Viral Diversity and Mortality of Phyto- and Bacterioplankton in the Arctic Ocean

摘要：北冰洋浮游植物和细菌的病毒多样性和死亡率本提案要求资助研究北冰洋病毒攻击导致的细菌和浮游植物死亡率的幅度和变异性。还将调查研究区域病毒的分布、丰度以及病毒组成和多样性的变化。除了对自然病毒群落进行检查外，还将对样品进行筛选，以确定是否存在来自污水的细菌病毒，作为陆地生物污染物渗透到北冰洋的指标。美国海军潜艇将在采样点完成海水样本的收集、固定和浓缩。后续分析将在实验室进行。在固定样品上用电子显微镜测定浮游植物和细菌的病毒总数和病毒感染频率。为了检验自然群落的多样性，将对病毒浓缩液进行纯化并进行脉冲场凝胶电泳，以根据基因组大小分离病毒DNA。根据不同条带的数量、位置和染色强度，可以估计每个样本中病毒的多样性、基因组大小和丰度。污水衍生的病毒将通过使用聚合酶链反应扩增目标DNA序列和通过空斑测定来检测。通过与其他研究人员的协调，病毒丰度、病毒感染频率和病毒群落多样性将与水文和生物变量相关，以确定那些可能在调节病毒传播中起重要作用的变量。

Proposal No: OPP-9623130 Principal Investigator: Azam, Farooq ABSTRACT Viral Diversity and Mortality of Phyto-and Bacterioplankton in the Arctic Ocean This proposal requests funding to investigate the magnitude and variability of bacteria and phytoplankton mortality due to viral attack in the Arctic Ocean. Viral distribution, abundance and variations in composition and diversity of viruses in the study area will also be investigated. In addition to the examination of natural viral communities, samples will be screened for the presence of sewage-derived bacterial viruses as an indicator of the penetration of terrestrial biological pollutants into the Arctic Ocean. Collection, fixation and concentration of sea water samples will be accomplished at sampling sites by deployment from a U.S. Navy submarine. Subsequent analyses will be conducted in the laboratory. Total counts of viruses and the frequency of viral infection in phytoplankton and bacteria will be determined by electron microscopy on fixed samples. To examine natural community diversity, concentrates of viruses will be purified and subjected to pulsed-field gel electrophoresis to separate viral DNA based on genome size. From the number, positions, and staining intensity of different bands, the diversity, genome sizes, and abundance of viruses in each sample will be estimated. Sewage derived viruses will be detected by amplification of target DNA sequences using the polymerase chain reaction and by plaque assay. Through coordination with other researchers, viral abundance, the frequency of viral infections, and viral community diversity will be related to hydrographic and biological variables in order to determine those which may be important in regulating viral propagation.