Ontology highlight
ABSTRACT:
PROVIDER: PRJNA841432 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR19360586.fastq.gz | Fastqsanger.gz |
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Project description:Herbivorax saccincola A7 is an alkali-thermophilic lignocellulolytic bacterium that was known to possess a strong xylan degradation ability. They can utilize a wide range of carbon sources, however the response and regulation mechanisms to different carbon sources, in terms of genes expression, are still not identified. In this study, H. saccincola A7 was cultured with cellobiose, cellulose, xylan, cellulose-xylan mixture, and empty fruit bunch (EFB) as the sole carbon source. We carried out a comparative expression analysis, focusing on biomass degradation-related genes. The cellulosomal scaffolding proteins and cellulosomal enzymes of H. saccincola A7 were drastically induced when a cellulose-containing substrate was used as a carbon source. On the other hand, xylan induced only some non-cellulosomal enzymes that might be necessary for xylan degradation. The identification of the carbohydrate-sensing mechanism in H. saccincola A7 appeared a similar system as previously described in Clostridium thermocellum. The lignocellulose degradation ability of H. saccincola A7 may be controlled enzyme expression by cellulose-containing substrate, not by xylan. Thus, recognition in the existence of cellulosic substrates is effective in enhancing a high xylan degradation ability in H. saccincola A7.
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Project description:A laboratory colony of Phlebotomus perniciosus sand flies was maintained. Sand flies were infected with cultured Leishmania infantum promastigotes in stationary phase. Ten infected sand flies were dissected after 5 days and promastigotes within the gut pooled. The cells were immediately washed in PBS once and lysed in TRIzol reagent (Life Technologies). RNA isolation was completed according to the manufacturer's instructions, obtaining 63ng. RNA-seq libraries were generated using the spliced leader sequence for second strand synthesis (Cuypers et al., 2017; Haydock et al., 2015), thus allowing for specific amplification of sequences from L. infantum promastigotes, thus avoiding contamination with material from the sand fly gut. Single-end sequencing was performed in an Illumina HiSeq2500 instrument and data analysis was conducted using bowtie2, samtools, featureCounts and Geneious. The main findings are: i) substantial differences in differential gene expression between sand fly-derived (sfPro) and cultured (acPro) promastigotes; and ii) over-expression of genes involved in metacyclogenesis in sfPro vs. acPro, including gp63 genes, autophagy genes, etc.
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Project description:Brown rot fungi have great potential in biorefinery wood conversion systems, because they are the primary wood decomposers in coniferous forests and have an efficient lignocellulose degrading system. Their initial wood degradation mechanism is thought to consist of an oxidative radical-based system that acts sequentially with an enzymatic saccharification system, but the complete molecular mechanism of this system has not yet been elucidated. Some studies have shown that wood degradation mechanisms of brown rot fungi have diversity in their substrate selectivity. Gloeophyllum trabeum, one of the most studied brown rot species, has broad substrate selectivity and even can degrade some grasses. However, the basis for this broad substrate specificity is poorly understood. In this study, we performed RNA-seq analyses on G. trabeum grown on media containing glucose, cellulose, or Japanese cedar (Cryptomeria japonica) as the sole carbon source. Beyond the gene expression on glucose, 1129 genes were upregulated on cellulose and 1516 genes were upregulated on cedar. Carbohydrate Active enZyme (CAZyme) genes upregulated on cellulose and cedar media by G. trabeum included GH12, GH131, CE1, AA3_1, AA3_2, AA3_4 and AA9, which is a newly reported expression pattern for brown rot fungi. The upregulation of both terpene synthase and cytochrome P450 genes on cedar media suggests the potential importance of these genes in the production of secondary metabolites associated with the chelator-mediated Fenton reaction. These results provide new insights into the inherent wood degradation mechanism of G. trabeum and the diversity of brown rot mechanisms.
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Project description:The silkworm (Bombyx mori) has long been considered a source of food and medicine due to its high nutritional, medicinal, and economic value in East Asia. However, in some sensitive individuals, silkworm consumption can cause allergenic reactions such as vomiting, asthma, and anaphylaxis. Therefore, the development of a reliable method for silkworm detection is required to avoid such allergenic incidents. In this study, two different methods (liquid chromatography combined with mass spectrometry [LC-MS/MS] and real-time polymerase chain reaction [PCR]) were developed to determine an efficient technique for silkworm detection in foods. The developed methods demonstrated high sensitivity in detecting the silkworm in processed foods. Silkworm-spiked model cookies were used to confirm the sensitivity of both LC-MS/MS (0.0005%) and real-time PCR (0.001%). These methods were found to be useful for detecting the silkworm in foods and avoiding allergenic reactions. To the best of our knowledge, this is the first study to compare LC-MS/MS and real-time PCR for silkworm detection in complex processed foods.
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