QTL Analysis of ß-Glucan Content and Other Grain Traits in a Recombinant Population of Spring Barley.

Barley with high grain ß-glucan content is valuable for functional foods. The identification of loci for high ß-glucan content is, thus, of great importance for barley breeding. Segregation mapping for the content in ß-glucan and other barley grain components (starch, protein, lipid, ash, phosphorous, calcium, sodium) was performed using the progeny of the cross between Glacier AC38, a mutant with high amylose, and CDC Fibar, a high ß-glucan waxy cultivar. The offspring of this cross showed transgressive segregation for ß-glucan content. Linkage analysis based on single-nucleotide polymorphism (SNP) molecular markers was used for the genotyping of the parents and recombinant inbred lines (RILs). Two Quantitative Trait Loci (QTL) for ß-glucan content and several QTL for other grain components were found. The former ones, located on chromosomes 1H and 7H, explained 27.9% and 27.4% of the phenotypic variance, respectively. Glacier AC38 provided the allele for high ß-glucan content at the QTL on chromosome 1H, whereas CDC Fibar contributed the allele at the QTL on chromosome 7H. Their recombination resulted in a novel haplotype with higher ß-glucan content, up to 18.4%. Candidate genes are proposed for these two QTL: HvCslF9, involved in ß-glucan biosynthesis, for the QTL on chromosome 1H; Horvu_PLANET_7H01G069300, a gene encoding an ATP-Binding Cassette (ABC) transporter, for the QTL on chromosome 7H.

A Travel through Landscapes of Seed Dormancy.

Basic features of seed dormancy are illustrated. The seed overall regulatory network governs seed metabolism and development, and it is coordinated by plant hormones. A functional model focused on abscisic acid (ABA), the foremost plant hormone in dormancy, is used as a framework to critically discuss the literature. Gibberellins (GAs) have a main role in germination, and the ABA-GAs balance is a typical feature of the seed state: ABA dominates during dormancy and GAs prevail through germination. Thus, the literature converges toward envisaging the development switch between dormancy and germination as represented by the ABA/GAs ratio. The ABA-GAs antagonism is based on mutual inhibition, a feature of the regulatory network architecture that characterizes development trajectories based on a regulatory circuit with a bistable switch. Properties of such kind of regulatory architecture are introduced step by step, and it is shown that seed development-toward either dormancy or germination-is more properly represented by a tristable regulatory circuit, whose intermediate metastable states ultimately take one or the other development trajectory. Although the ABA/GAs ratio can conveniently represent the state of the seed overall regulatory network along the seed development trajectory, specific (unknown) dormancy factors are required to determine the development trajectory. The development landscape is shown to provide a well-suited representation of seed states travelling along developmental trajectories, particularly when the states are envisioned as regulatory circuits. Looking at seed dormancy in terms of regulatory circuits and development landscapes offers a valuable perspective to improve our understanding of this biological phenomenon.

Indirect Measurement of ß-Glucan Content in Barley Grain with Near-Infrared Reflectance Spectroscopy.

ß-Glucan is a component of barley grains with functional properties that make it useful for human consumption. Cultivars with high grain ß-glucan are required for industrial processing. Breeding for barley genotypes with higher ß-glucan content requires a high-throughput method to assess ß-glucan quickly and cheaply. Wet-chemistry laboratory procedures are low-throughput and expensive, but indirect measurement methods such as near-infrared reflectance spectroscopy (NIRS) match the breeding requirements (once the NIR spectrometer is available). A predictive model for the indirect measurement of ß-glucan content in ground barley grains with NIRS was therefore developed using 248 samples with a wide range of ß-glucan contents (3.4%-17.6%). To develop such calibration, 198 unique samples were used for training and 50 for validation. The predictive model had R2 = 0.990, bias = 0.013% and RMSEP = 0.327% for validation. NIRS was confirmed to be a very useful technique for indirect measurement of ß-glucan content and evaluation of high-ß-glucan barleys.

In Dormant Red Rice Seeds, the Inhibition of Early Seedling Growth, but Not of Germination, Requires Extracellular ABA.

