Determine the genotype of the local fungal isolation Aspergillus niger 5 producers of citric acid

The current study included determining the genotype of local fungal isolation Aspergillus niger 5 producer of citric acid and preserved in the laboratory of fungi in the Department of Biology/College of Science/ University of AL-Qadisiya by using PCR-RFLP technique and get the sequence of DNA bases for region ITS1-5.8S rRNA gene-ITS2 and determining variation areas, genetic distances and Phylogenetic tree analysis for a local fungal isolation and compare it with some of the world fungal isolates registered in the National Centre for Information biotechnology database (NCBI) by using the MEGA 6 program. The results of the extraction of DNA of five replicates of A. niger 5 contains a single and clear DNA bands and after the measured by Nanodrop spectrophotometer was concentrations of DNA ranging from (12.6-15.0) ng/µL and purity ranged between (1.44-1.69). and after amplification the DNA by using the PCR technique was the result of amplification a molecular size 546 base pairs. The results of enzymatic cutting of the result of amplification by using RsaI restriction enzyme showed a single two bands with a molecular size (480,66) base pairs and that the genotype of the fungus under study is of the type (A). also found 10 variation areas after A lineup sequences of nucleotides for isolate under study and compare it with some of the world fungal isolates of A. niger registered in the (NCBI) and the Phylogenetic tree analysis showed the presence of A. niger 5 with six world isolates (HQ392477.1 Malaysia, JX291197.1 Malaysia, JX036483.1 China, HQ392475.1 Malaysia, KJ881376.1 India, JX501376.1 Malaysia) in one secondary group.