Plant Protoplasts as a Tool in Systems Biology

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Dovzhenko A.*, Palme K.**

Single plant cell analysis is a new frontier in plant gene function studies, and, through new discoveries derived from cellular heterogeneity, single cell analysis has the potential to decipher complicated molecular networks using systems biology approaches. Here we present examples on how single plant cells such as protoplasts can be used as unique models for plant systems biology. We have developed high-throughput assays using protoplasts from Arabidopsis thaliana and Nicotiana tabacum. Cells were immobilized and cultured in multi-well plates, underwent various treatments, and subjected to a novel automated image analysis platform. We demonstrate that by combining automated microscopy with TIRF and FLIM imaging techniques and computational analysis protoplasts are useful tool for multi-parametric in vivo cellular analysis. Assays to efficiently select functional artificial miRNAs and to identify factors regulating cell fate and development have been established. Future perspectives of this work will be discussed.