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Milled rice is an essential part of the regular diet for approximately half of the world's population. Its remarkable commercial value and consumer acceptance are mostly due to its promising cooking qualities, appealing sensory properties, and longer shelf life. However, the significant loss of the nutrient-rich bran layer during milling makes it less nutritious than the whole grain. Thus, enhancing the nutritive value of milled rice is vital in improving the health and wellbeing of rice consumers, particularly for those residing in the low-economic zones where rice is the primary source of calories and nutrition. This article provides a critical review on multiple frontiers of recent interventions, such as (1) infusing the genetic diversity to enrich amylose and resistant starch to reduce glycaemic index, (2) enhancing the minerals and vitamins through complementary fortification and biofortification as short and long-term interventions, and (3) developing transgenic solutions to improve the nutrient levels of milled rice. Additionally, the review highlights the benefits of functional ingredients of milled rice to human health and the potential of enhancing them in rice to address the triple burden of malnutrition. The potential merit of milled rice concerning food safety is also reviewed in this article.

Herein, optimized headspace solid phase microextraction with gas chromatography–tandem mass spectrometry (HS-SPME-GC-MS/MS) was used to estimate the 2-acetyl-1-pyrroline (2-AP) in raw and cooked rice samples of ten different traditional and improved varieties. Furthermore, HS-SPME-GC-MS-based volatile profiling was subjected to untargeted analyses to identify major odorants in raw and cooked rice samples, and to understand chemical proximities among volatile profiles. Results showed that 2-AP content was remarkably increased in cooked rice compared to raw. Among the varieties studied, Pusa-1652 (Improved Kala Namak) and Kala Namak-2 were superior in the 2-AP content than Basmati varieties. Additionally, Govind Bhog, Kala Jeera and Jeera-32 had 2-AP content equivalent to or superior to Basmati rice varieties. Altogether, 18 and 22 volatiles were identified in the raw and cooked rice samples studied, respectively. Of these, ethyl butyrate, ethyl 3-methylbutanoate, 2-undecanone, ethyl benzoate, ethyl benzeneacetate, 2-methylnaphthalene, and 1-methylnaphthalene were characteristically detected in the cooked rice. The high amount of 2-ethyl-1-hexanol was uniquely found in raw rice samples, which can be a marker compound for freshly milled rice. Along with 2-AP, butanoic acid and benzoic acid derivatives, phenylethyl alcohol, ethyl 3-hydroxybutyrate, and indole may be responsible for the overall perceived characteristic Basmati-like aroma in cooked rice.

BACKGROUND Conventional wheat-based foods contain high concentrations of readily digestible starch that commonly give these foods a high postprandial glycemic response and may contribute to the development of type 2 diabetes and cardiovascular disease. OBJECTIVES The aim of this study was to determine if bread made from high-amylose wheat (HAW) and enriched in resistant starch dampens postprandial glycemia compared with bread made from conventional low-amylose wheat (LAW). METHODS This single-center, randomized, double-blinded, crossover controlled study involved 7 consecutive weekly visits. On separate mornings, 20 healthy nondiabetic men and women (mean age 30 ± 3 y; body mass index 23 ± 0.7 kg/m2) consumed a glucose beverage or 4 different breads (each 121 g); LAW-R (refined), LAW-W (wholemeal), HAW-R, or HAW-W. The starch contents of the LAW and HAW breads were 24% and 74% amylose, respectively. Venous blood samples were collected at regular intervals before and for 3 h after the breakfast meal to measure plasma glucose, insulin, ghrelin, and incretin hormone concentrations, and the incremental area under the curve (AUC) was calculated (mmol/L × 3 h). Satiety and cravings were also measured at 30-min intervals during the postprandial period. RESULTS HAW breads had a glycemic response (AUC) that was 39% less than that achieved with conventional wheat breads (HAW 39 ± 5 mmol/L × 3 h; LAW 64 ± 5 mmol/L × 3 h; P

