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Impacts of different fungicide treatments on grapevine trunks affected by esca disease

ABSTRACT: Fungicides and the grapevine wood mycobiome. A case study on tracheomycotic ascomycete Phaeomoniella chlamydospora reveals potential for two novel control strategies.

PROVIDER: PRJEB32853 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR3348620_1.fastq.gz	Fastqsanger.gz
	ERR3348620_2.fastq.gz	Fastqsanger.gz
	ERR3348630_1.fastq.gz	Fastqsanger.gz
	ERR3348630_2.fastq.gz	Fastqsanger.gz
	ERR3348640_1.fastq.gz	Fastqsanger.gz

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Gene expression during wood pathogen infection in the presence of a root biocontrol agent in grapevine.

Project description:Transcriptional changes occurring at the infection site of 2 weeks old Cabernet sauvignon grapevine cuttings infected with a wood pathogen (Phaeomoniella chlamydospora) in the presence of a root-inoculated biocontrol agent (Pythium oligandrum). Gene expression profiling was done using the Nimblegen whole genome array with 3 biological replicates of 3 pooled wood chunks harvested 0 and 14 d after treatment (pathogen infection, biocontrol agent inoculation, mock treatment).

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Project description:The goals of this study are to compare NGS-derived transcriptome profiling (RNA-seq) from grapevine wood infected by a fungal pathogen in the presence of a root biological control agent. One of the goals was to obtain molecular data about the fungus pathogen (Phaeomoniella chlamydospora) during grapevine wood infection. Grapevine pathogen-infected wood mRNA profiles of 2-month-old plantlets (14 days post infection) were generated by deep sequencing, in triplicate, using Illumina Hiseq2500. The sequence reads that passed quality filters were analyzed by TopHat followed by Cufflinks. qRTaPCR validation was performed using SYBR Green assays. Using an optimized data analysis workflow, we mapped sequence reads to the grapevine genome (build IGGP 12x) and identified pathogen transcripts. RNAseq analyses, using a ribosomal RNA depletion technology for library preparation, provided identification of genes expressed by P. chlamydospora during infection: as for genes related to effector biosynthesis enzymes, carbohydrate-active enzymes and transcription regulators involved in known regulation pathways in fungi. Insights about P. oligandrum modulation of grapevine infection by this pathogen were also found. Our study represents the first detailed analysis of grapevine wood infection by a fungal pathogen generated by RNA-seq technology. The optimized data analysis workflows reported here should provide a framework for comparative investigations of expression profiles. Our results show that NGS offers a comprehensive evaluation of mRNA content within grapevine wood tissue. We conclude that RNA-seq based transcriptome characterization would permit the dissection of complex biologic interactions.

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Project description:Winemakers have long used copper as a conventional fungicide treatment on grapevine and only recently, the use of biocontrol agents depicted a promising alternative. Most of the studies that investigate the impact of fungicide treatments, focus on specific pathogenic microbes. In the present work instead, a field experiment conducted in South Africa shows the seasonal microbial change occurring on grapevine leaves, periodically treated with two different fungicide treatments: copper sulphate and Lactobacillus plantarum MW-1. In this work, NGS data were combined with strain-specific and community qPCRs to reveal the shift of the microbial communities throughout the growing season and highlight the impact of fungicides on the microbiota. Only the family of Lactobacillaceae systematically changed between treatments, while the bacterial community remained relatively stable over time. MW-1 was exclusively detected on biocontrol-sprayed leaves. Conversely, the fungal community was largely shaped by sampling time, underlining the succession of different dominant taxa over the months. Between treatments, only a few fungal taxa appeared to change significantly and the fungal load was also comparable. Monitoring the dynamics of the microbial community under different fungicide treatments may advise the best timing to apply treatments to the plants, toward the realization of more sustainable agricultural practices.

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