the development and evaluation of single cell suspension from wheat and barley as a model system; a first step towards functional genomics application的开发和评价单个细胞悬液从小麦和大麦作为模型系统;.pdf

Dong et al. BMC Plant Biology 2010, 10:239 /1471-2229/10/239 RESEARCH ARTICLE Open Access The development and evaluation of single cell suspension from wheat and barley as a model system; a first step towards functional genomics application 1,2 3 1* Jing Dong , Steve Bowra , Eva Vincze Abstract Background: The overall research objective was to develop single cell plant cultures as a model system to facilitate functional genomics of monocots, in particular wheat and barley. The essential first step towards achieving the stated objective was the development of a robust, viable single cell suspension culture from both species. Results: 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 1 -pyrroline-5-carboxylate synthetase (P5CS) and Δ -pyrroline-5-carboxylate reductase (PC5R) in response to abiotic stress. Conclusions: 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 s