Accumulation of mixed linkage (1 → 3) (1 → 4)-β-d-glucan during grain filling in barley: A vibrational spectroscopy study

The accumulation of mixed linkage barley (1 → 3) (1 → 4)-β-d-glucan (BG) during grain filling at eight stages was studied using standard reference methods and infrared spectroscopy. Two mutant barley genotypes having higher (starch mutant lys5f) and lower (high lysine mutant lys3a) BG content than the normal control Cork were studied. The Cork and lys3a genotypes showed a linear BG accumulation throughout the grain filling to reach a maximum of approximately 6 and 4% BG (w/w) dry matter, respectively. However, lys5f mutant exhibited an exponential increase in BG synthesis to a maximum of approximately 18% BG (w/w) dry matter 30 days after flowering (DAF), seemingly compensating for a decreased synthesis of starch.     

Barley (1→3; 1→4)-β-glucan and arabinoxylan content are related to kernel hardness and water uptake

Harder kernels in barley are thought to be a factor affecting the modification of the endosperm during malting by restricting water and enzyme movement within the endosperm. The objective of this study was to investigate the relationship between kernel hardness, water uptake and the endosperm composition in barley. A range of barley samples from 2003 and 2004 crops were analyzed for kernel hardness by the Single Kernel Characterization System, water uptake during steeping and chemical composition of the endosperm including (1→3; 1→4)-β-glucan, arabinoxylan and total protein. Both (1→3; 1→4)-β-glucan and arabinoxylan content of the endosperm were correlated significantly with kernel hardness in barley samples from both 2003 (r=0.873 and 0.601, respectively, p<0.01) and 2004 seasons (r=0.764 and 0.501, respectively, p<0.01). Hardness of the kernel was highly correlated with its water uptake in both 2003 and 2004 samples (r=−0.853 and −0.752, respectively, p<0.01). β-Glucan content of the endosperm was also correlated significantly with the kernel water uptake for both years (2003: r=−0.752, p<0.01; 2004: r=−0.551, p<0.01). Arabinoxylan content of the endosperm was correlated significantly with the kernel water uptake for the 2003 barley but not for 2004 barley (2003: r=−0.523, p<0.01; 2004: r=−0.151, p>0.01). Protein content of the endosperm was not correlated with the kernel hardness in either year. These results demonstrate that endosperm cell wall components may have significant impact on kernel hardness as well as water uptake of barley.     

大麦β 葡聚糖的提取及功能研究进展

大麦是世界上第四大禾谷类作物,其籽粒中的β-葡聚糖是重要的功效成分之一。本文综述了大麦籽粒β-葡聚糖的特性、含量变化和结构特征、提取方法和工艺,以及在大麦饲料、啤酒麦芽、饮料食品、保健医药中的功能研究进展。     

Characterization of Neoagarooligosaccharide Hydrolase BpGH117 from a Human Gut Bacterium Bacteroides plebeius

α-Neoagarobiose (NAB)/neoagarooligosaccharide (NAO) hydrolase plays an important role as an exo-acting 3,6-anhydro-α-(1,3)-L-galactosidase in agarose utilization. Agarose is an abundant polysaccharide found in red seaweeds, comprising 3,6-anhydro-L-galactose (AHG) and D-galactose residues. Unlike agarose degradation, which has been reported in marine microbes, recent metagenomic analysis of Bacteroides plebeius, a human gut bacterium, revealed the presence of genes encoding enzymes involved in agarose degradation, including α-NAB/NAO hydrolase. Among the agarolytic enzymes, BpGH117 has been partially characterized. Here, we characterized the exo-acting α-NAB/NAO hydrolase BpGH117, originating from B. plebeius. The optimal temperature and pH for His-tagged BpGH117 activity were 35 °C and 9.0, respectively, indicative of its unique origin. His-tagged BpGH117 was thermostable up to 35 °C, and the enzyme activity was maintained at 80% of the initial activity at a pre-incubation temperature of 40 °C for 120 min. K_m and V_max values for NAB were 30.22 mM and 54.84 U/mg, respectively, and k_cat/K_m was 2.65 s⁻¹ mM⁻¹. These results suggest that His-tagged BpGH117 can be used for producing bioactive products such as AHG and agarotriose from agarose efficiently.     

