Analysis of attached bacteria on algal cells by fluorescence in situ hybridization with oligonucleotide prove

国内外对蓝藻水华的研究较多。然而，目前我们对藻华发生的机理还不是很清楚，也不能预测藻华何时发生。我认为附着在藻细胞上的细菌在控制藻类生长和藻细胞分解方面发挥了重要作用。然而，关于藻细胞上附着的细菌与藻类的关系的研究很少。本研究旨在阐明藻类细胞上的细菌种类及其对藻类水华的影响。首先，利用16S rDNA方法研究了鹿儿湖浮游细菌的季节动态和地理动态。聚合酶链式反应-变性梯度凝胶电泳法(PCR-DGGE)结果表明，不同采样点之间的细菌群落变化不显著，但不同采样点之间的细菌群落变化不明显。然后，用pcr-dgge me…比较了湖水和去除悬浮固体(包括藻细胞)的预滤液中细菌的多样性。更多的方法。在预滤过的湖水中有条带较多的趋势，在湖水中有少量高密度条带的趋势。这一结果被认为是某些细菌种类在悬浮固体上占优势。有一种细菌属于放线杆菌，在总湖水和预滤过的水中经常出现优势菌，然后根据优势放线菌的16S rDNA序列设计了一条引物，对附着在藻细胞上的细菌进行分析。用该引物组和27F通用引物组对采自鹿津湖水中的几个细菌菌落进行了聚合酶链式反应。这套引物发现了两种细菌，并测定了这两种细菌的16S-rDNA序列。然而，不幸的是，这些细菌属于变形杆菌Y亚科。现在，我再次确定了其他几个引物序列。在本研究中，确定了多聚甲醛和荧光染料对鱼体的细菌固定条件。较少

There are many researches for algal bloom by Cyanobacteria. However, we have not quite understood the mechanism of algal bloom and we can not predict when algal blooms happen, yet. I suppose that attached bacteria on algal cells play an important role on control of algal growth and decomposition of algal cells. However, there are few studies about the relationship between attached bacteria on algal cells and algae. In this study, I focused to elucidate the bacterial species on the algal cells, and their effects to the algal bloom.First, I studied seasonal and geographical dynamics of bacterioplankton in Lake Kasumigaura by 16S rDNA methods. The results of PCR-denaturing gradient gel electrophoresis (PCR-DGGE) showed that changes in the bacterial community were prominent between different sampling times, but not between different sampling sites. Next, I compared diversity of bacteria in lake water and water removed suspended solids (include algal cells) by pre-filtrate using PCR-DGGE me … More thods. There is tendency that many bands are in pre-filtrated lake water, on the other hand small number of high density bands are in lake water. This result was regarded that certain bacterial species are predominant on suspended solids. One species bacteria belong to Actinobacteria came frequently predominant in total lake water and pre-filtrated water.Then I made a primer from 16S rDNA sequence of the Actinobacteria species came predominant to analyze the bacteria attached algal cells. PCR was carried out by using primer set of this primer and 27F universal primer on several bacterial colonies that were selected from Kasumigaura lake water. This primer set found 2 species bacteria and I determined 16S-rDNA sequence of this 2 species. However, unfortunately theses bacterial species belong to Proteobacteria Y Subdivision. Now, I am determining several other primer sequences, once again. And, I determined bacterial fixing condition by paraformaldehyde and fluorescent dye for FISH, in this study. Less