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‘Real time’ genetic manipulation: a new tool for ecological field studies
Author(s) -
Schäfer Martin,
Brütting Christoph,
Gase Klaus,
Reichelt Michael,
Baldwin Ian,
Meldau Stefan
Publication year - 2013
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/tpj.12301
Subject(s) - biology , gene silencing , rna interference , gene , phytoene desaturase , gene expression , ectopic expression , epigenetics , transgene , genetics , small hairpin rna , genetically modified crops , microbiology and biotechnology , ecology , gene knockdown , rna
Summary Field experiments with transgenic plants often reveal the functional significance of genetic traits that are important for the performance of the plants in their natural environments. Until now, only constitutive overexpression, ectopic expression and gene silencing methods have been used to analyze gene‐related phenotypes in natural habitats. These methods do not allow sufficient control over gene expression for the study of ecological interactions in real time, of genetic traits that play essential roles in development, or of dose‐dependent effects. We applied the sensitive dexamethasone ( DEX )‐inducible p O p6/ L h GR expression system to the ecological model plant N icotiana attenuata and established a lanolin‐based DEX application method to facilitate ectopic gene expression and RNA interference‐mediated gene silencing in the field and under challenging conditions (e.g. high temperature, wind and UV radiation). Fully established field‐grown plants were used to silence phytoene desaturase and thereby cause photobleaching only in specific plant sectors, and to activate expression of the cytokinin ( CK ) biosynthesis gene isopentenyl transferase ( ipt ). We used ipt expression to analyze the role of CK s in both the glasshouse and the field to understand resistance to the native herbivore T upiocoris notatus , which attacks plants at small spatial scales. By spatially restricting ipt expression and elevating CK levels in single leaves, damage by T . notatus increased, demonstrating the role of CK s in this plant–herbivore interaction at a small scale. As the arena of most ecological interactions is highly constrained in time and space, these tools will advance the genetic analysis of dynamic traits that matter for plant performance in nature.