In situ Analyses of the Distributions and Phylogeny of Cultivatable and Non-cultivatable Planctomycetales Using Phylogenetic Group-Specific RNA Probes

微生物中的大多数直接计数细菌 生态系统不能用传统的方法来识别或培育， 微生物技术因此，培养的细菌可能不会 代表了真正的微生物多样性。 的 浮游菌目是真细菌的十大分支之一 由16S核糖体RNA定义。 不寻常的核糖体RNA 这群人的签名使他们非常适合 利用核糖体RNA探针技术进行微生物生态学研究。 系统发育群特异DNA研究进展 探测器使得研究其分布成为可能， 浮游菌目在自然生境中的多样性， 培养要求的偏差。 两个探头用于 浮游菌目（PL1R和PL2R）和“通用”rRNA探针 将被用来测量浮游菌目 混合微生物DNA中的核糖体RNA基因 人口数量。 荧光标记的DNA探针将用于 测量浮游菌目对附着在 粘附在接触载玻片上的微生物种群。 系统发育 组特异性基因扩增引物将用于 鉴定浮游菌目的特殊成分 通过对rRNA基因的克隆和测序， 这些 结果将与已知浮菌目的数据库进行比较 以确定以前未被种植的物种 存在于人口中。

The majority of directly countable bacteria in microbial ecosystems cannot be identified or cultivated using traditional microbiological techniques thus, cultured bacteria may not be representative of actual microbial diversity. The Planctomycetales are one of the ten major eubacterial divisions defined by 16S ribosomal RNA. the unusual ribosomal RNA signatures of this group make them exceptionally well-suited for microbial ecology studies using ribosomal RNA probe technologies. The recent development of phylogenetic group-specific DNA probes has made it possible to study the distribution and diversity of the Planctomycetales in natural habitats without the biases of cultivation requirements. Two probes for the Planctomycetales (PL1R and PL2R) and a "universal" rRNA probe will be used to measure the fraction of Planctomycetales ribosomal RNA genes in DNA extracted from mixed microbial populations. Flourescently labeled DNA probes will be used to measure the contribution of the Planctomycetales to attached microbial populations adhering to contact slides. Phylogenetic group-specific gene amplification primers will be used to identify particular constituents of the Planctomycetales population by the cloning and sequencing of rRNA genes. These results will be compared to a data base of known Planctomycetales sequences to determine whether previously uncultivated species are present in the population.