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Orthophosphate, phytate, and total phosphorus determination in cereals by flow injection analysis.

A flow injection spectrophotometric procedure with enzymatic hydrolysis was developed for determination of orthophosphate, phytate and total phosphorus in cereal samples. Phosphorus species were extracted from cereals with 0.05 mol L(-1) potassium hydrogen phthalate buffer solution at pH 5.7. Orthophosphate was directly determined in the extracts by molybdenum blue spectrophotometric method. The phytate was hydrolyzed by the enzyme phytase coupled to a solid phase packed into an enzymatic reactor, and the resulting hydrolyzed orthophosphate was also determined by spectrophotometry at 650 nm. After optimization for phosphorus species extraction and enzymatic hydrolysis, a linear calibration graph was obtained up to 196 x 10(-6) mol L(-1) orthophosphate (P conc = -2.67 + 0.52x, r = 0.9998). Measurements are characterized by relative standard deviation of 1.6% for a standard of 72 x 10(-6) mol L(-1) orthophosphate and no baseline drift was observed during 4 h operation periods. It provides 72 measurements per hour, with 2.4 x 10(-)6) mol L(-1) and 7.9 x 10(-6) mol L(-1) as detection and quantification limits, respectively.

Edible Grain↗

Rapid multimineral determination in infant cereal matrices using wavelength dispersive X-ray fluorescence.

A rapid and simple method for the determination of a series of macroelements (sodium, magnesium, phosphorus, chlorine, potassium, and calcium) and trace elements (manganese, iron, and zinc) by wavelength dispersive X-ray fluorescence has been developed and validated for infant cereal matrices. Reference values were obtained by inductively coupled plasma optical emission spectroscopy and by potentiometry. The 88 investigated samples were commercially available products. Pellets of 4 g were prepared under 10 tonnes of pressure. For each sample, 3 pellets were prepared and analyzed. Limits of quantification and repeatabilities were evaluated. Calibrations were established with 43 samples, and method validation was made using a second set of 45 samples. An evaluation of this alternative method was done by comparison with data obtained from the reference methods. The results show the good performances of the alternative method to routine infant cereals analysis.

Edible Grain↗

Elucidating the transformation pattern of the cereal allelochemical 6-methoxy-2-benzoxazolinone (MBOA) and the trideuteriomethoxy analogue [D3]-MBOA in soil.

To deduce the structure of the large array of compounds arising from the transformation pathway of 6-methoxybenzoxazolin-2-one (MBOA), the combination of isotopic substitution and liquid chromatography analysis with mass spectrometry detection was used as a powerful tool. MBOA is formed in soil when the cereal allelochemical 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) is exuded from plant material to soil. Degradation experiments were performed in concentrations of 400 microg of benzoxazolinone/g of soil for MBOA and its isotopomer 6-trideuteriomethoxybenzoxazolin-2-one ([D3]-MBOA). Previously identified metabolites 2-amino-7-methoxyphenoxazin-3-one (AMPO) and 2-acetylamino-7-methoxyphenoxazin-3-one (AAMPO) were detected. Furthermore, several novel compounds were detected and provisionally characterized. The environmental impact of these compounds and their long-range effects are yet to be discovered. This is imperative due to the enhanced interest in exploiting the allelopathic properties of cereals as a means of reducing the use of synthetic pesticides.

Benzoxazoles↗

Biodegradation of oxalic acid from spinach using cereal radicles.

A high level of oxalate intake constitutes a health risk for infants and metabolically disposed adults. Spinach, acclaimed for its many health benefits, is among the vegetables richest in oxalate. Blanching reduces oxalate unsatisfactorily and unspecifically. An alternative, biological method is proposed on the basis of rye seedlings or radicles (also barley and wheat) containing an oxalate-specific oxalate oxidase by nature. Dissolved oxalate (0.25 mM) was rapidly degraded in the presence of radicles (e.g., 70% within 100 min). With commercial deep-frozen spinach, near-complete degradation of soluble oxalate was achieved at pH 3.5. The total level of oxalate was reduced by half. Similarly high rates occurred from 18 to 35 degrees C. Even at 55 degrees C appreciable rates were observed. The seedling as a whole is effective, too, and enrichment with cereal-specific healthy components would occur. Removal of oxalate from other vegetables, juices, cycled process waters, or feeds is conceivable with fresh or heat-dried cereal seedlings or radicles.

