Abstract
Despite the strong ecological importance of ectomycorrhizal (ECM) fungi, their vertical distribution remains poorly understood. To our knowledge, ECM structures associated with trees have never been reported in depths below 2 meters. In this study, fine roots and ECM root tips were sampled down to 4-m depth during the digging of two independent pits differing by their water availability. A meta-barcoding approach based on Illumina sequencing of internal transcribed spacers (ITS1 and ITS2) was carried out on DNA extracted from root samples (fine roots and ECM root tips separately). ECM fungi dominated the root-associated fungal community, with more than 90% of sequences assigned to the genus Pisolithus. The morphological and barcoding results demonstrated, for the first time, the presence of ECM symbiosis down to 4-m. The molecular diversity of Pisolithus spp. was strongly dependent on depth, with soil pH and soil water content as primary drivers of the Pisolithus spp. structure. Altogether, our results highlight the importance to consider the ECM symbiosis in deep soil layers to improve our understanding of fine roots functioning in tropical soils.
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Acknowledgments
We would like to thank the staff of the Itatinga Experimental Station (ESALQ-USP), as well as all the students present at the station during the sampling, and Eder Araujo da Silva and Floragro for their technical support. The site belongs to the SOERE F-ORE-T network, which is supported annually by ECOFOR, AllEnvi and the French national research infrastructure ANAEE (http://www.anaeefrance.fr/fr/). This work was performed in collaboration with the GeT core facility, Toulouse, France (http://get.genotoul.fr/), and was supported by France Génomique National infrastructure, funded as part of “Investissement d’avenir” program managed by Agence Nationale pour la Recherche (contract ANR-10-INBS-09).
Funding
This research was funded by the Centre de Coopération Internationale en Recherche Agronomique pour le Développement (CIRAD) and the project FAPESP 2016/18944-3 “Climate change and energy efficiency in agriculture: a focus on water stress, organic management and soil biology”
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A.R., J.P.L., J.P.B., F.D.A., E.C., C.P., J.L.M.G, P.H., C.J., designed the study; A.R., C.P., G.R.L., A.P.A.P., A.G., M.C.S. A.L.P., P.T., P.H., performed research; F.M., H.S, A.R., A.G., M.S., analyzed data; A.R. wrote the paper with contributions of C.P. and of all authors; all the authors helped to interpret the results of the study, reviewed the manuscript and contributed substantially to the revisions.
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Table S1.
Physical and chemical properties of the soil in the W+ and W- pits. The data comes from Pradier et al. (2017). The soil samples were collected at the same time, from the same pits, as the fine roots and ECM root tips analyzed in this study. Different letters indicate significant differences between depths. P-values in bold with an asterisk indicate significant effects (Tukey test, P < 0.05). pH: pH measured in CaCl2, %H: humidity percentage, %C and %N: total C and N contents, P: P extracted with an anion exchange resin, K: K extracted with a cation exchange resin. (DOCX 46 kb)
Figure S1.
Soil water content (v/v%) in the W+ and W- plots down to 4 meters. The 4 months around the sampling period are outlined. SWC was interpolated by a contour line interpolation using marching square algorithm (R software version 3.2.5 and the plotly package version 4.5.6). (PDF 146 kb)
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All codes and representative OTU sequences in HTML format. The purpose of this document is to provide the reader with details about the bioinformatic methods used to prepare this paper. The code snippets and shell commands presented here were executed on a Debian GNU/Linux 8, and might have to be adapted to your particular system. (HTML 206 kb)
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Robin, A., Pradier, C., Sanguin, H. et al. How deep can ectomycorrhizas go? A case study on Pisolithus down to 4 meters in a Brazilian eucalypt plantation. Mycorrhiza 29, 637–648 (2019). https://doi.org/10.1007/s00572-019-00917-y
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DOI: https://doi.org/10.1007/s00572-019-00917-y