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Engineering Curcumin Biosynthesis in Poplar Affects Lignification and Biomass Yield.

ABSTRACT: Lignocellulosic biomass is recalcitrant toward deconstruction into simple sugars mainly due to the presence of lignin. By engineering plants to partially replace traditional lignin monomers with alternative ones, lignin degradability and extractability can be enhanced. Previously, the alternative monomer curcumin has been successfully produced and incorporated into lignified cell walls of Arabidopsis by the heterologous expression of DIKETIDE-CoA SYNTHASE (DCS) and CURCUMIN SYNTHASE2 (CURS2). The resulting transgenic plants did not suffer from yield penalties and had an increased saccharification yield after alkaline pretreatment. Here, we translated this strategy into the bio-energy crop poplar. Via the heterologous expression of DCS and CURS2 under the control of the secondary cell wall CELLULOSE SYNTHASE A8-B promoter (ProCesA8-B), curcumin was also produced and incorporated into the lignified cell walls of poplar. ProCesA8-B:DCS_CURS2 transgenic poplars, however, suffered from shoot-tip necrosis and yield penalties. Compared to that of the wild-type (WT), the wood of transgenic poplars had 21% less cellulose, 28% more matrix polysaccharides, 23% more lignin and a significantly altered lignin composition. More specifically, ProCesA8-B:DCS_CURS2 lignin had a reduced syringyl/guaiacyl unit (S/G) ratio, an increased frequency of p-hydroxyphenyl (H) units, a decreased frequency of p-hydroxybenzoates and a higher fraction of phenylcoumaran units. Without, or with alkaline or hot water pretreatment, the saccharification efficiency of the transgenic lines was equal to that of the WT. These differences in (growth) phenotype illustrate that translational research in crops is essential to assess the value of an engineering strategy for applications. Further fine-tuning of this research strategy (e.g., by using more specific promoters or by translating this strategy to other crops such as maize) might lead to transgenic bio-energy crops with cell walls more amenable to deconstruction without settling in yield.

SUBMITTER: De Meester B

PROVIDER: S-EPMC9289561 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Quinoa high nutritive value increases interest worldwide, especially as a crop that could potentially feature in different cropping systems, however, climate change, particularly rising temperatures, challenges this and other crop species. Currently, only limited knowledge exists regarding the grain yield and other key traits response to higher temperatures of this crop, especially to increased night temperatures. In this context, the main objective of this study was to evaluate the effect of increased night temperature on quinoa yield, grain number, individual grain weight and processes involved in crop growth under the environmental conditions (control treatment) and night thermal increase at two phases: flowering (T1) and grain filling (T2) in southern Chile. A commercial genotype, Regalona, and a quinoa accession (Cod. BO5, N°191, grain bank from Semillas Baer, hereby referred to as Accession) were used, due to their adaptability to Southern Chilean conditions and contrasting grain yield potential, grain weight and size of plants. Temperature was increased ≈4°C above the ambient from 8 pm until 9 am the next morning. Control treatments reached a high grain yield (600 and 397 g m-2, i.e., Regalona and Accession). Temperature increase reduced grain yield by 31% under T1 treatment and 12% when under T2 in Regalona and 23 and 26% in Accession, respectively. Aboveground biomass was negatively affected by the thermal treatments and a positive linear association was found between grain yield and aboveground biomass across treatments. By contrast, the harvest index was unaffected either by genotype, or by thermal treatments. Grain number was significantly affected between treatments and this key trait was linearly associated with grain yield. On the other hand, grain weight showed a narrow range of variation across treatments. Additionally, leaf area index was not affected, but significant differences were found in SPAD values at the end of T1 treatment, compared to control. Little change was found in the harvest index, individual grain weight, grain protein content or water soluble carbohydrates in response to the increased night temperature in this crop.

Project description:BackgroundAs a leading biomass feedstock, poplar plants provide enormous lignocellulose resource convertible for biofuels and bio-chemicals. However, lignocellulose recalcitrance particularly in wood plants, basically causes a costly bioethanol production unacceptable for commercial marketing with potential secondary pollution to the environment. Therefore, it becomes important to reduce lignocellulose recalcitrance by genetic modification of plant cell walls, and meanwhile to establish advanced biomass process technology in woody plants. Brassinosteroids, plant-specific steroid hormones, are considered to participate in plant growth and development for biomass production, but little has been reported about brassinosteroids roles in plant cell wall assembly and modification. In this study, we generated transgenic poplar plant that overexpressed DEETIOLATED2 gene for brassinosteroids overproduction. We then detected cell wall feature alteration and examined biomass enzymatic saccharification for bioethanol production under various chemical pretreatments.ResultsCompared with wild type, the PtoDET2 overexpressed transgenic plants contained much higher brassinosteroids levels. The transgenic poplar also exhibited significantly enhanced plant growth rate and biomass yield by increasing xylem development and cell wall polymer deposition. Meanwhile, the transgenic plants showed significantly improved lignocellulose features such as reduced cellulose crystalline index and degree of polymerization values and decreased hemicellulose xylose/arabinose ratio for raised biomass porosity and accessibility, which led to integrated enhancement on biomass enzymatic saccharification and bioethanol yield under various chemical pretreatments. In contrast, the CRISPR/Cas9-generated mutation of PtoDET2 showed significantly lower brassinosteroids level for reduced biomass saccharification and bioethanol yield, compared to the wild type. Notably, the optimal green-like pretreatment could even achieve the highest bioethanol yield by effective lignin extraction in the transgenic plant. Hence, this study proposed a mechanistic model elucidating how brassinosteroid regulates cell wall modification for reduced lignocellulose recalcitrance and increased biomass porosity and accessibility for high bioethanol production.ConclusionsThis study has demonstrated a powerful strategy to enhance cellulosic bioethanol production by regulating brassinosteroid biosynthesis for reducing lignocellulose recalcitrance in the transgenic poplar plants. It has also provided a green-like process for biomass pretreatment and enzymatic saccharification in poplar and beyond.

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Engineering Curcumin Biosynthesis in Poplar Affects Lignification and Biomass Yield.

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