The present study was aimed at exploring the effect of soil application of different concentrations of orthophosphate (P) (0, 10, 20, 30, and 40 mg kg−1) on rice agronomic and yield parameters, arsenic (As) species accumulation, and polyphenol levels in the grain of rice grown under As spiked soil (10 mg kg−1). The contents of As species (As(V), As (III), MMA and DMA) and polyphenols in rice grain samples were estimated using LC-ICP-MS and LC-MS/MS, respectively. P treatments significantly reduced the toxic effects of As on agronomic parameters such as root weight and length, shoot and spike length, straw, and grain yield. Among the treatments studied, only the treatment of 30 mg kg−1 P helps to decrease the elevated levels of As (V), As (III), and DMA in rice grains due to As application. The study revealed that 30 mg kg−1 was the optimal P application amount to minimize AS accumulation in rice grains and As-linked toxicity on agronomic parameters and chlorophyll biosynthesis. Furthermore, the levels of trans-ferulic acid, chlorogenic acid, caffeic acid, and apigenin-7-glucoside increased in response to accumulation of As in the rice grain. In conclusion, the precise use of phosphorus may help to mitigate arsenic linked phytotoxicity and enhance the food safety aspect of rice grain.

Previous research has not considered the effect of high amylose wheat noodles on postprandial glycaemia. The aim of the study is to investigate the effect of consumption of high amylose noodles on postprandial glycaemia over 2-h periods by monitoring changes in blood glucose concentration and calculating the total area under the blood glucose concentration curve. Twelve healthy young adults were recruited to a repeated measure randomised, single-blinded crossover trial to compare the effect of consuming noodles (180 g) containing 15%, 20% and 45% amylose on postprandial glycaemia. Fasting blood glucose concentrations were taken via finger-prick blood samples. Postprandial blood glucose concentrations were taken at 15, 30, 45, 60, 90 and 120 min. Subjects consuming high amylose noodles made with flour containing 45% amylose had significantly lower blood glucose concentration at 15, 30 and 45 min (5.5 &plusmn, 0.11, 6.1 &plusmn, 0.11 and 5.6 &plusmn, 0.11 mmol/L, p = 0.01) compared to subjects consuming low amylose noodles with 15% amylose (5.8 &plusmn, 0.12, 6.6 &plusmn, 0.12 and 5.9 &plusmn, 0.12 mmol/L). The total area under the blood glucose concentration curve after consumption of high amylose noodles with 45% amylose was 640.4 &plusmn, 9.49 mmol/L/min, 3.4% lower than consumption of low amylose noodles with 15% amylose (662.9 &plusmn, 9.49 mmol/L/min), p = 0.021. Noodles made from high amylose wheat flour attenuate postprandial glycaemia in healthy young adults, as characterised by the significantly lower blood glucose concentration and a 3.4% reduction in glycaemic response.

Wheat storage proteins have been historically examined and periodically established to be the major determinant of wheat quality. Gluten proteins largely contribute to the formation of viscoelastic network in a dough, enabling processing of wheat to food products including bread. More recently starch, the major component constituting 60–70% of wheat grain, is understood to play key roles in flour quality, dough functionality and end product and nutritional quality. Starch is composed of two neutral macromolecules of glucose, amylose and amylopectin. The structural differences between amylose and amylopectin are predominantly dependent on the extent and distribution of α-1,4 and α-1,6 linkages that connect the glucose units to form these two polymers. The functional properties of starch as governed by its structure, molecular organisation, granule morphology and size distribution influence dough behaviour during processing, differentially impacting the end product qualities. Also, varyingly important are the roles of starch granule associated proteins, comprised of both surface proteins and granule-integral proteins with enzyme functions, in driving starch responses in a complex dough matrix system. This chapter aims to provide an extensive re-view on how starch, associated proteins and starch-protein interactions influence functional properties of food systems.

Non-communicable diseases such as diabetes, cardiovascular diseases and certain cancers are now the leading cause of death and disability, and their prevalence is rising worldwide. Poor diet is a major modifiable risk factor but changing eating habits has had limited success. Enhancing the nutritional quality of staple foods offers a complimentary intervention strategy for alleviating the burden of diet-related chronic disease. Wholegrain cereals, such as wheat, are prime targets. Their nutritional credentials and health-promoting potential are well established. Wheat is the major source of protein, minerals and vitamins, and dietary fibre for most people. Importantly, wheat is a versatile ingredient for producing foods that have high consumer appeal. Their popularity is expanding globally and small improvements in wheat grain composition conceivably translate to substantial dietary change across entire populations. A newly developed, high amylose wheat line illustrates the capacity for developing healthier processed staple food products with potential for remediating chronic disease risk. Dietary fibre density of this novel grain is markedly improved due to a >10 fold increase in resistant starch content. The sensory attributes of finished products are not compromised and preliminary studies demonstrate that consumption of the new wheat significantly improves indices of metabolic and digestive health.

