The potential of pmoA amplicon pyrosequencing for methanotroph diversity studies

We analyzed the potential of pmoA amplicon pyrosequencing compared to Sanger sequencing with paddy soils as model environment. We definded OTU cutoff values of 7 and 18 percent reflecting methanotrophic species and major phylogenetic pmoA lineages, respectively. Major lineages were already well covered by clone libraries, nevertheless, pyrosequencing provided higher diversity at the species level.

本研究以稻田土壤为模式环境，对比分析了pmoA扩增子焦磷酸测序（pmoA amplicon pyrosequencing）与桑格测序（Sanger sequencing）的应用潜力。我们设定了7%与18%的操作分类单元（Operational Taxonomic Unit, OTU）截断值，二者分别对应甲烷氧化菌物种及主要的系统发育pmoA进化枝。尽管克隆文库已较好覆盖了主要进化枝，但焦磷酸测序在物种水平上展现出更高的多样性。