Detection of diverse marine algal viruses in the South Sea regions of Korea by PCR amplification of the DNA polymerase and major capsid protein genes

Several molecular techniques have been used to study viruses under different environmental conditions and to examine the genetic diversity of natural virus communities. Here, natural marine virus samples were collected from six different southern coastal regions of Korea and subjected to PCR amplification with five different degenerate primers specific for either the DNA polymerase or capsid protein gene of algal viruses. PCR products ranging from 300 to 700 bp were observed on agarose gel analysis, and major PCR bands were purified and cloned. PCR using primers specific for the viral DNA polymerase gene and for the coat protein gene yielded 332 and 366 clones, respectively. Of the clones analyzed, 147 (44%) DNA polymerase gene clones and 326 (89%) coat protein clones showed similarity to known virus genes. Clustering and assembly revealed 23 and 38 unique sequences for the DNA polymerase and coat protein genes, respectively. BLASTX and phylogenetic analyses of these sequences revealed close relationships with various virus groups, including one of the major algal virus groups, Phycodnaviridae . Pairwise nucleotide comparisons among sequences classified into the same group revealed the extent of genetic diversity in both polymerase and coat protein gene sequences. These results indicated the presence of genetic diversity within similar virus groups in the marine system. Additionally, PCR products with the same sequences were recovered from different locations, indicating the presence of the same virus in different geographic locations in the southern coastal region of the Korean Peninsula.