The phytohormone abscisic acid (ABA) inhibits seed germination and seedling growth and is required for the inception of dormancy. Xanthoxal (also known as xanthoxin) is the first specific biosynthetic precursor of ABA. In this study, a modified method to produce xanthoxal is described. I tested the ability of either xanthoxal or ABA to reinstate dormancy in dormant red rice seeds whose dormancy was broken by fluridone (an inhibitor of the synthesis of carotenoids and, subsequently, ABA). Xanthoxal was shown to have a stronger inhibitory effect on germination than ABA when exogenously provided. Although this could indicate an additional effect of xanthoxal above that expected if xanthoxal were simply converted to ABA in the seed, alternative hypotheses cannot be excluded. One alternative is that exogenous xanthoxal may be trapped inside the cells to a greater extent than exogenous ABA, resulting in an intracellular level of ABA higher than that reached with a direct application of ABA. As a further alternative, exogenous xanthoxal may interfere with ABA action in the apoplast. In this study, following germination, early seedling growth was delayed only if ABA was applied. This suggests that inhibition of early seedling growth, but not of germination, requires extracellular ABA.

GWAS for Starch-Related Parameters in Japonica Rice (Oryza sativa L.).

Rice quality is mainly related to the following two starch components, apparent amylose content (AAC) and resistant starch (RS). The former affects grain cooking properties, while RS acts as a prebiotic. In the present study, a Genome Wide Association Scan (GWAS) was performed using 115 rice japonica accessions, including tropical and temperate genotypes, with the purpose of expanding the knowledge of the genetic bases affecting RS and AAC. High phenotypic variation was recorded for the two traits, which positively correlated. Moreover, both the parameters correlated with seed length (positive correlation) and seed width (negative correlation). A correlational selection according to human preferences has been hypothesized for the two starch traits and grain size. In addition, human selection has been proposed as the causal agent even for the different phenotypes related to starch and grain size showed by the tropical and temperate japonica accessions utilized in this study. The present GWAS led to the identification of 11 associations for RS on seven chromosomes and five associations for AAC on chromosome 6. Candidate genes and co-positional relationships with quantitative trait loci (QTLs) previously identified as affecting RS and AAC were identified for 6 associations. The candidate genes and the new RS- and/or AAC-associated regions detected provide valuable sources for future functional characterizations and for breeding programs aimed at improving rice grain quality.

Drought stress influenced sesamin and sesamolin content and polyphenolic components in sesame (Sesamum indicum L.) populations with contrasting seed coat colors.

Ten sesame genotypes planted under two irrigation regimes of 60% and 90%, as the maximum allowable depletion (MAD), were used to investigate the effects of drought stress on certain quantitative and qualitative characters of sesame seeds with four contrasting coat colors. The polyphenolic components, sesamin, sesamolin, total flavonoid content (TFC), total phenolic content (TPC), radical scavenging activity (RSA), seed yield, and oil content of the seeds were also examined. Results revealed that drought decreased seed yield, oil content, sesamin, and quercetin but increased TFC, TPC, and RSA as well as most of polyphenolic components and sesamolin. The drought-tolerant genotypes including Markazi1 exhibited higher chlorogenic, ellagic, and p-coumaric acids as well as TFC, RSA, and rutin. While the dark-seeded sesame genotypes contained higher caffeic, ferulic, ellagic acids as well as TPC and RSA, the light-seeded ones were richer in sesamin and sesamolin as well as p-coumaric and gallic acids. The findings of the study provided basic information on the changes in some seed secondary metabolites when sesame was subjected to drought stress. The results also confirmed not only the presence of considerable amounts of antioxidants in sesame seeds but also differences in secondary metabolite levels among the sesame seeds with different seed coat colors.

The Caryopsis of Red-Grained Rice Has Enhanced Resistance to Fungal Attack.

Seed persistence in the soil is threatened by microorganisms, but the seed coat helps protect the seed from them. Although modern rice (Oryza sativa L.) cultivars have a whitish caryopsis, some varieties have a red caryopsis coat, a trait typical of wild Oryza species. The red colour is due to the oxidation of proanthocyanidins, a class of flavonoids that is found in the outer layers of the seed in many species. We aimed to assess whether these natural compounds (proanthocyanidins and proanthocyanidin-derived pigment) have some protective effect against microbial attacks. Dehulled caryopses of white-grained and red-grained rice genotypes were employed to assay fungal infection. Specifically, three white-grained rice cultivars (Perla, Augusto, and Koral) and three red-grained rice varieties (Perla Rosso, Augusto Rosso, and Koral Rosso) were used. In a first test, the caryopses were infected with Epicoccum nigrum at 10 °C, and seedling growth was then assessed at 30 °C. In a second test, the degree of infection by the mycotoxigenic fungus Fusarium sporotrichioides was assayed by measuring the accumulation of T-2/HT-2 toxins in the caryopses. Infection was performed at 10 °C to prevent rice germination while allowing fungal growth. In both the tests, red caryopses showed reduced, or delayed, infection with respect to white ones. One black-grained cultivar (Venere) was assayed for the accumulation of T-2/HT-2 toxins as well, with results corresponding to those of the red-grained rice varieties. We argue that the red pigment accumulating in the caryopsis coat, and/or the proanthocyanidins associated with it, provides a protective barrier against challenging microorganisms.

