Design and Evaluation of DNA-Barcoding High Throughput Methods for Analyzing Diatom Diversity: a Test Case Along a South-North Gradient in Central Europe (Rivers Neiße/Oder)

水质和季节波动影响硅藻的数量和丰度。生物检测往往是不完整的，或者对方法改进的要求定位错误。我们的目标是在德国（Neiße/Oder）沿南北梯度建立和评估常规dna条形码高通量检测硅藻生物多样性的方法。在接下来的6年里，我们将在3个不同的季节，在沿水系的7个地点采集49份浮游生物样本。从春季采集的混合水样中提取dna条形码，通过454技术进行扩增和测序，并与参考数据库对齐，将与所有地点的光学显微镜和扫描电镜分析结果进行比较。我们将评估：(1)评估硅藻多样性的dna条形码方法是否可能标准化（定量和/或定性检测）；(2)可以获得对模型河流系统中不同季节南北向梯度硅藻的发生，分布和频率的新见解；(3)该方法适用于常规水质监测；(4)检测到的基因型变异可以分配给代金单样品。(5)尽管形态上具有同源性，但由于基因型变异，可以描述新的硅藻物种。

Water quality and seasonal fiuctuations influence diatom occurrences and abundances. Often organism detection is incomplete or wrong pinpointing to the demand of method amelioration. We aim to establish and evaluate routine DNA-Barcoding high throughput methods for the detection of diatom biodiversity along a south-north gradient in Germany (Neiße/Oder). In 2 Vi years 49 plankton samples will be taken at 7 locations along the river system in 3 different seasons. DNA-Barcodes from mixed water samples of the spring collection, amplified and sequenced via 454 technology and aligned to a reference database, will be compared to results from analyses via light microscopy and SEM from all sites. We will evaluate, whether (1) standardisation of DNA-Barcoding methods for assessing diatom diversity is possible (quantification and/or qualitative detection), (2) new insights into occurrence, distribution and frequency of diatoms along a south-north gradient in the model river system during different seasons can be obtained, (3) the method is applicable for routine water monitoring, (4) the detected genotypic variation can be assigned to voucher samples, and (5) novel diatom species can be described due to genotypic variation despite morphologically homologous traits.