Sequence and Phylogenetic Analysis of SSU rRNA Gene of Five Microsporidia

Abstract

The complete small subunit rRNA (SSU rRNA) gene sequences of five microsporidia including Nosema heliothidis, and four novel microsporidia isolated from Pieris rapae, Phyllobrotica armta, Hemerophila atrilineata, and Bombyx mori, respectively, were obtained by PCR amplification, cloning, and sequencing. Two phylogenetic trees based on SSU rRNA sequences had been constructed by using Neighbor-Joining of Phylip software and UPGMA of MEGA4.0 software. The taxonomic status of four novel microsporidia was determined by analysis of phylogenetic relationship, length, G?C content, identity, and divergence of the SSU rRNA sequences. The results showed that the microsporidia isolated from Pieris rapae, Phyllobrotica armta, and Hemerophila atrilineata have close phylogenetic relationship with the Nosema, while another microsporidium isolated from Bombyx mori is closely related to the Endoreticulatus. So, we temporarily classify three novel species of microsporidia to genus Nosema, as Nosema sp. PR, Nosema sp. PA, Nosema sp. HA. Another is temporarily classified into genus Endoreticulatus, as Endoreticulatus sp. Zhenjiang. The result indicated as well that it is feasible and valuable to elucidate phylogenetic relationships and taxonomic status of microsporidian species by analyzing information from SSU rRNA sequences of microsporidia.

Description of microsporidia in simulids: molecular and morphological characterization of microsporidia in the larvae of Simulium pertinax Kollar (Diptera: Simuliidae)

“…This identifi cation corroborated previous results [22][23][24][25][26] regarding the phylogeny of these parasites.…”

Complete Sequence and Gene Organization of the Nosema heliothidis Ribosomal RNA Gene Region

“…The spore band was collected and washed several times with sterile water (Johny et al 2006; Tsai et al 2003). Genomic DNA was extracted and the PCR conditions, primer sets, and sequencing methods followed those described previously (Dong et al 2010; Zhu et al 2010). The length and G+C content of the sequences of N. heliothidis rRNA sequences were analyzed with the Editseq program of DNAStar software.…”

Identification of a microsporidian isolate from Cnaphalocrocis Medinalis and its pathogenicity to Bombyx mori

“…The SSU rRNA gene sequence of CmM2 was compared with the homologous sequences, which were obtained from GenBank through search engine (http://www.ncbi.nlm.nih.gov/BLAST). Some SSU rRNA gene sequences of Nosema , Amblyospora , Endoreticulatus , Vairimorpha , Pleistophora , and Glugea were downloaded, the phylogenetic tree based on SSU rRNA gene sequence of CmM2 and 22 sequences of other strains of microsporidia was constructed, 1000 bootstrap replications were generated to test the robustness of the trees, Beauveria bassiana (EU334679) was used as an out‐group, as implemented by the Neighbor‐Joining of MEGA5.0 software . The similarity and genetic distance of SSU rRNA gene sequences of some sequences was analyzed by MegAlign of DNAStar software.…”