Di Meo C A, Wilbur A E, Holben W E, Feldman R A, Vrijenhoek R C, Cary S C
Graduate College of Marine Studies, University of Delaware, Lewes, Delaware 19958, USA.
Appl Environ Microbiol. 2000 Feb;66(2):651-8. doi: 10.1128/AEM.66.2.651-658.2000.
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.
生活在含硫深海热液喷口和冷泉中的巨型管虫,因其对化学自养内共生体的营养依赖而受到限制。在最近一项使用16S核糖体DNA的系统发育研究中,我们发现来自喷口和冷泉栖息地的内共生体形成了两个不同的进化枝,每个进化枝内的变异很小。在本研究中,我们使用了两种不同的方法来评估生物地理上不同的巨型管虫共生体之间的遗传变异。我们获得了与六种不同属的巨型管虫相关的共生体rRNA操纵子的非编码内部转录间隔区(ITS)区域的DNA序列。来自不同栖息地和广泛分布的喷口地点采集的宿主属内共生体的ITS序列惊人地保守。由于ITS区域不足以区分来自不同栖息地或地点的内共生体,我们使用了一种DNA指纹技术,即重复外基因回文PCR(REP-PCR),来揭示细菌内共生体基因组中重复序列分布的差异。大多数内共生体显示出独特的REP-PCR模式。由这些指纹图谱生成的分支图反映了可能受多种因素影响的关系,包括宿主属、地理位置和海底类型。