Department of Biology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
PLoS One. 2011;6(11):e26806. doi: 10.1371/journal.pone.0026806. Epub 2011 Nov 2.
Marine sponges can associate with abundant and diverse consortia of microbial symbionts. However, associated bacteria remain unexamined for the majority of host sponges and few studies use phylogenetic metrics to quantify symbiont community diversity. DNA fingerprinting techniques, such as terminal restriction fragment length polymorphisms (T-RFLP), might provide rapid profiling of these communities, but have not been explicitly compared to traditional methods.
METHODOLOGY/PRINCIPAL FINDINGS: We investigated the bacterial communities associated with the marine sponges Hymeniacidon heliophila and Haliclona tubifera, a sympatric tunicate, Didemnum sp., and ambient seawater from the northern Gulf of Mexico by combining replicated clone libraries with T-RFLP analyses of 16S rRNA gene sequences. Clone libraries revealed that bacterial communities associated with the two sponges exhibited lower species richness and lower species diversity than seawater and tunicate assemblages, with differences in species composition among all four source groups. T-RFLP profiles clustered microbial communities by source; individual T-RFs were matched to the majority (80.6%) of clone library sequences, indicating that T-RFLP analysis can be used to rapidly profile these communities. Phylogenetic metrics of community diversity indicated that the two sponge-associated bacterial communities include dominant and host-specific bacterial lineages that are distinct from bacteria recovered from seawater, tunicates, and unrelated sponge hosts. In addition, a large proportion of the symbionts associated with H. heliophila were shared with distant, conspecific host populations in the southwestern Atlantic (Brazil).
CONCLUSIONS/SIGNIFICANCE: The low diversity and species-specific nature of bacterial communities associated with H. heliophila and H. tubifera represent a distinctly different pattern from other, reportedly universal, sponge-associated bacterial communities. Our replicated sampling strategy, which included samples that reflect the ambient environment, allowed us to differentiate resident symbionts from potentially transient or prey bacteria. Pairing replicated clone library construction with rapid community profiling via T-RFLP analyses will greatly facilitate future studies of sponge-microbe symbioses.
海洋海绵可以与丰富多样的微生物共生体共生。然而,对于大多数宿主海绵,相关细菌仍未被研究,并且很少有研究使用系统发育指标来量化共生体群落的多样性。DNA 指纹图谱技术,如末端限制性片段长度多态性(T-RFLP),可能提供这些群落的快速分析,但尚未明确与传统方法进行比较。
方法/主要发现:我们通过结合重复克隆文库和 16S rRNA 基因序列的 T-RFLP 分析,调查了墨西哥湾北部海洋海绵 Hymeniacidon heliophila 和 Haliclona tubifera、共生被囊动物 Didemnum sp. 以及环境海水相关的细菌群落。克隆文库表明,与两种海绵相关的细菌群落的物种丰富度和物种多样性均低于海水和被囊动物群落,所有四个来源群的物种组成存在差异。T-RFLP 图谱按来源聚类微生物群落;单个 T-RF 与克隆文库序列的大多数(80.6%)匹配,表明 T-RFLP 分析可用于快速分析这些群落。群落多样性的系统发育指标表明,两种海绵相关的细菌群落包括与从海水、被囊动物和无关海绵宿主中回收的细菌不同的优势和宿主特异性细菌谱系。此外,与 H. heliophila 相关的共生体的很大一部分与西南大西洋(巴西)的遥远同种宿主种群共享。
结论/意义:与 H. heliophila 和 H. tubifera 相关的细菌群落的低多样性和物种特异性特征与其他据称普遍存在的海绵相关细菌群落形成鲜明对比。我们的重复采样策略包括反映环境的样本,使我们能够区分驻留共生体和潜在的瞬态或猎物细菌。通过 T-RFLP 分析将重复克隆文库构建与快速群落分析相结合,将极大地促进未来对海绵-微生物共生关系的研究。