Characterization of Culturable and Nonculturable Bacteria: A 16S rRNA Sequence Approach

这个项目将使用分子生物学的工具来解决一个长期存在的生物海洋学问题。其主要目标是(A)了解哪种细菌主导自然浮游种群，(B)产生一组异养寡养细菌的纯培养库，以及(C)确定哪些培养物代表自然种群的主要组成部分。到目前为止，大多数自然产生的细菌还没有得到系统发育的特征(或“鉴定”)，因为它们抵制培养它们的尝试，排除了常规鉴定。该项目将使用最先进的RNA测序和DNA克隆技术来绕过这个问题，以表征微生物。这样的工作现在可以在新收集的材料上进行，而不需要培养微生物。该项目将提供(1)明确识别优势海洋细菌，以适合于比较可培养和不可培养种群的物种组成，(2)选择与优势“真实”海洋种群关系最密切的菌株，适合实验室研究，(3)发展分子探针，以确定其他地点的种群分布，从而测量广泛不同环境内和环境之间的多样性；(4)这些探针可能用于单细胞杂交研究，以快速显微鉴定来自任何海洋环境的保存样本中的微生物。

This project will use tools of molecular biology to approach a long-standing biological oceanographic problem. Its major goals are to (a) learn what kinds of bacteria dominate natural planktonic populations, (b) generate a bank of pure cultures of heterotrophic oligotrophs, and (c) determine which, if any, cultures are representative of major components of the natural populations. Until now, most of the naturally-occurring bacteria have not been phylogenetically characterized (or "identified") because they have resisted attempts to culture them, precluding conventional identification. This project will circumvent the problem with state-of-the-art techniques of RNA sequencing, and DNA cloning to characterize microorganisms. Such work can now be done on freshly collected material without culturing the microorganisms. This project will provide for (1) unambiguous identification of the dominant marine bacteria, in a way appropriate for comparing species compositions of both culturable and nonculturable populations, (2) selection of isolates that are most closely related to those dominating "real" marine populations, suitable for lab studies, (3) the development of molecular probes to define population distribu- tions in other locales and thus measure diversity within and between broadly different environments, (4) the possible use of these probes for single-cell hybridization studies for rapid microscopic identification of microorganisms in preserved samples from any marine environment.