Biodegradation, Environmental↗

Front-face fluorescence spectroscopy as a rapid and nondestructive tool for differentiating various cereal products: a preliminary investigation.

The potential of intrinsic fluorescence spectroscopy was investigated for differentiating between processed grains (flours, pasta, and semolinas) of different wheat cereal products. A total of 59 samples (e.g., 20 complete Kamut, semicomplete Kamut, and soft wheat flours, 28 pasta, and 11 semolinas manufactured from complete Kamut, semicomplete Kamut, and hard wheat flours) were analyzed by front-face fluorescence spectroscopy. Tryptophan fluorescence spectra were scanned between 305 and 400 nm on samples following excitation at 290 nm. The principal component analysis (PCA) performed on flour spectra clearly differentiated complete Kamut and semicomplete Kamut samples from those produced from complete and semicomplete soft wheat flours. The PCA performed on pasta spectra discriminated samples manufactured from complete Kamut and complete hard wheat flours from those made with semicomplete Kamut and semicomplete hard wheat flours. The best discrimination was obtained from tryptophan spectra recorded on semolinas since the four groups were well discriminated. Correct classification amounting to 61.9% was obtained for pasta spectra. A better classification was obtained for flour and semolina spectra since correct classification amounted to 86.7% and 87.9%, respectively. Front-face fluorescence spectroscopy has the potential to be a rapid, low-cost, and efficient method for the authentication of cereal products.

Edible Grain↗

Treatment of cereal products with a tailored preparation of trichoderma enzymes increases the amount of soluble dietary fiber.

Nutritionists recommend increasing the intake of soluble dietary fiber (SDF), which is very low in most cereal-based products. Conversion of insoluble DF (IDF) into SDF can be achieved by chemical treatments, but this affects the sensorial properties of the products. In this study, the possibility of getting a substantial increase of SDF from cereal products using a tailored preparation of Trichoderma enzymes is reported. Enzymes were produced cultivating Trichoderma using durum wheat fiber (DWF) and barley spent grain (BSG) as unique carbon sources. Many Trichoderma strains were screened, and the hydrolysis conditions able to increase by enzymatic treatment the amount of SDF in DWF and BSG were determined. Results demonstrate in both products that it is possible to triple the amount of SDF without a marked decrease of total DF. The enzymatic treatment also causes the release of hydroxycinnamic acids, mainly ferulic acid, that are linked to the polysaccharides chains. This increases the free phenolic concentration, the water-soluble antioxidant activity, and, in turn, the phenol compounds bioavailability.

Coumaric Acids↗

Fluorescence of cereal flours.

Front-surface fluorescence spectra of cereal flours are easily measured with a commercial spectrofluorometer and a simple cell. The emission spectra are intense in the range 290-600 nm in which three emission bands are active. The most intense is due to the aromatic amino acid residues present in the proteins of flours. At least two other fluorophores are active in the above spectral range and compete for light with the chromophores present in the flours. Four absorption bands are revealed in the same spectral region by reflectance spectra, the most intense being due to the amino acidic fluorophores. Thus, the measured emission is the result of absorption, scattering, emission, and reabsorption processes. Information on the microenvironment of some fluorophores can be obtained. The possibility to recognize different species of cereals, even of different cultivars, is shown.

Edible Grain↗

Analysis of volatile compounds and their contribution to flavor in cereals.

Recent developments in methods for isolation and measurement of volatiles from cereals are reviewed. The main types of isolation methods, namely, direct extraction, distillation, and headspace, have recently been complemented by solid phase microextraction. Direct solvent extraction provides efficient recovery of compounds with a broad range of polarities and volatilities, whereas headspace techniques provide relatively clean extracts. Supercritical fluid and solid phase microextractions have not yet been fully evaluated for cereals. GC and GC/MS remain the dominant techniques for measurement of the extracted compounds, although new electronic noses show promise. Relating these results to human perception requires careful control to ensure valid comparisons and, in this respect, aroma extract dilution analysis is a useful procedure.

Chromatography, Gas↗

Antioxidant activity and total phenolics in selected cereal grains and their different morphological fractions.

