Seed coats of six pea genotypes contrasting in dormancy were studied by laser desorption/ionization mass spectrometry (LDI-MS). Multivariate statistical analysis discriminated dormant and non-dormant seeds in mature dry state. Separation between dormant and non-dormant types was observed despite important markers of particular dormant genotypes differ from each other. Normalized signals of long-chain hydroxylated fatty acids (HLFA) in dormant JI64 genotype seed coats were significantly higher than in other genotypes. These compounds seem to be important markers likely influencing JI64 seed imbibition and germination. HLFA importance was supported by study of recombinant inbred lines (JI64xJI92) contrasting in dormancy but similar in other seed properties. Furthemore HLFA distribution in seed coat was studied by mass spectrometry imaging. HLFA contents in strophiole and hilum are significantly lower compared to other parts indicating their role in water uptake. Results from LDI-MS experiments are useful in understanding (physical) dormancy (first phases of germination) mechanism and properties related to food processing technologies (e.g., seed treatment by cooking).
- MeSH
- Mass Spectrometry MeSH
- Pisum sativum metabolism physiology MeSH
- Fatty Acids analysis MeSH
- Seeds metabolism physiology MeSH
- Plant Dormancy * MeSH
- Publication type
- Journal Article MeSH
MAIN CONCLUSION: Seed-processing technologies such as polishing and washing enhance crop seed quality by limited removal of the outer layers and by leaching. Combined, this removes chemical compounds that inhibit germination. Industrial processing to deliver high-quality commercial seed includes removing chemical inhibitors of germination, and is essential to produce fresh sprouts, achieve vigorous crop establishment, and high yield potential in the field. Sugar beet (Beta vulgaris subsp. vulgaris var. altissima Doell.), the main sugar source of the temperate agricultural zone, routinely undergoes several processing steps during seed production to improve germination performance and seedling growth. Germination assays and seedling phenotyping was carried out on unprocessed, and processed (polished and washed) sugar beet fruits. Pericarp-derived solutes, known to inhibit germination, were tested in germination assays and their osmolality and conductivity assessed (ions). Abscisic acid (ABA) and ABA metabolites were quantified in both the true seed and pericarp tissue using UPLC-ESI(+)-MS/MS. Physical changes in the pericarp structures were assessed using scanning electron microscopy (SEM). We found that polishing and washing of the sugar beet fruits both had a positive effect on germination performance and seedling phenotype, and when combined, this positive effect was stronger. The mechanical action of polishing removed the outer pericarp (fruit coat) tissue (parenchyma), leaving the inner tissue (sclerenchyma) unaltered, as revealed by SEM. Polishing as well as washing removed germination inhibitors from the pericarp, specifically, ABA, ABA metabolites, and ions. Understanding the biochemistry underpinning the effectiveness of these processing treatments is key to driving further innovations in commercial seed quality.
Persistent seed banks are predicted to have an important impact on population genetic processes by increasing effective population size and storing past genetic diversity. Accordingly, persistent seed banks may buffer genetic effects of disturbance, fragmentation and/or selection. However, empirical studies surveying the relationship between aboveground and seed bank genetics under changing environments are scarce. Here, we compared genetic variation of aboveground and seed bank cohorts in 15 populations of the partially cleistogamous Viola elatior in two contrasting early and late successional habitats characterized by strong differences in light-availability and declining population size. Using AFLP markers, we found significantly higher aboveground than seed bank genetic diversity in early successional meadow but not in late successional woodland habitats. Moreover, individually, three of eight woodland populations even showed higher seed bank than aboveground diversity. Genetic differentiation among populations was very strong (фST = 0.8), but overall no significant differentiation could be detected between above ground and seed bank cohorts. Small scale spatial genetic structure was generally pronounced but was much stronger in meadow (Sp-statistic: aboveground: 0.60, seed bank: 0.32) than in woodland habitats (aboveground: 0.11; seed bank: 0.03). Our findings indicate that relative seed bank diversity (i.e. compared to aboveground diversity) increases with ongoing succession and despite decreasing population size. As corroborated by markedly lower small-scale genetic structure in late successional habitats, we suggest that the observed changes in relative seed bank diversity are driven by an increase of outcrossing rates. Persistent seed banks in Viola elatior hence will counteract effects of drift and selection, and assure a higher chance for the species' long term persistence, particularly maintaining genetic variation in declining populations of late successional habitats and thus enhancing success rates of population recovery after disturbance events.
Seed development in flowering plants is a critical part of plant life for successful reproduction. The formation of viable seeds requires the synchronous growth and development of the fruit and the three seed structures: the embryo, the endosperm, the seed coat. Molecular communication between these tissues is crucial to coordinate these developmental processes. The phytohormone auxin is a significant player in embryo, seed and fruit development. Its regulated local biosynthesis and its cell-to-cell transport capacity make of auxin the perfect candidate as a signaling molecule to coordinate the growth and development of the embryo, endosperm, seed and fruit. Moreover, newly formed seeds need nutrients and form new carbon sink, generating high sugar flow from vegetative tissues to the seeds. This review will discuss how auxin and sugars may be considered as signaling molecules to coordinate seed and fruit development.
