Volume 9 - 2018 | https://doi.org/10.3389/fmicb.2018.00568 Disease-suppressive soils are soils in which specific soil-borne plant pathogens cause only limited disease although the pathogen and susceptible host plants are both present Suppressiveness is in most cases of microbial origin We conducted a comparative metabarcoding analysis of the taxonomic diversity of fungal and bacterial communities from suppressive and non-suppressive (conducive) soils as regards Fusarium wilts sampled from the Châteaurenard region (France) Bioassays based on Fusarium wilt of flax confirmed that disease incidence was significantly lower in the suppressive soil than in the conducive soil we succeeded in partly transferring Fusarium wilt-suppressiveness to the conducive soil by mixing 10% (w/w) of the suppressive soil into the conducive soil Fungal diversity differed significantly between the suppressive and conducive soils Among dominant fungal operational taxonomic units (OTUs) affiliated to known genera 17 OTUs were detected exclusively in the suppressive soil These OTUs were assigned to the Acremonium the relative abundance of specific members of the bacterial community was significantly higher in the suppressive and mixed soils than in the conducive soil OTUs found more abundant in Fusarium wilt-suppressive soils were affiliated to the bacterial genera Adhaeribacter Several of the fungal and bacterial genera detected exclusively or more abundantly in the Fusarium wilt-suppressive soil included genera known for their activity against F this study supports the potential role of known fungal and bacterial genera in Fusarium wilt suppressive soils from Châteaurenard and pinpoints new bacterial and fungal genera for their putative role in Fusarium wilt suppressiveness We used next-generation sequencing to conduct a comparative metabarcoding analysis of the microbiomes of both the Fusarium wilt suppressive soil of Châteaurenard and a non-suppressive soil from a nearby field The analysis was based on 454-pyrosequencing of the fungal internal transcribed spacer (ITS) region and the bacterial 16S rRNA gene The aim was to detect and identify fungal and bacterial genera associated with Fusarium-wilt-suppressive soils not yet identified and get new insights into potentially novel microbial genera and mechanisms involved in Fusarium wilt suppressiveness The Fusarium-wilt-suppressive soil was harvested from a field that had remained a fallow for several years in the Châteaurenard region France (43°53′15″N The conducive soil was sampled nearby from a temporarily uncultivated field following muskmelon cropping and before lettuce cropping (43°51′53″N The two soils were geographically very close (3 km apart) Physicochemical profiles were respectively 17.1% clay nitrogen content and pH were related to the current uses of the soils To confirm the biological nature of the soil suppressiveness the suppressive soil was autoclaved three times on three consecutive days for 20 min at 120°C and stored at room temperature for 1 week before use Soil disease suppressiveness was tested under greenhouse conditions with flax as the host plant and the flax pathogen F lini MIAE00347 (Collection of Microorganisms of Interest for Agriculture and Environment The plants were grown in different types of soil: suppressive soil (S) conducive soil amended with 10% (w/w) of suppressive soil (referred to as mixed soil or M) Soils were inoculated with 103 conidia of the pathogen per mL of soil variety Opaline) seeds were sown on the same day three replicates were performed with 20 plants/replicate Non-inoculated soils were used as controls Plants were grown in a growth chamber with 70% relative humidity a 16/8 h daylight/dark photoperiod at 17°C (day) and 15°C (night) the first week and 25°C (day) and 22°C (night) afterwards Diseased plants were recorded 25 days after inoculation Data were subjected to one-way analysis of variance (ANOVA) Pairwise comparisons were performed using Fisher's test (P < 0.05) Flax seeds were sown in 9/9/9.5 cm pots containing 300 g of soil Twenty plants per pot were grown under the same conditions of light and humidity as above and suppressive soil inoculated with 103 conidia of F lini MIAE00347 per mL of soil (referred to as pathogen-inoculated suppressive soil or IS soil) The inoculum dose used was relatively low (103 conidia per mL) to ensure that plants do not die too quickly in the conducive soil and prevent to collect the active rhizospheric soil needed to conduct microbial communities comparative analyses Three replicates per modality were performed the rhizospheric soil was collected from each replicate The root systems were isolated and shaken to remove free soil particles The soil around the roots influenced by root development and plant exudates was considered as rhizospheric soil It was sieved to 2 mm to remove fine roots and organic debris Large pieces of roots were removed manually with tweezers Two grams of soil samples from each modality were placed in 2-ml cryotubes and frozen at −20°C for further DNA extractions The protocol used for DNA extractions was described by Plassart et al. (2012) Extraction was performed by mechanical lysis using FastPrep®-24 (MP-Biomedicals USA) with a lysis buffer and two purification steps with PVPP (polyvinyl polypyrrolydone) minicolumns (BIORAD France) and Geneclean Turbo kit (Q-Biogene Total DNA was extracted from each replicate of the four modalities Each purified DNA sample was quantified by a fluorometric assay using the PicoGreen dsDNA Assay Kit (Invitrogen Life Technologies) and the StepOnePlus™System (Applied Biosystems®) according to the manufacturer's instructions DNA extracted from each soil sample served as a template in PCR reactions to amplify a fungal barcode and a bacterial barcode Three replicates were used for each of the four soil modalities PCR reactions were performed in quadruplicate for each replicate For fungal identification, the variable internal transcribed spacer 1 (ITS1) region was amplified using specific fungal primers ITS1F (5′-CTTGGTCATTTAGAGGAAGTAA-3′) and ITS2 (5′-GCTGCGTTCTTCATCGATGC-3′; White et al., 1990; Gardes and Bruns, 1993; Buée et al., 2009) The primers were tagged with four-base-pair multiplex identifiers (MIDs) at the 5′ and 3′ positions to specifically identify each sample as recommended by the manufacturer (Beckman Coulter Genomics) PCR conditions were as follows: 95°C for 5 min For bacterial identification, a 16S rRNA gene fragment (partial V3, V4 and partial V5) was amplified using primers 530F (5′-ACTCCTACGGGAGGCAGCAG-3′; Acosta-Martínez et al., 2008) and 803R (5′-CTACCNGGGTATCTAAT-3′; Zancarini et al., 2013) Ten-base-pair MIDs at the 5′ and 3′ positions were added to the primers to specifically identify each sample PCR conditions were as follows: 95°C for 10 min All amplifications were performed in a Mastercycler (Eppendorf amplicons of the four replicated PCRs were pooled and purified using a MinElute PCR Purification Kit (Qiagen France) following the manufacturer's protocol Amplicon concentrations were then estimated by fluorometric assay (PicoGreen dsDNA Assay Kit) an equimolar pooling of all samples was prepared (total DNA amount: ~3.2 μg per library) Adapter sequences were added by ligation as recommended by the manufacturer and 454-pyrosequencing was carried out by Beckman Coulter Genomics (Danvers USA) on a Genome Sequencer FLX 454 (Life Sciences/Roche Applied Biosystems) The first clustering included all sequences and the second clustering was conducted with the batch of consensus sequences from the previous clustering We kept from these sequences only the amplified regions of sequences using our primer set and deleted those with too many mismatches with our primer set (more than 3 mismatches with one primer) all these “artificial reads” with a reliable taxonomy were clustered using our specific program of clustering at various threshold levels (100% to 90% Regarding the results and the amplified regions the 93.8% threshold was the best suited to efficiently define the genus level as it was the closest one to the genus level defined by the SILVA taxonomy of analyzed sequences global analysis of the soil samples was computed by merging all homogenized high-quality reads from each sample into one global file and defining OTUs as previously described before subsequent analyses All raw sequences collected in this study have been deposited in the European Bioinformatics Institute nucleotide sequence database system under the accession number PRJEB24081 SINA uses the search result to derive a classification with the LCA (lowest common ancestor) method Each query sequence is assigned the shared part of the classifications of the search results OTU abundance levels in the different soils were compared based on Kruskal-Wallis tests using XLSTAT software (Addinsoft®) The plants grown in the non-inoculated soils showed no wilt disease symptoms Typical symptoms of Fusarium wilt were observed in pathogen-inoculated soils The percentage of healthy plants was significantly higher (P < 0.05) in soil S (46.6 ± 10.6) and soil M (43.5 ± 19.4) than in soil C (15.3 ± 5.7) The percentage of healthy plants was significantly lower (P < 0.05) in heat-treated suppressive soil (9.2 ± 0.3) than in soils S further confirming the biological nature of the soil suppressiveness Fungal and bacterial targeted regions were successfully amplified by PCR and sequenced for all soils The raw sequence libraries were filtered to remove reads originating from sequencing errors or putative chimeric sequences and 67,675 reads (89% of raw sequences) passed all quality controls The number of high-quality reads ranged from 3,451 to 10,821 for fungal datasets and from 