Summary Mycoheterotrophic plants depend entirely on fungal associations for organic nutrients. While most mycoheterotrophic plants are associated with the mycorrhizal partners of surrounding green plants, some mycoheterotrophs are believed to obtain carbon from decaying litter or dead wood by parasitising saprotrophic fungi, based on culture experiments and 13C and 15N isotopic signatures. The carbon age (the time since carbon was fixed from atmospheric CO2 by photosynthesis) can be estimated by measuring the concentration of 14C arising from the bomb tests of the 1950s and 1960s. Given that mycorrhizal fungi obtain photosynthate from their plant partners, and saprotrophic wood‐decaying fungi obtain carbon from older sources, radiocarbon could represent a new and powerful tool to investigate carbon sources of mycoheterotrophic plants. We showed that the Δ14C values of mycoheterotrophs exploiting ectomycorrhizal fungi were close to 0‰, similar to those of autotrophic plants. By contrast, the Δ14C values of mycoheterotrophs exploiting saprotrophic fungi ranged from 110.7‰ to 324.8‰, due to the 14C‐enriched bomb carbon from dead wood via saprotrophic fungi. Our study provides evidence supporting that some mycoheterotrophic orchids depend on forest woody debris. Our study also indicates that radiocarbon could be used to predict the trophic strategies of mycoheterotroph‐associated fungal symbionts.

The long-distance migrations by marine fishes are difficult to track by field observation. Here, we propose a new method to track such migrations using stable nitrogen isotopic composition at the base of the food web (d 15 N Base ), which can be estimated by using compound-specific isotope analysis. d 15 N Base exclusively reflects the d 15 N of nitrate in the ocean at a regional scale and is not affected by the trophic position of sampled organisms. In other words, d 15 N Base allows for direct comparison of isotope ratios between proxy organisms of the isoscape and the target migratory animal. We initially constructed a d 15 N Base isoscape in the northern North Pacific by bulk and compound-specific isotope analyses of copepods (n = 360 and 24, respectively), and then we determined retrospective d 15 N Base values of spawning chum salmon (Oncorhynchus keta) from their vertebral centra (10 sections from each of two salmon). We then estimated the migration routes of chum salmon during their skeletal growth by using a state-space model. Our isotope tracking method successfully reproduced a known chum salmon migration route between the Okhotsk and Bering seas, and our findings suggest the presence of a new migration route to the Bering Sea Shelf during a later growth stage.

The majority of chlorophyllous orchids form mycorrhizal associations with so‐called rhizoctonia fungi, a phylogenetically heterogeneous assemblage of predominantly saprotrophic fungi in Ceratobasidiaceae, Tulasnellaceae, and Serendipitaceae. It is still a matter of debate whether adult orchids mainly associated with rhizoctonia species are partially mycoheterotrophic. Here, we investigated the nutritional modes of green and albino variants of Goodyera velutina, an orchid species considered to be mainly associated with Ceratobasidium spp., by measuring their 13C and 15N abundances, and by molecular barcoding of their mycorrhizal fungi. Molecular analysis revealed that both green and albino variants of G. velutina harbored a similar range of mycobionts, mainly saprotrophic Ceratobasidium spp., Tulasnella spp., and ectomycorrhizal Russula spp. In addition, stable isotope analysis revealed that albino variants were significantly enriched in 13C but not so greatly in 15N, suggesting that saprotrophic Ceratobasidium spp. and Tulasnella spp. are their main carbon source. However, in green variants, 13C levels were depleted and those of 15N were indistinguishable from the co‐occurring autotrophic plants. Therefore, we concluded that the albino G. velutina variants are fully mycoheterotrophic plants whose C derives mainly from saprotrophic rhizoctonia, while the green G. velutina variants are mainly autotrophic plants, at least at our study site, in spite of their additional associations with ectomycorrhizal fungi. This is the first report demonstrating that adult nonphotosynthetic albino variants can obtain their nutrition mainly from nonectomycorrhizal rhizoctonia.

Human activities have had the strongest impacts on natural ecosystems since the last glacial period, including the alteration of interspecific relationships such as food webs. In this paper, we present a historical record of major alterations of trophic structure by revealing millennium-scale dietary shifts of brown bears (Ursus arctos) on the Hokkaido islands, Japan, using carbon, nitrogen, and sulfur stable isotope analysis. Dietary analysis of brown bears revealed that salmon consumption by bears in the eastern region of Hokkaido significantly decreased from 19% to 8%. In addition, consumption of terrestrial animals decreased from 56% to 5% in western region, and 64% to 8% in eastern region. These dietary shifts are likely to have occurred in the last approximately 100–200 years, which coincides with the beginning of modernisation in this region. Our results suggest that human activities have caused an alteration in the trophic structure of brown bears in the Hokkaido islands. This alteration includes a major decline in the marine-terrestrial linkage in eastern region, and a loss of indirect-interactions between bears and wolves, because the interactions potentially enhanced deer predation by brown bears.

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Most land plants, from liverworts to angiosperms, form mutualistic mycorrhizal symbioses with fungal partners. However, several plants known as mycoheterotrophs exploit fungal partners by reversing the polarity of carbon movement, which usually moves from plant to fungus. We investigated the physiological ecology of a photosynthetic orchid, Apostasia nipponica, which belongs to the first branching group within the Orchidaceae, to improve our understanding of mycoheterotrophic evolution in orchids. The fungal symbionts and nutrition modes of A. nipponica were investigated using molecular barcoding and carbon-13 (13 C) and nitrogen-15 (15 N) measurements, respectively. Community profiling based on a metabarcoding technique revealed that A. nipponica associates with specific Ceratobasidium spp. within ectomycorrhizas-forming clades, whereas isotope analysis revealed that A. nipponica was similar to fully mycoheterotrophic orchids in its 13 C signature and was even more enriched in 15 N than most of the fully mycoheterotrophic orchids that exploit ectomycorrhizal fungi. Our molecular and mass-spectrometric approaches demonstrated, for the first time, that a member of the Apostasioideae, the earliest-diverging lineage of the Orchidaceae, gains carbon through both photosynthesis and fungal cheating (i.e. partial mycoheterotrophy) during the adult stage.

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Abstract1. Isotope analysis has high potential for understanding fish ecology and food-web structure in aquatic ecosystems. The utility of isotope analysis will be greatly improved if we can reconstruct the chronology of several isotopes at multiple growth stages of individual fish. However, no practical methods exist for reconstructing the chronology of light-element isotopes (e.g. δ 13 C, δ 15 N, δ 34 S, andHere, we present and test a new analytical approach for reconstructing the isotopic ratios of light isotopes at multiple life-stages in teleost fishes.2. We sampled an anadromous salmon species, masu salmon Oncorhynchus masou (n = 3), along with water from its natal stream and from the ocean. We subdivided the vertebral centra of the salmon equally into 10 sections and extracted bone collagen from each sample. We then measured the stable sulphur isotope ratios of each vertebral section and compared them with δ 4. Our results show that the vertebral centra of teleost fishes record isotopic information from juvenile to adult life stages. We suggest that our method can provide reproducible isotopic chronology, even in teleost fishes smaller than 50 cm. This method can be used in isoscape studies and in studies of the ecology of marine teleost fishes. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. K E Y W O R D S