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Institute
In this work, a protocol for portable nanopore sequencing of DNA from pollen collected from honey bees, bumble bees, and wild bees was developed. DNA metabarcoding is applied to identify genera within the mixed DNA samples. The DNA extraction and ITS and ITS2 PCR parameters tested for this purpose were applied to the collected pollen sample and the amplicons were then decoded using the Flongle sequencer adapter from Oxford Nanopore Technologies. It is shown that the main pollinator resources at the different sites can be identified in percentage proportions. The protocol generated in this study can be used for further ecological questions.
In the present bachelor thesis, nanopore sequencing and Illumina sequencing was compared using pollen DNA collected from honeybees and bumble bees. Therefore, nanopore sequencing was performed with the MinION sequencers and the generated reads were analysed with bash programming. A quantitative and qualitative (based on ITS2 sequences) BLAST run was performed. The results confirme the error probability of nanopore sequencing that is described in the literature. Nevertheless, with both sequencing methods similar sample preferences of the bees could have been observed, allowing ecological conclusions.
Development of a genetic biomonitoring test for the investigation of pollinator-plant-interactions
(2021)
There is a world-wide decline in biodiversity recorded. Especially insects and accompanying pollinators are threatened. When the foraging behaviour of pollinators is understood in detail, future crop and floral pollination services can be sustained and it is possible to establish projects for the conservation of pollinators and plant biodiversity. With the use of nanopore sequencing methods it is possible to detect pollen species that were collected by pollinators by their genetic information. In this study, a protocol for portable nanopore sequencing of DNA from pollen that was collected by honey bees, bumble bees and wild bees is being designed. DNAmetabarcoding is used to identify species within the mixed DNA sample. The ITS2-region will be used as a barcode. We will investigate pollen preferences of three pollinator species by placing their hives or nests at the same. Based on the results, landscape management schemes are developed that target pollen preferences and nutritional requirements of managed and wild social bee species as well as solitary wild bees.