High amylose wheat (HAW) starch has been the focus of a number of nutritional studies, but there is limited information around its effect on the mechanical properties of wheat flour dough. This study investigated the size, shape and packing volume of HAW starch and their effect on the microstructure and rheology of dough. Four flour blends were formulated by adding vital wheat gluten and either HAW or commercial wheat starch to HAW flour to achieve a constant 14% protein content, but varied amounts of HAW starch. A large number of small and irregularly shaped HAW starch granules resulted in a high packing volume per gram of starch. Confocal laser scanning microscopy of optimally mixed doughs correlated the degree of starch granule aggregation with the level of HAW starch in the bi-continuous dough network. Small deformation rheology demonstrated that increased quantities of HAW starch in the dough increased the elastic modulus G′ values. Uniaxial extension measurements highlighted a synergy between HAW starch and sources of gluten proteins resulting in increased strain hardening. The impact of HAW starch on dough rheology was attributed to its irregular shape and large number of small granules leading to greater granule-granule interactions.

Improving wheat quality and safety is very important for wheat breeders and related industries. While considerable research has been carried out internationally, collaborations among laboratories have been limited, and each group often uses different materials, methods and gene nomenclature.

Summary Resistant starch (RS), a type of dietary fibre, plays an important role in human health; however, the content of RS in most modern processed starchy foods is low. Cereal starch, when structurally manipulated through a modified starch biosynthetic pathway to greatly increase the amylose content, could be an important food source of RS. Transgenic studies have previously revealed the requirement of simultaneous down-regulation of two starch branching enzyme (SBE) II isoforms both located on the long arm of chromosome 2, namely SBEIIa and SBEIIb, to elevate the amylose content in wheat from ~25% to ~75%. The current study revealed close proximity of genes encoding SBEIIa and SBEIIb isoforms in wheat with a genetic distance of 0.5 cM on chromosome 2B. A series of deletion and single nucleotide polymorphism (SNP) loss of function alleles in SBEIIa, SBEIIb or both was isolated from two different wheat populations. A breeding strategy to combine deletions and SNPs generated wheat genotypes with altered expression levels of SBEIIa and SBEIIb, elevating the amylose content to an unprecedented ~85%, with a marked concomitant increase in RS content. Biochemical assays were used to confirm the complete absence in the grain of expression of SBEIIa from all three genomes in combination with the absence of SBEIIb from one of the genomes.

Summary Manipulation of SSI expression in wheat using RNAi technology leads to the production of wheat grains with novel starch structure and properties., Studies in Arabidopsis and rice suggest that manipulation of starch synthase I (SSI) expression in wheat may lead to the production of wheat grains with novel starch structure and properties. This work describes the suppression of SSI expression in wheat grains using RNAi technology, which leads to a low level of enzymatic activity for SSI in the developing endosperm, and a low abundance of SSI protein inside the starch granules of mature grains. The amylopectin fraction of starch from the SSI suppressed lines showed an increased frequency of very short chains (degree of polymerization, dp 6 and 7), a lower proportion of short chains (dp 8–12), and more intermediate chains (dp 13–20) than in the grain from their negative segregant lines. In the most severely affected line, amylose content was significantly increased, the morphology of starch granules was changed, and the proportion of B starch granules was significantly reduced. The change of the fine structure of the starch in the SSI-RNAi suppression lines alters the gelatinization temperature, swelling power, and viscosity of the starch. This work demonstrates that the roles of SSI in the determination of starch structure and properties are similar among different cereals and Arabidopsis.

The relationships between starch structure and functionality are important in underpinning the industrial and nutritional utilisation of starches. In this work, the relationships between the biosynthesis, structure, molecular organisation and functionality have been examined using a series of defined genotypes in barley with low (20%), standard (20-30%), elevated (30-50%) and high (50%) amylose starches. A range of techniques have been employed to determine starch physical features, higher order structure and functionality. The two genetic mechanisms for generating high amylose contents (down-regulation of branching enzymes and starch synthases, respectively) yielded starches with very different amylopectin structures but similar gelatinisation and viscosity properties driven by reduced granular order and increased amylose content. Principal components analysis (PCA) was used to elucidate the relationships between genotypes and starch molecular structure and functionality. Parameters associated with granule order (PC1) accounted for a large percentage of the variance (57%) and were closely related to amylose content. Parameters associated with amylopectin fine structure accounted for 18% of the variance but were less closely aligned to functionality parameters.

