Development of Ribosomal RNA-Based Oligonucleotide Probes for Green Ultraphytoplankton

在低营养的海水中，微小的光合浮游生物对初级生产力起着重要的作用。然而，这些存在的超浮游生物的特征很差。从这些种群中培养的藻类可能不能代表实际的优势生物。用高效液相色谱、流式细胞术和电子显微镜描述深水群落结构的尝试提供了一些重要信息，但存在定量和/或定性问题。本项目计划开发一种不同的方法来解决基于核糖体rna的寡核苷酸探针识别和定量光合作用超微浮游生物的问题。将合成与特定核糖体rna序列杂交的短链寡核苷酸（放射性标记的或荧光的）。由于细胞中rRNA的丰度，这些寡核苷酸可以用来探测整个细胞的混合群体。针对特定目的绿藻（特别是已知与一些小型球藻浮游植物有关的Mamiellales）的探针将被生产出来，并用一些球藻和鞭毛虫藻进行测试。这些探针应该允许在野外样品中枚举属于特定目的藻类。

Microscopic, photosynthetic plankton are responsible for a significant portion on the primary productivity in low nutrient ocean waters. However, these ultraplankton which are present are poorly characterized. Algae cultured from these populations may not be representative of the actual dominant organisms. Attempts at describing the structure of deep-water communities by high performance liquid chromatography, flow cytometry, and electron microscopy have provided some important information, but suffer from quantitative and/or qualitative problems. This project plans to develop a different method to address the problem of identifying and quantifying photosynthetic ultraplankton by ribosomal RNA-based oligonucleotide probes. Short strands of oligonucleotides (radiolabelled or fluorescent) which hybridize to specific ribosomal-RNA sequences will be synthesized. Because of the abundance of rRNA in cells, these oligonucleotides can be used to probe mixed populations of whole cells. Probes specific for certain orders of green algae (especially the Mamiellales, which are known to be related to some small coccoid phytoplankton) will be produced and tested with a number of coccoid and flagellate algae. These probes should allow the enumeration of algae belonging to specific orders in field samples.