Transcriptomic responses to conspecific and congeneric competition in co‐occurring Trifolium

1. Species coexistence requires differential response to inter-and intraspecific competition, typically conceptualized as niche differentiation. Coexistence of close relatives therefore poses an interesting scenario with regards to niche differentiation since these species generally have many traits in common due to shared ancestry. Native Californian Trifolium assemblages are locally diverse and represent a unique system for understanding competitive interactions among close relatives. 2. We conducted two similar greenhouse studies in which Trifolium fucatum was grown alone, with a con-specific competitor, and with a congeneric competitor (Trifolium macraei). In the first study, we assessed biomass production in T. fucatum, and in the second study we conducted an RNAseq analysis of T. fucatum roots to test for differentially expressed genes that may mediate competitive interactions and potentially coexistence. 3. Compared to plants grown alone, competition (i.e. growth in the same pot) with a conspecific resulted in a greater reduction in biomass than competition with a congener, as predicted by theory. However, competition with a congener resulted in twice as many differentially expressed genes as competition with a conspecific. 4. Annotations of identified genes differentiating congeneric from conspecific competition suggest several functions attributed to interactions with third-party organisms, including nodulation with rhizobial mutualists, and defence responses against pathogens and herbivores. In addition, salt-responsive genes and an iron transporter were differentially expressed in congeneric competition, and comparisons of sodium and iron concentrations in field soils where these species are found showed that T. fucatum occurs in higher sodium and iron microsites than T. macraei. Thus, the transcriptome highlighted abiotic niche axes worth pursuing in future ecological experiments as potential mediators of coexistence. 5. We also found a subset of genes that responded similarly to both congeneric and conspecific competition in both direction and magnitude, indicating some conserved responses to competition, regardless of neighbour identity. 6. Synthesis. Transcriptomic analyses represent novel tools for identifying the molecular mechanisms underlying interactions among species. Working iteratively with ecological experimentation and observation, transcriptomes may allow us to identify novel dimensions of the n-dimensional niche that determine species' distributions and their ability to coexist.