Phylogenetic Analysis Based on Internal Transcribed Spacer Spacer Region (ITS1-5.8S-ITS2) of Aspergillus niger Producing Phytase from Indonesia

Aspergillus niger is a filamentous fungus which has been used in industries for the production of metabolites and enzymes like phytase. The present study aimed at performing a phylogenetic analysis of A. niger isolates from soybean and Sorghum flour based on ITS region. Genomic DNA of phytase positive fungal isolates was isolated and a set of primers ITS1F and ITS4R were used to amplify the ITS region and sequenced. Similarity search was performed at National Center for Biotechnology Information (NCBI) for comparison with the reference sequences and identification. Multiple sequences alignment were performed using Clustal W and four isolates designated SB1, SB2, BS, and WF were compared with 18 A. niger sequences from gene bank. Penicillium chryosogenum strain WG11799 (JX406503.1) and Penicillium simplicissimum strain Dal-10 (GU004303.1) were used as an outgoing group. Phylogenetic relationship was estimated using MEGA 7, a phylogenetic tree was inferred using Neighbor-joining methods in 1000 bootstrap replicates run and evolutionary distance was estimated using maximum likelihood model. Similarity analysis showed the isolates SB1 and SB2 were 97% and 99% respectively homology with A. niger voucher MRC200804 (MF078659.1), BS was 99% homology with A. niger IHB F 2919 (KM817216.1 and WF isolate was 98% homology with A. niger SS5 (KJ432863.1). Phylogenetic analysis showed the four strains formed one main clade, which divided to form two subclades; the first consisted of isolate BS and WF and other A. niger species that were closely related but different from another isolate SB1 and SB2 in the second subclade.