In this hands-on course, students learn to apply a variety of molecular and ecophysiological laboratory techniques that are increasingly being used in today’s global change research. This practical course integrates various research techniques in several smaller global change related research topics combining the theory of several courses of the curriculum. At the unset of the course, several lines of organisms are experimentally subjected to environmental challenges. The experiments implement a panoply of techniques allowing a multidisciplinary approach in order to monitor the morphological as well as biochemical and physiological changes. The student learns the possibilities and restrictions of the most important current experimental techniques which are used on a routine base in current research.
This course consists of 28 days divided over two equal parts corresponding to the semesters in which it is scheduled. In semester 1, the experiment is performed, samples are harvested and the techniques requiring fresh material are performed. In semester 2, we focus on techniques using frozen tissue obtained in semester 1.
In semester 1 the organisms are experimentally subjected to multiple environmental challenges. During the experiments, organism health is monitored and at the end of the experiment, organisms are harvested for analysis of the stress response. For the analysis of plant material, we focus on the in vivo measurement of the gas exchange, stomatal conductance, photosynthesis (via chlorophyll fluorescence) and light interception. These data are coupled to morphological growth characteristics by performing a kinematic growth analysis and growth imaging using microscopy. The stress response is monitored by determination of specific stress enzyme activities and antioxidant capacities using spectrophotometry and HPLC.
In semester 2 we continue with the material harvested in part 1. Physiological, biochemical and molecular experiments are done to further characterize the origin of the morphological and physiological changes observed in the experiments performed during semester 1. The techniques applied are e.g. ELISA and mass spectrometry coupled (UP)LC and GC to allow the identification and quantification of specific primary and secondary metabolites produced as an organism’s specific protection strategy, Gene profiling, transformation, DNA sequencing and verification in combination with the use of reporter constructs to identify specific genes and proteins encoding these strategies. Additionally, performance tests will be conducted using fish to investigate how these organisms cope with a changing environment. These tests will include adverse outcome pathways in zebrafish embryo tests, swimming and respirometry trials in fish, and enzymatic and gene expression analysis.