Lipid Inhibitory Effect of (−)-loliolide Isolated from Sargassum horneri in 3T3-L1 Adipocytes: Inhibitory Mechanism of Adipose-Specific Proteins

Sargassum horneri (S. horneri) is a well-known brown seaweed widely distributed worldwide. Several biological activities of S. horneri have been reported. However, its effects on lipid metabolism and the underlying mechanisms remain elusive. In the present study, we examined the inhibitory effect of the active compound “(−)-loliolide ((6S,7aR)-6-hydroxy-4,4,7a-trimethyl-5,6,7,7a-tetrahydro-1-benzofuran-2(4H)-one (HTT))” from S. horneri extract on lipid accumulation in differentiated adipocytes. MTT assays demonstrated that (−)-loliolide is not toxic to 3T3-L1 adipocytes in a range of concentrations. (−)-loliolide significantly reduced intracellular lipid accumulation in the differentiated phase of 3T3-L1 adipocytes as shown by Oil Red O staining. Western blot analysis revealed that (−)-loliolide increased the expression of lipolytic protein phospho-hormone-sensitive lipase (p-HSL) and thermogenic protein peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1). Additionally, (−)-loliolide decreased expression of adipogenic and lipogenic proteins, including sterol regulatory element-binding protein-1 (SREBP-1), peroxisome proliferator-activated receptor-γ (PPAR-γ), CCAAT/enhancer-binding protein-α (C/EBP-α), and fatty acid-binding protein 4 (FABP4) in 3T3-L1 adipocytes. These results indicate that (−)-loliolide from S. horneri could suppress lipid accumulation via regulation of antiadipogenic and prolipolytic mechanisms in 3T3-L1 cells. Considering the multifunctional effect of (−)-loliolide, it can be useful as a lipid-lowering agent in the management of patients who suffer from obesity.     

Near infrared spectra indicate specific mutant endosperm genes and reveal a new mechanism for substituting starch with (1→3,1→4)-β-glucan in barley

Near Infrared Reflectance spectroscopy was tested as a screening method to characterise high lysine mutants from a barley collection by classification through Principal Component Analysis (PCA). Mean spectra of the samples within each cluster identified gene-specific patterns in the 2270–2360 nm region. The characteristic spectral signatures representing the lys5 locus (Risø mutants 13 and 29) were found to be associated with large changes in percentage of starch and (1→3,1→4)-β-glucan. These alleles compensated for a low level of starch (down to 30%) by a high level of (1→3,1→4)-β-glucan (up to 15–20%), thus, maintaining a constant production of polysaccharides at 50–55%, within the range of normal barley.The spectral tool was tested by an independent data set with six mutants with unknown polysaccharide composition. Spectral data from four of these were classified within the high (1→3,1→4)-β-glucan BG lys5 cluster in a PCA. Their high (1→3,1→4)-β-glucan and low starch content was verified. It is concluded that genetic diversity such as from gene regulated polysaccharide and storage protein pathways in the endosperm tissue can be discovered directly from the phenotype by chemometric classification of a spectral library, representing the digitised phenome from a barley gene bank.     

A Collaborative Evaluation of LC-MS/MS Based Methods for BMAA Analysis: Soluble Bound BMAA Found to Be an Important Fraction