The roles of starch branching enzyme (SBE, EC 2.4.1.18) IIa and SBE IIb in defining the structure of amylose and amylopectin in barley (Hordeum vulgare) endosperm were examined. Barley lines with low expression of SBE IIa or SBE IIb, and with the low expression of both isoforms were generated through RNA-mediated silencing technology. These lines enabled the study of the role of each of these isoforms in determining the amylose content, the distribution of chain lengths, and the frequency of branching in both amylose and amylopectin. In lines where both SBE IIa and SBE IIb expression were reduced by >80%, a high amylose phenotype (>70%) was observed, while a reduction in the expression of either of these isoforms alone had minor impact on amylose content. The structure and properties of the high amylose starch resulting from the concomitant reduction in the expression of both isoforms of SBE II in barley were found to approximate changes seen in amylose extender mutants of maize, which result from lesions eliminating expression of the SBE IIb gene. Amylopectin chain length distribution analysis indicated that both SBE IIa and SBE IIb isoforms play distinct roles in determining the fine structure of amylopectin. A significant reduction in the frequency of branches in amylopectin was noticed only when both SBE IIa and SBE IIb were reduced, whereas there was a significant increase in the branching frequency of amylose when SBE IIb alone was reduced. Functional interactions between SBE isoforms are suggested, and a possible inhibitory role of SBE IIb on other SBE isoforms is discussed.

Wheat starch is considered to have a low paste viscosity relative to other starches. Consequently, wheat starch is not preferred for many applications as compared to other high paste viscosity starches. Increasing the viscosity of wheat starch is expected to increase the functionality of a range of wheat flour-based products in which the texture is an important aspect of consumer acceptance (e.g., pasta, and instant and yellow alkaline noodles). To understand the molecular basis of starch viscosity, we have undertaken a comprehensive structural and rheological analysis of starches from a genetically diverse set of wheat genotypes, which revealed significant variation in starch traits including starch granule protein content, starch-associated lipid content and composition, phosphate content, and the structures of the amylose and amylopectin fractions. Statistical analysis highlighted the association between amylopectin chains of 18-25 glucose residues and starch pasting properties. Principal component analysis also identified an association between monoesterified phosphate and starch pasting properties in wheat despite the low starch-phosphate level in wheat as compared to tuber starches. We also found a strong negative correlation between the phosphate ester content and the starch content in flour. Previously observed associations between internal starch granule fatty acids and the swelling peak time and pasting temperature have been confirmed. This study has highlighted a range of parameters associated with increased starch viscosity that could be used in prebreeding/breeding programs to modify wheat starch pasting properties.

Some of the starch consumed by humans is not digested in the small intestine. Such starch, known as resistant starch, is fermented in the large intestine and leads to the production of short chain fatty acids. Increased consumption of resistant starch is associated with improved cardio-vascular health. A high proportion of amylose in the starch consumed is correlated with increased resistant starch but other unknown aspects of starch structure may also influence the digestibility of starch. Detailed investigation of the starch biosynthetic pathway has revealed that reducing the activity of specific isoforms of branching enzymes and starch synthases can lead to increased amylose. Methods to alter the expression of and detect mutations in targeted genes involved are discussed.

Starch synthases (SS) are responsible for elongating the alpha-1,4 glucan chains of starch. A doubled haploid population was generated by crossing a line of wheat, which lacks functional ssIIa genes on each genome (abd), and an Australian wheat cultivar, Sunco, with wild type ssIIa alleles on each genome (ABD). Evidence has been presented previously indicating that the SGP-1 (starch granule protein-1) proteins present in the starch granule in wheat are products of the ssIIa genes. Analysis of 100 progeny lines demonstrated co-segregation of the ssIIa alleles from the three genomes with the SGP-1 proteins, providing further evidence that the SGP-1 proteins are the products of the ssIIa genes. From the progeny lines, 40 doubled haploid lines representing the eight possible genotypes for SSIIa (ABD, aBD, AbD, ABd, abD, aBd, Abd, abd) were characterized for their grain weight, protein content, total starch content and starch properties. For some properties (chain length distribution, pasting properties, swelling power, and gelatinization properties), a progressive change was observed across the four classes of genotypes (wild type, single nulls, double nulls and triple nulls). However, for other grain properties (seed weight and protein content) and starch properties (total starch content, granule morphology and crystallinity, granule size distribution, amylose content, amylose-lipid dissociation properties), a statistically significant change only occurred for the triple nulls, indicating that all three genes had to be missing or inactive for a change to occur. These results illustrate the importance of SSIIa in controlling grain and starch properties and the importance of amylopectin fine structure in controlling starch granule properties in wheat.