2,505 to 8,842 for bacterial datasets a total of 41,412 high-quality fungal ITS sequences were clustered into 1,798 OTUs for all soil samples and represented 31.7% of the detected richness including 1,162 singletons (50.9% of the richness) were observed for 30,060 high-quality bacterial 16S rDNA sequences Rarefaction curves were drawn for fungal and bacterial datasets based on OTUs (Supplementary Figure 1) Fungal curves showed that: (i) the number of OTUs at 97% similarity increased with the number of sequences and saturation was not reached for all soils; (ii) based on soil replicates the number of sequences was sufficient to obtain a representative coverage of the major fungal groups; and (iii) differences among soils were recorded in the slope and level of the curves (Supplementary Figure 1A) rarefaction curves also revealed that the number of OTUs increased with the number of reads and reads were in sufficient numbers to allow for an accurate description of the major bacterial genera in each of the soil samples (Supplementary Figure 1B) no significant differences were detected in the slope and level of bacterial curves in any of the soils showing less heterogeneity of bacterial richness among the four modalities Fungal and bacterial richness and diversity indices of the conducive soil (C) and pathogen-inoculated suppressive soil (IS) Relative abundance levels of fungal classes (A) and bacterial phyla (B) Concerning bacterial communities, the soils had the same overall bacterial composition, with Proteobacteria, Bacteroidetes, Actinobacteria, Firmicutes, and Acidobacteria as major phyla (Figure 1B) Proteobacteria was the dominant bacterial phylum and represented 46–55% of all bacterial DNA sequences and Nitrospirae) were also identified; they accounted for ca with more Gamma-Proteobacteria in soil M than in soils S and IS (P < 0.05) Actinobacteria were more represented in soils S and IS than in soil C (P < 0.05) but no distinction of these two groups was visible along the second axis Principal component plots of the conducive soil (C) and pathogen-inoculated suppressive soil (IS) generated from fungal (A) and bacterial (B) OTU matrices of ITS1 and 16S relative abundance levels Regarding OTU composition, 189 fungal OTUs and 414 bacterial OTUs were shared between soils C and S (Figure 3) To further focus on the microbial genera associated with disease suppressiveness we compared the fungal and bacterial community compositions between soils S and M on the one hand and between soils S and IS on the other hand 122 fungal OTUs and 100 bacterial OTUs were shared between soils S and M but absent from soil C 214 fungal OTUs and 145 bacterial OTUs shared between soils S and IS were potentially promoted by the presence of the pathogen Venn diagrams indicating the numbers of shared and unique OTUs between the conducive soil (C) and pathogen-inoculated suppressive soil (IS) in fungal (A) and bacterial communities (B) Heat map comparison of the dominant fungal genera detected in the soils according to each modality The four different modalities (conducive soil and pathogen-inoculated suppressive soil) were organized based on the UPGMA dendrogram of UNIFRAC weighted and normalized distances between corresponding soil samples The legend shows the Z-scores (relative abundance levels are expressed as median-centered Z-scores between all samples and colors are scaled to standard deviations) OTUs with a star (*) indicate groups that were not assigned to a precise fungal genus Underlined OTUs indicate significant differences in the relative abundance levels of particular fungal genera in the four modalities Heat map comparison of the dominant bacterial genera detected in the soils according to each modality OTUs with a star (*) indicate groups that were not assigned to a precise bacterial genus Underlined OTUs indicate significant differences in the relative abundance levels of particular bacterial genera in the four modalities Microbial genera including OTUs associated with suppressive soil (S) and mixed soil (M) and known to include biological control agents against fungal diseases these other species probably contribute only weakly to soil suppressiveness of Fusarium wilt or do so via other modes of action which remain to be identified This could explain why its pathogenic activity remains controlled in the suppressive soil of Châteaurenard oxysporum by reducing Fe availability in the rhizosphere or producing antifungal phenazines the Pseudomonas genus is ubiquitous and was indeed detected in all soils but given their discrimination level due to the length of obtained reads OTUs did not distinguish the Pseudomonas strains directly involved Unfortunately the sustainability of this suppressiveness seems difficult to acquire as durably as in Châteaurenard soil It is therefore likely that the balance reached by Châteaurenard soil microbial communities results from a long natural evolutionary process; the suppressiveness of this soil is called “native,” this is the reason why it is so stable In the cases of “acquired” soil suppressiveness the agricultural practices used to manage microbial community patterns are still too recent to fix their assemblage (structure) in a sustainable way but all the results reported in the literature show that this is an alternative to be favored and EC: carried out the experiments and acquired the data; VE-H and ST: carried out bioinformatics analyzes; VE-H and CS: animated scientific discussions over the duration of the project and contributed equally to the writing of the article This work was supported by a grant to KS-H from the Conseil Régional de Bourgogne (CRB 2011-9201AAO048S05563). It was also supported by the European Commission within the framework of the EcoFINDERS project (FP7-264465) and the Plant Health and Environment department (INRA http://www.spe.inra.fr/en) The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest The authors wish to thank the colleagues of the Genosol platform (https://www2.dijon.inra.fr/plateforme_genosol/en/homepage) for technical discussions and relevant advice regarding the metabarcoding approach Buchwalter for language edition of this manuscript The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmicb.2018.00568/full#supplementary-material Acosta-Martínez Microbial communities and enzymatic activities under different management in semiarid soils Fusarium-wilt suppressive soils from the Châteaurenard region: review of a 10-year study CrossRef Full Text | Google Scholar Fusarium wilt suppressive soils: an example of disease-suppressive soils CrossRef Full Text | Google Scholar Microbiological control of soil-borne phytopathogenic fungi with special emphasis on wilt-inducing Fusarium oxysporum Rhizobium japonicum as a biocontrol agent of soybean root rot disease caused by Fusarium solani and Macrophomina phaseolina Assessment of the relationship between geologic origin of soil rhizobacterial community composition and soil receptivity to tobacco black root rot in Savoie region (France) Effect of clay mineralogy on iron bioavailability and rhizosphere transcription of 2,4-diacetylphloroglucinol biosynthetic genes in biocontrol Pseudomonas protegens Root-colonizing fungi from oilseed rape and their inhibition of Verticillium dahliae CrossRef Full Text | Google Scholar Gapped BLAST and PSI-BLAST: a new generation of protein database search programs PubMed Abstract | CrossRef Full Text Involvement of soil abiotic factors in the mechanisms of soil suppressiveness to fusarium wilts CrossRef Full Text | Google Scholar Identification of Rhizobium isolates possessing antagonistic activity against Fusarium oxysporum f.sp ciceris the causal agent of Fusarium wilt of chickpea MN: The American Phytopathological Society Google Scholar Biocontrol of gray mold in tomato plants by Clonostachys rosea CrossRef Full Text | Google Scholar 454 Pyrosequencing analyses of forest soils reveal an unexpectedly high fungal diversity Microbial and biochemical basis of a Fusarium wilt-suppressive soil and co-occurrence of soil microbial communities in abandoned sites of a rare earth elements mine Manipulating the soil microbiome to increase soil health and plant fertility CrossRef Full Text | Google Scholar Chitinase and beta-1,3-glucanase enzyme production by the mycoparasite Clonostachys rosea f The ribosomal database Project: improved alignments and new tools for rRNA analysis “Fungal genes and metabolites associated with the biocontrol of soil-borne plant pathogenic fungi,” in Fungal Metabolites Ramawat (Basel: Springer International Publishing) Google Scholar Ascomycete communities associated with suppression of Sclerotium rolfsii in compost CrossRef Full Text | Google Scholar Adaptation and response to mycotoxin presence in pathogen-pathogen interactions within the Fusarium genus CrossRef Full Text | Google Scholar Induced resistance by Penicillium oxalicum against Fusarium oxysporum f sp lycopersici: histological studies of infected and induced tomato stems Hydrophobins are required for conidial hydrophobicity and plant root colonization in the fungal biocontrol agent Clonostachys rosea Microbial antagonism at the root level is involved in the suppression of Fusarium wilt by the combination of nonpathogenic Fusarium oxysporum Fo47 and Pseudomonas putida WCS358 Search and clustering orders of magnitude faster than BLAST The relation of different crop roots exudates to the survival and suppressive effect of Stenotrophomonas maltophilia (PD4560) biocontrol agent of bacterial xilt of potato Cross-biome comparison of microbial association networks Microbial diversity in soil: selection of microbial populations by plant and soil type and implications for disease