The effect of WRI1 and DGAT1 overexpression on lipid metabolism in developing pennycress embryos
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ABSTRACT:
ORGANISM(S): Thlaspi arvense
PROVIDER: GSE283815 | GEO | 2026/03/19
REPOSITORIES: GEO
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Project description:The aviation industry’s growing interest in renewable jet fuel has encouraged the exploration of alternative oilseed crops. Replacing traditional fossil fuels with a sustainable, domestically sourced crop can substantially reduce carbon emissions, thus mitigating global climate instability. Pennycress (Thlaspi arvense L.) is an emerging oilseed intermediate crop that can be grown during the offseason between maize (Zea mays) and soybean (Glycine max) to produce renewable biofuel. Pennycress is being domesticated through breeding and mutagenesis, providing opportunities for trait enhancement. Here, we employed metabolic engineering strategies to improve seed oil composition and bolster the plant's economic competitiveness. FATTY ACID ELONGATION 1 (FAE1) was targeted using CRISPR-Cas 9 gene editing to eliminate very long chain fatty acids (VLCFAs) from pennycress seed oil, thereby enhancing its cold flow properties. Through an integrated multi-omics approach, we investigated the impact of eliminating VLCFAs in developing and mature plant embryos. Our findings revealed improved cold-germination efficiency in fae1, with seedling emergence occurring up to three days earlier at 10 ºC. However, these alterations led to a trade-off between storage oil content and composition. Additionally, these shifts in lipid biosynthesis were accompanied by broad metabolic changes, such as the accumulation of glucose and ADP-glucose quantities consistent with increased starch production. Furthermore, shifts to shorter FA chains triggered the upregulation of heat shock proteins, underscoring the importance of VLCFAs in stress signaling pathways. Overall, this research provides crucial insights for optimizing pennycress seed oil while preserving essential traits for biofuel applications.
2024-11-29 | GSE283023 | GEO
Project description:Thlaspi arvense (Pennycress) is an emerging feedstock for biofuel production because of its high seed oil content enriched in erucic acid. A transcriptomic and a lipidomic study were performed to analyze the dynamics of gene expression, glycerolipid content and acyl-group distribution during seed maturation. Genes involved in fatty acid biosynthesis were expressed at the early stages of seed maturation. Genes encoding enzymes of the Kennedy pathway like TaDGAT1, TaLPAT or TaGPAT increased their expression with maturation, coinciding with the increase in TAG species containing 22:1. Positional analysis showed that the most abundant TAG species contained 18:2 at sn-2 position in all maturation stages, suggesting no specificity of the TaLPAT for VLCFAs. TaDGAT2 mRNA was more abundant at the initial maturation stages, coincident with the rapid incorporation of 22:1 to TAG, suggesting a coordination between DGAT enzymes for TAG biosynthesis. Genes encoding TaPDAT1, TaLPCAT or TaPDCT, involved in acyl-editing or PC-derived DAG/TAG biosynthesis showed also higher expression at the early maturation stages, coinciding with a higher proportion of TAG species containing C18 fatty acids. These results suggested a higher contribution of these two pathways at the early stages of seed maturation. LC-MS analysis of the content and acyl-group distribution of DAG and PC pools was compatible with the acyl content in TAG at the different maturation stages. Our data point to a model in which a strong temporal coordination between pathways and isoforms in each pathway, both at the expression and acyl-group incorporation, contributes to high erucic TAG accumulation in Pennycress.
2024-04-25 | GSE256460 | GEO
Project description:Understanding the mechanisms of transcriptional regulation for genes involved with FA biosynthesis and TAG accumulation in seeds can represent an important step in the development of crops with increased oil content. Among well studied transcription factors (TFs), WRINKLED1 (WRI1), a member of the APETALA2 (AP2) family of TFs, has been characterized as a central regulator of FA biosynthesis in seeds of many plant species. Despite its importance for the process, the detailed mechanisms of WRI1 function in regulating FA biosynthesis in seeds is still not well understood. This study aimed to characterize the gene networks controled by WRI1 during soybean embryo development and its function as central regulator of FA biosynthesis and TAG accumulation in seeds. In this part of the study, we identified genes misregulated in the embryos of the Arabidopsis mutant wrinkled1-1, when compared to wild-type.
2024-10-16 | GSE253109 | GEO
Project description:Pennycress (Thlaspi arvense) is a winter oilseed domesticated recently to be incorporated as an intermediate crop between the existing cropping systems of the US Midwest. We show that a natural accession of pennycress, 2032, is more susceptible to the necrotrophic fungal pathogens Sclerotinia sclerotiorum and Alternaria japonica than the reference pennycress accession MN106. A previously identified marker associated with earliness in pennycress was found to be present in in a gene homologous to Arabidopsis Jumonji 14 (JMJ14). It has been reported that AtJMJ14 promotes disease resistance and represses flowering, and greenhouse studies of breeding populations confirmed this phenomenon in pennycress. Plants with the 2032 TaJMJ14 allele were more susceptible to fungi and flowered early. CRISPR-Cas9 editing was used to generate additional TaJMJ14 alleles. A 9 bp deletion in TaJMJ14 showed trends of early flowering and S. sclerotiorum susceptibility, whereas a complete loss-of-function allele led to infertility. We further investigated the transcriptomes of MN106 and 2032 plants in the early stages of S. sclerotiorum and A. japonica infection to identify potential resistance and susceptibility genes. Differences in the expression of pathogen-associated molecular pattern-triggered immunity (PTI)-associated genes led us to discover that 2032 plants have defects in elicitor-triggered oxidative bursts. The transcriptional responses unique to each accession lay a foundation for future gene-editing and breeding approaches to keep the beneficial early flowering phenotype conferred by 2032 but uncouple it from disease susceptibility.
