bioRxiv Science⌕ Search

Biology subjects

Lepiniec, L.

Publications and source records attributed to Lepiniec, L..

5 recordsLinked to original sources

Elevated temperature drives the biosynthesis of novel acylated glucosinolates in Arabidopsis thaliana seeds

Glucosinolates (GSLs) are major defensive compounds massively accumulated in Brassicaceae seeds, including that of the model plant Arabidopsis thaliana. While most studies have focused on the role of GSL in responses to biotic stress, the potential regulation and function of GSLs in responses to abiotic stresses have been neglected, particularly in seeds. In this study, multi-omic analyses revealed a previously uncharacterized GSL modification pathway induced by elevated temperature (ET) during A. thaliana seed development. Activation of this pathway leads to the production of several novel thioglucose-acylated GSLs, including sinapoylated and benzoylated derivatives. A reverse genetics approach demonstrated that the SERINE CARBOXYPEPTIDASE-LIKE 17 (SCPL17) and BENZOYLOXYGLUCOSINOLATE 1 (BZO1) enzymes are required for the acylation of GSL thioglucose moieties. Furthermore, the accumulation of acylated GSLs in seeds of 85 A. thaliana accessions grown under standard condition was shown to correlate with the average annual temperature of their origin site, suggesting that thioglucose-acylated GSLs production may reflect long-term thermal adaptation across natural populations. Taken together, these results demonstrate that thioglucose acylation by SCPL17 and BZO1 represents a new layer of GSL diversification in A. thaliana seeds that contributes to both ET response and long-term environmental adaptation.

plant biology↗

Multi-omic analyses unveil temporal and spatial distribution of specialized metabolites in seeds of Camelina sativa

Seeds of Brassicaceae produce a large diversity of beneficial and antinutritional specialized metabolites (SMs) that influence their quality and provide resistance to stresses. While the distribution of these compounds has been described in leaves and roots tissues, limited information is available about their spatio-temporal accumulation in seeds. Camelina sativa (camelina) is an oilseed Brassicaceae cultivated for human and animal nutrition, and for industrial uses. While we previously explored SM diversity and plasticity, no information is available about SM distribution and expression of related proteins and genes in camelina seeds. In this study, we used a multi-omic approach, integrating untargeted metabolomics, data-independent acquisition proteomics, and transcriptomics to investigate the synthesis, modifications and degradations of SMs accumulated in the different seed tissues (i.e. seed coat, endosperm, and embryo) at 6 developmental and 2 germination stages. Our results showed distinct patterns of SMs and their related pathways, highlighting significant contrasts in seed composition and spatial distribution for the defence-related and antinutritional glucosinolate (GSL) compounds among camelina, Arabidopsis thaliana, and Brassica napus, three closely-related Brassicaceae species. Notably, the variation in GSL spatial distributions was primarily driven by differences in their structure and transport mechanisms. Long chain C8-C11 methylsulfinylalkyl GSLs were predominantly accumulated in the seed coat and endosperm, while mid- and short-chain C3-C7 methylsulfinylalkyl GSLs were accumulated in the embryo. Characterizing the spatial dynamics of seed SMs provides valuable insights that can guide the development of crops with optimized distribution of beneficial and toxic metabolites, improving seed nutritional profiles for feed and food.

plant biology↗

Sterol 3-beta-Glucosyltransferase TRANSPARENT TESTA15 Controls Seed Development and Flavanol Accumulation through its Role in Vacuole Biogenesis and Maintenance in Arabidopsis

The Arabidopsis sterol 3-beta-glucosyltransferase UGT80B1/TRANSPARENT TESTA15 (TT15) catalyzes sterol glucoside biosynthesis. Its loss of function causes reduced seed size, defective flavanol, polysaccharide and lipid polyester deposition at the seed coat and reduced seed dormancy. How TT15 controls seed development and physiology is unknown. Here we show that tt15 mutants exhibit seed lethality with incomplete penetrance and maternal determinism that is correlated with endosperm cellularization defects, together with an increased sensitivity of seed germination to exogenous abscisic acid and paclobutrazol. We also reveal that flavanol deposition in the vacuole during tt15 seed development triggers premature endothelium cell death. An autoimmune-like syndrome characterized by callose and H2O2 accumulation was detected in endothelium at the seed abaxial pole. Similar phenotypes were observed with tt9/gfs9, a mutant defective in endomembrane trafficking and homotypic vacuole fusion. Double mutant analysis showed that tt9 partially rescued tt15 endothelium phenotypes. Consistent with seed mutant phenotypes, TT15 promoter activity was detected in endothelium and endosperm and TT15 protein was located mainly at the vacuolar membrane (tonoplast). Using fluorescence recovery after photobleaching, we demonstrated that tonoplast fluidity was increased in tt15 roots. Altogether our data suggest that TT15 regulates seed development and flavanol accumulation by modulating vacuole biogenesis and maintenance.

plant biology↗

Untargeted metabolomic analyses reveal the diversity and plasticity of the specialized metabolome in seeds of different Camelina sativa varieties

Despite the essential role of Specialized Metabolites (SMs) in the interaction of plants with the environment, studying the ability of crop seeds to produce these protective compounds has been neglected. Furthermore, seeds produce a myriad of SMs providing an interesting model to investigate their diversity and plasticity. Camelina sativa gained a lot of interest in the past few years as rustic oil seed crop. A characterization of seed SM landscapes in six camelina genotypes grown in the field and harvested during five growing seasons has been undertaken in this work. This allowed a comprehensive annotation of seed SMs combining analyses that cluster SMs based on their chemical structures and co-accumulation patterns. These data showed broad effects of the environment on the stimulation of the seed-specialized metabolome. Among well annotated compounds, flavonols were identified as the metabolic class characterised by high plasticity, revealing significant variable accumulation according to the year and/or the genotype. Lastly, a deeper characterisation of primary metabolites and lipids in two selected genotypes has been performed. We showed that, in addition to flavonols, alkaloids and glucosinolates displayed a higher phenotypic plasticity with respect to most of the primary metabolites, including some sugars and major storage compounds such as fatty acids, proteins and most lipid classes (e.g. DAG, TAG), but similar plasticity compared to free aminoacids and carboxylic acids. This work highlighted major and unexplored effects of the environment on the seed specialized metabolome demonstrating that seeds exhibit a dynamic and plastic metabolism, with an impact on seed quality. Significance statementSeeds produce a myriad of Specialized Metabolites (SMs) with an essential role in the interaction of plants with the environment. We characterized SM landscapes, primary metabolites and lipid composition in the seeds of camelina genotypes grown in the open field in five consecutive growing seasons. Our results showed the predominant effect of the environment on the regulation of the seed - specialized metabolome, with a potential impact on seed quality of camelina that may also occur in other oilseed crops.

plant biology↗

Natural variation in expression of the HECT E3 ligase UPL3 influences seed size and crop yields in Brassica napus by altering regulatory gene expression.

Identifying genetic variation that increases crop yields is a primary objective in plant breeding. We have used association analyses of Brassica napus (oilseed rape/canola) accessions to identify variation in the expression of a HECT E3 ligase gene, BnaUPL3.C03, that influences seed size and final yield. We establish a mechanism in which UPL3 mediates the proteasomal degradation of LEC2, a master regulator of seed maturation. Reduced UPL3 expression increases LEC2 protein levels and prolongs expression of lipid biosynthetic genes and seed maturation. Natural variation in BnaUPL3.C03 expression has not yet been exploited in current Brassica napus breeding lines and can therefore be used as a new approach to maximize future yields in this important oil crop.

plant biology↗