INFECTION OF EYED EGGS OF RAINBOW TROUT WITH FLAVOBACTERIUM COLUMNARE

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。我们将使用培养和分子方法相结合的方法，对黄松叶片在出苗、成熟、衰老和分解过程中的细菌演替进行初步研究。我们将通过在R2a琼脂上涂抹洗叶液，从活的和死亡的、分解的叶子中分离出可培养的细菌。我们还将从叶片表面提取细菌DNA，并通过聚合酶链式反应扩增细菌16S rRNA基因。将不同菌种的16S rRNA产物进行变性梯度凝胶电泳(DGGE)。从DGGE凝胶中获得最丰富的16S rRNA产物将被克隆和测序。从DGGE凝胶和培养的细菌中搜索16S rRNA序列，在16S rRNA数据库中进行临时鉴定，并确定是否可以培养活的和死亡的松叶上丰富的细菌物种。荧光显微镜将被用来估计不同时期松树叶片上的细菌总数。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We will use a combination of culturing and molecular methods to conduct a preliminary study of bacterial succession on the leaves of Pinus ponderosa, as leaves emerge, mature, senesce, and decompose. We will isolate culturable bacteria from living and dead, decomposing leaves by plating leaf washes on R2A agar. We will also isolate bacterial DNA from leaf surfaces and amplify bacterial 16S rRNA genes by PCR. The 16S rRNA products of different bacterial species will be separated denaturing gradient gel electrophotesis(DGGE). The most abundant 16S rRNA products from DGGE gels will be cloned and sequenced. 16S rRNA sequences from DGGE gels and cultured bacteria will be searched against 16S rRNA databases to make provisional identifications, and determine if the abundant bacterial species on living and dead pine leaves can be cultured. Epifluorescence microscopy will be used to estimate total bacterial numbers on pine leaves of different stages.