Metal accumulation in seeds is a prerequisite for germination and establishment of plants but also for micronutrient delivery to humans. To investigate metal transport processes and their interactions in seeds, we focused on METAL TOLERANCE PROTEIN8 (MTP8), a tonoplast transporter of the manganese (Mn) subclade of cation diffusion facilitators, which in Arabidopsis (Arabidopsis thaliana) is expressed in embryos of seeds. The x-ray fluorescence imaging showed that expression of MTP8 was responsible for Mn localization in subepidermal cells on the abaxial side of the cotyledons and in cortical cells of the hypocotyl. Accordingly, under low Mn availability, MTP8 increased seed stores of Mn, required for efficient seed germination. In mutant embryos lacking expression ofVACUOLAR IRON TRANSPORTER1(VIT1), MTP8 built up iron (Fe) hotspots inMTP8-expressing cells types, suggesting that MTP8 transports Fe in addition to Mn. Inmtp8 vit1double mutant seeds, Mn and Fe were distributed in all cell types of the embryo. An Fe transport function of MTP8 was confirmed by its ability to complement Fe hypersensitivity of a yeast mutant defective in vacuolar Fe transport. Imbibingmtp8-1mutant seeds in the presence of Mn or subjecting seeds to wet-dry cycles showed that MTP8 conferred Mn tolerance. During germination, MTP8 promoted reallocation of Fe from the vasculature. These results indicate that cell type-specific accumulation of Mn and Fe in seeds depends on MTP8 and that this transporter plays an important role in the generation of seed metal stores as well as for metal homeostasis and germination efficiency under challenging environmental conditions.
- MeSH
- Arabidopsis embryology genetics metabolism MeSH
- Models, Biological MeSH
- Gene Knockout Techniques MeSH
- Homeostasis * MeSH
- Germination * genetics MeSH
- Manganese metabolism MeSH
- Mutation genetics MeSH
- Promoter Regions, Genetic genetics MeSH
- Arabidopsis Proteins metabolism MeSH
- Cation Transport Proteins metabolism MeSH
- Gene Expression Regulation, Plant MeSH
- Saccharomyces cerevisiae metabolism MeSH
- Seeds embryology genetics MeSH
- Spectrometry, X-Ray Emission MeSH
- Genetic Complementation Test MeSH
- Gene Expression Regulation, Developmental MeSH
- Iron metabolism MeSH
- Publication type
- Journal Article MeSH
The maintenance of genome integrity over cell divisions is critical for plant development and the correct transmission of genetic information to the progeny. A key factor involved in this process is the STRUCTURAL MAINTENANCE OF CHROMOSOME5 (SMC5) and SMC6 (SMC5/6) complex, related to the cohesin and condensin complexes that control sister chromatid alignment and chromosome condensation, respectively. Here, we characterize NON-SMC ELEMENT4 (NSE4) paralogs of the SMC5/6 complex in Arabidopsis (Arabidopsis thaliana). NSE4A is expressed in meristems and accumulates during DNA damage repair. Partial loss-of-function nse4a mutants are viable but hypersensitive to DNA damage induced by zebularine. In addition, nse4a mutants produce abnormal seeds, with noncellularized endosperm and embryos that maximally develop to the heart or torpedo stage. This phenotype resembles the defects in cohesin and condensin mutants and suggests a role for all three SMC complexes in differentiation during seed development. By contrast, NSE4B is expressed in only a few cell types, and loss-of-function mutants do not have any obvious abnormal phenotype. In summary, our study shows that the NSE4A subunit of the SMC5-SMC6 complex is essential for DNA damage repair in somatic tissues and plays a role in plant reproduction.