The purpose of this study was to examine the antioxidant properties of water and 80% methanolic extracts of cereal grains and their different morphological fractions. Wheat (Triticum aestivum L.) cv. Almari and cv. Henika, barley (Hordeum vulgare L.) cv. Gregor and cv. Mobek, rye (Secale cereale L.) cv. Dańkowskie Zlote, oat (Avena sativa L.) cv. Slawko and buckwheat (Fagopyrum esculentum Moench) cv. Kora were used. PC (L-alpha-phosphatidylcholine) liposome system and spectrophotometric assay of total antioxidant activity (TAA) were used to evaluate the antioxidative activity of extracts. Among the water extracts, only the one prepared from buckwheat exhibited antioxidant activity at the concentration analyzed. The following hierarchy of antioxidant activity was provided for 80% methanolic extracts originated from whole grain: buckwheat > barley > oat > wheat congruent with rye. The antioxidant activity was observed in extract prepared from separated parts of buckwheat and barley. In respect to hulls, the antioxidant hierarchy was as follows: buckwheat > oat > barley. The correlation coefficient between total phenolic compounds and total antioxidative activity of the extracts was -0.35 for water extracts and 0.96, 0.99, 0.80, and 0.99 for 80% methanolic extracts originated from whole grains, hulls, pericarb with testa fractions and endosperm with embryo fractions, respectively.

Antioxidants↗

Content of phenolic acids and ferulic acid dehydrodimers in 17 rye (Secale cereale L.) varieties.

The contents of pnenolic acids and ferulic acid dehydrodimers were quantified by HPLC analysis after alkaline hydrolysis in kernels of 17 rye (Secale cereale L.) varieties grown in one location in Denmark during 1997 and 1998. Significant variations (P < 0.05) with regard to the concentration of the analyzed components were observed among the different rye varieties and also between different harvest years. However, the content of phenolic acids in the analyzed rye varieties was narrow compared to cereals such as wheat and barley. The concentration of ferulic acid, the most abundant phenolic acid ranged from 900 to 1170 microgram g(-1) dry matter. The content in sinapic acid ranged from 70 to 140 microgram g(-1) dry matter, p-coumaric acid ranged from 40 to 70 microgram g(-1) dry matter, and caffeic, p-hydroxybenzoic, protocatechuic, and vanillic acids were all detected in concentrations less than 20 microgram g(-1) dry matter. The most abundant ferulic acid dehydrodimer 8-O-4 -DiFA was quantified in concentrations from 130 to 200 microgram g(-1) dry matter followed by 8,5 -DiFA benzofuran form (50-100 microgram g(-1) dry matter), 5,5 -DiFA (40-70 microgram g(-1) dry matter), and 8,5 -DiFA (20-40 microgram g(-1) dry matter).

Bread↗

Analysis of programmed cell death in wheat endosperm reveals differences in endosperm development between cereals.

Although maize endosperm undergoes programmed cell death during its development, it is not known whether this developmental feature is common to cereals or whether it arose inadvertently from the selection process that resulted in the enlarged endosperm of modern maize. Examination of wheat endosperm during its development revealed that this tissue undergoes a programmed cell death that shares features with the maize program but differs in some aspects of its execution. Cell death initiated and progressed stochastically in wheat endosperm in contrast to maize where cell death initiates within the upper central endosperm and expands outward. After a peak of ethylene production during early development, wheat endosperm DNA underwent internucleosomal fragmentation that was detectable from mid to late development. The developmental onset and progression of DNA degradation was regulated by the level of ethylene production and perception. These observations suggest that programmed cell death of the endosperm and regulation of this program by ethylene is not unique to maize but that differences in the execution of the program appear to exist among cereals.

Apoptosis↗

High expression level of a gene coding for a chloroplastic amino acid selective channel protein is correlated to cold acclimation in cereals.

