High fructan barley lines produced by selective breeding may alter ß-glucan and amylopectin molecular structure.

Six cross-bred barley lines developed by a breeding strategy with the target to enhance the fructan synthesis activity and reduce the fructan hydrolysis activity were analyzed together with their parental lines, and a reference line (Gustav) to determine whether the breeding strategy also affected the content and molecular structure of amylopectin and ß-glucan. The highest fructan and ß-glucan content achieved in the novel barley lines was 8.6 % and 12 %, respectively (12.3-fold and 3.2-fold higher than in Gustav). The lines with low fructan synthesis activity had higher starch content, smaller building blocks in amylopectin, and smaller structural units of ß-glucans than the lines with high-fructan synthesis activity. Correlation analysis confirmed that low starch content was associated with high amylose, fructan, and ß-glucan content, and larger building blocks in amylopectin.

Achieving of high-diet-fiber barley via managing fructan hydrolysis.

High fructan content in the grain of cereals is an important trait in agriculture such as environmental resilience and dietary fiber food production. To understand the mechanism in determining final grain fructan content and achieve high fructan cereal, a cross breeding strategy based on fructan synthesis and hydrolysis activities was set up and have achieved barley lines with 11.8% storage fructan in the harvested grain. Our study discovered that high activity of fructan hydrolysis at later grain developmental stage leads to the low fructan content in mature seeds, simultaneously increasing fructan synthesis at early stage and decreasing fructan hydrolysis at later stage through crossing breeding is an efficient way to elevate grain diet-fiber content. A good correlation between fructan and beta glucans was also discovered with obvious interest. Field trials showed that the achieved high fructan barley produced over seven folds higher fructan content than control barley and pull carbon-flux to fructan through decreasing fructan hydrolysis without disruption starch synthesis will probably not bring yield deficiency.

Comparative compositions of metabolites and dietary fibre components in doughs and breads produced from bread wheat, emmer and spelt and using yeast and sourdough processes.

Wholemeal flours from blends of bread wheat, emmer and spelt were processed into bread using yeast-based and sourdough fermentation. The bread wheat flour contained significantly higher concentrations of total dietary fibre and fructans than the spelt and emmer flours, the latter having the lowest contents. Breadmaking using sourdough and yeast systems resulted in changes in composition from flour to dough to bread including increases in organic acids and mannitol in the sourdough system and increases in amino acids and sugars (released by hydrolysis of proteins and starch, respectively) in both processing systems. The concentrations of fructans and raffinose (the major endogenous FODMAPs) were reduced by yeast and sourdough fermentation, with yeast having the greater effect. Both systems resulted in greater increases in sugars and glycerol in emmer than in bread wheat and spelt, but the significance of these differences for human health has not been established.

Dietary fiber components, microstructure, and texture of date fruits (Phoenix dactylifera, L.).

Date fruits vary widely in the hardness of their edible parts and they are classified accordingly into soft, semi-dry, and dry varieties. Fruit texture, a significant parameter in determining consumer acceptance, is related to the tissue structure and chemical composition of the fruit, mainly the ratio of sucrose to reducing sugars. This study aimed to understand the relationship between the chemical composition, microstructure, and texture profile of 10 major Emirati date fruits. The soluble sugars, glucose and fructose, represent ca 80 g/100 g of the fruits on the basis of dry weight (DW) while the dietary fiber contents varied 5.2-7.4 g/100 dg D.W. with lignin being the main determinant of the variability. The textures of the samples were studied using instrumental texture profile analysis. While no correlation was found between the soluble sugar and texture parameters in this study, the different fiber constituents correlated variably with the different parameters of date fruit texture. Lignin, arabinoxylan, galactomannan, and pectin were found to correlate significantly with fruit hardness and the related parameters, gumminess and chewiness. Both lignin and arabinoxylan correlated with resilience, and arabinoxylan exhibited a strong correlation with cohesiveness.

Effect of Different Extrusion Parameters on Dietary Fiber in Wheat Bran and Rye Bran.

