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)

水质和季节变化会影响硅藻的数量和数量。针对方法改进的要求，生物体检测往往是不完整或错误的。我们的目标是建立和评估常规的DNA条形码高通量方法，用于检测德国南北梯度(Neiçe/Oder)的硅藻生物多样性。在2个Vi年中，将在3个不同的季节在沿河系统的7个地点采集49个浮游生物样本。来自泉水采集的混合水样的DNA条形码，通过454技术进行放大和测序，并与参考数据库进行比对，将与通过光学显微镜和扫描电子显微镜从所有地点进行分析的结果进行比较。我们将评估：(1)用于评估硅藻多样性的DNA条形码方法是否可能标准化(定量和/或定性检测)；(2)是否可以获得对不同季节模式河系中沿南北梯度的硅藻的发生、分布和频率的新见解；(3)该方法适用于常规水质监测；(4)检测到的基因变异可以归类于凭证样本；以及(5)尽管具有形态上的同源特征，但由于基因变异，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.