Laura Nyström

The HEALTHGRAIN Cereal Diversity Screen : Concept, Results, and Prospects

One hundred and fifty bread wheat lines and 50 other lines of small-grain cereals (spelt, durum wheat, Triticum monococcum, Triticum dicoccum, oats, rye, and barley) were selected for diversity in their geographical origin, age, and characteristics. They were grown on a single site in Hungary in 2004-2005, harvested, milled, and analyzed for a range of phytochemicals (tocols, sterols, phenolic acids, folates, alkylresorcinols) and fiber components that are considered to have health benefits. Detailed analyses of these components in the different species are reported in a series of accompanying papers. The present paper discusses the comparative levels of the bioactive components in the different species, showing differences in both ranges and mean amounts. Furthermore, detailed comparisons of the bread wheat lines show that it is possible to identify lines in which high levels of phytochemicals and dietary fiber components are combined with good yield and processing quality. This means that commercially competitive lines with high levels of bioactive components are a realistic goal for plant breeders.

Phytochemicals and dietary fiber components in rye varieties in the HEALTHGRAIN diversity screen

Ten rye varieties grown in one location were analyzed for their contents of dietary fiber (arabinoxylan and beta-glucan) and phytochemicals (folate, tocols, phenolic acids, alkylresorcinols, and sterols). The varieties included old and modern varieties from five European countries. Significant differences were observed in the contents of all phytochemicals in whole grains and in the fiber contents in the flour and bran. The old French varieties Haute Loire and Queyras had high contents of most phytochemicals, whereas the Polish varieties Dankowskie-Zlote and Warko were relatively poor in phytochemicals. The varieties with a high content of folate tended to have low alkylresorcinol contents and vice versa. Furthermore, high contents of arabinoxylans were associated with high contents in tocols and sterols. The 10 selected rye samples comprising old populations and old and modern varieties from different ecological regions of Europe demonstrate high natural variation in their composition and show that landraces and old populations are useful genetic resources for plant breeding. The contents of single phytochemicals can likely be affected by breeding, and they may be adjusted by the right selection of genotype.

Phytochemical and fiber components in oat varieties in the healthgrain diversity screen

The levels and compositions of a range of phytochemicals (sterols, tocols, avenanthramides, folates, phenolic acids) and dietary fiber components were determined in five oat cultivars (four husked and one naked) grown on a single site in 2005. The total levels of tocols, phenolic acids, and avenanthramides varied by over 2-fold between cultivars, but less variation occurred in total sterols and total folates. Limited variation was also observed in the dietary fiber content and composition of the four husked lines. These results indicate that it may be possible to selectively breed for lines with high contents of dietary fiber and specific groups of phytochemicals.

Phytosterols in Wheat Genotypes in the HEALTHGRAIN Diversity Screen

The phytosterol contents of 130 winter wheat, 20 spring wheat, 10 durum wheat, 5 spelt, 5 einkorn, and 5 emmer wheat genotypes, grown at the same location in the same year, were analyzed with gas chromatography. Considerable variation was observed in total phytosterol contents in all wheat types. The total sterol contents ranged from 670 to 959 microg/g of dm in winter wheat and from 797 to 949 microg/g of dm in spring wheat. The highest sterol contents were found in spelt, durum wheat, and einkorn wheat. The proportions of the main phytosterols also varied substantially among the different genotypes. The most abundant phytosterol in all wheat genotypes was sitosterol (40-61% of total sterols), whereas the highest variation was seen in total stanols (7-31% of total sterols). The comprehensive data set produced in this study constitutes a valuable basis for plant breeding and selection of phytosterol-rich genotypes.

Contents of dietary fibre components and their relation to associated bioactive components in whole grain wheat samples from the HEALTHGRAIN diversity screen

A large and diverse material collection of whole grain wheat samples (n=129) was analysed for total dietary fibre (TDF) content and composition, including fructan (11.5-15.5%). Correlations between the dietary fibre components, associated bioactive components (e.g. tocols, sterols, phenolic acids and folates) and agronomic properties previously determined on the same samples were found with multivariate analysis (PCA). Samples from the same countries had similar characteristics. The first PC described variation in components concentrated in the starchy endosperm (e.g. starch, β-glucan and fructan) and the dietary fibre components concentrated in the bran (e.g. TDF, arabinoxylan and cellulose). The second PC described the variation in kernel weight and other bran components such as alkylresorcinols, tocols and sterols. Interestingly, there was no correlation among these different groups of bran components, which reflected their concentration in different bran tissues. The results are of importance for plant breeders who wish to develop varieties with health-promoting effects.

