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SRX5010918: GSM3473828: BI16-16; Solanum lycopersicum; RNA-Seq
1 ILLUMINA (Illumina HiSeq 4000) run: 15.6M spots, 795.4M bases, 264.1Mb downloads

Submitted by: NCBI (GEO)
Study: Adaptation of necrotrophic infection strategies by Botrytis cinerea, Fusarium acuminatum, and Rhizopus stolonifer as a function of  tomato fruit ripening
show Abstracthide Abstract
Worldwide, 20-25% of all harvested fruit and vegetables are lost annually in the field and throughout the postharvest supply handling chain due to spoilage by fungal pathogens. Most impactful postharvest pathogens exhibit necrotrophic lifestyles, resulting in rotting of the host tissues and complete loss of marketable commodities. Necrotrophic fungi can readily infect ripe fruit leading to the rapid establishment of disease symptoms. However, these pathogens generally fail to infect unripe fruit, or remain quiescent until host and environmental conditions stimulate a successful infection. Current research on necrotrophic infections of fruit was mainly focused on the host by characterizing genetic and physicochemical factors that inhibit or promote the disease. However, the pathogenicity and virulence strategies employed by necrotrophic pathogens in ripe and unripe fruit are mostly understudied. Here, we provide a first comparative transcriptomics study of fungal postharvest pathogens: Botrytis cinerea, Rhizopus stolonifer and Fusarium acuminatum, all of which display necrotrophic behavior when infecting fruit. We de novo assembled and annotated the transcriptomes of R. stolonifer, and F. acuminatum and performed a differential gene expression analysis comparing the three fungal transcriptomes during fruit infection with that of fungal in-vitro growth. Analysis of the differentially expressed genes for enrichment of functional annotations revealed shared strategies of the three fungi during infection of compatible (ripe fruit) and incompatible (unripe fruit) hosts. We furthermore selected candidate genes that are involved in these strategies to characterize their expression during infection of unripe and ripe-like fruit of the non-ripening (nor) tomato mutant, both of which are physiologically and biochemically similar to unripe wildtype fruit. By enabling a better understanding of fungal necrotrophic infection  strategies, we move closer to generating accurate models of fruit diseases and development of early detection tools and effective management strategies. Overall design: Differential expression analysis of pathogen genes in green or red fruit compared to in vitro. Classification of differentially expressed genes by gene ontology, Pfam domains, and other functional categories.
Sample: BI16-16
SAMN10430654 • SRS4044829 • All experiments • All runs
Library:
Instrument: Illumina HiSeq 4000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Two grams of tissue per sample were ground in liquid nitrogen and 10mL of the RNA extraction buffer (CTAB 2% v/v, PVP 2%v/v, 100mMTris pH 8, 2MNaCl, 25mM EDTA, 0.5 g/L spermidine, 10mM β-mercaptoethanol) were added. The samples were immediately incubated for 5min at 65◦C. Two extractions with one equal volume of chloro- form:isoamyl alchohol (24:1, v/v) followed by centrifugation at 4000 rpm for 45 min at 4◦C were performed. The supernatant was recovered and 1/10 volume of 1MKOAc was added followed by centrifugation at 4000 rpm for 20 min at 4◦C. The super- natant was collected and 1/4 volume of 10M LiCl was added. Samples were incubated overnight at−20◦C and then centrifuged at 4000 rpm for 45 min at 4◦C. The supernatant was discarded and the RNA pellet was further purified using the RNeasy Plant Mini Kit (Qiagen®). DNAse treatment (RNase-Free DNase Set,Qiagen®) was done in column during the purification step. The RNA was resuspended in 35µL of nuclease-free water. The RNA concentration and purity were measured using NanoDrop 2000c Spectrophotometer (Thermo Scientific, Inc.). The RNA integrity was checked by agarose gel electrophoresis. Libraries were prepared using the Illumina TruSeq RNA Sample Preparation Kit v2 (Illumina, CA).
Experiment attributes:
GEO Accession: GSM3473828
Links:
Runs: 1 run, 15.6M spots, 795.4M bases, 264.1Mb
Run# of Spots# of BasesSizePublished
SRR819135115,596,453795.4M264.1Mb2018-12-02

ID:
6764375

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