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DNA fragmentation in cereal roots indicative of programmed root cortical cell death.

In cereals, a progressively increasing root cortical cell death (RCD) occurs from the root tip and upwards when measured with vital staining methods. In this study, nuclear DNA fragmentation was studied in seminal root segments of wheat and barley in order to investigate if the cell death resembled apoptosis. The fraction of cells with TUNEL-positive nuclei increased gradually with increasing root age in both the cortex and the stele. Southern analysis showed a typical ladder pattern, indicating nucleosomal fragmentation already in 2-day-old root segments, and this became more pronounced in older root segments. DNA fragmentation appeared to be more extensive in wheat than in barley roots. These results confirm earlier studies, where RCD has been found to be earlier initiated and to proceed at a faster rate in wheat. The characteristic DNA fragmentation found in the roots indicates programmed cell death with mechanistic similarities to apoptosis. Ultrastructural examination of nuclei in cortex cells with transmission electron microscopy revealed an increased chromatin condensation in older roots, particularly in wheat. In addition, we found nucleosomal DNA ladders in young leaf tissue from wheat but not from barley.

Journal Article↗

From common signalling components to cell specific responses: insights from the cereal aleurone.

Studies into the molecules underlying plant signal transduction events continue to reveal the involvement of highly conserved factors such as Ca2+, calmodulin, cyclic GMP and phospholipases in a remarkably diverse array of physiological processes. The hormonal response systems in the aleurone cells of the cereal grain and in the stomatal guard cell are beginning to reveal how diversity of response can be hard wired into these cells despite the use of these common signalling intermediates. In both the aleurone and the guard cell ABA signalling operates through the action of phospholipase D and alterations in a Ca2+-dependent signalling system. The role of phospholipase D is highly analogous in these two divergent cell types, perhaps reflecting the closeness of this enzyme to a conserved ABA receptor. However, specificity in response becomes evident in elements downstream from PLD, such as in the Ca2+ signalling system. For example, ABA has opposite effects on cytoplasmic Ca2+ in the aleurone and guard cell. Combining the Ca2+-dependent signalling activities in networks with parallel regulatory activities such as cyclic GMP appears to underlie the flexible regulatory systems that are the hallmark of plant cell function.

Journal Article↗

Genetic diversity amongst landraces of rye (Secale cereale L.) from northern Europe.

Genetic variation and diversity were estimated for three improved varieties and 18 landraces of Secale cereale, originating from the northern Europe. This material was compared with eight improved varieties and 16 landraces from Sweden, which were analysed before. The analysis used starch gel electrophoresis with the enzymes ACO, GPI, MDH, PGD and PGM and resulted in one fixed locus and seven polymorphic loci. Most of the genetic diversity was found within the accessions and it was very small between accessions. The results show that the landraces from Germany and Norway have a low genetic variation compared to the other landraces in this study. This can possibly be explained with a small original sample size of some accessions. However, in the dendrogram these accessions are well separated. The landraces from Sweden and Finland showed a high genetic variation, which is almost the same for all these accessions. In the dendrogram all of the Finnish landraces and 11 of the Swedish ones were grouped together with a very small diversity index; they can almost be considered as part of the same accession. This is probably due to the fact that a high number of Finnish immigrants arrived in Sweden during the 17th century bringing their own rye material. The early Finnish rye landraces may have the same original source. The data also show that landraces as well as improved varieties have a high heterozygosity but they are separated in the dendrogram into different clusters. The currently used varieties show a large genetic distance variation, different from that in the landraces.

Chi-Square Distribution↗

Processing of wild cereal grains in the Upper Palaeolithic revealed by starch grain analysis.

Barley (Hordeum vulgare L.) and wheat (Triticum monococcum L. and Triticum turgidum L.) were among the principal 'founder crops' of southwest Asian agriculture. Two issues that were central to the cultural transition from foraging to food production are poorly understood. They are the dates at which human groups began to routinely exploit wild varieties of wheat and barley, and when foragers first utilized technologies to pound and grind the hard, fibrous seeds of these and other plants to turn them into easily digestible foodstuffs. Here we report the earliest direct evidence for human processing of grass seeds, including barley and possibly wheat, in the form of starch grains recovered from a ground stone artefact from the Upper Palaeolithic site of Ohalo II in Israel. Associated evidence for an oven-like hearth was also found at this site, suggesting that dough made from grain flour was baked. Our data indicate that routine processing of a selected group of wild cereals, combined with effective methods of cooking ground seeds, were practiced at least 12,000 years before their domestication in southwest Asia.