suppressiveness doi: 10.1146/annurev.phyto.42.012604.135455 ITS primers with enhanced specificity for basidiomycetes - application to the identification of mycorrhizae and rusts Gómez Expósito Current insights into the role of rhizosphere bacteria in disease suppressive soils Halfeld-Vieira Understanding the mechanism of biological control of passionfruit bacterial blight promoted by autochthonous phylloplane bacteria Biotechnological application and taxonomical distribution of plant growth promoting actinobacteria Significant and persistent impact of timber harvesting on soil microbial communities in Northern coniferous forests Involvement of clay type and pH in the mechanisms of soil suppressiveness to fusarium wilt of flax CrossRef Full Text Illumina MiSeq investigations on the changes of microbial community in the Fusarium oxysporum f.sp cubense infected soil during and after reductive soil disinfestation Biological control of Phytophthora palmivora causing root rot of pomelo using Chaetomium spp Microbial consortium-mediated reprogramming of defence network in pea to enhance tolerance against Sclerotinia sclerotiorum Soil health through soil disease suppression: which strategy from descriptors to indicators Characterization of the Penicillium chrysogenum antifungal protein PAF Microbial diversity and ecological networks as indicators of environmental quality Kirk, P. M. (2017). “Species Fungorum (version Jan 2016),” in Species 2000 & ITIS Catalogue of Life, eds Y. Roskov, L. Abucay, T. Orrell, D. Nicolson, N. Bailly, P. M. Kirk, T. Bourgoin, R. E. DeWalt, W. Decock, A. De Wever, E. Nieukerken, J. van Zarucchi, and L. Penev (Leiden: Species 2000: Naturalis). Available online at: www.catalogueoflife.org/col (Accessed December 20 Google Scholar Soil suppressiveness to fusarium disease: shifts in root microbiome associated with reduction of pathogen root colonization Analysis of microbial taxonomical groups present in maize stalks suppressive to colonization by toxigenic Fusarium spp.: a strategy for the identification of potential antagonists Kõljalg Towards a Unified Paradigm for Sequence-Based Identification of Fungi Kyselková “Pseudomonas and other microbes in disease-suppressive soils,” in Organic Fertilisation Lichtfouse (Dordrecht: Springer Science+Business Media) Kyselková Comparison of rhizobacterial community composition in soil suppressive or conducive to tobacco black root rot disease Biocontrol of Fusarium and Verticillium wilt of tomato by Penicillium oxalicum under greenhouse and field conditions Soil health paradigms and implications for disease management Suppression of Fusarium wilts by fluorescent pseudomonads: mechanisms and applications CrossRef Full Text | Google Scholar Antagonistic effect of nonpathogenic Fusarium oxysporum Fo47 and pseudobactin 358 upon pathogenic Fusarium oxysporum f Declined soil suppressiveness to Fusarium oxysporum by rhizosphere microflora of cotton in soil sickness Recherches sur la résistance des sols aux maladies Mise en évidence de la nature microbiologique de la résistance d'un sol au développement de la fusariose vasculaire du melon Enhancing predation of a subterranean insect pest: a conservation benefit of winter vegetation in agroecosystems CrossRef Full Text | Google Scholar Impact of brassicaceous seed meals on the composition of the soil fungal community and the incidence of Fusarium wilt on chili pepper Phenazine antibiotics produced by fluorescent pseudomonads contribute to natural soil suppressiveness to Fusarium wilt Manipulation of rhizosphere bacterial communities to induce suppressive soil Google Scholar Wheat genotype-specific induction of soil microbial communities suppressive to disease incited by Rhizoctonia solani anastomosis group AG-5 and AG-8 Enzyme production by the mycoparasite Verticillium biguttatum against Rhizoctonia solani Deciphering the rhizosphere microbiome for disease-suppressive bacteria Stenotrophomonas maltophilia: a new potential biocontrol agent of Ralstonia solanacearum Volatile organic compounds: a potential direct long-distance mechanism for antagonistic action of Fusarium oxysporum strain MSA35 Identification of microorganisms for biological control of powdery mildew in Cornus florida CrossRef Full Text | Google Scholar Temporal occurrence and impact of Scytalidium uredinicola CrossRef Full Text | Google Scholar Antagonism of the two-needle pine stem rust fungi Cronartium flaccidum and Peridermium pini by Cladosporium tenuissimum in vitro and in planta Mosquera-Espinosa The double life of Ceratobasidium: orchid mycorrhizal fungi and their potential for biocontrol of Rhizoctonia solani sheath blight of rice Biodiversity of genes encoding anti-microbial traits within plant associated microbes Biocontrol of Fusarium wilt disease in tomato by Paenibacillus ehimensis KWN38 Combining leeway on farm and supply basin scales to promote technical innovations in lettuce production Antagonism against Rhizoctonia solani and fungitoxic metabolite production by some Penicillium isolates An open source software package for automated extraction of ITS1 and ITS2 from fungal ITS sequences for use in high-throughput community assays and molecular ecology Oliveros, J. C. (2007). Venny. An Interactive Tool for Comparing Lists with Venn's diagrams. (Madrid: CNB-Csic), http://bioinfogp.cnb.csic.es/tools/venny/index.html Google Scholar Antifungal activity against plant pathogenic fungi of chaetoviridins isolated from Chaetomium globosum Fungal community structure in disease suppressive soils assessed by 28S LSU gene sequencing Combining biocontrol agents with different mechanisms of action in a strategy to control Botrytis cinerea on grapevine Evaluation of the ISO Standard 11063 DNA extraction procedure for assessing soil microbial abundance and community structure Zearalenone lactonohydrolase activity in Hypocreales and its evolutionary relationships within the epoxide hydrolase subset of a/b-hydrolases SINA: accurate high-throughput multiple sequence alignment of ribosomal RNA genes The SILVA ribosomal RNA gene database project: improved data processing and web-based tools The rhizosphere: a playground and battlefield for soilborne pathogens and beneficial microorganisms platform-independent community-supported software for describing and comparing microbial communities Soils naturally suppressive to banana Fusarium wilt disease harbor unique bacterial communities Deep 16S rRNA pyrosequencing reveals a bacterial community associated with banana Fusarium wilt disease suppression induced by Bio-organic fertilizer application Application of Chaetomium species (Ketomium®) as a new broad spectrum biological fungicide for plant disease control: a review article “Root interactions with non-pathogenic Fusarium what do you do in life when you do not infect a plant?,” in Belowground Defence Strategies in Plants Kazan (Basel: Springer International Publishing Switzerland) tillage and rotation practices influence soil health and suppressiveness to soilborne pests and pathogens of ginger Suarez-Estrella Biological control of plant pathogens by microorganisms isolated from agro-industrial composts Molecular biomass and metataxogenomic assessment of soil microbial communities as influenced by soil DNA extraction procedure Meta-barcoded evaluation of the ISO standard 11063 DNA extraction procedure to characterize soil bacterial and fungal community diversity and composition Synergistic effect of dazomet soil fumigation and Clonostachys rosea against cucumber Fusarium wilt “Fusarium-suppressive soils,” in Biology and control of soil-borne plant pathogens Google Scholar epidemiology and management of Rhizoctonia solani on potato CrossRef Full Text | Google Scholar Impact of interspecific interactions on antimicrobial activity among soil bacteria Soil health indicators and Fusarium wilt suppression in organically and conventionally managed greenhouse soils Microbial profiling of a suppressiveness-induced agricultural soil amended with composted almond shells Fungal communities associated with durum wheat production system: a characterization by growth stage CrossRef Full Text | Google Scholar Bioactive metabolites from Chaetomium globosum L18 an endophytic fungus in the medicinal plant Curcuma wenyujin Microbial populations responsible for specific soil suppressiveness to plant pathogens doi: 10.1146/annurev.phyto.40.030402.110010 Soil suppressiveness to Heterodera schachtii under different cropping sequences CrossRef Full Text | Google Scholar “Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics,” in PCR Protocols Google Scholar Demonstrating microbial co-occurrence pattern analyses within and between ecosystems Combining ecophysiological and microbial ecological approaches to study the relationship between Medicago truncatula genotypes and their associated rhizosphere bacterial communities Efficacy assessment of antifungal metabolites from Chaetomium globosum No.05 Bio-organic fertilizer application significantly reduces the Fusarium oxysporum population and alters the composition of fungi communities of watermelon Fusarium wilt rhizosphere soil Raaijmakers JM and Steinberg C (2018) Comparative Microbiome Analysis of a Fusarium Wilt Suppressive Soil and a Fusarium Wilt Conducive Soil From the Châteaurenard Region Received: 11 October 2017; Accepted: 13 March 2018; Published: 04 April 2018 Copyright © 2018 Siegel-Hertz, Edel-Hermann, Chapelle, Terrat, Raaijmakers and Steinberg. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) distribution or reproduction in other forums is permitted provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited in accordance with accepted academic practice distribution or reproduction is permitted which does not comply with these terms *Correspondence: Christian Steinberg, Y2hyaXN0aWFuLnN0ZWluYmVyZ0BpbnJhLmZy Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher 94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish Metrics details We report the first discoveries of high-pressure minerals in the historical L6 chondrite fall Château-Renard scanning electron microscopy (SEM) with energy-dispersive X-ray spectroscopy and electron backscatter diffraction and transmission electron microscopy (TEM) with selected-area electron diffraction A single polished section contains a network of melt veins from ~40 to ~200 μm wide with no cross-cutting features requiring multiple vein generations We find high-pressure minerals in veins greater than ~50 μm wide including assemblages of ringwoodite + wadsleyite ringwoodite + wadsleyite + majorite-pyropess In association with ahrensite + wadsleyite at both SEM and TEM scale we find a sodic pyroxene whose Raman spectrum is indistinguishable from that of jadeite but whose composition and structure are those of omphacite We discuss constraints on the impact record of this meteorite and the L-chondrites in general no verifiable occurrence of any HP mineral in this meteorite has been reported in the literature and structural constraints on a high-pressure pyroxene phase in Château-Renard and note the curious observation that this phase displays the Raman bands commonly associated with jadeite Château-Renard. Transmitted light mosaic image of Château-Renard polished section (L4361) showing the complex network of melt veins. Focus areas indicated by MV1-a, MV1-b, MV2, MV3, MV4 and MV5 are called out and enlarged in subsequent panels. (e) The EBSD pattern is indexed here as C2/c clinopyroxene but it is consistent with several different pyroxene polymorphs (MAD = 0.69°) Mineral abbreviations: Merr: merrilite; Opmh: omphacite Although no HP minerals have been previously reported in the Château-Renard meteorite we have observed within the melt veins a number of HP minerals and report on them here for the first time Within clasts and grains displaying olivine stoichiometry there are clear correlations between the Fe/(Mg + Fe) ratio (expressed here as mole percent of the Fe2SiO4 fayalite component we place the formal divide between ringwoodite (the Mg2SiO4 endmember) and ahrensite (the Fe2SiO4 endmember) at Fa50 We also observe a sodic pyroxene that shares some features with jadeite; we document below our investigation of the identity of this mineral and show that it is not and omphacite (#R061129) from RRUFF database are given for comparison which is quite Fe-rich for wadsleyite; it is likely that this analysis includes unresolved areas of less Fe-rich wadsleyite and more Fe-rich olivine We discuss in the next section of this paper whether these bands are in fact diagnostic of jadeite what other pyroxene-structured phases might have very similar Raman spectra and what additional observations are needed to confirm or reject the identification of jadeite so these points are most likely ahrensite as well These may be rims of olivine grains whose cores are out of the plane of the section so this question was further studied by transmission electron microscopy (TEM) Although the high accelerating potentials used during TEM may cause substantial beam damage and reduce the Na count in TEM-EDS analysis we stress that the compositional assignment to omphacite is here based on EPMA analysis using analytical conditions under which sodic pyroxenes are typically beam-stable (a) Bright field (BF) image of the omphacite (dark-grey contrast) SAED patterns of omphacite along the (b) [\(1\overline{34}\)] and (c) [012] zone axes (d) BF image of intergrown ahrensite and wadselyite (dark contrast) The striation in the grain is inferred to be due to stacking faults (e) SAED pattern of ahrensite [110] zone overlaps with wadsleyite [010] zone (f) Simulated pattern showing the same topotaxis of ahrensite (red spots) and wadsleyite (green spots) Its composition (En83Fs14) is distinct from the sodic pyroxene in the Si-rich pool. From these observations we conclude that the mineralogy of the groundmass is fully consistent with the high-pressure minerals found as large grains likely to have crystallized from the melt at near-equilibrium conditions leads to the same inferences about pressure and temperature conditions as the coarse grains despite the possibility of kinetic limitations on the achievement of equilibrium in the coarse grains This study presents the finding of four series of HP minerals (ringwoodite-ahrensite and sodic pyroxene) in Château-Renard; all these HP minerals are described from this meteorite for the first time Although we do observe an usual pyroxene coexisting with Na- and Si-rich melt it is not jadeite either by composition or by structure Omphacite is characteristic of eclogite-facies metamorphism and is probably an indicator of elevated pressure but any pressure constraints based on experimental thresholds for the formation of jadeite sensu stricto are not relevant Here we discuss the use of these observations to constrain the peak pressure and shock duration experienced by this meteorite (c) Time-Temperature diagram for back-transformation of HP ringwoodite to LP olivine for the case of Château-Renard The time for complete solidification of 0.7 msec suggests a maximum time remaining at high-T is the maximum to avoid back-transformation of all ringwoodite to olivine while at the same time the T should not exceed that of ca whereas wadsleyite would imply a lower P range One explanation for their coexistence in the same meteorite or even in the same melt vein would be spatially or temporally variable pressure during the shock event but we find the two phases intimately intergrown Their coexistence might therefore be interpreted as a very specific constraint on pressure followed by topotactic growth of wadsleyite from ringwoodite nucleation sites A different approach considers the phase relations in the binary Mg2SiO4-Fe2SiO4 system since the typical Fa content of L-chondrite olivines is nearly 25% Compositional segregation during growth from a superliquidus state originally in the ringwoodite field would not explain the intergrowth: early crystallization of ringwoodite would enrich residual melt in Fe moving it away from the stability field of wadsleyite if a solid-state transformation occurred at high enough temperature to allow Fe-Mg interdiffusion one could reconcile the observation of Fe-depleted forsterite olivine cores with moderately Fe-enriched wadsleyite and highly Fe-enriched wadsleyite-ahrensite intergrowths with a considerable range of pressures from 13–18 GPa (assuming equilibrium) The different HP polymorphs of olivine and the range of compsitions observed in the different melt veins could then be attributable to different cooling rates rather than to large variations in pressure Although preservation of the highest-P indicator minerals might be problematic the presence of sodic clinopyroxene and the absence of Ca-ferrite P ≤ 15.5 GPa in the pyroxene-bearing regions Given the diversity of mineral assemblages described within the single studied thin section it is clear that Château-Renard records variable apparent pressure and temperature conditions Possibly the different veins record different times along a common P-T path that they all experiened depending largely on the local compositions the presence of discrete veins directly proves heterogeneity of the temperature field which is likely the result of collapse of spatially variable porosity during shock compression or slip along localized shear bands (despite some shape preferred orientation of large clasts parallel to the vein elongation no convincing evidence of shear flow across the veins is observed) Shocking a heterogeneous medium also results in a heterogeneous pressure distribution Although it is likely that pressure gradients on the order of GPa/mm would relax considerably after passage of the shock wave and before pressure release it is hard to quantify the pressure differences that might persist over the potentially much shorter timescales involved in quenching the melt veins we lack a sound basis for asserting that a global peak P-T condition or global P-T path can be defined for the meteorite the different veins may be recording altogether different shock events Although the omphacite we have discovered could not explain the near-albite compositions reported for most jadeite occurences in L6 chondrites our data do reveal that other sodic pyroxenes with stability fields different from those of pure jadeite may present Raman bands indistinguishable from those of jadeite A compositional analysis is clearly required to confirm a Raman identification of jadeite before the stability field of jadeite can be used as a pressure minimum for a meteorite Such jadeite presumably forms by solid-state decomposition of albite the omphacite that we find in melt veins in Château-Renard combines components derived from more than one precursor phase — Na from plagioclase and Ca Hence is seems necessary that the omphacite grew from a melt whose formation digested both plagioclase and clinopyroxene The melt then cooled enough to begin crystallizing while it remained at high enough pressure to stabilize omphacite We seek to quantify the necessary duration of the shock pulse by combining the Avrami equation (a general formalism for solid-state phase transformation) with an Arrhenius-type temperature dependence of the transformation rate constant. The result of this calculation (see Supplementary material) is summarized in Fig. 8 they would cool to ~1000 °C within 0.7 ms and then cool much more slowly from this