Proteomics reveals commitment to germination in barley seeds is marked by loss of stress response proteins and mobilisation of nutrient reservoirs.

Germination is a critical process in the reproduction and propagation of flowering plants, and is also the key stage of industrial grain malting. Germination commences when seeds are steeped in water, followed by degradation of the endosperm cell walls, enzymatic digestion of starch and proteins to provide nutrients for the growing plant, and emergence of the radicle from the seed. Dormancy is a state where seeds fail to germinate upon steeping, but which prevents inappropriate premature germination of the seeds before harvest from the field. This can result in inefficiencies in industrial malting. We used Sequential Window Acquisition of all THeoretical ions Mass Spectrometry (SWATH-MS) proteomics to measure changes in the barley seed proteome throughout germination. We found a large number of proteins involved in desiccation tolerance and germination inhibition rapidly decreased in abundance after imbibition. This was followed by a decrease in proteins involved in lipid, protein and nutrient reservoir storage, consistent with induction and activation of systems for nutrient mobilisation to provide nutrients to the growing embryo. Dormant seeds that failed to germinate showed substantial biochemical activity distinct from that of seeds undergoing germination, with differences in sulfur metabolic enzymes, endogenous alpha-amylase/trypsin inhibitors, and histone proteins. We verified our findings with analysis of germinating barley seeds from two commercial malting facilities, demonstrating that key features of the dynamic proteome of germinating barley seeds were conserved between laboratory and industrial scales. The results provide a more detailed understanding of the changes in the barley proteome during germination and give possible target proteins for testing or to inform selective breeding to enhance germination or control dormancy. SIGNIFICANCE: Germination is critical to the reproduction and propagation of flowering plants, and in industrial malting. Dormancy, where seeds fail to germinate upon steeping, can result in inefficiencies in industrial malting. Our DIA/SWATH-MS proteomics analyses identified key changes during germination, including an initial loss of proteins involved in desiccation tolerance and germination inhibition, followed by decreases in lipid, protein and nutrient reservoir storage. These changes were consistent between laboratory and industrial malting scales, and therefore demonstrate the utility of laboratory-scale barley germination as a model system for industrial malt house processes. We also showed that dormant seeds that failed to germinate showed substantial biochemical activity distinct from that of seeds undergoing germination, consistent with dormancy being an actively regulated state. Our results provide a more detailed understanding of the changes in the barley proteome during germination and give possible target proteins for testing or to inform selective breeding to enhance germination or control dormancy.

Water droplet surface tension method - An innovation in quantifying saponin content in quinoa seed.

Quinoa surface borne saponins are bitter tasting anti-nutritional compounds that must be removed before consumption of the seed. To determine saponin content, the currently available standard afrosimetric foam test method only determines the presence of saponin via a rating of either 'acceptable' or 'unacceptable'. A water droplet surface tension (WDST) based innovative method was developed that can quantify saponin content in aqueous solutions with greater accuracy. The method comprised four steps: solution preparation, droplet creation, image capture and image analysis using Axisymmetric Drop Shape Analysis computer software. The method applied satisfactorily to saponin content up to 0.2 mg.ml-1 as higher concentrations did not further reduce the surface tension. Results demonstrated that saponin concentration may be measured in the range 0.05 to 0.15 mg.ml-1 (0.05 - 0.15% saponin by weight of seed). Validation of the WDST method on commercial and experimental samples offers quinoa processors an accurate inexpensive way of measuring saponin concentration to satisfy current seed quality specifications.

Acute effect of sorghum flour-containing pasta on plasma total polyphenols, antioxidant capacity and oxidative stress markers in healthy subjects: A randomised controlled trial.

BACKGROUND & AIMS: It has been previously reported that pasta containing wholegrain sorghum flour exhibits high content of polyphenols and antioxidant capacity and hence might enhance antioxidant status and reduce markers of oxidative stress in vivo; however no clinical studies have yet been reported. Therefore, the present study assessed the effect of pasta containing red or white wholegrain sorghum flour on plasma total polyphenols, antioxidant capacity and oxidative stress markers in humans. The study was registered with the Australian New Zealand Clinical Trials Registry (ACTRN: 12612000324819). METHODS: In a randomised crossover design, healthy subjects (n = 20) consumed three test meals of control pasta (CP), 30% red sorghum pasta (RSP) or 30% white sorghum pasta (WSP), 1-2 wk apart. The test meals were consumed as breakfast after an overnight fast. Blood samples were obtained at fasting and 2 h after consumption and analysed for total polyphenols, antioxidant capacity, superoxide dismutase (SOD) activity, protein carbonyl and 8-isoprostanes. RESULTS: Compared to baseline, the 2 h post-prandial levels following the RSP meal of plasma polyphenols, antioxidant capacity and SOD activity were significantly (P < 0.001) higher while the protein carbonyl level was significantly lower (P = 0.035). Furthermore, net changes in polyphenols, antioxidant capacity and SOD activity were significantly (P < 0.001) higher while protein carbonyl were significantly (P = 0.035) lower following consumption of the RSP meal than the CP meal. CONCLUSION: The results demonstrated that pasta containing red wholegrain sorghum flour enhanced antioxidant status and improved markers of oxidative stress in healthy subjects.

Effect of sorghum flour addition on in vitro starch digestibility, cooking quality, and consumer acceptability of durum wheat pasta.

Whole grain sorghum is a valuable source of resistant starch and polyphenolic antioxidants and its addition into staple food like pasta may reduce the starch digestibility. However, incorporating nondurum wheat materials into pasta provides a challenge in terms of maintaining cooking quality and consumer acceptability. Pasta was prepared from 100% durum wheat semolina (DWS) as control or by replacing DWS with either wholegrain red sorghum flour (RSF) or white sorghum flour (WSF) each at 20%, 30%, and 40% incorporation levels, following a laboratory-scale procedure. Pasta samples were evaluated for proximate composition, in vitro starch digestibility, cooking quality, and consumer acceptability. The addition of both RSF and WSF lowered the extent of in vitro starch digestion at all substitution levels compared to the control pasta. The rapidly digestible starch was lowered in all the sorghum-containing pastas compared to the control pasta. Neither RSF or WSF addition affected the pasta quality attributes (water absorption, swelling index, dry matter, adhesiveness, cohesiveness, and springiness), except color and hardness which were negatively affected. Consumer sensory results indicated that pasta samples containing 20% and 30% RSF or WSF had acceptable palatability based on meeting one or both of the preset acceptability criteria. It is concluded that the addition of wholegrain sorghum flour to pasta at 30% incorporation level is possible to reduce starch digestibility, while maintaining adequate cooking quality and consumer acceptability.