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Applied and Environmental Microbiology, February 2000, p. 651-658, Vol. 66, No. 2
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Genetic Variation among Endosymbionts of Widely Distributed Vestimentiferan Tubeworms

Carol A. Di Meo,1 Ami E. Wilbur,2 William E. Holben,3 Robert A. Feldman,4 Robert C. Vrijenhoek,5,dagger and S. Craig Cary1,*

Graduate College of Marine Studies, University of Delaware, Lewes, Delaware 199581; Florida Marine Research Institute, St. Petersburg, Florida 337012; Division of Biological Sciences, The University of Montana, Missoula, Montana 59812-10023; Molecular Dynamics, Sunnyvale, California 94086-45204; and Center for Theoretical and Applied Genetics, Cook College, Rutgers University, New Brunswick, New Jersey 08903-02315

Received 29 July 1999/Accepted 30 November 1999

Vestimentiferan tubeworms thriving in sulfidic deep-sea hydrothermal vents and cold seeps are constrained by their nutritional reliance on chemoautotrophic endosymbionts. In a recent phylogenetic study using 16S ribosomal DNA, we found that endosymbionts from vent and seep habitats form two distinct clades with little variation within each clade. In the present study, we used two different approaches to assess the genetic variation among biogeographically distinct vestimentiferan symbionts. DNA sequences were obtained for the noncoding, internal transcribed spacer (ITS) regions of the rRNA operons of symbionts associated with six different genera of vestimentiferan tubeworms. ITS sequences from endosymbionts of host genera collected from different habitats and widely distributed vent sites were surprisingly conserved. Because the ITS region was not sufficient for distinguishing endosymbionts from different habitats or locations, we used a DNA fingerprinting technique, repetitive-extragenic-palindrome PCR (REP-PCR), to reveal differences in the distribution of repetitive sequences in the genomes of the bacterial endosymbionts. Most of the endosymbionts displayed unique REP-PCR patterns. A cladogram generated from these fingerprints reflected relationships that may be influenced by a variety of factors, including host genera, geographic location, and bottom type.

* Corresponding author. Mailing address: Graduate College of Marine Studies, University of Delaware, Cannon Lab, 700 Pilottown Rd., Lewes, DE 19958. Phone: (302) 645-4078. Fax: (302) 645-4007. E-mail: caryc{at}udel.edu.

dagger Present address: Monterey Bay Aquarium Research Institute, Moss Landing, CA 95039-0628.

Applied and Environmental Microbiology, February 2000, p. 651-658, Vol. 66, No. 2
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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