2025-04-21 | GSE294781 | GEO
Project description:Understanding the mechanisms of transcriptional regulation for genes involved with FA biosynthesis and TAG accumulation in seeds can represent an important step in the development of crops with increased oil content. Among well studied transcription factors (TFs), WRINKLED1 (WRI1), a member of the APETALA2 (AP2) family of TFs, has been characterized as a central regulator of FA biosynthesis in seeds of many plant species. Despite its importance for the process, the detailed mechanisms of WRI1 function in regulating FA biosynthesis in seeds is still not well understood. This study aimed to characterize the gene networks controled by WRI1 during soybean embryo development and its function as central regulator of FA biosynthesis and TAG accumulation in seeds. In this part of the study, we identified genes regulated by the soybean WRI1s (GmWRI1s) by overexpressing GmWRI1-A (Glyma.08G227700) in Arabidopsis leaf mesophyll protoplasts, and identifying the mRNAs that are induced 16h after transfection.
2024-10-16 | GSE253107 | GEO
Project description:Thlaspi arvense cultivar:Pennycress Raw sequence reads
| PRJNA808106 | ENA
Project description:Triacylglycerol (TAG) is the main storage lipid in plant seeds and the major form of plant oil used for food and, increasingly, for industrial and biofuel applications. Several transcription factors, including FUSCA3 (At3g26790, FUS3), are associated with regulation of embryo 3maturation and oil biosynthesis in seeds. However, the ability of FUS3 to increase TAG biosynthesis in other tissues has not been quantitatively examined. Here, we evaluated the ability of FUS3 to activate TAG accumulation in non-seed tissues. Overexpression of FUS3 driven by an estradiol-inducible promoter increased oil contents in Arabidopsis seedlings up to 6% of dry weight; more than 50 fold over controls. Eicosenoic acid, a characteristic fatty acid of Arabidopsis seed oil, accumulated to over 20% of fatty acids in cotyledons and leaves. These large increases depended on added sucrose, although without sucrose TAG increased 3-4 fold. Inducing the expression of FUS3 in tobacco BY2 cells also increased TAG accumulation, and co-expression of FUS3 and diacylglycerol acyltransferase 1 (DGAT1) further increased TAG levels to 4% of dry weight. BY2 cell growth was not altered by FUS3 expression, although Arabidopsis seedling development was impaired, consistent with the ability of FUS3 to induce embryo characteristics in non-seed tissues. Microarrays of Arabidopsis seedlings revealed that FUS3 overexpression increased expression of a higher proportion of genes involved in TAG biosynthesis than genes involved in fatty acid biosynthesis or other lipid pathways. Together these results provide additional insights into FUS3 functions in TAG metabolism and suggest complementary strategies for engineering vegetative oil accumulation.
2016-04-19 | GSE80360 | GEO
Project description:Brassica napus, Camelina sativa and field pennycress (Thlaspi arvense) are three important oilseed crops in the Brassicaceae family. As sessile organisms, these crops are continuously exposed to various stresses when grown in the field. Interestingly, the responses of these three crops to different environmental stimuli vary to a great extent, but there is very limited knowledge about the molecular basis of these discrepancies. In this study, we employed untargeted metabolomics to compare the metabolic profile of these crops, and examined the potential causal genes through further integration with transcriptomic analysis. Our data revealed distinctive overall metabolic profiles among these three crops, where in particular, a variety of phenylpropanoids showed differential accumulation and the corresponding putative genes’ expression varied significantly. Our results here will provide valuable insight into the understanding of metabolic variations among these three important oilseed crops, and provide potential targets for the future breeding of stress tolerant crops.
2025-07-23 | GSE279986 | GEO
Project description:Global transcript profiling of Arabidopsis dgat1-1 seed at different stages of embryo development was used to identify differentially expressed genes in the mutant compared to wild-type (Col-0) seed. These genes provided information about the remodeling of lipid metabolism and TAG synthesis in response to the lack of DGAT1 activity.
2019-08-20 | GSE123303 | GEO
Project description:Biomass crops engineered to accumulate energy-dense triacylglycerols (TAG or “vegetable oils”) in their vegetative tissues have emerged as potential feedstocks to meet the growing demand for renewable diesel and sustainable aviation fuel (SAF). Unlike oil palm and oilseed crops, the current commercial sources of TAG, vegetative tissues, such as leaves and stems, only transiently accumulate TAG. In this report, we used grain (Texas430 or TX430) and sugar-accumulating “sweet” (Ramada) genotypes of sorghum, a high-yielding, environmentally resilient biomass crop, to accumulate TAG in leaves and stems. We initially tested several gene combinations for a “push-pull-protect" strategy. The top TAG-yielding constructs contained five oil transgenes for a sorghum Wrinkled1 transcription factor (“push”), a Cuphea viscosissima diacylglycerol acyltransferase (DGAT; "pull"), a modified sesame oleosin (“protect”) and two combinations of specialized Cuphea lysophosphatidic acid acyltransferases and medium-chain acyl-acyl carrier protein thioesterases. Though intended to generate oils with medium-chain fatty acids, engineered lines accumulated oleic acid-rich oil to amounts of up to 2.5% DW in leaves and 2.0% DW in stems in the greenhouse, 36-fold and 49-fold increases relative to wild-type plants, respectively. Under field conditions, the top-performing event accumulated TAG to amount of up to 5.5% DW in leaves and 3.5% DW in stems, 78-fold and 58-fold increases, respectively, relative to wild-type TX430. Transcriptomic and fluxomic analyses revealed potential bottlenecks for increased TAG accumulation. Overall, our studies highlight the utility of a lab-to-field pipeline coupled with systems biology studies to deliver high vegetative oil sorghum for SAF and renewable diesel production.
2024-07-31 | GSE272636 | GEO