Seed Dormancy Involves a Transcriptional Program That Supports Early Plastid Functionality during Imbibition.

Red rice fully dormant seeds do not germinate even under favorable germination conditions. In several species, including rice, seed dormancy can be removed by dry-afterripening (warm storage); thus, dormant and non-dormant seeds can be compared for the same genotype. A weedy (red) rice genotype with strong dormancy was used for mRNA expression profiling, by RNA-Seq, of dormant and non-dormant dehulled caryopses (here addressed as seeds) at two temperatures (30 °C and 10 °C) and two durations of incubation in water (8 h and 8 days). Aim of the study was to highlight the differences in the transcriptome of dormant and non-dormant imbibed seeds. Transcript data suggested important differences between these seeds (at least, as inferred by expression-based metabolism reconstruction): dry-afterripening seems to impose a respiratory impairment onto non-dormant seeds, thus glycolysis is deduced to be preferentially directed to alcoholic fermentation in non-dormant seeds but to alanine production in dormant ones; phosphoenolpyruvate carboxykinase, pyruvate phosphate dikinase and alanine aminotransferase pathways appear to have an important gluconeogenetic role associated with the restoration of plastid functions in the dormant seed following imbibition; correspondingly, co-expression analysis pointed out a commitment to guarantee plastid functionality in dormant seeds. At 8 h of imbibition, as inferred by gene expression, dormant seeds appear to preferentially use carbon and nitrogen resources for biosynthetic processes in the plastid, including starch and proanthocyanidins accumulation. Chromatin modification appears to be a possible mechanism involved in the transition from dormancy to germination. Non-dormant seeds show higher expression of genes related to cell wall modification, suggesting they prepare for acrospire/radicle elongation.

QTLs for Woolly Poplar Aphid (Phloeomyzus passerinii L.) Resistance Detected in an Inter-Specific Populus deltoides x P. nigra Mapping Population.

The genus Populus represents one of the most economically important groups of forest trees. It is composed by approximately 30 species used for wood and non-wood products, phytoremediation and biomass. Poplar is subjected to several biological and environmental threats although, compared to annual crops, we know far less about the genetic bases of biotic stress resistance. Woolly poplar aphid (Phloeomyzus passerinii) is considered a main pest of cultivated poplars in European and American countries. In this work we present two high density linkage maps in poplar obtained by a genotyping by sequencing (GBS) approach and the identification of QTLs involved in Ph. passerinii resistance. A total of 5,667 polymorphic markers (5,606 SNPs and 61 SSRs) identified on expressed sequences have been used to genotype 131 plants of an F1 population P ×canadensis obtained by an interspecific mate between Populus deltoides (resistant to woolly poplar aphid) and Populus nigra (susceptible to woolly poplar aphid). The two linkage maps, obtained following the two-way pseudo-testcross mapping strategy, have been used to investigate the genetic bases of woolly poplar aphid resistance. One major QTL and two QTLs with minor effects (mapped on LGV, LGXVI and LG XIX) explaining the 65.8% of the genetic variance observed in the progeny in response to Ph. passerinii attack were found. The high density coverage of functional markers allowed the identification of three genes belonging to disease resistance pathway as putative candidates for P. deltoides resistance to woolly poplar aphid. This work is the first report on genetic of woolly poplar aphid genetic resistance and the resistant loci associated markers identified represent a valuable tool in resistance poplar breeding programs.

Deep sequencing transcriptional fingerprinting of rice kernels for dissecting grain quality traits.

