Chapter title |
Real-Time Genetic Manipulations of the Cytokinin Pathway: A Tool for Laboratory and Field Studies
|
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Chapter number | 10 |
Book title |
Auxins and Cytokinins in Plant Biology
|
Published in |
Methods in molecular biology, March 2017
|
DOI | 10.1007/978-1-4939-6831-2_10 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6829-9, 978-1-4939-6831-2
|
Authors |
Martin Schäfer, Stefan Meldau |
Editors |
Thomas Dandekar, Muhammad Naseem |
Abstract |
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. |
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