Real-Time Genetic Manipulations of the Cytokinin Pathway: A Tool for Laboratory and Field Studies
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Although many established tools for cytokinin (CK) pathway manipulations are well suitable for the analysis of molecular interactions, their use on a whole plant scale is often limited by the induction of severe developmental defects. To circumvent this problem, different methods were developed that allow for a more precise manipulation of the CK pathway. Here we present one of these systems, the pOp6/LhGR system for chemically inducible gene expression. This system allows regulation on a spatial, temporal, and quantitative scale and therefore provides a superior tool for analyzing the role of CKs in the interactions of plants with their environment. The pOp6/LhGR system was tested for RNAi-mediated gene silencing and heterologous gene expression and was successfully used for CK pathway manipulations in different model organisms (Arabidopsis thaliana, Nicotiana tabaccum, Nicotiana attenuata, Citrus sinensis × C. trifoliate). Here we describe specific aspects of the screening procedure and present an experimental setup that can not only be used in the laboratory but is also applicable under field conditions.
Key wordsCytokinins Isopentenyl transferase pOp6 LhGR Dexamethasone Stress response Ecology Fieldwork
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