BACKGROUND: Rice represents one the most important foods all over the world. In Europe, Italy is the first rice producer and Italian production is driven by tradition and quality. All main rice grain quality traits, like cooking properties, texture, gelatinization temperature, chalkiness and yield, are related to the content and composition of starch and seed-storage proteins in the endosperm and to grain shape. In addition, a number of nutraceutical compounds and allergens are known to have a significant effect on grain quality determination. To investigate the genetic bases underlying the qualitative differences that characterize traditional Italian rice cultivars, a comparative RNA-Seq-based transcriptomic analysis of developing caryopsis was conducted at 14 days after flowering on six popular Italian varieties (Carnaroli, Arborio, Balilla, Vialone Nano, Gigante Vercelli and Volano) phenotypically differing for qualitative grain-related traits. RESULTS: Co-regulation analyses of differentially expressed genes showing the same expression patterns in the six genotypes highlighted clusters of loci up or down-regulated in specific varieties, with respect to the others. Among them, we detected loci involved in cell wall biosynthesis, protein metabolism and redox homeostasis, classes of genes affecting in chalkiness determination. Moreover, loci encoding for seed-storage proteins, allergens or involved in the biosynthesis of specific nutraceutical compounds were also present and specifically regulated in the different clusters. A wider investigation of all the DEGs detected in pair-wise comparisons revealed transcriptional variation, among the six genotypes, for quality-related loci involved in starch biosynthesis (e.g. GBSSI, starch synthases and AGPase), genes encoding for transcription factors, additional seed storage proteins, allergens or belonging to additional nutraceutical compounds biosynthetic pathways and loci affecting grain size. Putative functional SNPs associated to amylose content in starch, gelatinization temperature and grain size were also identified. CONCLUSIONS: The present work represents a more extended phenotypic characterization of a set of rice accessions that present a wider genetic variability than described nowadays in literature. The results provide the first transcriptional picture for several of the grain quality differences observed among the Italian rice varieties analyzed and reveal that each variety is characterized by the over-expression of a peculiar set of loci affecting grain appearance and quality. A list of candidates and SNPs affecting specific grain properties has been identified offering a starting point for further works aimed to characterize genes and molecular markers for breeding programs.

Improvement of marker-based predictability of Apparent Amylose Content in japonica rice through GBSSI allele mining.

BACKGROUND: Apparent Amylose Content (AAC), regulated by the Waxy gene, represents the key determinant of rice cooking properties. In occidental countries high AAC rice represents the most requested market class but the availability of molecular markers allowing specific selection of high AAC varieties is limited. RESULTS: In this study, the effectiveness of available molecular markers in predicting AAC was evaluated in a collection of 127 rice accessions (125 japonica ssp. and 2 indica ssp.) characterized by AAC values from glutinous to 26%. The analyses highlighted the presence of several different allelic patterns identifiable by a few molecular markers, and two of them, i.e., the SNPs at intron1 and exon 6, were able to explain a maximum of 79.5% of AAC variation. However, the available molecular markers haplotypes did not provide tools for predicting accessions with AAC higher than 24.5%. To identify additional polymorphisms, the re-sequencing of the Waxy gene and 1kbp of the putative upstream regulatory region was performed in 21 genotypes representing all the AAC classes identified. Several previously un-characterized SNPs were identified and four of them were used to develop dCAPS markers. CONCLUSIONS: The addition of the SNPs newly identified slightly increased the AAC explained variation and allowed the identification of a haplotype almost unequivocally associated to AAC higher than 24.5%. Haplotypes at the waxy locus were also associated to grain length and length/width (L/W) ratio. In particular, the SNP at the first intron, which identifies the Wxa and Wxb alleles, was associated with differences in the width of the grain, the L/W ratio and the length of the kernel, most likely as a result of human selection.

Haplotype variability and identification of new functional alleles at the Rdg2a leaf stripe resistance gene locus.

The barley Rdg2a locus confers resistance to the leaf stripe pathogen Pyrenophora graminea and, in the barley genotype Thibaut, it is composed of a gene family with three highly similar paralogs. Only one member of the gene family (called as Rdg2a) encoding for a CC-NB-LRR protein is able to confer resistance to the leaf stripe isolate Dg2. To study the genome evolution and diversity at the Rdg2a locus, sequences spanning the Rdg2a gene were compared in two barley cultivars, Thibaut and Morex, respectively, resistant and susceptible to leaf stripe. An overall high level of sequence conservation interrupted by several rearrangements that included three main deletions was observed in the Morex contig. The main deletion of 13,692 bp was most likely derived from unequal crossing over between Rdg2a paralogs leading to the generation of a chimeric Morex rdg2a gene which was not associated to detectable level of resistance toward leaf stripe. PCR-based analyses of genic and intergenic regions at the Rdg2a locus in 29 H. vulgare lines and one H. vulgare ssp. spontaneum accession indicated large haplotype variability in the cultivated barley gene pool suggesting rapid and recent divergence at this locus. Barley genotypes showing the same haplotype as Thibaut at the Rdg2a locus were selected for a Rdg2a allele mining through allele re-sequencing and two lines with polymorphic nucleotides leading to amino acid changes in the CC-NB and LRR encoding domains, respectively, were identified. Analysis of nucleotide diversity of the Rdg2a alleles revealed that the polymorphic sites were subjected to positive selection. Moreover, strong positively selected sites were located in the LRR encoding domain suggesting that both positive selection and divergence at homologous loci are possibly representing the molecular mechanism for the generation of high diversity at the Rdg2a locus in the barley gene pool.

