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

通过寡核苷酸荧光原位杂交分析藻细胞上附着的细菌证明

• 批准号: 12640692
• 负责人: TOMIOKA Noriko
• 金额: $ 1.6万
• 依托单位: National Institute for Environmental Studies
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12640692/
• 关键词: FISH; DGGE; clone analysis; eutropic lake; hybridization; アオコ

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

蓝藻水华的研究很多。然而，目前人们对水华的发生机制还不十分了解，也无法预测水华的发生时间。笔者认为藻类细胞上的细菌在控制藻类生长和分解藻类细胞方面起着重要作用。然而，关于藻细胞上附着的细菌与藻类之间的关系的研究却很少。本研究以霞浦湖浮游细菌为研究对象，首先利用16 SrDNA技术研究了霞浦湖浮游细菌的季节和地理动态。PCR-变性梯度凝胶电泳（PCR-DGGE）结果表明，不同采样时间的细菌群落结构变化显著，但不同采样点的细菌群落结构变化不显著。其次，利用PCR-DGGE技术比较了湖水和预滤去除悬浮物（包括藻类细胞）后的水中细菌的多样性 ...更多信息 方法过滤前的湖水中有多条谱带的趋势，而过滤后的湖水中有少量高密度谱带。这一结果被认为是某些细菌物种在悬浮固体上占优势。本研究以总湖水和预滤水中的放线菌为优势菌，根据该优势菌的16 SrDNA序列设计引物，对藻细胞上的细菌进行了分析。用该引物和27 F通用引物对霞浦湖水中的几个细菌菌落进行PCR扩增。用该引物对2种细菌进行了16 S-rDNA序列测定。然而，不幸的是，这些细菌物种属于变形菌Y亚门。现在，我再一次确定其他几个引物序列。同时，本研究还确定了FISH用多聚甲醛和荧光染料固定细菌的条件。少