Background Glycogen average chain length (ACL) has been linked with bacterial durability, but this was on the basis of observations across different species. We therefore wished to investigate the relationship between bacterial durability and glycogen ACL by varying glycogen average chain length in a single species. It has been shown that progressive shortening of the N-terminus of glycogen branching enzyme (GBE) leads to a lengthening of oligosaccharide inter-α-1,6-glycosidic chain lengths, so we sought to harness this to create a set of Escherichia coli DH5α strains with a range of glycogen average chain lengths, and assess these strains for durability related attributes, such as starvation, cold and desiccation stress resistance, and biofilm formation. Results A series of Escherichia coli DH5α mutants were created with glgB genes that were in situ progressively N-terminus truncated. N-terminal truncation shifted the distribution of glycogen chain lengths from 5-11 DP toward 13-50 DP, but the relationship between glgB length and glycogen ACL was not linear. Surprisingly, removal of the first 270 nucleotides of glgB (glgBΔ270) resulted in comparatively high glycogen accumulation, with the glycogen having short ACL. Complete knockout of glgB led to the formation of amylose-like glycogen containing long, linear α1,4-glucan chains with significantly reduced branching frequency. Physiologically, the set of mutant strains had reduced bacterial starvation resistance, while minimally increasing bacterial desiccation resistance. Finally, although there were no obvious changes in cold stress resistance or biofilm forming ability, one strain (glgBΔ180) had significantly increased biofilm formation in favourable media. Conclusions Despite glgB being the first gene of an operon, it is clear that in situ mutation is a viable means to create more biologically relevant mutant strains. Secondly, there was the suggestion in the data that impairments of starvation, cold and desiccation resistance were worse for the strain lacking glgB, though the first of these was not statistically significant. The results provide prima facie evidence linking abiotic stress tolerance with shorter glycogen ACL. However, further work needs to be done, perhaps in a less labile species. Further work is also required to tease out the complex relationship between glycogen abundance and glycogen structure. Electronic supplementary material The online version of this article (doi:10.1186/s12866-015-0421-9) contains supplementary material, which is available to authorized users.

Studies of maize starch branching enzyme mutants suggest that the amylose extender high amylose starch phenotype is a consequence of the lack of expression of the predominant starch branching enzyme II isoform expressed in the endosperm, SBEIIb. However, in wheat, the ratio of SBEIIb and SBEIIa expression are inversely related to the expression levels observed in maize and rice. Analysis of RNA at 15 days post anthesis suggests that there are about 4-fold more RNA for SBE IIa than for SBE IIb. The genes for SBE IIa and SBE IIb from wheat are distinguished in the size of the first three exons, allowing isoform-specific antibodies to be produced. These antibodies were used to demonstrate that in the soluble fraction, the amount of SBE IIa protein is two to three fold higher than SBIIb, whereas in the starch granule, there is two to three fold more SBE IIb protein amount than SBE IIa. In a further difference to maize and rice, the genes for SBE IIa and SBE IIb are both located on the long arm of chromosome 2 in wheat, in a position not expected from rice-maize-wheat synteny.

The synthesis of starch in wheat and barley is an important topic for research because of the exten sive utility of starch from these crop species in human and animal foods, and in industrial processes. Wheat and barley starches are highly characteristic due to their granular architecture and multi-modal granule size distribution. This granule architecture is important because it defines the ways in which wheat and barley starches behave during food processing. The core starch biosynthetic genes of wheat have been cloned and shown to exist as homeologous sets of genes represented on each of the three wheat genomes. While hexaploidyrepresents a major impediment to the selection of altered starch phenotypes by phenotypic screening, the availability of methods for identifying the products of homeologus genes from each of the wheat genes has provided methods for the selection of triple null lines from waxy, starch synthase ha, and branching enzyme I genes. In barley, direct phenotypic selection has resulted in the identification of waxy, amol and SSIIa mutations. In this paper, we review the state of knowledge of starch synthesis in wheat and barley and discuss the relationships between individual genes and their roles in starch biosynthesis.

