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      Carbon Transfer from the Host to Tuber melanosporum Mycorrhizas and Ascocarps Followed Using a 13C Pulse-Labeling Technique

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          Abstract

          Truffles ascocarps need carbon to grow, but it is not known whether this carbon comes directly from the tree (heterotrophy) or from soil organic matter (saprotrophy). The objective of this work was to investigate the heterotrophic side of the ascocarp nutrition by assessing the allocation of carbon by the host to Tuber melanosporum mycorrhizas and ascocarps. In 2010, a single hazel tree selected for its high truffle ( Tuber melanosporum) production and situated in the west part of the Vosges, France, was labeled with 13CO 2. The transfer of 13C from the leaves to the fine roots and T. melanosporum mycorrhizas was very slow compared with the results found in the literature for herbaceous plants or other tree species. The fine roots primarily acted as a carbon conduit; they accumulated little 13C and transferred it slowly to the mycorrhizas. The mycorrhizas first formed a carbon sink and accumulated 13C prior to ascocarp development. Then, the mycorrhizas transferred 13C to the ascocarps to provide constitutive carbon (1.7 mg of 13C per day). The ascocarps accumulated host carbon until reaching complete maturity, 200 days after the first labeling and 150 days after the second labeling event. This role of the Tuber ascocarps as a carbon sink occurred several months after the end of carbon assimilation by the host and at low temperature. This finding suggests that carbon allocated to the ascocarps during winter was provided by reserve compounds stored in the wood and hydrolyzed during a period of frost. Almost all of the constitutive carbon allocated to the truffles (1% of the total carbon assimilated by the tree during the growing season) came from the host.

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          Large-scale forest girdling shows that current photosynthesis drives soil respiration.

          The respiratory activities of plant roots, of their mycorrhizal fungi and of the free-living microbial heterotrophs (decomposers) in soils are significant components of the global carbon balance, but their relative contributions remain uncertain. To separate mycorrhizal root respiration from heterotrophic respiration in aboreal pine forest, we conducted a large-scale tree-girdling experiment, comprising 9 plots each containing about 120 trees. Tree-girdling involves stripping the stem bark to the depth of the current xylem at breast height terminating the supply of current photosynthates to roots and their mycorrhizal fungi without physically disturbing the delicate root-microbe-soil system. Here we report that girdling reduced soil respiration within 1-2 months by about 54% relative to respiration on ungirdled control plots, and that decreases of up to 37% were detected within 5 days. These values clearly show that the flux of current assimilates to roots is a key driver of soil respiration; they are conservative estimates of root respiration, however, because girdling increased the use of starch reserves in the roots. Our results indicate that models of soil respiration should incorporate measures of photosynthesis and of seasonal patterns of photosynthate allocation to roots.
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            13CO2 pulse-labelling of photoassimilates reveals carbon allocation within and between tree rings.

            Post-photosynthetic fractionation processes during translocation, storage and remobilization of photoassimilate are closely related to intra-annual sigma13C of tree rings, and understanding how these processes affect tree-ring sigma13C is therefore indispensable for improving the quality of climate reconstruction. Our first objective was to study the relationship between translocation path and phloem grain. We pulse-labelled a branch of Larix gmelinii (Rupr.) Rupr. and later analysed the sigma13C distribution in the stem. A 13C spiral translocation path closely related to the spiral grain was observed. Our second objective was to study the use of remobilized storage material for earlywood formation in spring, which is a suspected cause of the autocorrelation (correlation of ring parameters to the climate in the previous year) observed in (isotope) dendroclimatology. We pulse-labelled whole trees to study how spring, summer and autumn photoassimilate is later used for both earlywood and latewood formation. Analysis of intra-annual sigma13C of the tree rings formed after the labelling revealed that earlywood contained photoassimilate from the previous summer and autumn as well as from the current spring. Latewood was mainly composed of photoassimilate from the current year's summer/autumn, although it also relied on stored material in some cases. These results emphasize the need for separating earlywood and latewood for climate reconstruction work with narrow boreal tree rings.
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              Seasonal course of translocation, storage and remobilization of 13C pulse-labeled photoassimilate in naturally growing Larix gmelinii saplings.