Exposure to β-N-methylamino-l-alanine (BMAA) might be linked to the incidence of amyotrophic lateral sclerosis, Alzheimer’s disease and Parkinson’s disease. Analytical chemistry plays a crucial role in determining human BMAA exposure and the associated health risk, but the performance of various analytical methods currently employed is rarely compared. A CYANOCOST initiated workshop was organized aimed at training scientists in BMAA analysis, creating mutual understanding and paving the way towards interlaboratory comparison exercises. During this workshop, we tested different methods (extraction followed by derivatization and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) analysis, or directly followed by LC-MS/MS analysis) for trueness and intermediate precision. We adapted three workup methods for the underivatized analysis of animal, brain and cyanobacterial samples. Based on recovery of the internal standard D₃BMAA, the underivatized methods were accurate (mean recovery 80%) and precise (mean relative standard deviation 10%), except for the cyanobacterium Leptolyngbya. However, total BMAA concentrations in the positive controls (cycad seeds) showed higher variation (relative standard deviation 21%–32%), implying that D₃BMAA was not a good indicator for the release of BMAA from bound forms. Significant losses occurred during workup for the derivatized method, resulting in low recovery (<10%). Most BMAA was found in a trichloroacetic acid soluble, bound form and we recommend including this fraction during analysis.     

Changes Caused by Genotype and Environmental Conditions in Beta-Glucan Content of Spring Barley for Dietetically Beneficial Human Nutrition

Over the 5-year period (2000-2004), a significantly higher beta-glucan content was detected in the waxy varieties Washonubet, Wabet, and Wanubet (6.8-7.6%) and lines formed by crossing these varieties with malting varieties (5.8-7.1%). Conversely, the non-waxy hulled malting-type varieties Kompakt (4.0%) and Krona (4.3%) had significantly lower contents of beta-glucan. The observations also showed that concentrations of beta-glucans in 2000-2004 were significantly affected not only by varieties, but also environmental conditions in the growing periods and interactions of these two factors. Higher precipitation during the flowering time and grain filling period and lower temperatures during the flowering time in 2002 had negative effects on concentration of beta-glucans. Conversely, drier and warmer weather in 2003 enhanced the content of beta-glucans. The results show that it is possible to increase the content of beta-glucan in spring barley grain by implementing selective breeding practices. Compared to the parental malting varieties, the mean content of beta-glucans in F(4)-F(8) generations was increased by 1.8 and 2.0% by recombination in lines Kompakt x Wabet and Wanubet x Krona, respectively. Significant effect of environmental conditions and their interactions with varieties indicated the necessity to assess standard qualities of barley as a food material.     

Physicochemical and functional characterization of wheat milling co-products: Fine grinding to achieve high fiber antioxidant-rich fractions

Milling of wheat produces co-products rich in dietary fiber, micronutrients and phytochemicals which can be used to integrate healthy functional foods. In the study different co-products including bran, shorts, and red dog were identified by physicochemical and functional analyses. The results showed that the fat, protein and starch contents decreased in order of red dog > shorts > bran (P < 0.05). The ash, neutral detergent fiber (NDF), acid detergent fiber (ADF), hemicellulose, water and oil holding capacities (WHC, OHC) were in order bran > shorts > red dog, respectively (P < 0.05). Antioxidant capacity was in order red dog > shorts > bran (P < 0.05). The bran was selected as the co-product with the highest fiber that was finely grounded to four different fractions (>355, 250–355, 180–250, <180 μm) and they were characterized more detail. The fat, protein and starch contents decreased with increasing bran particle size (P < 0.05). The ash, crude fiber, NDF, ADF, hemicellulose and WHC and OHC increased with the increasing bran particle size (P < 0.05). DPPH radical scavenging activity increased with increasing particle size (P < 0.05). The bran fractions 250–355 and >355 μm can be used as high fiber ingredients rich in antioxidants to generate functional foods.     

Efficient genetic transformation of Jatropha curcas L. by microprojectile bombardment using embryo axes