Effects of barley ß-glucan-enriched flour fractions on the glycaemic index of bread.

The aim of this research was to evaluate ß-glucan-enriched flours, obtained from barleys with either normal or waxy starch, for their effects on the glycaemic index (GI) and the quality of bread. Rheological results confirmed that when barley flour was included in the dough the overall quality of bread slightly worsened. However, positive consequences on glycaemia were obtained with the normal starch barley: the GI of all-wheat bread (82.8 ± 7.2) was significantly reduced (57.2 ± 7.9) when 40% of wheat flour was substituted with ß-glucan-enriched barley flour (6.0% ± 0.1 ß-glucan in the final flour blend). In contrast, this positive effect was significantly reduced (GI: 70.1 ± 9.1) when 40% of wheat flour was substituted with the ß-glucan-enriched flour of a waxy barley (CDC Alamo; 6.6 ± 0.2 ß-glucan in the final flour blend), suggesting that the ability of ß-glucans to lower the GI was affected by the barley starch-type.

Constitutive differences between steely and mealy barley samples associated with endosperm modification.

BACKGROUND: Structurally different areas may occur in the endosperm of the barley grain, and they can be visually classified as either mealy or steely. Barleys with a high proportion of grains that are mostly steely often show uneven physical-chemical modification of the endosperm during malting. To study the relationship between steeliness and endosperm modification, two samples of barley cv. Scarlett with contrasting malting quality were analysed. RESULTS: The proportions of steely grains were 77% and 46% in the two samples, which were then defined as steely sample and mealy sample, respectively. The steely sample showed slower modification during malting (in terms of beta-glucan degradation, friability increase, and Calcofluor staining), lower hot water extract (HWE) and acrospire growth, and higher extract viscosity. Endosperm permeation to large molecules (tested with the fluorescein isothiocyanate-dextran conjugate, FITC-D) closely followed cell wall modification in the steely sample, but this was not so in the mealy sample. CONCLUSIONS: Higher steeliness was associated with higher levels of C hordeins in the grain of barley cv. Scarlett. It is proposed that such hordeins can increase the permeability to large molecules (FITC-D) but slow modification. Like steeliness and the level of C hordeins, permeability to FITC-D appears to be more linked to environmental rather than genetic effects. Although a more general association of C hordeins with steeliness of malting barley still has to be ascertained, the negative role of C hordeins in malting quality has been confirmed.

A theoretical framework for beta-glucan degradation during barley malting.

During malting, barley germinates and produces hydrolytic enzymes that de-structure the endosperm, making the grains soft and friable. This process starts close to the embryo and spreads throughout the whole grain. It is leaded by the degradation of cell walls, which are mainly constituted of beta-glucans. Fast and extended breakdown of beta-glucans occurs by means of an expanding reaction front driven by beta-glucanase, and appears to follow pseudo-first-order kinetics. Endosperm permeabilization to macromolecules is closely linked to the dismantling of cell walls, thus that access to beta-glucans by beta-glucanase itself is limited. It is shown that the kinetics of beta-glucan degradation during malting are consequent to this condition, and can be explained according to an anomalous evolution of the reverse quasi-steady-state approximation (rQSSA) for enzymatic reactions. In fact, kinetics based on the rQSSA include a transient phase wherein fast substrate depletion is indeed of pseudo-first-order. In the germinating barley, the conditions in which the physical modification of the endosperm occurs are shown to be suitable for the fast transient to persist in dynamic equilibrium while it progressively expands throughout the grain, depleting most beta-glucans and, then, establishing the overall kinetics of beta-glucan breakdown.

On the role of abscisic acid in seed dormancy of red rice.

