A persistent major mutation in canonical jasmonate signaling is embedded in an herbivory-elicited gene network.

Abstract

When insect herbivores attack plants, elicitors from oral secretions and regurgitants (OS) enter wounds during feeding, eliciting defense responses. These generally require plant jasmonate (JA) signaling, specifically, a jasmonoyl-L-isoleucine (JA-Ile) burst, for their activation and are well studied in the native tobacco <i>Nicotiana attenuata</i>. We used intraspecific diversity captured in a 26-parent MAGIC population planted in nature and an updated genome assembly to impute natural variation in the OS-elicited JA-Ile burst linked to a mutation in the JA-Ile biosynthetic gene <i>NaJAR4</i>. Experiments revealed that <i>NaJAR4</i> variants were associated with higher fitness in the absence of herbivores but compromised foliar defenses, with two <i>NaJAR</i> homologues (4 and 6) complementing each other spatially and temporally. From decade-long seed collections of natural populations, we uncovered enzymatically inactive variants occurring at variable frequencies, consistent with a balancing selection regime maintaining variants. Integrative analyses of OS-induced transcriptomes and metabolomes of natural accessions revealed that <i>NaJAR4</i> is embedded in a nonlinear complex gene coexpression network orchestrating responses to OS, which we tested by silencing four hub genes in two connected coexpressed networks and examining their OS-elicited metabolic responses. Lines silenced in two hub genes (<i>NaGLR</i> and <i>NaFB67</i>) co-occurring in the <i>NaJAR4/6</i> module showed responses proportional to JA-Ile accumulations; two from an adjacent module (<i>NaERF</i> and <i>NaFB61</i>) had constitutively expressed defenses with high resistance. We infer that mutations with large fitness consequences can persist in natural populations due to compensatory responses from gene networks, which allow for diversification in conserved signaling pathways and are generally consistent with predictions of an omnigene model.