Archaeology↗

Genetics and geography of wild cereal domestication in the near east.

About 12,000 years ago, humans began the transition from hunter-gathering to a sedentary, agriculture-based society. From its origins in the Near East, farming expanded throughout Europe, Asia and Africa, together with various domesticated plants and animals. Where, how and why agriculture originated is still debated. But newer findings, on the basis of genome-wide measures of genetic similarity, have traced the origins of some domesticated cereals to wild populations of naturally occurring grasses that persist in the Near East. A better understanding of the genetic differences between wild grasses and domesticated crops adds important facets to the continuing debate on the origin of Western agriculture and the societies to which it gave rise.

Agriculture↗

Effects of GI and content of indigestible carbohydrates of cereal-based evening meals on glucose tolerance at a subsequent standardised breakfast.

OBJECTIVE: To evaluate the impact of four low-glycaemic index (GI) and one high-GI cereal-based evening meals on glucose tolerance at a subsequent standardised breakfast. DESIGN: Wheat kernels, barley kernels, spaghetti, spaghetti with added wheat bran and white wheat bread (WWB) were consumed in the evening in a random order at five different occasions. At the subsequent breakfast, blood glucose, serum insulin, plasma short chain fatty acid, plasma free fatty acid (FFA) and breath hydrogen were measured. SETTING: The study was performed at Applied Nutrition and Food Chemistry, Lund University, Sweden. SUBJECTS: Fifteen healthy volunteers were recruited. One subject was later excluded owing to abnormal blood glucose values. RESULTS: The blood glucose response (0-120 min) to the standardised breakfast was significantly lower after consuming barley kernels in the evening compared with evening meals with WWB (P=0.019) or spaghetti+wheat bran (P=0.046). There were no significant differences in insulin concentrations at breakfast. Breath hydrogen excretion at breakfast was significantly higher after an evening meal with barley kernels compared with WWB, wheat kernels or spaghetti (P=0.026, 0.026 and 0.015, respectively), and the concentration of plasma propionate at breakfast was significantly higher following an evening meal with barley kernels compared with an evening meal with WWB (P=0.041). In parallel, FFA concentrations were significantly lower after barley kernels compared with WWB (P=0.042) or spaghetti evening meals (P=0.019). CONCLUSIONS: The improved glucose tolerance at breakfast, following an evening meal with barley kernels appeared to emanate from suppression of FFA levels, mediated by colonic fermentation of the specific indigestible carbohydrates present in this product, or, to the combination of the low-GI features and colonic fermentation.

Adult↗

Cereal grains and legumes in the prevention of coronary heart disease and stroke: a review of the literature.

UNLABELLED: A number of reviewers have examined studies investigating the relationship between coronary heart disease and stroke prior to 2000. Since then, several key studies have been published. Five studies have examined the relationship between wholegrain consumption, coronary heart disease (CHD) and cardiovascular (CVD) disease and found protection for either or both diseases. The researchers concluded that a relationship between wholegrain intake and CHD is seen with at least a 20% and perhaps a 40% reduction in risk for those who eat wholegrain food habitually vs those who eat them rarely. Notwithstanding the fact that fibre is an important component of wholegrains, many studies have not shown an independent effect of fibre alone on CHD events. Thus in terms of CHD prevention, fibre is best obtained from wholegrain sources. Wholegrain products have strong antioxidant activity and contain phytoestrogens, but there is insufficient evidence to determine whether this is beneficial in CHD prevention. Soluble fibre clearly lowers cholesterol to a small but significant degree and one would expect that this would reduce CHD events. There have been a small number of epidemiological studies showing soy consumption is associated with lower rates of heart disease. Countering the positive evidence for wholegrain and legume intake has been the Nurses Health Study in 2000 that showed women who were overweight or obese consuming a high glycaemic load (GL) diet doubled their relative risk of CHD compared with those consuming a low GL diet. Although the literature relating GL with CHD events is somewhat mixed, the relationship with risk factors such as HDL cholesterol, triglyceride and C reactive protein is relatively clear. Thus, carbohydrate-rich foods should be wholegrain and, if they are not, then the lowest glycaemic index (GI) product should be used. Promotion of carbohydrate foods should be focused on wholegrain cereals because these have proven to be associated with health benefits. There is insufficient evidence about whether the addition of other components of wholegrains such as polyphenolics or minerals (such as magnesium or zinc) would improve the health benefits of refined grain foods and this needs investigation. Whether adding bran to refined carbohydrate foods can improve the situation is also not clear, and it was found that added bran lowered heart disease risk in men by 30%. This persisted after full adjustment (including GL) suggesting, at least in men, that fibre may be more important than GI. Thus there are two messages: The intake of wholegrain foods clearly protects against heart disease and stroke but the exact mechanism is not clear. Fibre, magnesium, folate and vitamins B6 and vitamin E may be important. The intake of high GI carbohydrates (from both grain and non-grain sources) in large amounts is associated with an increased risk of heart disease in overweight and obese women even when fibre intake is high but this requires further confirmation in normal-weight women. RECOMMENDATION: Carbohydrate-rich foods should be wholegrain and if they are not, then the lowest GI product available should be consumed. Glycemic index is largely irrelevant for foods that contain small amounts of carbohydrate per serve (such as most vegetables).