Wheat bran and rye bran are mostly used as animal feed today, but their high content of dietary fiber and bioactive components are beneficial to human health. Increased use of bran as food raw material could therefore be desirable. However, bran mainly contains unextractable dietary fiber and deteriorates the sensory properties of products. Processing by extrusion could increase the extractability of dietary fiber and increase the sensory qualities of bran products. Wheat bran and rye bran were therefore extruded at different levels of moisture content, screw speed and temperature, in order to find the optimal setting for increased extractability of dietary fiber and positive sensory properties. A water content of 24% for wheat bran and 30% for rye bran, a screw speed of 400 rpm, and a temperature of 130 °C resulted in the highest extractability of total dietary fiber and arabinoxylan. Arabinoxylan extractability increased from 5.8% in wheat bran to 9.0% in extruded wheat bran at those settings, and from 14.6% to 19.2% for rye bran. Total contents of dietary fiber and arabinoxylan were not affected by extrusion. Content of ß-glucan was also maintained during extrusion, while its molecular weight decreased slightly and extractability increased slightly. Extrusion at these settings is therefore a suitable process for increasing the use of wheat bran and rye bran as a food raw material.

Natural variation in grain composition of wheat and related cereals.

The wheat grain comprises three groups of major components, starch, protein, and cell wall polysaccharides (dietary fiber), and a range of minor components that may confer benefits to human health. Detailed analyses of dietary fiber and other bioactive components were carried out under the EU FP6 HEALTHGRAIN program on 150 bread wheat lines grown on a single site, 50 lines of other wheat species and other cereals grown on the same site, and 23-26 bread wheat lines grown in six environments. Principal component analysis allowed the 150 bread wheat lines to be classified on the basis of differences in their contents of bioactive components and wheat species (bread, durum, spelt, emmer, and einkorn wheats) to be clearly separated from related cereals (barley, rye, and oats). Such multivariate analyses could be used to define substantial equivalence when novel (including transgenic) cereals are considered.

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, ß-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.

Relationship between the contents of bioactive components in grain and the release dates of wheat lines in the HEALTHGRAIN diversity screen.

The EU FP6 HEALTHGRAIN has generated an extensive database on the contents of phytochemicals (alkylresorcinols, tocols, sterols, phenolic acids, folates) and dietary fiber components in the grain of wheat, including analyses of 150 lines grown on a single site in Hungary in 2005 and a smaller set of lines grown under five (three lines) or six (23 lines) different environments (in Hungary in 2005, 2006, and 2007 and in France, Poland, and the United Kingdom in 2007). The lines analyzed included land races and varieties bred between the mid-19th and early 21st centuries. These results have been analyzed to determine whether the contents of these groups of bioactive components in the grain have decreased with the development of intensive plant breeding in the second part of the 20th century. No decreases in the contents of any groups of bioactive components were observed in relation to release date, showing that selection for increased yield and protein quality has been effectively neutral for other grain components.

The HEALTHGRAIN wheat diversity screen: effects of genotype and environment on phytochemicals and dietary fiber components.

Analysis of the contents of bioactive components (tocols, sterols, alkylresorcinols, folates, phenolic acids, and fiber components) in 26 wheat cultivars grown in six site x year combinations showed that the extent of variation due to variety and environment differed significantly between components. The total contents of tocols, sterols, and arabinoxylan fiber were highly heritable and hence an appropriate target for plant breeding. However, significant correlations between the contents of bioactive components and environmental factors (precipitation and temperature) during grain development also occurred, with even highly heritable components differing in amount between grain samples grown in different years on different sites.

Effects of genotype and environment on the content and composition of phytochemicals and dietary fiber components in rye in the HEALTHGRAIN diversity screen.

The effects of genotype and environment on the content of bioactive components in rye were determined with four varieties being grown on one site for three years and on three additional sites in the third year and a fourth variety being included in all trials except year 1. Clear differences were observed in the extent to which the contents of dietary fiber components (arabinoxylan, beta-glucan, total dietary fiber) and phytochemicals (folates, alkylresorcinols, sterols, tocols, phenolic acids) varied between varieties and between the same varieties grown in different sites (United Kingdom, France, Hungary, Poland) and years (2005-2007 in Hungary), with sterols being the most stable and phenolic acids the least. However, no single variety could be selected as having the highest overall level of bioactive components or as being more stable in comparison across environments.

Effects of environment and variety on alkylresorcinols in wheat in the HEALTHGRAIN diversity screen.

