My name is Annelies Beniers. Since 2006 I work at Plant Research International at Business Unit Agrosystems Research in the quantitative Nematology. In 2011, I started besides my job a Master thesis with as specialization Plant Pathology and Entomology.
For my Master thesis, I compared two novel methods for their ability to quantify the number of viable eggs of the potato cyst nematode (Globodera rostochiensis and Globodera pallida). One is based on the loss of membrane integrity upon death and uses trehalose (a disaccharide) as a marker (Van den Elsen et al., 2012), the second test exploits the rapid degeneration of mRNA upon decease with a RNA-specific Real-time PCR assay (Van der Lee, 2011). First, I developed standard curves for both methods. Then, the robustness of the trehalose measurements were checked, as well as the effect of adding cysts with dead content, the use of non-crushed cysts and the effect of a prior fumigant treatment with allylisothiocyanate. To improve the reliability of the Real-time RT-PCR assay an internal virus control of the RNA extraction was developed and to improve the efficiency of the RNA extraction a high throughput method using a robot was tested. I liked to work in the lab and to learn new techniques. The people were very nice of the chair group and helped me a lot with the devices I had to use for my experiments. I’m grateful I had this opportunity.