Molecular Characteristics of Eisenia fetida (Haplotaxid; Lumbricidae)and Electrophoretic Pattern of Glycolipoprotein Complex of G-۹۰

  BACKGROUND: A large part of our country's waste is organic matter, so researchers are studying fertilizer production from organic waste in various ways, including compost and vermicompost. On the other hand, glycolipoprotein extract of Eisenia fetida (G-۹۰) has been defined to show numerous biological activities, e.g., anticoagulation, fibrinolysis, and anti-oxidative, etc. OBJECTIVES: The purpose of the present study was to determine phylogenetic relationship of the Iranian isolate of Eisenia fetida (E. fetida) with the other available taxa and to determine the G-۹۰ protein complex for evaluation of its biological activities. METHODS: A piece of ۱×۱cm clitellum was separated and used for DNA extraction after homogenate preparation (by two different methods). The second internal transcribed spacer of the nuclear ribosomal DNA (rDNA-ITS۲) and cytochrome C oxidase subunit ۱ of the mitochondrial DNA (mtDNA-COX۱) from adult E. fetida were amplified using polymerase chain reaction (PCR). Subsequently, PCR products were sequenced and their phylogenetic relationships were determined. Further-more, The G-۹۰ protein complex was extracted from adult worms and electrophoretic pattern of proteins was obtained by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: The electrophoretic pattern of glycolipoprotein G-۹۰, a protein complex, showed ۱۰ bands with molecular weights of ۱۴-۱۳۰ kDa. ITS۲ and COX۱ sequences were ۵۱۶ bp and ۲۷۷ bp long, respectively. The amplified DNA sequences from both ribosomal and mitochondrial sequences had ۸۸%-۹۹% and ۹۹% similarity to relevant sequences in GenBank, respectively. CONCLUSIONS: mtDNA-COX۱ showed no considerable sequence variations as compared to other isolates in Gen-Bank, while rDNA-ITS۲ exhibited more variations, in comparison with other isolates, indicating more variations among some of these isolates. Our results revealed that rDNA-ITS۲ of our E. fetida was in a separate subclade, showing the greatest similarity with EF۵۳۴۷۰۹.۱ isolate (۹۹%) provided in GenBank, followed by JX۵۳۱۶۱۸.۱ (۹۴%), and KU۷۰۸۴۶۹.۱ (۸۸%). Our COX۱ sequence demonstrated the high similarity of ۹۹% when compared with five isolates from GenBank (MH۴۷۵۶۷۴.۱, MH۴۷۵۶۷۳.۱, MH۴۷۵۶۷۲.۱, MH۴۷۵۶۷۰.۱, and MH۴۷۵۶۶۶.۱). The G-۹۰ glycolipoprotein showed different proteins that can be assessed for their potential biological activities in medicinal properties

کلیدواژه ها:

earthworm ، Eisenia fetida ، G-۹۰ ، Molecular characteristics