Denaturing gradient gel electrophoresis analysis of gut bacterial community for the Japanese earthworms

Abstract

The gut bacterial community structure for Pheretima hilgendorfi and P. heteropoda (Family Megascolecidae), and Allolobophora japonica (Family Lumbricidae) collected from agricultural grasslands in Japan was analyzed by denaturing gradient gel electrophoresis of polymerase chain reaction-amplified 16S rRNA gene fragments (PCR-DGGE) and compared with those in the surrounding soils. Denaturing gradient gel electrophoresis (DGGE) profiles indicated that each earthworm species had their own specific bacterial communities, and multidimentional scaling analysis grouped the DGGE profiles into three groups: gut samples from P. hilgendorfi and P. heteropoda, gut samples from A. japonica and samples from the surrounding soils. Nine dominant bands were identified by their direct sequencing and cloning. Major three bands from P. hilgendorfi and P. heteropoda were closely related to Bacillus species belonging to the phylum Firmicutes. Major four and two bands from A. japonica were closely related to the phyla Proteobacteria and Bacteroidetes, respectively.

Keywords:

• Allolobophora japonica
• earthworm gut
• PCR-DGGE
• Pheretima heteropoda
• Pheretima hilgendorfi.

Acknowledgments

The authors are grateful to Dr. Takafumi Gotoh and the staff of Kuju Agricultural Research for their assistance in collecting the earthworms and to Emeritus Prof. Dr. Yoshio Nakamura for his assistance in identifying the earthworm species.