The genetic similarity of different generations of Neocallimastix frontalis SK

The genetic similarity of different generations of Neocallimastix frontalis SK was examined by random amplified polymorphic DNA (RAPD) profiling and internal transcribed spacer 1 (ITS1) sequence analysis. N. frontalis SK was subcultured every 2-4 days, and SK-1, SK-3M, and SK-1Y represented N. frontalis SK cultures after one subculture, 50 subcultures, and 150 subcultures. The DNA polymorphisms of the different N. frontalis SK generations were compared by RAPD profiling. The RAPD results gave the same patterns for SK-1, SK-3M and SK-1Y using 12 selected random primers. The partial 18S rDNA, 5.8S rDNA, and ITS1 regions of different generations of N. frontalis SK were amplified and sequenced. The results of alignment and pairwise similarity indicated that the analyzed rRNA sequences of SK-1, SK-3M and SK-1Y were totally identical. This study thus demonstrated genetically identical DNA polymorphisms by RAPD profiling and an unvaried ITS1 region for N. frontalis SK when the strain is subcultured frequently. This suggests that this strain is homokaryotic and grows via an asexual life cycle in vitro.