temperature (roughly the average of the peak vein temperature and the matrix temperature) over the ensuing several seconds We conclude that the pressure was maintained in the stability field of ringwoodite for at least one second enabling continued cooling of the melt veins and preservation of ringwoodite A pressure wave of this duration requires an impactor at least meters in scale preserves in some veins peak P-T conditions intermediate between Sahara 98222 and Tenham (and distinct from both) and no HP minerals were observed (though they may be present at scales below our observations) The range in apparent peak P conditions within a single thin section is problematic for this discussion unless attributed to differences in preservation that (1) a characteristic peak P can be defined for each meteorite (2) these peak pressures differ among the meteorites the single-impact scenario then implies that each meteorite was derived from a distinct location relative to the impact point with shock pressure systematically decreasing with increasing distance from the point of impact a single-parent body and multiple-impact scenario allows for a range of impact velocities and impactor densities and sizes as well as different locations in the target or one in which some of the impact events recorded by the shock metamorphic assemblage in some L-chondrites followed the disruption of an original single body there need not be any relationship among the observations in different meteorites other differences among the MVs might offer alternative explanations for the differences in HP mineral formation or preservation; e.g. the thickest MV is enriched in silicate clasts whereas the thin MVs tend to be richer in sulfides (mainly troilite) our inability to find HP minerals in a thin melt vein suggests that we should entertain the hypothesis that our section contains two generations of MVs representing different shock events we should also attempt to define which came first and how much time elapsed between these two events No cross-cutting relationship is observed to prove either scenario HP mineral -bearing veins formed after the narrow then the second event likely reached higher peak pressure sufficient to form the HP assemblage of majorite-pyropess garnet + ringwoodite As the first-generation MVs would have already been consolidated during the first impact they would have lacked the porosity or focused shear deformation needed to generate local high temperature and HP mineral growth during the second impact A critical challenge for a multiple-impact scenario is the persistence through the first impact event of porosity sufficient to provide locations of focused heating leading to MV formation during the second impact a possible resolution is a two-pass interaction in which an impactor fragmented during a close approach to the parent body; the fragments continued to travel on related orbits and each impacted sequentially on the next close approach This scenario can yield multiple impact events on the parent body within a period of a few seconds Château-Renard is a highly shocked L6 ordinary chondrite containg numerous HP minerals These HP minerals occur within thick melt veins (>50 μm in width) whereas the thinner MV that we inspected apparently lacks HP minerals The HP assemblages are (1) ringwoodite + wadsleyite; (2) ringwoodite + wadsleyite + majorite-pyropess; (3) ahrensite + wadsleyite; and (4) sodic pyroxene + ahrensite + wadsleyite + clinoenstatite The absence of periclase + (retrogressed) bridgmanite or periclase + stishovite suggests an upper bound for the peak pressure of ~23–25 GPa whereas the presence of ringwoodite and majorite suggest peak pressures in the range of 17–23 GPa co-occurrence of ringwoodite-ahrensite solutions with wadsleyite implies a modestly lower P range 14–18 GPa (or less accounting for Fe-rich compositions) along with rate-controlled nucleation of the HP polymorphs of olivine using binary jadeite-diopside phase diagrams to estimate the pressure implied by the occurrence of omphacitic pyroxene suggests peak P ≤ 15.5 GPa The inconsistency of ≥1.5 GPa in these pressure estimates suggests that either spatial heterogeneity or some combination of these are recorded by the various HP mineral assemblages in the investigated section the temperatures estimated for majorite growth (≥1800 °C) in the MV centers and wadsleyite formation (≤1500 °C) at the MV edges require a temperature gradient during HP mineral growth Without cross-cutting relations between the various melt veins it is challenging to distinguish among the various explanations for preservation of heterogeneous conditions Eight historical covered thin sections (A981 and J2892) and one polished thick section (L4361) of Château-Renard from the collection of the Natural History Museum Melt veins were examined under the optical microscope and we then conducted a careful search for shock effects in mineral grains and HP minerals The section was carbon-coated and investigated with the NHMV JEOL JSM-6610 LV equipped with a highly sensitive backscattered electron detector and an energy-dispersive X-ray spectrometer (EDS) Analyses were conducted using a 15 kV accelerating voltage and a ∼20 nA probe current yielding analytical volumes with diameters less than 3 μm Additional SEM analyses were performed at the California Institute of Technology (Caltech) GPS using a Zeiss 1550VP field-emission scanning electron microscope equipped with an angle-sensitive backscattered electron detector 80 mm2 active area Oxford X-Max Si-drift-detector EDS SEM imaging and EDS analyses used a 15 kV accelerating potential and a 120 μm field aperture in high-current mode (∼4 nA probe current) yielding imaging resolution better than 2 nm and an activation volume for EDS analysis ∼1–2 μm3 on silicates Single crystal EBSD analyses at a sub-micrometer scale were performed at 20 kV and 6 nA in focused beam mode with a 70° tilted stage on uncoated specimens in “variable pressure” mode (25 Pa of N2 gas in the chamber to reduce specimen charging) and EBSD were conducted using the SmartSEM Major element compositions of matrix and MV minerals were determined using a JEOL JXA8530F Field Emission EPMA instruments (FE-EPMA) equipped with five wavelength-dispersive spectrometers (WDS) and one energy-dispersive spectrometer (EDS) at both the NHMV and the Institut für Mineralogie Mineral analyses were performed with an accelerating voltage of 15 kV and counting times of 10 s on-peak and 5 s on each background position were used Natural mineral standards used were albite (Na and Ni-oxide (Ni) with ZAF matrix correction We used a FEI Nova 600 Nanolab DualBeam focused ion beam (FIB) and scanning electron microscope (SEM) for the sample preparation and lift-out The sample thinning was finalized with an 8 kV The analytical transmission electron microscopy (ATEM) analysis was performed on FEI Tecnai TF20 with super-twin objective lens The EDS data were collected in TEM mode using a EDAX SiLi detector with 10 eV/channel and 51.2 µs process time to achieve 500 cps signal and 20–50% deadtime The FIB and TEM facilities are in the Kavli Nanoscience Institute at Caltech Raman spectra for preliminary phase identification were conducted on the polished thin section using a dispersive confocal Raman microscope Renishaw inVia Reflex at the National Hellenic Research Foundation Analyses used a a 514 nm Ar-ion laser and a ×100 objective lens and spectra were collected in the Stokes region for Raman shifts from 200–1600 cm−1 Additional Raman analyses were performed at the Open University using a Horiba Jobin-Yvon LabRam HR Raman Microscope equipped with both 514 nm and 633 nm lasers The laser beam was spread across ~1–2 μm spots at relative low incident power (ca 5 mW) in order to avoid sample destruction For each spot analysis on the Open University system we averaged spectra over 5 consecutive 60 sec accumulation times Gaussian-Lorentzian peak fitting (Spectragryph version 1.0.5) was used to remove background and estimate the peak centers Collected spectra were compared with published data from RRUFF and the Handbook of Raman Spectra The locations of each Raman spot analysis were recorded and co-located EPMA analytical points were collected in order to couple structural and compositional characterization at common spots where κ is the thermal diffusivity and λ is a dimensionless coefficient that accounts for the boundary conditions and latent heat λ is obtained by substitution of the following two boundary conditions at the moving front We estimated the time over which HP minerals might persist without back-transformation to their low-pressure equivalents We combined the Avrami equation (which describes how solids transform from one phase to another at constant temperature) and an Arrhenius relationship for the transformation rate constant Assuming that the shock event that disrupted the L-chondrite parent body happened at 470 Ma majorite and wadsleyite over this time requires that the temperature be maintained below the conversion boundary curves for this time period in t-T space the upper bound temperature corresponds to ca The datasets generated during and/or analysed during the current study are included in this published article (and its Supplementary Information files) but also are available from the corresponding author on reasonable request Shock metamorphism as a fundamental process in the evolution of planetary bodies: information from meteorites Timescales of shock processes in chondritic and martian meteorites Shock events in the Solar System: The message from minerals in terrestrial planets and asteroids The majorite-pyrope plus magnesiowustite assemblage: constraints on the history