Pennycress is a potentially lucrative biofuel crop due to its high content of long-chain unsaturated fatty acids, and because it uses non-conventional pathways to achieve efficient oil production. However, metabolic engineering is required to improve pennycress oilseed content and make it an economically viable source of aviation fuel. Research is warranted to determine if further upregulation of these non-conventional pathways could improve oil production within the species even more, which would indicate these processes serve as promising metabolic engineering targets and could provide the improvement necessary for economic feasibility of this crop. To test this hypothesis, we performed a comparative biomass, metabolomic, and transcriptomic analyses between a high oil accession (HO) and low oil accession (LO) of pennycress to assess potential factors required to optimize oil content. An evident reduction in glycolysis intermediates, improved oxidative pentose phosphate pathway activity, malate accumulation in the tricarboxylic acid cycle, and an anaplerotic pathway upregulation were noted in the HO genotype. Additionally, higher levels of threonine aldolase transcripts imply a pyruvate bypass mechanism for acetyl-CoA production. Nucleotide sugar and ascorbate accumulation also were evident in HO, suggesting differential fate of associated carbon between the two genotypes. An altered transcriptome related to lipid droplet (LD) biosynthesis and stability suggests a contribution to a more tightly-packed LD arrangement in HO cotyledons. In addition to the importance of central carbon metabolism augmentation, alternative routes of carbon entry into fatty acid synthesis and modification, as well as transcriptionally modified changes in LD regulation, are key aspects of metabolism and storage associated with economically favorable phenotypes of the species.
SummaryPennycress (Thlaspi arvense L.), a member of the Brassicaceae family, produces seed oil high in erucic acid, suitable for biodiesel and aviation fuel. Although pennycress, a winter annual, could be grown as a dedicated bioenergy crop, an increase in its seed oil content is required to improve its economic competitiveness. The success of crop improvement relies upon finding the right combination of biomarkers and targets, and the best genetic engineering and/or breeding strategies. In this work, we combined biomass composition with metabolomic and transcriptomic studies of developing embryos from 22 pennycress natural variants to identify targets for oil improvement. The selected accession collection presented diverse levels of fatty acids at maturity ranging from 29% to 41%. Pearson correlation analyses, weighted gene co‐expression network analysis and biomarker identifications were used as complementary approaches to detect associations between metabolite level or gene expression and oil content at maturity. The results indicated that improving seed oil content can lead to a concomitant increase in the proportion of erucic acid without affecting the weight of embryos. Processes, such as carbon partitioning towards the chloroplast, lipid metabolism, photosynthesis, and a tight control of nitrogen availability, were found to be key for oil improvement in pennycress. Besides identifying specific targets, our results also provide guidance regarding the best timing for their modification, early or middle maturation. Thus, this work lays out promising strategies, specific for pennycress, to accelerate the successful development of lines with increased seed oil content for biofuel applications.
ResumenEste artículo presenta el trabajo realizado en práctica pedagógica en modalidad integral, durante el primer semestre del 2013, con 12 estudiantes del componente Biología Molecular de la Licenciatura en Biología de la Universidad Pedagógica Nacional (UPN), extensión Centro Valle de Tenza (CVT) ubicado en el municipio de Sutatenza (Boyacá).El trabajo consto de cuatro fases orientadas por la Línea de Investigación Enseñanza de la Biotecnología en Colombia de la UPN, las cuales tuvieron como propósito contribuir a la enseñanza de la Biología Molecular a través de prácticas de laboratorio (virtuales, convencionales y artesanales), asumiéndose como estrategia didáctica.En la primera fase se encontró; que para los estudiantes las prácticas de laboratorio les permiten un acercamiento y fortalecimiento de los elementos teóricos, llevándose la teoría a la práctica. Sin embargo, fueron casi nulas en su formación previa a la universitaria. En la segunda fase se realizaron actividades que buscaron el fortalecimiento del Laboratorio de Biología del CVT, destacándose la realización del Manual de Bioseguridad del Laboratorio de Biología y el apoyo al trabajo de la Ludoteca UPN-CVT.La tercera y cuarta fase consistieron en la implementación y análisis de los tres tipos de prácticas de laboratorio (virtuales, convencionales y artesanales), que se configuraron como una tripleta funcional, donde sus ventajas se complementan y hacen significativo el proceso de aprendizaje de temáticas en torno a la Biología Molecular en contextos rurales.
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