Abscisic acid (ABA) is commonly assumed to be the primary effector of seed dormancy, but conclusive evidence for this role is lacking. This paper reports on the relationships occurring in red rice between ABA and seed dormancy. Content of free ABA in dry and imbibed caryopses, both dormant and after-ripened, the effects of inhibitors, and the ability of applied ABA to revert dormancy breakage were considered. The results indicate: (i) no direct correlation of ABA content with the dormancy status of the seed, either dry or imbibed; (ii) different sensitivity to ABA of non-dormant seed and seed that was forced to germinate by fluridone; and (iii) an inability of exogenous ABA to reinstate dormancy in fluridone-treated seed, even though applied at a pH which favoured high ABA accumulation. These considerations suggest that ABA is involved in regulating the first steps of germination, but unidentified developmental effectors that are specific to dormancy appear to stimulate ABA synthesis and to enforce the responsiveness to this phytohormone. These primary effectors appear physiologically to modulate dormancy and via ABA they effect the growth of the embryo. Therefore, it is suggested that ABA plays a key role in integrating the dormancy-specific developmental signals with the control of growth.

Characterization of antioxidant compounds of red and white rice and changes in total antioxidant capacity during processing.

Although most rice cultivars have whitish kernel, some varieties have a red testa. Aim of this work was to compare the total antioxidant capacity (TAC) and the antioxidant chemical composition (namely tocols, gamma-oryzanols, and polyphenols) of red and white rices. In addition, the effect of milling and cooking on antioxidants was investigated in both rices. Dehulled red rice showed a TAC more than three times greater than dehulled white rice and its high TAC was essentially characterized by the presence of proanthocyanidins (PA) and associated phenolics. Milling caused a significant loss of TAC, even if red rice maintained a higher TAC. Cooking caused a further loss of antioxidants, but when there was a full uptake of cooking water by the grains ("risotto") this loss was limited. Thus, the consumption of whole or partially milled rice cooked as risotto would be preferred to preserve its nutritional properties.

Ethylene production is associated with germination but not seed dormancy in red rice.

BACKGROUND AND AIMS: The relationship between ethylene production and both seed dormancy and germination was investigated using red rice (weedy rice) as a model species. METHODS: Both fully dormant and after-ripened (non-dormant) naked caryopses were incubated with or without inhibitors of ethylene synthesis [aminoethoxyvinylglycine (AVG)] and perception [silver thiosulfate (STS)], or in the presence of the natural ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC). The kinetics of ethylene emissions were measured with a sensitive laser-photoacoustic system. KEY RESULTS: Dormant red rice caryopses did not produce ethylene. In non-dormant caryopses, ethylene evolution never preceded the first visible stage of germination (pericarp splitting), and ethylene inhibitors completely blocked ethylene production, but not pericarp splitting. Accordingly, endogenous ACC appeared to be lacking before pericarp splitting. However, early seedling growth (radicle or coleoptile attaining the length of 1 mm) followed ethylene evolution and was delayed by the inhibitors. Wounding the dormant caryopses induced them to germinate and produce ethylene, but their germination was slow and pericarp splitting could be speeded up by ethylene. CONCLUSIONS: The findings suggest that, in red rice, endogenous ethylene stimulates the growth of the nascent seedling, but does not affect seed dormancy or germination inception. Correspondingly, this phytohormone does not play a role in the dormancy breakage induced by wounding, but accelerates germination after such breakage has occurred.

In vivo modeling of beta-glucan degradation in contrasting barley (Hordeum vulgare L.) genotypes.

An important determinative of malt quality is the malt beta-glucan content, which in turn depends on the initial barley beta-glucan content as well as the beta-glucan depolymerization by beta-glucanase (EC 3.2.1.73) during malting. Another enzyme, named beta-glucan solubilase, has been suggested to act prior to beta-glucanase; its existence, however, has not been unequivocally proven. We monitored changes in beta-glucan levels and in the development of beta-glucan-degrading enzymes during malting of five lots of contrasting barley genotypes. Two models of in vivo kinetics for beta-glucan degradation were then compared as follows: (i) a biphasic model based on the sequential action of beta-glucan solubilase and beta-glucanase and (ii) a monophasic model assuming that all beta-glucans are depolymerized by beta-glucanase without the previous intervention of another enzyme. Confirmatory regression analysis was used to test the fit of the models to the observed data. Our results show that beta-glucan degradation is mostly monophasic, although some enzyme other than beta-glucanase seems to be required for the early solubilization of a small fraction of insoluble beta-glucans (on average, 7% of total beta-glucans). Furthermore, the genotype-dependent kinetic rate constant (indicating beta-glucan degradability), in addition to beta-glucanase activity, is suggested to play a major role in malting quality.