A cold-regulated gene (cor tmc-ap3) coding for a putative chloroplastic amino acid selective channel protein was isolated from cold-treated barley leaves combining the differential display and the 5'-RACE techniques. Cor tmc-ap3 is expressed at low level under normal growing temperature, and its expression is strongly enhanced after cold treatment. A positive correlation between the expression of cor tmc-ap3 and frost tolerance was found both among barley cultivars and among cereal species. The COR TMC-AP3 protein was expressed in vitro, purified and used to raise a polyclonal antibody. Western analysis showed that the cor tmc-ap3 gene product is localized to the chloroplastic outer envelope fraction, supporting its putative function. The frost-resistant winter cultivar Onice accumulated COR TMC-AP3 more rapidly and at a higher level than the frost-susceptible spring cultivar Gitane. After 28 days of cold acclimation the winter cultivar had about 2-fold more protein than the spring genotype. All these results suggest that an increased amount of a chloroplastic amino acid selective channel protein could be required for cold acclimation in cereals. Hypotheses about the role of COR TMC-AP3 during the hardening process are discussed.

Acclimatization↗

Phytotoxicity of pathogenic fungi and their mycotoxins to cereal seedling viability.

Aspergillus flavus, Alternaria alternata and Fusarium oxysporum were the pathogenic fungi most reduced cereal (barley, sorghum and wheat) seedlings. The pathogens have the ability to produce aflatoxin B1 and G1, diacetoxyscirpenol, kojic acid and tenuazonic acid that reduced seedling viability. The inhibition dose for 50% reduction (LD50) was recorded by aflatoxins at 0.83 mg L-1 for barley, 1.74 mg L-1 for wheat and 2.75 mg L-1 for sorghum. Diacetoxyscirpenol produced its inhibition at 1.26 mg L-1 for barley, 3.98 mg L-1 for wheat and 10 mg L-1 for sorghum. Kojic acid induced 50% inhibition at 63 mg L-1 for barley, 105 mg L-1 for wheat and 251 mg L-1 for sorghum. However, tenuazonic acid was less toxic where the toxicity ranged between 79-550 mg L-1. The germination inhibition was more pronounced in barley followed by wheat and was negligible in sorghum for all tested mycotoxins. This inhibition was attributed to the reduction in the seedling amylase activity, where amylase was also reduced in the same trend: barley > wheat > sorghum. Grain treated with carboxin-captan and thiophanatemethyl-thiram at 1 g kg-1 grain increased the seedlings vigour of wheat in sterilized soil by 45 and 22%, barley by 24 and 33% and sorghum by 15 and 30%, respectively. These fungicides also had a positive effect on cereal when the soil was inoculated with A. flavus, A. alternata and F. oxysporum, but the improvement was still below normal.

Amylases↗

Statistical methods for QTL mapping in cereals.

This paper gives an overview of the statistical theory suitable for mapping quantitative trait loci in experimental populations derived from inbred parents, with a particular emphasis on methodology for cereal crops. The basic theory is described, and some new areas of statistical research appropriate for mapping in cereal crops are discussed.

Chromosome Mapping↗

Phylogenetic analysis reveals that a dwarfing disease on different cereal crops in China is due to rice black streaked dwarf virus (RBSDV).

A viral disease with dwarfing symptoms is associated with severe damage of different cereal crops including rice, maize, wheat and sorghum grown in China. It is believed that the pathogenic agent of the disease on rice and sorghum is rice black streaked dwarf virus (RBSDV), however, the cause of maize dwarf disease in China is still inconclusive. In this report, dsRNA was isolated from virus particles obtained from the diseased plants of rice, maize, wheat and sorghum from two Chinese provinces. Full-length cDNAs of genome segments 9 (S9) and 10 (S 10) were obtained through a RT-PCR approach. Sequence analysis showed that the S9 sequences of Chinese isolates and Japanese RBSDV isolate were very similar to each other (89.1-89.6% identity at the nucleotide level, 92.3-92.9% and 95.8-98.6% identity at the amino acid level for ORF1 and ORF2, respectively). In addition, the S10 sequences of Chinese isolates and Japanese RBSDV were very similar to each other (93.0-95.4% identical nucleotides and 96.2-97.0% identical amino acids, respectively). However, there were lower similarities for S9 and S10 sequences between Chinese isolates and an Italian Maize Rough Dwarf Virus (MRDV) isolate. Phylogenetic analysis indicates that Chinese viral isolates found to infect rice, maize, wheat and sorghum and leading to similar cereal dwarfing manifestations could be grouped to the same virus species, RBSDV.