An efficient and reproducible protocol was established for genetic transformation in Jatropha curcas through microprojectile bombardment. Decotyledonated embryos from mature seeds were pre-cultured for 5 days and elongated embryonic axis was subjected to bombardment for the optimization of physical parameters. The frequency of transient gus expression and survival of putative transformants were taken into consideration for the assessment of physical parameters. Statistical analysis reveal that microcarrier size, helium pressure and target distance had significant influence on transformation efficiency. Among different variables evaluated, microcarrier size 1 μm, He pressure 1100 and 1350 psi with a target distance of 9 and 12 cm respectively were found optimum by co-relating microcarrier size, helium pressure and target distance on the frequency of gus expression and survival of putative transformants. Selection of putative transformants was done with increasing concentrations (5-7 mg L⁻¹) of hygromycin. The integration of desired gene into Jatropha genome was confirmed with PCR amplification of 0.96 and 1.28 kb bands of hptII and gus gene respectively from the T₀ transgenics and Southern blot analysis using PCR amplified DIG labeled hptII gene as a probe. A successful attempt of genetic transformation was made with optimized conditions using particle gene gun and establishing a stable transformation in J. curcas with 44.7% transformation efficiency. The procedure described will be very useful for the introgression of desired genes into J. curcas and the molecular analysis of gene function.     

Comparison of predictions of baking volume using large deformation rheological properties

Three large deformation rheological tests, the Kieffer dough extensibility system, the D/R dough inflation system and the 2 g mixograph test, were carried out on doughs made from a large number of winter wheat lines and cultivars grown in Poland. These lines and cultivars represented a broad spread in baking performance in order to assess their suitability as predictors of baking volume. The parameters most closely associated with baking volume were strain hardening index, bubble failure strain, and mixograph bandwidth at 10 min. Simple correlations with baking volume indicate that bubble failure strain and strain hardening index give the highest correlations, whilst the use of best subsets regression, which selects the best combination of parameters, gave increased correlations with R²=0.865 for dough inflation parameters, R²=0.842 for Kieffer parameters and R²=0.760 for mixograph parameters.     

Effects of genotype and environment on avenanthramides and antioxidant activity of oats grown in northwestern China

To investigate the effects of genotype and growing environment on avenanthramides and antioxidant activity of oats, the total phenolic content (TPC), avenanthramide compositions, and antioxidant activity were determined for whole oat from 39 cultivars grown in four locations in northwestern China (Inner Mongolia, Qinghai, Shanxi and Gansu). The results demonstrated that environment (E), genotype (G) and the interaction of these factors (E × G) significantly affected TPC, concentration of avenanthramide 2c (2c, ranged from 6.24 to 136.20 mg kg⁻¹), avenanthramide 2p (2p, 6.07–112.25 mg kg⁻¹), avenanthramide 2f (2f, 7.26–222.77 mg kg⁻¹) and antioxidant activity of oats. The effect of E was considerably greater than that of G or E × G. Based on G plus G × E interaction biplots (GGE biplots) analysis, the four locations in northwestern China can be divided into either two or three regions. Qinghai was determined to be distinct from the other locations. Cultivar Bayou 9 and oats grown in Gansu exhibited the highest TPCs, concentrations of 2c, 2p, 2f and antioxidant activity. The results suggest that oats containing more avenanthramides and exhibiting high levels of antioxidant activity could be obtained by selecting an appropriate genotype and growth location.     

The agronomic performance and nutritional content of oat and barley varieties grown in a northern maritime environment depends on variety and growing conditions

Warmer temperatures and increasing interest in high provenance food and drink products are creating new opportunities for cereal growing in northern Europe. Nevertheless, cultivation of oats and barley in these areas for malting and milling remains a challenge, primarily because of the weather, and there are few reports of their nutritional content from this region. In this study, trials in Orkney compared agronomic characteristics and nutritional content of recommended UK oat and barley varieties with Scandinavian varieties over three years. For a subset of varieties, nutritional content was compared with samples cultivated in more southerly sites. For Orkney, barley was considered a more suitable crop than oats because varieties matured earlier. In both crops, Scandinavian varieties matured earlier than UK varieties and some produced comparable yields. The range of values for macronutrients and minerals in oats and barley in Orkney were similar to those reported previously for other locations, but there were some significant differences attributable to variety and year. Compared with grain samples from more southerly locations, oats in Orkney had a significantly lower β-glucan and higher sodium content. The lower β-glucan may have resulted from higher rainfall and lower temperatures during the months of grain filling and maturation.