Dietary fiber shortens gut transit time, but data on the effects of fiber components (including resistant starch, RS) on intestinal contractility are limited. We have examined RS effects in male Sprague–Dawley rats fed either a high-amylose maize starch (HAMS) or a wholemeal made from high-amylose wheat (HAW) on ileal and colonic contractility ex vivo and expression of genes associated with smooth muscle contractility. Rats were fed diets containing 19 % fat, 20 % protein, and either low-amylose maize starch (LAMS), HAMS, wholemeal low-amylose wheat (LAW) or HAW for 11 week. Isolated ileal and proximal colonic sections were induced to contract electrically, or by receptor-independent (KCl) or receptor-dependent agents. Colonic gene expression was assessed using an Affymetrix microarray. Ileal contractility was unaffected by treatment. Maximal proximal colonic contractility induced electrically or by angiotensin II or carbachol was lower for rats fed HAMS and LAW relative to those fed LAMS (P

Shortly after genetic modification of plants became possible in the early 1980’s, attempts were made to modify starch structure by altering the expression of starch biosynthetic genes or by introducing novel enzymes from other organisms. Much of this work was done initially in potato and later work focussed on the model plant Arabidopsis and the regulation of starch breakdown in leaves. In this chapter, we will review recent advances in generating novel starch compositions and structures using new breeding technologies, focusing mainly on cereal starches but including other starches where appropriate advances have been made. We will first describe attempts to alter starch structure in order to improve starch functionality, then explain how increasing knowledge of the regulation of starch biosynthesis is being used to increase starch production, and finish by summarizing new methods for increasing the genetic diversity in crops as well as methods for fine-tuning gene expression in plants in order to bring improved starch-based products with value-added consumer benefits to the marketplace.

Genes and cDNAs for starch-branching enzyme II (SBEII) have been isolated from libraries constructed from Aegilops tauschii and wheat (Triticum aestivum) endosperm, respectively. One class of genes has been termedwSBEII-DA1 and encodes the N terminus reported for an SBEII from wheat endosperm. On the basis of phylogenetic comparisons with other branching enzyme sequences, wSBEII-DA1 is considered to be a member of the SBEIIa class. ThewSBEII-DA1 gene consists of 22 exons with exons 4 to 21 being identical in length to the maize (Zea mays) SBEIIb gene, and the gene is located in the proximal region of the long arm of chromosome 2 at a locus designated sbe2a. RNA encoding SBEIIa can be detected in the endosperm from 6 d after flowering and is at its maximum level from 15 to 18 d after anthesis. Use of antibodies specific for SBEIIa demonstrated that this protein was present in both the soluble and granule bound fractions in developing wheat endosperm. We also report a cDNA sequence for SBEIIa that could arise by variant transcription/splicing. A second gene, termedwSBEII-DB1, was isolated and encodes an SBEII, which shows greater sequence identity with SBEIIb-type sequences than with SBEIIa-type sequences. Comparisons of SBEII gene structures among wheat, maize, and Arabidopsis indicate the lineage of the SBEII genes.

Starch biosynthesis in plants involves the concerted action of a number of enzymes, including ADPglucose pyrophosphorylase, starch synthases, branching enzymes and debranching enzymes. We report on the cloning and characterisation of genes encoding these enzymes from wheat and on their chromosomal locations. The prospects for manipulating wheat starch structure and functionality using these genes is discussed.

Resistant starch (RS), fed as high amylose maize starch (HAMS) or butyrylated HAMS (HAMSB), opposes dietary protein-induced colonocyte DNA damage in rats. In this study, rats were fed Western-type diets moderate in fat (19%) and protein (20%) containing digestible starches [low amylose maize starch (LAMS) or low amylose whole wheat (LAW)] or RS [HAMS, HAMSB, or a whole high amylose wheat (HAW) generated by RNA interference] for 11 wk (n = 10/group). A control diet included 7% fat, 13% protein, and LAMS. Colonocyte DNA single-strand breaks (SSB) were significantly higher (by 70%) in rats fed the Western diet containing LAMS relative to controls. Dietary HAW, HAMS, and HAMSB opposed this effect while raising digesta levels of SCFA and lowering ammonia and phenol levels. SSB correlated inversely with total large bowel SCFA, including colonic butyrate concentration (R(2) = 0.40; P = 0.009), and positively with colonic ammonia concentration (R(2) = 0.40; P = 0.014). Analysis of gut microbiota populations using a phylogenetic microarray revealed profiles that fell into 3 distinct groups: control and LAMS; HAMS and HAMSB; and LAW and HAW. The expression of colonic genes associated with the maintenance of genomic integrity (notably Mdm2, Top1, Msh3, Ung, Rere, Cebpa, Gmnn, and Parg) was altered and varied with RS source. HAW is as effective as HAMS and HAMSB in opposing diet-induced colonic DNA damage in rats, but their effects on the large bowel microbiota and colonocyte gene expression differ, possibly due to the presence of other fiber components in HAW.

