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Nicotiana attenuata, a wild tobacco species native of the southwestern USA that grows in the immediate postfire environment, is one of the important host plants for herbivore populations recolonizing recently burned habitats in the Great Basin Desert. Based on more than 20 years of field research on this eco-genomics model system established in our group, we have developed a genetic and analytical toolbox that allows us to assess the importance of particular genes and metabolites for the survival of this plant in its native habitat. This toolbox has been extensively applied to study the activation of jasmonate signaling after the attack of different herbivore species. Here, we provide detailed guidelines for the analysis, under field conditions, of induced changes in jasmonate pools during insect herbivory. The procedures range from selection and field release of well-characterized transgenic lines for testing the physiological consequences of manipulating jasmonate biogenesis, metabolism, or perception to the metabolic elicitation of chewing herbivore attack and the quantification of the resulting changes in jasmonate fluxes.

The introduction of genetically modified plants into natural habitats represents a valuable means to determine organismic level functions of a gene and its effects on a plant's interaction with other organisms. Nicotiana attenuata, a wild tobacco species native of the southwestern USA that grows in the immediate postfire environment, is one of the important host plants for herbivore populations recolonizing recently burned habitats in the Great Basin Desert. Here, we provide detailed guidelines for the analysis, under field conditions, of jasmonate-dependent defense and its impact on the plant's native herbivore community. The procedures are based on the field release of transgenic lines silenced for jasmonate biogenesis, metabolism, or perception to conduct association studies between defense trait expression (secondary metabolite and trypsin proteinase inhibitor accumulation) and insect infestations. Additionally, because some insects have evolved mechanisms to "eavesdrop" on jasmonate signaling when selecting their host plants, we describe how leafhoppers of the species Empoasca, which selectively colonize jasmonate-deficient plants, can be used as "bloodhounds" for identifying natural variations in jasmonate signaling among natural N. attenuata populations.