Alkylresorcinols (AR), phenolic lipids found in high amounts in whole grain wheat and rye, can be used as biomarkers for these cereals. The content (on a dry matter basis) and homologue composition of AR were determined in 26 wheat varieties grown in Hungary in 2005-2007, as well as in the United Kingdom, France, and Poland in 2007. There was a significant effect of year, location, and variety on both total AR and individual AR homologue content (p<0.001). A warm and dry climate generally resulted in higher AR contents, whereas high precipitation especially during plant development and grain-filling resulted in lower contents. There was a significant negative correlation between AR content and thousand-kernel weight (p<0.001), which may be explained by the warm and dry climate giving smaller kernels with higher AR content. The difference between varieties was generally consistent, with highest and lowest AR contents for the same varieties during different years and at different locations (p<0.001). Total AR content was correlated with the relative proportions of the different homologues, with a relatively lower concentration of homologues C17:0 and C19:0 and a lower C17:0/C12:0 ratio at higher overall contents. The results show that AR content is a highly heritable phytochemical component but that it is also affected by the environment.

Down-regulation of the CSLF6 gene results in decreased (1,3;1,4)-beta-D-glucan in endosperm of wheat.

(1,3;1,4)-beta-d-Glucan (beta-glucan) accounts for 20% of the total cell walls in the starchy endosperm of wheat (Triticum aestivum) and is an important source of dietary fiber for human nutrition with potential health benefits. Bioinformatic and array analyses of gene expression profiles in developing caryopses identified the CELLULOSE SYNTHASE-LIKE F6 (CSLF6) gene as encoding a putative beta-glucan synthase. RNA interference constructs were therefore designed to down-regulate CSLF6 gene expression and expressed in transgenic wheat under the control of a starchy endosperm-specific HMW subunit gene promoter. Analysis of wholemeal flours using an enzyme-based kit and by high-performance anion-exchange chromatography after digestion with lichenase showed decreases in total beta-glucan of between 30% and 52% and between 36% and 53%, respectively, in five transgenic lines compared to three control lines. The content of water-extractable beta-glucan was also reduced by about 50% in the transgenic lines, and the M(r) distribution of the fraction was decreased from an average of 79 to 85 x 10(4) g/mol in the controls and 36 to 57 x 10(4) g/mol in the transgenics. Immunolocalization of beta-glucan in semithin sections of mature and developing grains confirmed that the impact of the transgene was confined to the starchy endosperm with little or no effect on the aleurone or outer layers of the grain. The results confirm that the CSLF6 gene of wheat encodes a beta-glucan synthase and indicate that transgenic manipulation can be used to enhance the health benefits of wheat products.

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.

Alkylresorcinols in wheat varieties in the HEALTHGRAIN Diversity Screen.

The contents of alkylresorcinols (AR) were analyzed in 131 winter wheats, 20 spring wheats, 10 durum wheats, 5 spelt wheats, and 10 early cultivated forms of wheat (5 diploid einkorn and 5 tetraploid emmer), which are part of the HEALTHGRAIN diversity screen. AR were analyzed by gas chromatography (GC), which provides both total contents and relative homologue compositions, as well as with a Fast Blue colorimetric method that provides only total contents but which is fast and easily screens a large number of samples. There was considerable variation in the total AR content analyzed with GC: winter wheat (220-652 microg/g of dm), spring wheat (254-537 microg/g of dm), durum wheat (194-531 microg/g of dm), spelt (490-741 microg/g of dm), einkorn (545-654 microg/g of dm), and emmer wheat (531-714 microg/g of dm). The relative AR homologue composition was different for different types of wheat, with a C17:0 to C21:0 ratio of 0.1 for winter, spring, and spelt wheats, 0.04 for einkorn and emmer wheat, and 0.01 for durum wheat. The total AR content analyzed with the Fast Blue method was lower than that analyzed with GC but there was a good correlation between the two methods (R(2) = 0.76).

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 dietary fiber components in barley varieties in the HEALTHGRAIN Diversity Screen.

Ten different barley varieties grown in one location were studied for their content of tocols, folate, plant sterols, alkylresorcinols, and phenolic acids, as well as dietary fiber components (arabinoxylan and beta-glucan). The samples included hulled and hull-less barley types and types with normal, high-amylose, and waxy starch. The aim was to study the composition of raw materials, and therefore the hulls were not removed from the hulled barleys. A large variation was observed in the contents of all phytochemicals and dietary fibers. Two varieties from the INRA Clermont Ferrand barley program in France (CFL93-149 and CFL98-398) had high content of tocopherols and alkylresorcinols, whereas the variety Dicktoo was highest in dietary fiber content and phenolics. Positive correlations were found between 1000 kernel weight, alkylresorcinols, and tocols, as well as between dietary fiber content and phenolic compounds. The results demonstrate that the levels of phytochemicals in barley can likely be affected by breeding and that the contents of single phytochemicals may easily be adjusted by a right selection of a 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.