of shock veins in chondrites Natural (Mg,Fe)SiO3-ilmenite and-perovskite in the Tenham meteorite Natural NaAlSi3O8-hollandite in the shocked Sixiangkou meteorite Formation of high-pressure minerals in shocked L6 chondrite Yamato 791384: constraints on shock conditions and parent body size Natural high-pressure polymorph of merrillite in the shock veins of the Suizhou meteorite Estimating shock pressures based on high-pressure minerals in shock-induced melt veins of L-chondrites mechanisms of formation of high‐pressure minerals in shock melt veins of L6 chondrites and pressure‐temperature conditions of the shock events Tissint Martian meteorite as evidence for the largest impact excavation Back-transformation of high-pressure phases in a shock melt vein of an H-chondrite during atmospheric passage: Implications for the survival of high-pressure phases after decompression Back-transformation of high-pressure minerals in shocked chondrites: Low-pressure mineral evidence for strong shock Fe)2SiO4 phases in the Tenham chondritic meteorite Thermal and shock metamorphism of the Tenham chondrite: A TEM examination Pyroxene-garnet transformation in Coorara meteorite Discovery of the Fe-analogue of akimotoite in the shocked Suizhou L6 chondrite High‐pressure minerals in shocked meteorites Chromite-plagioclase assemblages as a new shock indicator; implications for the shock and thermal histories of ordinary chondrites A chemical-petrologic classification for the chondritic meteorites Thermal histories of meteorites by the 39Ar-40Ar method In Meteorite research (407-417) Springer Netherlands (1969) L‐chondrite asteroid breakup tied to Ordovician meteorite shower by multiple isochron 40Ar‐39 Ar dating Records of the Moon‐forming impact and the 470 Ma disruption of the L chondrite parent body in the asteroid belt from U‐Pb apatite ages of Novato (L6) Refined Ordovician timescale reveals no link between asteroid breakup and biodiversification Thermodynamics and behavior of γ-Mg2SiO4 at high pressure: Implications for Mg2SiO4 phase equilibrium A pressure-induced phase transition in MgSiO3-rich garnet revealed by Raman spectroscopy Pyroxene polymorphs in melt veins of the heavily shocked Sixiangkou L6 chondrite Vibrational spectroscopy of pyrope-majorite garnets: Structural implications a new shock-metamorphic mineral from the Tissint meteorite: implications for the Tissint shock event on Mars High-pressure Raman spectroscopic study of Fo90 hydrous wadsleyite Phase transitions in the Earth’s mantle and mantle mineralogy Mantle petrology: field observations and high pressure experimentation 6 High-pressure minerals in shocked L6-chondrites: constraints on impact conditions Jadeite in Chelyabinsk meteorite and the nature of an impact event on its parent body Pressure‐temperature phase diagram for the Allende meteorite The system Mg2SiO4-Fe2SiO4 at high pressures and temperatures: Precise determination of stabilities of olivine Intracrystalline transformation of olivine to wadsleyite and ringwoodite under subduction zone conditions Ultrafast growth of wadsleyite in shock-produced melts and its implications for early solar system impact processes The breakdown of olivine to perovskite and magnesiowüstite Phase transition processes of olivine in the shocked Martian meteorite Tissint: Clues to origin of ringwoodite- bridgmanite-and magnesiowüstite-bearing assemblages Transformation of enstatite—diopside—jadeite pyroxenes to garnet Phase relations on the diopside–jadeite–hedenbergite join up to 24GPa and stability of Na-bearing majoritic garnet The pressures and temperatures of meteorite impact: Evidence from micro-Raman mapping of mineral phases in the strongly shocked Taiban ordinary chondrite “Spherulite-like” jadeite growth in shock-melt veins of the Novosibirsk H5/6 chondrite Shock-induced P-T conditions and formation mechanism of akimotoite-pyroxene glass assemblages in the Grove Mountains (GRV) 052082 (L6) meteorite high-pressure clinopyroxene in the Tissint martian meteorite Multiple shock events and diamond formation on Mars Proceedings of the Lunar and Planetary Science Conference 44 Mechanisms of ringwoodite formation in shocked meteorites: Evidence from L5 chondrite Dhofar 1970 Two distinct assemblages of high‐pressure liquidus phases in shock veins of the Sixiangkou meteorite 40Ar/39Ar ages of impacts involving ordinary chondrite meteorites Shock‐induced transition of NaAlSi3O8 feldspar into a hollandite structure in a L6 chondrite Discovery of natural MgSiO3 tetragonal garnet in a shocked chondritic meteorite Anatomy of black veins in Zagami: clues to the formation of high-pressure phases Shock melt veins of Tenham chondrite as a possible paleomagnetic recorder: Rock magnetism and high‐pressure minerals Download references This research received support from SYNTHESYS (www.synthesys.info) [AT-TAF-4526] a European Union-funded Integrated Activities grant JH was supported by the Caltech-JPL President and Director’s Fund acknowledges funding received from the European Union’s Horizon 2020 research and innovation programme under grant agreement No -704696 RESOLVE acknowledges funding by the UK Science and Technology Facilities Council (ST/L000776/1 & ST/P000657/1) Brandstaetter is kindly acknowledged for fruitful discussions during the stay of I.B greatfully acknowledges the National Hellenic Research Foundation and E Department of Natural Resources Management and Agricultural Engineering Division of Geological and Planetary Sciences observed and analysed the meteorite sections constructed the thermal and transformation time modeling The authors declare no competing interests Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations Download citation DOI: https://doi.org/10.1038/s41598-018-28191-6 Anyone you share the following link with will be able to read this content: a shareable link is not currently available for this article Sign up for the Nature Briefing newsletter — what matters in science Please enable JS and disable any ad blocker Sign up for our daily Newsletter and stay up to date with all the latest news You are receiving this pop-up because this is the first time you are visiting our site You are using software which is blocking our advertisements (adblocker) we are relying on revenues from our banners So please disable your adblocker and reload the page to continue using this site.Thanks Starting a production of red kiwis in France means waiting 6 years before the first harvest Orchards of this new variety therefore struggle to see the light of day a major player of the agrifood sector located in Châteaurenard (Bouches-du-Rhône) has an ecological solution that could adapt to several crops and help gain fruit and energy independence development and production manager for the Peruzzo group reveals his analysis and solutions on the subject.  The Peruzzo group discovered the red kiwi during a trip 3 years ago is very popular among consumers for its sweet red fruit taste the group has been developing its cultivation in France with a plan to plant 10 hectares of red kiwis and 13 hectares of yellow kiwis in 2022 followed by 8 hectares of red kiwis and 4 of yellow kiwis in photovoltaic greenhouse in 2023 Peruzzo wants to offer to farmers and new producers a “turnkey” ecological project that would require very little investment with the possibility to produce in agrivoltaic greenhouses or under photovoltaic shades But the solution struggles to see the light of day.  “We have discovered this fruit with an exceptional flavor and we want to develop its production in France but we are confronted with the regulations in force which slow down considerably the ‘turnkey’ process we want to offer to the producers it takes at least two years to obtain a permit to build a greenhouse with no guarantee that it will be obtained and then another four years until the first harvest so a total of at least six years during which the land cannot be used and there is no income The Local Urbanism Plans and Relocation Pilot Project Policies are also causing some delays it is more relevant to focus on our independence in terms of energy We are faced with major difficulties to implement these projects We have received calls from about 50 producers who want to plant red and yellow kiwis and so far The delays have scared off the other producers,” explains Sylvain Colleville While investment costs for the structures have increased by 40% for fruit growers along with the costs of phytosanitary products to protect the fruit from weather hazards agrivoltaic solutions could be the answer to both problems; an advantageous solution for both agriculture and the development of green energy.  