- MeSH
- Arabidopsis embryology genetics immunology MeSH
- Gene Duplication MeSH
- Genome, Plant MeSH
- DNA Repair * genetics MeSH
- Protein Subunits metabolism MeSH
- DNA Damage * genetics MeSH
- Cell Cycle Proteins genetics metabolism MeSH
- Arabidopsis Proteins genetics metabolism MeSH
- Pollen genetics MeSH
- Gene Expression Regulation, Plant MeSH
- Seeds genetics metabolism MeSH
- Transcriptome genetics MeSH
- Up-Regulation genetics MeSH
- Ovule genetics MeSH
- Protein Binding MeSH
- Gene Expression Regulation, Developmental MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
The plastid-localized phosphoglucose isomerase isoform PGI1 is an important determinant of growth in Arabidopsis thaliana, likely due to its involvement in the biosynthesis of plastidial isoprenoid-derived hormones. Here, we investigated whether PGI1 also influences seed yields. PGI1 is strongly expressed in maturing seed embryos and vascular tissues. PGI1-null pgi1-2 plants had ∼60% lower seed yields than wild-type plants, with reduced numbers of inflorescences and thus fewer siliques and seeds per plant. These traits were associated with low bioactive gibberellin (GA) contents. Accordingly, wild-type phenotypes were restored by exogenous GA application. pgi1-2 seeds were lighter and accumulated ∼50% less fatty acids (FAs) and ∼35% less protein than wild-type seeds. Seeds of cytokinin-deficient plants overexpressing CYTOKININ OXIDASE/DEHYDROGENASE1 (35S:AtCKX1) and GA-deficient ga20ox1 ga20ox2 mutants did not accumulate low levels of FAs, and exogenous application of the cytokinin 6-benzylaminopurine and GAs did not rescue the reduced weight and FA content of pgi1-2 seeds. Seeds from reciprocal crosses between pgi1-2 and wild-type plants accumulated wild-type levels of FAs and proteins. Therefore, PGI1 is an important determinant of Arabidopsis seed yield due to its involvement in two processes: GA-mediated reproductive development and the metabolic conversion of plastidial glucose-6-phosphate to storage reserves in the embryo.
- MeSH
- Arabidopsis enzymology metabolism MeSH
- Gibberellins metabolism MeSH
- Glucose-6-Phosphate metabolism MeSH
- Glucose-6-Phosphate Isomerase genetics metabolism MeSH
- Membrane Proteins genetics metabolism MeSH
- Oxidoreductases Acting on CH-NH Group Donors genetics metabolism MeSH
- Plastids metabolism MeSH
- Arabidopsis Proteins genetics metabolism MeSH
- Gene Expression Regulation, Plant MeSH
- Seeds enzymology metabolism MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
BACKGROUND: Within last few years, the occurrence of food allergens and corresponding food allergies has been increasing, therefore research into the individual allergens is required. In the present work, the effect of cereal processing on the amounts of allergenic proteins is studied by modern proteomic-based approaches. The most important wheat and barley allergens are low-molecular-weight (LMW) proteins. Therefore we investigated the relative quantitative changes of these proteins after food technological processing, namely wheat couscous production and barley malting. RESULTS: A comparative study using mass spectrometry in connection with the technique of isobaric tag for relative and absolute quantification (iTRAQ) revealed that the amount of wheat allergenic LMW proteins decreased significantly during couscous production (approximately to 5-26% of their initial content in wheat flour). After barley malting, the amounts of the majority of LMW proteins decreased as well, although to a lesser extent than in the case of wheat/couscous. The level of two allergens even slightly increased. CONCLUSION: Suggested proteomic strategy proved as universal and sensitive method for fast and reliable identification of various cereal allergens and monitoring of their quantitative changes during food processing. Such information is important for consumers who suffer from allergies.
- MeSH
- Allergens adverse effects analysis chemistry MeSH
- Antigens, Plant adverse effects analysis chemistry MeSH
- Dietary Proteins adverse effects analysis chemistry MeSH
- Spectrometry, Mass, Electrospray Ionization MeSH
- Hordeum chemistry MeSH
- Edible Grain adverse effects chemistry MeSH
- Food Inspection methods MeSH
- Humans MeSH
- Food Handling * MeSH
- Molecular Weight MeSH
- Peptide Mapping MeSH
- Food Hypersensitivity etiology prevention & control MeSH
- Proteomics MeSH
- Triticum chemistry MeSH
- Plant Proteins adverse effects analysis chemistry MeSH
- Seedlings chemistry MeSH
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Comparative Study MeSH
- Geographicals
- Czech Republic MeSH
Using Arabidopsis thaliana wild type (WT) plants and diacylglycerol kinase knockouts (single mutants - dgk3, dgk1, dgk6; double mutants - dgk3dgk7, dgk5dgk6, dgk1dgk2) we observed that the inhibitor of brassinosteroid (BR) biosynthesis, brassinazole (BRZ), drastically decreased germination of dgk mutants under salt stress, while BRZ co-administration with 24-epibrassinolide (EBL) partially improved germination rates. We also observed a statistically significant decrease in alternative and cytochrome respiratory pathways in response to BRZ treatment under salinity conditions. We showed that production of the lipid second messenger phosphatidic acid (PA) is impaired in dgk mutants in response to EBL treatment and inhibitor of diacylglycerol kinase (DGK) - R59022. This study demonstrates that dgk mutants possess lower germination rates, lower total respiration rates, an alternative respiratory pathway and PA content under optimal and high salinity conditions in response to EBL treatment comparing to WT plants.
- MeSH
- Arabidopsis chemistry metabolism MeSH
- Brassinosteroids pharmacology MeSH
- Diacylglycerol Kinase antagonists & inhibitors deficiency metabolism MeSH
- Phosphatidic Acids chemistry metabolism MeSH
- Salinity MeSH
- Seeds drug effects growth & development metabolism MeSH
- Triazoles pharmacology MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH