Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1173486 | ENA |
REPOSITORIES: ENA
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Project description:Paulownia elongata is a fast-growing tree species native to China that is grown in different climates, types of soils, and can be easily re-grown. The versatility of the P. elongata species makes it an ideal candidate for biofuel production. High soil salinity is known to inhibit plant growth dramatically or lead to death. Salinity in soil is a detrimental abiotic stress affecting crop production worldwide and a hindrance for potential crop candidates used for biofuel production. The purpose of this study was to characterize the salt-induced transcriptome of P. elongata. Transcriptome differences in response to salt stress were determined by RNA sequencing (RNA-seq) using next generation sequencing and bioinformatics analysis. A total of 646 genes were found to have significant altered expression in response to salt stress, and expression levels of a selective subset of these genes were chosen and confirmed using quantitative real-time PCR. To the best of our knowledge, this is the first report of salt-induced transcriptome analysis in P. elongata. The current study indicates that the differential expression of certain genes may have an important role in the adaptation of P. elongata in response to salt stress. Functional characterization of these genes will assist in future development of salt tolerance in P. elongata, which could be used to enhance biofuel production.
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Project description:Background: Evolutionary engineering is a powerful approach to isolate suppressor mutants and industrially relevant genotypes. Until recently, DNA microarray analysis was the only affordable genome-wide approach to identify the responsible mutations. This situation has changed due to the rapidly decreasing costs of whole genome (re)sequencing. DNA microarray-based mRNA expression analysis and whole genome resequencing were combined in a study on lactate transport in Saccharomyces cerevisiae. Jen1p is the only S. cerevisiae lactate transporter reported in literature. To identify alternative lactate transporters, a jen1Δ strain was evolved for growth on lactate. Results: Two independent evolution experiments yielded Jen1p-independent growth on lactate (μmax 0.14 and 0.18 h-1 for single-cell lines IMW004 and IMW005, respectively). Whereas mRNA expression analysis did not provide leads, whole-genome resequencing showed different single nucleotide changes (C755G/Leu219Val and C655G/Ala252Gly) in the acetate transporter gene ADY2. Analysis of mRNA levels and depth of coverage of DNA sequencing combined with karyotyping, gene deletions and diagnostic PCR showed that in IMW004 an isochromosome III (~475 kb), which contains two additional copies of ADY2C755G, was formed via crossover between YCLWΔ15 and YCRCΔ6. Introduction of the ADY2 alleles in a jen1 ady2 strain resulted in growth on lactate (μmax 0.14 h-1 for Ady2pLeu219Val and 0.12 h-1 for Ady2pAla252Gly). Conclusions: Whole-genome resequencing of yeast strains obtained from independent evolution experiments enabled rapid identification of a key gene that was not identified by mRNA expression analysis of the same strains. Reverse metabolic engineering showed that mutated alleles of ADY2 (C655G and C755G) encode efficient lactate transporters.
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