The Effect of In Vitro Digestion on Steryl Ferulates from Rice (Oryza sativa L.) and Other Grains

Polished and cargo rice, wild rice, rice bran, corn bran, and wheat bran were subjected to a static in vitro digestion model, to monitor changes in their steryl ferulate content and composition. Free sterols, possible hydrolysis products of steryl ferulates, were also measured. Additionally, steryl ferulate bioaccessibility was calculated as the percentage of steryl ferulates liberated from the grain matrix into the digestive juice. Steryl ferulate content ranged between 6.1 and 3900 μg/g and decreased by 1-63% due to digestion. A parallel increase in free sterols of more than 70% was observed in all samples. Additionally, bioaccessibility of steryl ferulates was found to be almost negligible. These findings suggest that intestinal enzymes immediately hydrolyze steryl ferulates, which are liberated from the grain matrix, and thus they are practically unavailable for absorption in the small intestine. This further indicates that the hydrolysis products of steryl ferulates could be bioactive in the gut.

Effects of genotype and environment on steryl ferulates in wheat and rye in the HEALTHGRAIN diversity screen.

The effects of genetic and environmental factors on the content and composition of steryl ferulates in wheat and rye were studied. The wheat and rye genotypes were grown at the same location in Hungary over three consecutive years (28 genotypes) or at four different locations across Europe during a single year (12 genotypes). The steryl ferulates were analyzed using HPLC. The genotype and growing location significantly affected the content and composition of wheat steryl ferulates, whereas the year of growth did not result in considerable variation. Less variation was observed in rye, due to fewer genotypes. Campestanyl and sitostanyl ferulates were the main species in both cereals. Knowledge of the natural variation in steryl ferulates and other bioactive compounds allows cultivators and plant breeders to select genotypes with high, stable levels of beneficial compounds. Thus, it is possible to enhance the intake of health-promoting compounds from natural sources.

Reactive oxygen species responsible for beta-glucan degradation

The presence of iron(II) in beta-glucan in solution causes the formation of hydroxyl radical, which further oxidises the polysaccharide. This degradation can be enhanced by the presence of a reducing agent, such as ascorbic acid. In this study we investigated the effect the iron(II) concentration on the hydroxyl radical-mediated degradation of beta-glucan and identified the intermediate species involved in the formation of hydroxyl radicals. An increase in the iron(II) concentration did not have a significant effect on the degradation in the presence of a reducing agent (ascorbic acid), while in the mere presence of iron(II) it accelerates the degradation. The addition of catalase and superoxide dismutase (SOD) prevented the hydroxyl radical driven-degradation of beta-glucan induced by iron(II) or ascorbic acid/iron(II), demonstrating the involvement of both superoxide and hydrogen peroxide in the hydroxyl radical formation. SOD, which catalyses the dismutation of superoxide into hydrogen peroxide, should have stimulated the formation of radicals, since these radicals are generated from the reaction between hydrogen peroxide and iron(II). In the present study, we hypothesise the mechanism of the inhibition of beta-glucan degradation by superoxide dismutase.

In situ enrichment of folate by microorganisms in beta-glucan rich oat and barley matrices.

The objective was to study folate production of yeast strains, bacteria isolated from oat bran, and selected lactic acid bacteria as well as one propionibacterium in oat and barley based models. Simultaneously, we aimed at sustaining the stability of viscosity, representing the physicochemical state of beta-glucan. Total folate contents were determined microbiologically and vitamers for selected samples by UHPLC. Folate in yeast cells comprised mainly 5-methyltetrahydrofolate and tetrahydrofolate. Folate production by microbes in YPD medium was different to that in cereal fermentations where vitamers included 5-methyltetrahydrofolate, 5,10-methenyltetrahydrofolate and formylated derivatives. Microbes producing significant amounts of folate without affecting viscosity were Saccharomyces cerevisiae ALKO743 and Candida milleri ABM4949 among yeasts and Pseudomonas sp. ON8 and Janthinobacterium sp. RB4 among bacteria. Net folate production was up to 120 ng/g after 24 h fermentation and could increase during 2-week storage. Glucose addition increased the proportion of 5-methyltetrahydrofolate. Streptococcus thermophilus ABM5097, Lactobacillus reuteri, and Propionibacterium sp. ABM5378 produced folate but in lower concentrations. Both endogenous and added microbes contribute to folate enhancement. Selection of microbes with folate producing capability and limited hydrolytic activity will enable the development of products rich in folate and beta-glucan.

Effects of Environment and Genotype on Phytosterols in Wheat in the HEALTHGRAIN Diversity Screen

The effects of environment on the content and composition of phytosterols were examined in 26 wheat genotypes grown at four locations in Europe during a single year and at one location over three consecutive years. Total phytosterol contents varied among the locations, whereas no effect was found for the harvesting year. A significant genetic variation was observed in total sterol contents (700-928 microg/g of dm). The genotype and environment resulted in statistically significant differences in the proportions of the main phytosterols. The high phytosterol contents were characterized by low proportions of sitosterol and high proportions of stanols. Small wheat kernels with proportionally high bran yield and lipid content contained higher levels of phytosterols than large kernels. Knowledge of the level and variability of phytosterols in wheat enables the selection of genotypes with high and stabile phytosterol contents for cultivation or plant breeding purposes.