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Uncultured fungus

ABSTRACT: Fungal microbiome of herbaceous plant roots collected on truffle grounds (Tuber melanosporum)

PROVIDER: PRJNA397712 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR5930420.fastq.gz	Fastqsanger.gz
	SRR5930430.fastq.gz	Fastqsanger.gz
	SRR5930440.fastq.gz	Fastqsanger.gz
	SRR5930450.fastq.gz	Fastqsanger.gz
	SRR5930460.fastq.gz	Fastqsanger.gz

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Bisulfite-Seq and RNA-seq profiling of truffle (Tuber melanosporum)

Project description:We investigated the ascomycete truffle Tuber melanosporum exploits DNA methylation and transcription to cope with the more than 45,000 repeated elements that are present in its genome. Whole-genome bisulfite sequencing and RNA-sequencing, were performed on different developmental stages of this symbiotic hypogeous fungus -fruitbody (FB), free-living mycelium (FLM), and ectomycorrhiza. Examination of DNA methylation and transcription of truffle in its free living mycelium (FLM), fruit body (FB), and ectomycorrhizal root tips (ECM)

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Project description:The aim of this analysis was to better understand the complex symbiotic stage of Tuber melanosporum by combining the use of laser capture microdissection and microarray gene expression analysis. We isolated the fungal/soil (i.e. the mantle) and the fungal/plant (i.e. the Hartig net) interfaces from transverse sections of T. melanosporum/Corylus avellana ectomycorrhizas and identified the transcriptional landscape associated with each compartment. We compared these data to the transcriptome of ectomycorrhizal root tips, free-living mycelium and fruiting bodies of Tuber melanosporum (Series GSE17529). The T. melanosporum custom-exon expression array (4 x 72K) manufactured by Roche NimbleGen Systems Limited (Madison, WI) contained five independent, nonidentical, 60-mer probes per gene model coding sequence. Included in the oligoarray were 12,232 annotated gene models, 3,913 random 60-mer control probes and labelling controls. Sequences used for the oligonucleotide design were from an early draft of the gene catalog containing several TE families. For 1,876 gene models, technical duplicates were included on the array. We performed 6 hybridizations (NimbleGen) with samples derived from microdissected mantles (3 biological replicates) and Hartig nets ((3 biological replicates each) from T. melanosporum/Corylus avellana ectomycorrhizal root tips . All samples were labeled with Cy3.

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Project description:Truffles of the Tuber species as expensive foods are predestined for food fraud. To find markers for distinguishing different truffle species the proteomes as phenotype determining basis were investigated. First, established sample preparation protocols subsequent to LC-MS/MS analysis for a bottom up proteomic approach were tested on a truffle sample. The sample preparation included protein extraction and in-solution tryptic digestion. For protein extraction sodium deoxycholate (SDC), sodium dodecylsulfate (SDS) and urea were tested, followed by direct tryptic digestion in solution (SDC extracted proteins) and a tryptic digestion utilizing the FASP approach with molecular cut-off filters (SDS and urea extracted proteins). By this approach the SDC sample preparation protocol was classified as the most suited one to prepare truffle samples. It resulted in a high protein identification rate, had a high reproducibility, was cost efficient and most easy to handle. By a quantitative proteomic approach using label-free and non-targeted bottom up LC-MS/MS the species T. magnatum (n=13), T. aestivum (n=16), T. uncinatum (n=4) and T. melanosporum (n=7) were compared and truffle species specific markers were identified.

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