Plants That Eat Animals: Apart from some spectacular exceptions, such as pitcher plants and Venus fly traps, most plants are thought to acquire nitrogen passively from microbial decomposition and the activities of nitrogen-fixing bacteria. Metarhizium species are common endophytes—fungi that live within plant tissues without causing disease. This genus is also found ubiquitously in soil, where they parasitize insects. In a series of microcosm experiments, Behie et al. (p. 1576 ) investigated whether these fungi could couple their endophytic life-styles with their parasitic modes and be a conduit by which plants could obtain nitrogen from animals. Radio-labeled moth larvae were added to the microcosms in which bean and grass plants were grown, and when the larvae were inoculated with fungi, it was only a matter of days before the nitrogen label was detected in the plants.

• Premise of the study: The soil‐inhabiting insect‐pathogenic fungus Metarhizium robertsii also colonizes plant roots endophytically, thus showing potential as a plant symbiont. Metarhizium robertsii is not randomly distributed in soils but preferentially associates with the plant rhizosphere when applied in agricultural settings. Root surface and endophytic colonization of switchgrass (Panicum virgatum) and haricot beans (Phaseolus vulgaris) by M. robertsii were examined after inoculation with fungal conidia.• Methods: We used light and confocal microscopy to ascertain the plant endophytic association with GFP‐expressing M. robertsii. Root lengths, root hair density, and lateral roots emerged were also observed.• Key results: Initially, M. robertsii conidia adhered to, germinated on, and colonized roots. Furthermore, plant roots treated with Metarhizium grew faster and the density of plant root hairs increased when compared with control plants. The onset of plant root hair proliferation was initiated before germination of M. robertsii on the root (within 1–2 d). Plants inoculated with M. robertsii ΔMAD2 (plant adhesin gene) took significantly longer to show root hair proliferation than the wild type. Cell free extracts of M. robertsii did not stimulate root hair proliferation. Longer‐term (60 d) associations showed that M. robertsii endophytically colonized cortical cells within bean roots. Metarhizium appeared as a mycelial aggregate within root cortical cells as well as between the intercellular spaces with no apparent damage to the plant.• Conclusions: These results suggest that M. robertsii is not only rhizosphere competent but also displays a beneficial endophytic association with plant roots that results in the proliferation of root hairs.

The study of symbiotic nitrogen transfer in soil has largely focused on nitrogen-fixing bacteria. Vascular plants can lose a substantial amount of their nitrogen through insect herbivory. Previously, we showed that plants were able to reacquire nitrogen from insects through a partnership with the endophytic, insect-pathogenic fungus Metarhizium robertsii . That is, the endophytic capability and insect pathogenicity of M. robertsii are coupled so that the fungus acts as a conduit to provide insect-derived nitrogen to plant hosts. Here, we assess the ubiquity of this nitrogen transfer in five Metarhizium species representing those with broad ( M. robertsii , M. brunneum , and M. guizhouense ) and narrower insect host ranges ( M. acridum and M. flavoviride ), as well as the insect-pathogenic fungi Beauveria bassiana and Lecanicillium lecanii . Insects were injected with ¹⁵ N-labeled nitrogen, and we tracked the incorporation of ¹⁵ N into two dicots, haricot bean ( Phaseolus vulgaris ) and soybean ( Glycine max ), and two monocots, switchgrass ( Panicum virgatum ) and wheat ( Triticum aestivum ), in the presence of these fungi in soil microcosms. All Metarhizium species and B. bassiana but not L. lecanii showed the capacity to transfer nitrogen to plants, although to various degrees. Endophytic association by these fungi increased overall plant productivity. We also showed that in the field, where microbial competition is potentially high, M. robertsii was able to transfer insect-derived nitrogen to plants. Metarhizium spp. and B. bassiana have a worldwide distribution with high soil abundance and may play an important role in the ecological cycling of insect nitrogen back to plant communities.

Metarhizium robertsii is a common soil fungus that occupies a specialized ecological niche as an endophyte and an insect pathogen. Previously, we showed that the endophytic capability and insect pathogenicity of Metarhizium are coupled to provide an active method of insect-derived nitrogen transfer to a host plant via fungal mycelia. We speculated that in exchange for this insect-derived nitrogen, the plant would provide photosynthate to the fungus. By using ¹³CO₂, we show the incorporation of ¹³C into photosynthate and the subsequent translocation of ¹³C into fungal-specific carbohydrates (trehalose and chitin) in the root/endophyte complex. We determined the amount of ¹³C present in root-associated fungal biomass over a 21-day period by extracting fungal carbohydrates and analysing their composition using nuclear magnetic resonance (NMR) spectroscopy. These findings are evidence that the host plant is providing photosynthate to the fungus, likely in exchange for insect-derived nitrogen in a tripartite, and symbiotic, interaction.