Coronary Disease↗

Smoking, cereal consumption, and supplementation affect cadmium content in breast milk.

Breast milk might be a source of potentially toxic metals such as cadmium (Cd). The purpose of the present study is to provide data pertaining to the influence of maternal lifestyles on Cd concentrations in breast milk in the Austrian setting. Breast milk was obtained from 124 Austrian women. Each participant provided 10 ml of milk. A second group of eight mothers were recruited to investigate changes in their milk cadmium levels at 1, 3, 5, 7 and 9 weeks post partum. The study participants filled a questionnaire concerning nutrition, supplementation, and smoking habits. The samples were analyzed using GF-Atomic Absorption Spectrophotometer (AAS). The mean Cd content in breast milk was among the lowest in Europe (0.086 +/- 0.085 microg/l, 95% CI: 0.07-0.10; n = 124). Increased Cd levels in breast milk were found to be significantly associated with frequent cereal consumption and smoking. Smokers had a two-fold higher concentration than did non-smokers (0.15 versus 0.07 microg/l; P = 0.000). In contrast, low Cd levels in breast milk were associated with the intake of supplements containing trace elements or vitamins and trace elements (P < 0.05). This protective effect of supplements on Cd levels was only observed in non-smokers. The Cd levels registered in the present investigation were far below critical levels. We conclude that the current maternal Cd levels in Austria signify no risk for the breastfed infant of a healthy mother. Further research will have to focus on the specific effects of supplementation and smoking on Cd concentrations.

Adult↗

Decline in ribonucleic acid and protein synthesis with loss of viability during the early hours of imbibition of rye (Secale cereale L.) embryos.

A decline in protein synthesis and slow germination accompanies loss of viability in embryos of rye (Secale cereale L.) grains. Associated with this, incorporation of precursors into all the major classes of RNA is decreased and the processing of precursor rRNA to 25S and 18S RNA is retarded. Embryos that just reach 0% viability still synthesize some low-molecular-weight non-nucleolar material, although they do not synthesize protein. It is suggested that early-synthesized RNA could play a major part in determining the extent of protein synthesis at early germination, and thereby regulate the rate at which germination can proceed.

Plant Proteins↗

Isolation and partial characterization of wheat-germ-agglutinin-like lectins from rye (Secale cereale) and barley (Hordeum vulgare) embryos.

Lectins have been isolated from embryos of Secale cereale (rye) and Hordeum vulgare (barley) by affinity chromatography on immobilized N-acetylglucosamine. Both lectins are dimeric proteins of two identical subunits of mol.wt. 18000. They resemble strongly wheat-germ agglutinin with respect to their chemical, physical, biological and immunological properties.

Amino Acids↗

Homology between legumin-like polypeptides from cereals and pea.