A line of wheat (Triticum aestivum L.), sgp-1, that does not express starch synthase II (SSII, also known as SGP-1) has previously been reported. In this study, F1 derived doubled haploid lines with homozygous wild type or mutant alleles for SGP-1 genes were identified from a cross between the original mutant and a wild type Australian cultivar. Analysis of the starch granules showed that in the mutant lines they are markedly distorted from 15 days postanthesis during grain development. Starch branching patterns showed an increase in the proportion of short chains (DP 6–10) at an earlier stage, but this increase became much more pronounced at 15 days postanthesis and persisted until maturity. There was also a consistent and drastic reduction throughout seed development in the relative amounts of starch branching enzyme II (SBEII, comprising SBEIIa and SBEIIb) and starch synthase I (SSI) bound to the starch granules. In the soluble phase, however, there was relatively little change in the amount of SBEIIb, SBEIIa or SSI protein. Therefore loss of SSII specifically leads to the loss of SBEIIb, SBEIIa and SSI protein in the granule-bound phase and the effect of this mutation is clearly manifest from the mid-stage of endosperm development in wheat.

Foods high in resistant starch have the potential to improve human health and lower the risk of serious noninfectious diseases. RNA interference was used to down-regulate the two different isoforms of starch-branching enzyme (SBE) II (SBEIIa and SBEIIb) in wheat endosperm to raise its amylose content. Suppression of SBEIIb expression alone had no effect on amylose content; however, suppression of both SBEIIa and SBEIIb expression resulted in starch containing >70% amylose. When the >70% amylose wheat grain was fed to rats in a diet as a wholemeal, several indices of large-bowel function, including short-chain fatty acids, were improved relative to standard wholemeal wheat. These results indicate that this high-amylose wheat has a significant potential to improve human health through its resistant starch content.

The role of starch branching enzyme-I (SBE-I) in determining starch structure in the endosperm has been investigated. Null mutations of SBE-I at the A, B and D genomes of wheat were identified in Australian wheat varieties by immunoblotting. By combining individual null mutations at the B and D genomes through hybridisation, a double-null mutant wheat, which lacks the B and D isoforms of SBE-I, was developed. Wheat mutants lacking all the three isoforms of SBE-I were generated from a doubled haploid progeny of a cross between the BD double-null mutant line and a Chinese Spring (CS) deletion line lacking the A genome isoform. Comparison of starch from this mutant wheat to that from wild type revealed no substantial alteration in any of the structural or functional properties analysed. Further analysis of this triple-null mutant line revealed the presence of another residual peak of SBE-I activity, referred to as SBE-Ir, in wheat endosperm representing < 3% of the activity of SBE-I in wild type endosperm.

Genes for an isoamylase-like debranching enzyme have been isolated from rice and Aegilops tauschii, the donor of the D genome to wheat. The structures of the genes are very similar to each other and to the maize SU1 isoamylase gene and consist of 18 exons spread over approximately 7.5 kb. Southern analysis and fluorescent in situ hybridization showed the Ae. tauschii gene to be located in the proximal region of the short arm of chromosome 7D, thus showing synteny with the localization of the rice isoamylase gene on rice chromosome 8. Analysis of the expression pattern of wheat sugary isoamylase genes indicates that they are strongly expressed in the developing endosperm 6 days after flowering. Three distinct Sugary-type cDNA sequences were isolated from the wheat endosperm that are likely to correspond to the products of the three genomes. The deduced amino acid sequence of rice and wheat Sugary-type isoamylase is compared with other sequences available in the database and the results demonstrate that there are three types of isoamylase sequences in plants: those containing 18 exons (the Sugary-type isoamylase gene), those containing 21 exons, and those containing only 1 exon. It is possible that different combinations of isoamylase genes are expressed in different tissues.Key words: isoamylase, rice, wheat, sugary, FISH.