photovoltaic greenhouses or shades allow for: The photovoltaic installations would help develop new fruit crops in France that would be sustainable and profitable for the producers while taking advantage of farmland to produce a lot of green energy not considering these solutions seems irresponsible to me The procedures to obtain building permits and hail protection nets must be simplified I would like us to come up with a solution so as not to lose these two years and I want to present this solution to French Minister of Agriculture Julien Denormandie,” explains Sylvain Colleville red kiwis are highly sensitive to bacteriosis and must be protected from the rain which makes their production in photovoltaic greenhouses all the more interesting For more information:Peruzzohttps://www.peruzzo-group.com   FreshPublishers © 2005-2025 FreshPlaza.com The Tour de la Provence 2020 is the 5th edition of this road cycling race which takes part on the UCI Europe Tour 2020 in category 2.1 more than 633 kilometers will occur between the Bouches-du-Rhône the Var and the Vaucluse departments and in addition to all of this Stage 1 – Châteaurenard > Saintes-Maries-de-la-Mer – 149 km – 13th februaryStage 2 – Aubagne > La Ciotat – 175 km – 14 februaryStage 3 – Istres > Mont Ventoux – Chalet Reynard – 138 km – 15th februaryStage 4 – Avignon > Aix-en-Provence – 171 km – 16th february The Fleet Air Arm Museum at HMAS Albatross will be the new home for some incredible pieces of Australian military history All articles from our websiteThe digital version of Today's PaperBreaking news alerts direct to your inboxAll articles from the other regional websites in your areaContinueA number of pieces of wreckage from an 882 Grumman Wildcat Mk V flown by an Australian which was shot down in southern France in 1944 have been unveiled at the museum The aircraft was being flown by the then Royal Australian Navy Sub Lieutenant Fred Sherborne  was shot down five miles south of Avignon on August 19 Museum manager and senior curator Terry Hetherington opened the boxes containing the special delivery from France on Tuesday View +11 PhotosYou would never have known inside the two nondescript boxes were some of the most significant Australian military historical artifacts “ Mr Hetherington joked as he unwrapped and described each piece Fleet Air Arm Museum manager and senior curator Terry Hetherington with some of the parts from Sub-Lieutenant Fred Sherborne’s downed 882 Grumman Wildcat from World War II.After taking fire Sherborne crash landed the aircraft in a field of aubergine and was harboured by the French Resistance until the Allies took the village of Chateaurenard Villagers used a tractor to hide his plane in the nearby Durance River so the Germans were not aware of its or SBLT Sherborne’s existence He was hidden by the villagers for two weeks hiding in a chook house where he was fed cheese wine and eggs before the village was secured SBLT Sherborne was feted as a symbol of heroism by the villagers during their victory parade Over time a number villagers souvenired parts off the plane The actual logbook entry.Through a lot of negotiations some of the recovered parts have been shipped to Australia and will now be on permanent display at Nowra’s Fleet Air Arm Museum Mr Hetherington said it was incredible to be holding such a significant part of Australian World War II history “Over the last couple of years we have been negotiating with the people in the village of Chateaurenard to send us some of the relics,” he said The relics sent to Australia include part of the wing flap part of the control cables out of the cockpit a control rod which would have run through the fuselage to the rudder or elevator complete with a label that says Wildcat V (5) and Sherborne and the name of the person who “salvaged”  the part which incredibly still has the Royal Navy identification numbers on it “The identification number really gives relevance and significance to the component as it genuinely identifies it together with other manufacturer’s plates and markings that it is from a Grumman Wildcat aircraft,” Mr Hetherington said There are also fittings from inside the engine bay that would control the pilot’s throttle cable to the carburetor (supercharger controls) hydraulic pipelines to operate either the wing flaps or brakes and a “mysterious piece” which Mr Hetherington says will take some research to try and identify what function it had within the aircraft “They even offered to send us a belt of live 50 calibre ammunition but we had to decline,” he said “To have pieces of the aircraft here is incredible “These components have never been in Australia It was one of the many aircraft given to the Royal Navy by the United States Navy The RN operated a lot of American planes.” To be holding such a significant part of Australian World War II history is incredible He said the aviation industry in Britain at the time was very pressed to continue the manufacture of its own aircraft “An agreement known as the lend lease act was struck between the US and British governments components and engines built in the US were transferred to the Royal Navy,” he said “At the time the Royal Navy had their aircraft painted in a dark grey and green paint finish as you can see from some of these parts “One of those aircraft came here to Nowra to the Royal Naval Air Station Nowra in 1944-45.” Also included among the parts was a lifesize poster of the the SBLT Sherborne taken in 1944 another when he was part of the victory celebrations and a photograph of a villager sitting on the downed aircraft Photographs of the aircraft after the crash landing show it was relatively complete “The only damage to the aircraft was to the propeller when it crash landed on its belly,” Mr Hetherington said “There was some damage to the aircraft from the ground fire Fred himself suffered a shoulder injury and cuts to his face from pieces of glass and perspex from his cockpit canopy and windscreen that hit him.” The family tells how one of the bullets grazed his temple The village has also retained some parts of the plane in its museum “The village has been very generous in providing these components so we can mount an exhibition to recognise and honour Fred Sherborne not just during his time in conflict during World War II but the time he served here in the RAN up until the late 1960s as well,” Mr Hetherington said “He served here at Albatross and was one of the founding members of the Fleet Air Arm He went on to command the Sea Fury Squadron in charge of all the squadrons on the base Sherborne’s log book shows he was involved in several operational engagements “Prior to this incident where he was shot down Fred’s squadron was involved in the attack on the German battleship the Tirpitz in a fiord in Norway,” Mr Hetherington said “Fred was mentioned in dispatches for his involvement in that particular action.” It also told the story of  his crash on August 19 50 miles from our lines.’ the logbook documents photos and logbook will be part of a special exhibition to be launched in October The items will then be on permanent display within the museum Fred Sherborne was one of very few Australians who flew as fighter pilots and other capacities Mr Hetherington said the relics were a significant piece of Australian history “During World War II there were very few Australians who flew in fighter operations,” he said “In World War I a lot of Australians travelled at their own expense to England and fought and flew with with the Royal Navy “Some of our collection contains artifacts from the Australian fighter ace Robert Little “Fred Sherborne was one of very few Australians who flew as fighter pilots and other capacities He had joined the Australian Navy as a seaman and was a gunner on a merchant ship in the Mediterranean When that vessel was sunk Fred went back to England to be reallocated to another ship “When he was in the office waiting to be reposted he was aware they were selecting people to go to Florida in the US to learn how to fly,” Mr Hetherington said “Fred jumped from one queue to the other and found himself in Florida as a young 23-year-old Leading Seaman When authorities discovered him they apparently said ‘you’re here now we might as well teach you how to fly’.” He became a highly proficient and respected fighter pilot in the 882 Royal Navy Squadron and also later in his career in the RAN “These artifacts from WWII and in particular parts of Fred Sherborne’s aircraft add another aspect to the story we tell here at the Fleet Air Arm Museum of how our people, aircrews our engineers and our support personnel have operated throughout the century where Australians have been flying aircraft with ships in peace and war,” Mr Hetherington said Fred Sherborne wrote the following in pencil on fullscap [sic] about his experiences in the south of France in August 1944 “The invasion of France was going on apace with four losses on our part and an orderly retreat by the Germans The assault carrier force to which I was attached had carried out numerous sorties tactical and armed recce’s were flown off and dive-bombing and strafing were a daily feature in the life of pilot in nos 7 and 3 fighter wings of the Assault Carrier Force there was not much of it and to our group who had just left the very heavily guarded and armed convoys and installations of Norway It was reckoned to be such a ‘piece of cake’ that one looked upon it all more as training than actual warfare and D plus 4 we had all been lulled into a false sense of security and took to the air with a rather condescending blase air On that day four aircraft were shot down of which I was one two being killed and the other being taken P of W All four were shot down within ten miles of one another and some 50 miles inside the German lines luckily on the armoured glass directly in front of my face which stopped the main force of the shell but I received a big piece on my forehead and smaller pieces round my eyes and nose My flight had just dropped bombs on and near gun posts just outside Orange in the face of light AA - not intense and carried down on the deck for my get away I climbed up to four thousand feet to carry out a strafing attack on two a/c which I had noticed on an airfield earlier in the recce As we entered the dive for the high speed strafe the 40mm and lighter stuff started coming up and just as I was about to fire my guns my machine luckily on the armoured glass directly in front of my face This stopped the main force of the shell but I received a big piece on my forehead and smaller pieces round my eyes and nose Just about this time the whizzer stopped turning and there was quiet all round All of this time I was jinking both to avoid more shells and to make the Jerry think