The presence of legumin-like constituents within the globulin fractions of wheat (Triticum aestivum), rye (Secale cereale) and corn (maize, Zea mays) was demonstrated. Two-dimensional analysis of wheat globulins in the presence and absence of a reducing agent revealed the existence of reducible approximately 60 kDa polypeptides. Western-blot analysis with 125I-labelled antibodies raised against the oat (Avena sativa) 12S globulin holoprotein or its alpha-subunits demonstrated, firstly, the immunological homology between the alpha- and beta-subunits of pea (Pisum sativum) legumin and oat 12S globulin, and secondly, the similar occurrence in wheat of antigenically homologous approximately 20kDa and approximately 40 kDa polypeptides that associate via disulphide linkage to form approximately 60 kDa dimers. Western blotting also showed the presence of disulphide-linked approximately 20 kDa and approximately 40 kDa legumin-like subunits within the globulin fractions of rye and corn.

Edible Grain↗

The regulation of phosphatidylcholine biosynthesis in rye (Secale cereale) roots. Stimulation of the nucleotide pathway by low temperature.

The incorporation of [14C]choline chloride and [14C]glycerol into segments taken from rye (Secale cereale L., cv. Rheidal) roots was greater in segments from roots grown at 5 degrees C than in segments taken from roots growing at 20 degrees C. The incorporation was measured at the temperature at which the root had been growing. Measurements in vitro of the enzymes of the nucleotide pathway showed activity of choline kinase (EC 2.7.1.32), choline-phosphate cytidylyltransferase (EC 2.7.7.15) and cholinephosphotransferase (EC 2.7.8.2) to be higher in homogenates from the cooler roots when assayed at 5 degrees C than the activities assayed at 20 degrees C in the 20 degrees C-root homogenates. Changes in vivo in the pool sizes of the CDP-base intermediates with temperature, relative differences in nucleotide-pathway-enzyme activities and a pulse-chase experiment with [14C]choline indicated that the rate-limiting step for phosphatidylcholine biosynthesis in this tissue, at both temperatures, was the reaction catalysed by cytidylyltransferase.

Choline↗

Isolation of Bacillus strains from the rhizosphere of cereals and in vitro screening for antagonism against phytopathogenic, food-borne pathogenic and spoilage micro-organisms.

Bacillus strains were isolated from the rhizosphere of cereals in order to be used as natural biocontrol agents (BCAs). They were screened for antagonism in vitro against various test micro-organisms. The isolates showing antagonism were identified to species level. A combination of techniques was employed for the isolation of Bacillus species. Using the direct method, only one of the 25 isolates screened showed antagonistic properties. This strain (IFS-01) was identified by means of API test strips and the ATB Plus computer programme. It proved to be Bacillus subtilis and consequently has been designated as Bacillus subtilis IFS-01. This strain produced either a broad spectrum antimicrobial compound or several compounds with different activities. The fungi and Gram-positive bacteria were more sensitive to the antagonistic isolate than the Gram-negative bacteria. A Bacillus strain producing BCAs which can be used as biopesticides or organic preservatives has been isolated and identified.

Antibiosis↗

Identifying key cereal aphid predators by molecular gut analysis.

We describe polymerase chain reaction (PCR) primers for gut analysis of aphid predators. The primers amplify aphid mitochondrial COII fragments ranging in size from 77 to 386 bp. Using these primers, we were able to distinguish six species of US Great Plains cereal aphids, including two congeners, Rhopalosiphum maidis (Fitch) and R. padi (L.), and to detect them in extracts of coccinellid and chrysopid predators. We devised a protocol for deriving half-lives of detectability for the DNA of a single aphid consumed by predators maintained under simulated field dietary and temperature conditions. Using this protocol and primers that amplify a 198-bp fragment, we determined statistically different half-lives of detectability for a single R. maidis of 3.95 h in Chrysoperla plorabunda (Fitch) and 8. 78 h in Hippodamia convergens Guerin. The detectability half-life for a 339-bp R. maidis fragment was statistically longer in C. plorabunda but not in H. convergens. The sensitivity of the assay for the 198-bp fragment is 10-7 aphid equivalents. For species-specific predator gut analysis, PCR is superior to monoclonal antibody technology, giving comparable detectability half-lives with lower expense, much shorter development times, and greater certainty of a successful outcome.

Animals↗

Transgenic cereal (tritordeum) plants obtained at high efficiency by microprojectile bombardment of inflorescence tissue.

