Ribosomal DNA Sequences in Marine Yeasts: A Model for Identification and Quantification of Marine Eukaryotes

本研究将利用分子技术快速准确地测定群落结构，确定两种截然不同的微真菌类群：担子菌酵母属和卵菌属盐生菌属的真菌生物多样性和种群生物量。这两个群体在以碎屑为基础的食物网中都扮演着重要的角色。研究计划将包括实验室和实地研究。实验室研究将完善已知物种数据库，作为确定野外群落结构的基础。新的程序将开发与使用激光检测红外标记引物的定量PCR （QPCR）的初步重点。实地调查将集中在珊瑚礁和红树林栖息地。利用经典微生物技术和分子方法相结合，确定已知和未知可培养真菌的群落结构和相对丰度。这些物种的身份将通过自动DNA序列分析和与数据库的核苷酸比对来确定。将定位物种特异性区域并开发引物，以测试PCR技术在估计群落结构方面的准确性和敏感性。通过PCR和QPCR，估计不可培养物种的发生和种群密度。本研究开发的技术可以应用于其他微观或宏观真核生物群落的种群分析。

9504105 Fell Using molecular techniques for rapid and accurate determination of community structure, this research will determine fungal biodiversity and population biomass of two distinctly different groups of micro-fungi: the basidiomycetous yeasts and the oomycetous genus Halophythophora. Both groups have important roles in detrital based food webs. The research program will include laboratory and field studies. Laboratory studies will complete the data bank of know species as a basis for determining community structure in the field. New procedures will be developed with preliminary emphasis on quantitative PCR (QPCR) using laser detected infrared labeled primers. Field research will center on reef and mangrove habitats. Using a combination of classical microbial techniques and molecular methods, the community structure and relative abundance of known and unknown culturable fungi species will be determined. The identity of these species will be ascertained by automated DNA sequence analysis and nucleotide alignment with the data bank. Species-specific regions will be located and primers developed to test the accuracy and sensitivity of PCR techniques in estimating community structure. Through the use of PCR and QPCR, the occurrence of unculturable species and population densities will be estimated. The techniques developed in this research can be applied to population analyses of other micro- or macro-eukaryote communities.