I had not been hit There was of course no hope of baling out nor was there a chance of picking a suitable field for a landing as it would have given my position away completely had I attempted to zoom switched off all the switches and trusted to luck and the hardiness of the machine This takes some time to relate but it all happened in seconds – Luckily as the speed dropped off I sensed rather than saw blood over my eyes made it fairly difficult to see properly Straightaway I pushed the nose into those on the near side of the field hoping that they would slow me down sufficiently and I found myself stopped on the ground right side up My crack on the head by the shell together with the crash-landing makes my next hour or two hazy but it helps show how one will work out lightning moves under great stress There was of course no hope of baling out nor was there a chance of picking a suitable field for a landing as it would have given my position away Firstly I took my parachute out of the cockpit and hid it My reason for doing that is now rather obscure but I hoped the Jerry might think they had the wrong machine and that the pilot of this one had baled out I next hurried – I couldn’t run as my leg had been twisted on landing - away from the a/c as fast as I could Very shortly I came upon a small creek and decided to walk up it even though it was at right angles to my track so that my scent would not be followed by the dogs which we had been told they possessed After some time of this I decided I felt bloody with spirits at a very low ebb and therefore decided to try my luck with a French peasant should I come across one well kept patches of land growing all types of stone fruits and which shortly I was going to curse good & roundly whom I hoped was a Frenchman and a partisan and waited in my creek until he worked down close to me trying to remain concealed as much as possible When he saw me he was most voluble and when he realized I was ‘Je suis Aviator Anglais’ he hurried me away to his tool shed where he locked me in and carried on with his work His colloquial French was a little too much for me who knew little French anyway so that I was not quite certain of his intentions However he seemed a decent type so I just sat and waited albeit a little frightened was still working and I observed that it was time to be hungry I thought it best not to open my emergency rations in case I should need them at a later date and so just continued to sit and become stiff and sore and sorry for myself arrived by at dusk and with him a girl Marie and another young man - ? Marie proceeded to wash the blood off my face and to carefully bathe my very sore eyes When she had completed this to her satisfaction He poured me a stiff tot of this clear liquid which proved to be Kir a potent drink made by the locals – I think out of pears Marie poured more of it into a bowl and bathed my forehead with it Fairly neat alcohol on an open wound is not the sort of thing one would use if there were any other antiseptic about but as things turned out and probably owing to my youth and good health The four empty sacks in the shed made the ground a little less hard but it was almost impossible to keep warm so that I was glad when the first signs of dawn approached having on only a pair of shorts and American summer weight flying over-alls I had kept my Mae West and that provided quite a lot of warmth Simeon came along around about 7o’clock and pulled my breakfast out of his haversack It consisted of a very hard type of meat roll hacking pieces off with a very sharp dagger come knife which I always carried Eating proved to be a major operation as I found my teeth jagged and broken by now also my eyes had both closed up completely and all round things were pretty grim Simone left some luscious peaches and a bottle of wine with me and started his day’s toil – wine eventually became my bug bear my guardians couldn’t get the idea that I wished to have some drinking water – The day passed uneventfully into the night and about nine o’clock I heard footsteps and slunk into the darkest corner behind some sacks in case it was the unfriends It proved to be Simeon who had now arranged to transport me back to?’ house where I was to hide up He double dinked me over some pretty rough tracks until we arrived at his place where his wife had a salad supper waiting and more wine I did justice to the meal and washed it down with more wine – their speech was quite intelligent to me but we got on together very well.  After the meal I was shown to my hide-out where I was doomed to remain for another 7 days This---was a loft above the fowl run but for my stay the fowls were induced to keep out of my part They disliked me intensely for doing them out of their nice loft and I in time couldn’t have cared less if I never saw a chook again in my whole life I saw Germans prying about on my second day in the fowl run but eventually they just gathered some grapes and pushed off – much to my relief My meals were provided by separate branches of the family whose husband was a POW at Leipzig luncheon and supper They all provided red wine and Marie whose favourite I seemed to be gave me anything I desired plus a white wine rather like a heavy hock She also bathed my eyes and head several times a day and looked after my general health (a type of malaria which I was prone to get) and became very restless and had a terrific temperature Malou then showed their stirling qualities one of them was continually with me until the fever started to wear off By this time we understood each other fairly well and it took me a great deal of pleading to stop them getting in a doctor I wanted to get as few people as possible know that I was about for fear of reprisals Every night I heard tanks firing and manoevering and as the days went by so the firing came nearer & nearer until finally I was told that the Germans were evacuating the Avignon area I could visualise days and weeks going by without having to see or hear a chook and the prospect was most pleasing I saw Germans prying about on my second day in the fowl run but eventually they just gathered some grapes and pushed off – much to my relief I was taken out of my hutch and allowed to dine in state with the family Halfway through dinner we heard footsteps so my place was cleared away and I was relegated to a position under the stairs It proved to be friends on a visit so I was brought forth and paraded like a champion bull at the show One of the lasses of the party was noticed to dash off after a while but she returned later with numerous other friends all carrying bottles of wine and so the party started Unfortunately in my weakened state I could not indulge too freely and in fact ‘pushed off’ to my coop after a very short time The party however continued for quite some time A villager sits on Fred Sherborne's downed Grumman Wildcat not long after it crashed near Avignon in southern France on August 19 1944.The Grumman F4F Wildcat is an American carrier-based fighter aircraft that began service with both the United States Navy and the British Royal Navy (as the Martlet) in 1940 First used in combat by the British in Europe the Wildcat was the only effective fighter available to the United States Navy and Marine Corps in the Pacific Theater during the early part of World War II in 1941 and 1942; the disappointing Brewster Buffalo was withdrawn in favor of the Wildcat and replaced as units became available the Wildcat was outperformed by the faster 331 mph (533 km/h) coupled with tactics such as the Thach Weave resulted in a claimed air combat kill-to-loss ratio of 5.9:1 in 1942 and 6.9:1 for the entire war Onboard merchant ship ‘Imperial Star’ when sunk by enemy action in Mediterranean on September 27 Selected for fighter pilot training with RN and USN and appointed Sub Lieutenant on September 26 Flew bombing missions against battleship ‘Tirpitz’ in Norway Posted to RN escort carrier HMS ‘Searcher’ with 882 ‘Wildcat’ Squadron for the invasion of Southern France Shot down five miles south of Avignon on August 19 Harboured by french resistance until Allies retook the village of Chateaurenard Feted as a symbol of heroism by the villagers during their victory parade some eight days later and re-engaged in Australian Fleet Air Arm in January 1948 Returned to UK for flying refresher training and posted to 805 ‘Sea Fury’ squadron 1948–1951 onboard ‘Sydney’ and at ‘Albatross’ Promoted to Lieutenant Commander in September 1951 Two years exchange with RN squadrons 1951-1953 Returned to ‘Albatross’ and ‘Sydney’ to command 805 Squadron 1953-1955 Promoted to Commander in June 1957 and appointed ‘Melbourne’ as Commander Air and Fleet Aviation Officer Posted to ‘Albatross’ as Commander Air August 1958 and tempy executive officer Appointed Staff Officer Air to Australian Navy representative in London January 1962 followed by Naval Attache to Indonesia in January 1966 Took command of ‘Kuttabul’ March 1969 and retired from RAN in October 1969 Robert Crawford is a senior journalist at the South Coast Register, for Australian Community Media . Everyone's got a story, what's yours? Email robert.crawford@austcommunitymedia.com.au Newsletters & AlertsView allDaily Today's top stories curated by our news team Get the editor's insights: what's happening & why it matters tips & travel writing to transport you around the globe Your weekday morning newsletter on national affairs reviews and expert insights every Thursday from CarExpert Your essential national news digest: all the big issues on Wednesday and great reading every Saturday Let the ACM network's editors and journalists bring you news and views from all over Your digital replica of Today's Paper