Transgenic cereal plants expressing the beta-glucuronidase (uidA) and neomycin phosphotransferase (neo) genes were obtained via microprojectile bombardment of immature inflorescence tissue of tritordeum (the fertile Hordeum x Triticum amphiploid, HchHchAABB). A total of 17 independent transgenic plants were recovered from 32 bombardments (on average four inflorescences per shot). Of the bombardment and culture parameters tested, explant preculture had the most influence on stable transformation frequency. The uidA and neo genes were supplied on two separate plasmids (co-transformation) and 88% of the transgenic plants recovered expressed both genes. Southern analysis confirmed the results of histochemical GUS and NPT II assays. Transgenic plants were grown to maturity and flowered and set seed. Pollen from four T0 GUS+ plants analysed showed GUS activity and T1 seedlings derived from one of the transgenic plants showed a segregation of 2.75:1 (GUS+:GUS-) for uidA gene activity.

Culture Techniques↗

Target genes and regulatory domains of the GAMYB transcriptional activator in cereal aleurone.

GAMYB is an MYB transcription factor which is expressed in cereal aleurone cells in response to gibberellin (GA). HvGAMYB binds to the TAACAAA box of a high-pl alpha-amylase gene promoter and transcriptionally activates its expression. In this study, we examined the role of HvGAMYB in activating expression of other GA-regulated genes encoding hydrolytic enzymes. In transient expression experiments, HvGAMYB transactivated expression of reporter genes fused to a low-pl alpha-amylase gene promoter, an EII (1-3, 1-4)-beta-glucanase gene promoter and a cathepsin B-like protease promoter. HvGAMYB DNA binding specificity was determined using a PCR-based random site selection using HvGAMYB fusion protein isolated from E. coli. The deduced consensus closely resembled gibberellin response elements in alpha-amylase promoters. Functional analysis of HvGAMYB by transient expression of C terminal HvGAMYB deletions in barley aleurone cells identified two transcriptional activation domains (TADs) which function in transcriptional regulation of both high- and low-pl alpha-amylase promoters. The same TADs were identified using a heterologous yeast expression system. Together, these results indicate that HvGAMYB has two TADs. These domains are C-terminal to its DNA-binding domain.

Base Sequence↗

Determination of phytase activity in cereal grains by direct incubation.

In contrast to supplemented microbial phytases, considerably lower cereal phytase activities were found after application of an enzyme solution extracted in citrate/NaOH buffer (pH 5.5) for 1 h as compared with the direct incubation of the plant material. Differences between both methods obtained were 70% for wheat and spelt and 50% for barley and rye. The determination of phytase activity of a wheat sample by direct incubation was affected by the sample size and amount of substrate (Na-phytate) added. Optimal conditions are a maximal level of phytase activity of < or = 0.05 U incubated for 60 min in a buffered solution (citrate/NaOH, pH 5.5) with 10 ml of 32.6 mmol/l Na-phytate at 37 composite function C. Possible reasons for the differences between both methods and for the factors affecting phytase activity by direct incubation are discussed.

6-Phytase↗

Cholesteryl ester enrichment of plasma low-density lipoproteins in hamsters fed cereal-based diets containing cholesterol.

Male Syrian hamsters were fed 0.02, 0.03, or 0.05% cholesterol to test the hypothesis that moderate cholesterol intake increases the cholesteryl ester content of the plasma low-density lipoproteins (LDL). Dietary cholesterol levels of 0.02%-0.05% were chosen to reflect typical human intakes of cholesterol. Hamsters were fed ad libitum a cereal-based diet (modified NIH-07 open formula) for 15 weeks. Increasing dietary cholesterol from 0.02% to 0.05% resulted in significantly increased plasma LDL and high-density lipoprotein cholesterol concentration, increased liver cholesterol concentration, and increased total aorta cholesterol content. The cholesteryl ester content of plasma LDL was determined as the molar ratio of cholesteryl ester to apolipoprotein B and to surface lipid (i.e., phospholipid + free cholesterol). Increasing dietary cholesterol from 0.02% to 0.05% resulted in significantly increased cholesteryl ester content of LDL particles. Furthermore, cholesteryl ester content of LDL was directly associated with increased total aorta cholesterol, whereas a linear relationship between plasma LDL cholesterol concentration and aorta cholesterol was not observed. Thus, the data suggest that LDL cholesteryl ester content may be an important atherogenic feature of plasma LDL.

Animals↗