool of the NCBI database to establish the identity of the isolate ADR1. Nucleotide sequences producing significant alignments after BLAST analysis with the 16S rRNA gene sequence of ADR1 were retrieved in FASTA format. These sequences were used to generate phylogenetic relationship of ADR1 with them by using the software, Phylogeny.fr [26,27]. The analysis was done by using advanced mode of this tool, which is an automated programme that performs step-by-step analysis starting from the multiple alignment of the sequences (MUSCLE 3.8.31) [28], alignment curation (Gblocks 0.91b), construction of phylogenetic tree (PhyML 3.1/3.0 aLRT) [29,30] to the visualization of phylogenetic tree (TreeDyn 198.3) [31]. The culture was characterized for its morphological features on different international *Streptomyces* protocol (ISP) media [32]. Isolate ADR1 was streaked on ISP1, ISP2, ISP3, ISP4, ISP5, ISP6 and ISP7 media plates and was incubated for 7 days at 28 ◦C for phenotypic and morphological observations. Single colony morphology of the culture was observed under Nikon stereo zoom microscope SMZ1270 at zooming ratio of 12.7:1 and resolution of 8×. Mycelial structure was observed under Nikon E600 microscope (Nikon, Tokyo, Japan) at a resolution of 100×.
