Phylogenetic identification of unknown microalgae which isolated from the Caspian Sea

Microalgae are rich sources of active metabolites, which today are considered as important sources of biofuels in addition to medicinal and food uses. In this study, ribosomal markers were used to determine the phylogenetic posation of microalgae species which isolated from the southern coast of the Caspian Sea.

For molecular studies, microalgae grown in suspension culture medium were precipitated using iron (III) chloride solution. After extraction of genomic DNA by freez- thawing method, the gene loci encoding ribosomal subunits (16srDNA) were amplified using 36 pairs oligoes by polymerase chain reaction (PCR), the amplified gene loci were inserted into the pTZ57R /T vector by TA cloning technique then transformed in the Dh5α bacterial and sequenced follow by purification.

Fourteen pairs of primers showed acceptable amplification of target gene in the microalgae genome. The obtained sequences were edited by Vector NTi ver. 11 software and then blasted at the NCBI. Sequences with more than 90% similarity were selected and the molecular distance of the data was estimated by MEGA 6 software and phylogenetic analyzes were performed by calculating the maximum short molecular trees (Maximum Parsimony, MP) for the sequence of the studied genes, and Then, Bootstrap 1000 index was used for analysis to test the accuracy of the nodes.

The results of phylogenetic tree and similarity matrix with Maximum Parsimony method for all 14 gene regions showed that the studied species has the lowest genetic distance with the algal strain Spirulina laxissima.