China↗

Aggregation pheromone of the cereal leaf beetle: field evaluation and emission from males in the laboratory.

The previously identified, male-specific compound of the cereal leaf beetle (CLB, Chrysomelidae; Oulema melanopus), (E)-8-hydroxy-6-methyl-6-octen-3-one, was studied further with respect to field activity and emission rate from male beetles. In a 5-week field experiment in Oregon, the compound was shown to function as an aggregation pheromone in attracting male and female CLBs migrating from overwintering sites in spring. Traps baited with the synthetic compound (500 microg per rubber septum) caught 3.3 times more CLBs than control traps. Lower doses of the pheromone (50 and 150 microg) were less attractive than the 500 microg dose. One relatively abundant, volatile compound from the host plant (oats), (Z)-3-hexenyl acetate, that elicited responses from beetle antennae was not attractive, either by itself or as a synergist of the pheromone. Both sexes were captured about equally for all treatments. We also measured daily pheromone emission by male beetles in the laboratory. Individual males feeding on oat seedlings under greenhouse conditions emitted as much as 6 microg per day, which is about 500 times higher than had been previously observed under incubator conditions. The pheromone emission rate was at least five times higher during the day than at night, and in one male, emission spanned a period of 28 d. The release rate of synthetic pheromone from the 500 microg septa was very similar to the maximum from single males; thus, future experiments should evaluate even higher doses. The field results indicate that the pheromone has potential as a monitoring tool for early detection of CLBs as they move from their overwintering sites into newly planted cereal crops in spring.

Animals↗

Programmed cell death in cereal aleurone.

Progress in understanding programmed cell death (PCD) in the cereal aleurone is described. Cereal aleurone cells are specialized endosperm cells that function to synthesize and secrete hydrolytic enzymes that break down reserves in the starchy endosperm. Unlike the cells of the starchy endosperm, aleurone cells are viable in mature grain but undergo PCD when germination is triggered or when isolated aleurone layers or protoplasts are incubated in gibberellic acid (GA). Abscisic acid (ABA) slows down the process of aleurone cell death and isolated aleurone protoplasts can be kept alive in media containing ABA for up to 6 months. Cell death in barley aleurone occurs only after cells become highly vacuolated and is manifested in an abrupt loss of plasma membrane integrity. Aleurone cell death does not follow the apoptotic pathway found in many animal cells. The hallmarks of apoptosis, including internucleosomal DNA cleavage, plasma membrane and nuclear blebbing and formation of apoptotic bodies, are not observed in dying aleurone cells. PCD in barley aleurone cells is accompanied by the accumulation of a spectrum of nuclease and protease activities and the loss of organelles as a result of cellular autolysis.

Apoptosis↗

Demography of the cereal rust mite Abacarus hystrix (Acari: Eriophyoidea) on quack grass.

Demography parameters of the cereal rust mite Abacarus hystrix (Nalepa) on quack grass were studied to investigate its potential capacity of population increase in conditions of initially low density. The experiment was maintained under laboratory conditions at a constant temperature of 19.5-20.5 degrees C and 94 +/- 1% RH. Life-history data were used to calculate duration of developmental stages, survival of adults and rates of population increase. A new method of estimation of age-dependent fecundity is proposed. On average, eggs required 7.98 (n = 33, 95% CI: 7.68-8.21) days to develop into adults. Life expectancy of females was longer than that of males (9.72 and 5.41 days, respectively). The mean sex ratio, expressed as the proportion of females, was 0.80 (n = 122, CI: 0.71-0.86). The reproductive output for females was age-dependent and daily egg production reached a peak (3.83 eggs/day, CI: 2.50-5.15) on the 5th day, and then decreased steeply. The net reproductive rate (R0) was 10.12 female progeny per female per generation, the generation time (T) was 11.31 days, the intrinsic rate of increase (r) was 0.20 female progeny per female per day, and the finite rate of increase (lambda) was 1.23 female progeny per day. These estimates showed that A. hystrix has a great potential capacity for rapid population increase when colonising new hosts and its density is low. Therefore, we conclude that the population of the cereal rust mite on quack grass may rapidly build up to very high densities and can be a reservoir population, which may easily disperse and infest other, including cultivated, grasses.

Animals↗