              Autocorrelation--correlation of tree-ring parameters such as ring width, density and isotope ratios to the environmental conditions of the previous year(s)--is associated with the use of previous photoassimilate for current year's tree ring formation. To clarify the seasonal course of carbon allocation patterns among needles, branches, stem and roots, we pulse-labeled 10 Larix gmelinii growing in a continuous permafrost zone with 13CO2. Photoassimilate incorporated in June was allocated mainly to above-ground parts, indicating active above-ground growth in spring. Very little was allocated to below-ground parts (2.6-7.9%), probably because root growth is inhibited by low soil temperatures in spring. Conversely, a higher proportion of July and August photoassimilate was allocated to below-ground parts (32-44 and 12-24%, respectively). About half the carbon in new needles was derived from stored material. The starch pool in non-needle parts, which can be used for xylem formation, drew approx. 43% of its carbon from the previous year's photoassimilate, indicating that carbon storage is a key mechanism behind autocorrelation in (isotope) dendroclimatology.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2013
                31 May 2013
                : 8
                : 5
                : e64626
                Affiliations
                [1 ]INRA, UMR 1136, Interactions Arbres/Microorganismes (IAM), Centre INRA de Nancy, Champenoux, France
                [2 ]Université de Lorraine, UMR 1136, Interactions Arbres/Microorganismes (IAM), Faculté des Sciences, Vandoeuvre les Nancy, France
                [3 ]INRA, UR 1138, Biogéochimie des Ecosystèmes Forestiers (BEF), Centre INRA de Nancy, Champenoux, France
                [4 ]INRA, UMR 1137, Ecologie et Ecophysiologie Forestières (EEF), Centre INRA de Nancy, Champenoux, France
                [5 ]Université de Lorraine, UMR 1137, Ecologie et Ecophysiologie Forestières (EEF), Faculté des Sciences, Vandoeuvre les Nancy, France
                [6 ]Université de Lorraine, UMR 1121 « Agronomie & Environnement » Nancy-Colmar, Vandœuvre les Nancy, France
                [7 ]INRA, UMR 1121 « Agronomie & Environnement » Nancy-Colmar, Centre INRA de Nancy, Vandœuvre les Nancy, France
                Roehampton University, United Kingdom
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: FLT BZ. Performed the experiments: BZ CP CR FLT. Analyzed the data: CP BZ FLT CR. Contributed reagents/materials/analysis tools: CH CB CP. Wrote the paper: FLT. Edited the manuscript: FLT CR CP BZ.

                Article
                PONE-D-12-36831
                10.1371/journal.pone.0064626
                3669392
                23741356
                49f933d3-9b21-4c16-9ad1-d09686fd6907
                Copyright @ 2013

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 22 November 2012
                : 16 April 2013
                Page count
                Pages: 9
                Funding
                For this work, the authors utilized the online continuous flow CN analyzer (Carlo Erba NA1500) coupled with an isotope ratio mass spectrometer (Finnigan delta S) and DNA sequencing facilities at INRA-Nancy financed by INRA and the Regional Council of Lorraine. The pulse-labeling experiment was supported by the SYSTRUF programme (An integrated approach for sustainable management of ecosystems producing Black Truffle, Tuber melanosporum) financed by the French ANR (Agence Nationale de la Recherche; programme SYSTERRA, ANR-09-STRA-10-02). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Agriculture
                Agricultural Production
                Agroecology
                Agronomic Ecology
                Forestry
                Soil Science
                Biology
                Microbiology
                Plant Science
                Botany
                Mycology
                Fungal Biochemistry
                Fungal Physiology
                Fungi
                Plant Ecology
                Plant-Environment Interactions
                Plant Biochemistry
                Plant Physiology
                Plants

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