The development and evaluation of single cell suspension from wheat and barley as a model system; a first step towards functional genomics application

We established growth conditions to allow routine culturing of somatic cells in 24 well microtiter plate format. Evaluation of the wheat and barley cell suspension as model cell system is a multi step process. As an initial step in the evaluation procedure we chose to study the impact of selected abiotic stress elicitors at the physiological, biochemical and molecular level. We report the results of osmotic stress imposed by NaCl and PEG. As proline is an important osmoprotectant of the cereal cells, colorimetric assay for proline detection was developed for small volumes (200 μl). We performed RT-PCR experiments to study the change in the expression of the genes encoding Δ1-pyrroline-5-carboxylate synthetase (P5CS) and Δ1-pyrroline-5-carboxylate reductase (PC5R) in response to abiotic stress.We found differences between the wheat and barley suspension cultures, barley being more tolerant to the applied osmotic stresses. We suggested a model to explain the obtained differences in stress tolerance between the two species. The suspension cell cultures have proven useful for determining changes in proline concentration and expression level of genes (P5CS, P5CR) under various treatments and we suggest that the cells can be used as a model host system to study gene expression and regulation in monocots.Plant cell culture has a very long history. It was in 1902, when German botanist Gottlieb Haberlandt published the article "Cultuversuche mit isolierten Pflanzenzellen" which described his vision of the totipotency of plant cells [1]. Since then plant cell suspension cultures have been used not only for clonal plant propagation but also to investigate the physiological, biochemical, and molecular aspects of various cellular functions. For instance, isolated plant cells have been extensively used to study photosynthesis [2], ion transport [3,4], secondary metabolite production [5], cell growth and differentiation [6] and programmed cell death [7]. Recently Arabidopsis and Z