Down-regulation of the CSLF6 gene results in decreased (1,3;1,4)-beta-D-glucan in endosperm of wheat

(1,3;1,4)-beta-d-Glucan (beta-glucan) accounts for 20% of the total cell walls in the starchy endosperm of wheat (Triticum aestivum) and is an important source of dietary fiber for human nutrition with potential health benefits. Bioinformatic and array analyses of gene expression profiles in developing caryopses identified the CELLULOSE SYNTHASE-LIKE F6 (CSLF6) gene as encoding a putative beta-glucan synthase. RNA interference constructs were therefore designed to down-regulate CSLF6 gene expression and expressed in transgenic wheat under the control of a starchy endosperm-specific HMW subunit gene promoter. Analysis of wholemeal flours using an enzyme-based kit and by high-performance anion-exchange chromatography after digestion with lichenase showed decreases in total beta-glucan of between 30% and 52% and between 36% and 53%, respectively, in five transgenic lines compared to three control lines. The content of water-extractable beta-glucan was also reduced by about 50% in the transgenic lines, and the M(r) distribution of the fraction was decreased from an average of 79 to 85 x 10(4) g/mol in the controls and 36 to 57 x 10(4) g/mol in the transgenics. Immunolocalization of beta-glucan in semithin sections of mature and developing grains confirmed that the impact of the transgene was confined to the starchy endosperm with little or no effect on the aleurone or outer layers of the grain. The results confirm that the CSLF6 gene of wheat encodes a beta-glucan synthase and indicate that transgenic manipulation can be used to enhance the health benefits of wheat products.

Pollen ultrastructure in anther cultures of Datura innoxia. III. Incomplete microspore division

During the microspore division in Datura innoxia, the mitotic spindle is oriented in planes both perpendicular (PE) and oblique (OB) to the spore wall against which the nucleus is situated. However, irrespective of polarity, the usual type of hemispherical wall is laid down at cytokinesis and isolates the generative cell from the rest of the pollen grain (type A). In PE spores the vegetative nucleus initially occupies a central position in the pollen grain, whereas in OB spores the vegetative nucleus lies at the periphery of the grain close to the generative cell. In anther cultures initiated just before the microspore division is due to take place, no marked change can be observed in either orientation or symmetry of the mitotic spindle when the spores divide. In some, however, cytokinesis is disrupted and deposition of the hemispherical wall arrested. In the absence of a complete wall, differentiation of the generative cell cannot take place and binucleate pollen grains are formed having 2 vegetative-type nuclei (type B). The 2 nuclei in the B pollens are always situated against the pollen-grain wall, suggesting that the disruption phenomenon is related to the OB spores. The incomplete wall always makes contact with the intine on the intine-side of the spindle. Wall material may be represented merely as short stubs projecting out from the intine into the cytoplasm, in which event the 2 nuclei lie close to each other and are separated by only a narrow zone of cytoplasm. In other grains the wall is partially developed between the nuclei and terminates at varying distances from the tonoplast; in these, the nuclei are separated by a wider zone of cytoplasm. The significance of these binucleate grains in pollen embryogenesis is discussed.

Pollen ultrastructure in anther cultures of Datura innoxia. II. The generative-cell wall.

In young pollen grains of Datura innoxia, a wall of the usual hemispherical type separates the 2 gametophytic cells initially and, in the electron microscope, appears as an electron-translucent matrix which is contiguous with the intine. Before detachment of the generative cell from the intine, the matrix decreases in thickness and in places is dispersed altogether leaving the plasmalemmae on either side of it in close apposition. A particularly prominent zone, triangular in profile, is left where the wall joins with the intine. After detachment of the cell, remnants of the matrix can be seen distributed irregularly around the cell and it is supposed that these are partly derived from material in the triangular zone as the cell is drawn away from the intine. The wall residues persist throughout the maturation phase of the pollen and are considered to be either callose resulting from incomplete digestion of the initial wall, or some other polysaccharide material which is unevenly laid down along the wall and concentrated at the junction with the intine. In pollen induced into embryogenesis by anther culture, wall material is also distributed irregularly around the detached cell in a series of discrete zones, but these are more extensive than in vivo, closer together and in many instances highly dilated. The wall profiles thus have a beaded appearance, the 'beads' being connected together by short links of the 2 apposed plasmalemmae. The contents of the swollen zones have a similar electron density to that of the matrix in vivo but also show traces of a fibrillar component. It is postulated that this unusual swelling is a prelude to dispersal of the wall by disruption of the plasmalemmal links and to the establishment of cytoplasmic continuity between the 2 cells. The significance of such binucleate pollen grains in the formation of non-haploid embryos is discussed.

The effect of incremental replacement of wheat with soya hulls in diets for Jersey cows on lactational performance, diet digestibility and feeding behaviour

Eight Jersey cows were used in two balanced 4 x 4 Latin Squares to investigate the effects of replacement of dietary starch with non-forage fibre on productivity, diet digestibility and feeding behaviour. Total-mixed rations consisted of maize silage, grass silage and a soyabean meal-based concentrate mixture, each at 250g/kg DM, with the remaining 250g consisting of cracked wheat/soya hulls (SH) in the ratios of 250:0, 167:83; 83:167 and 0:250 g, respectively, for treatments SH0, SH83, SH167 and SH250. Starch concentrations were 302, 248, 193 and 140g/kg DM, and NDF concentrations were 316, 355, 394 and 434g/kg DM, for treatments SHO, SH83, SH167 and SH250, respectively. Total eating time increased (p < 0.05) as SH inclusion increased, but total rumination time was unaffected. Digestibility of DM, organic matter and starch declined (p < 0.01) as SH inclusion increased, whilst digestibility of NDF and ADF increased (p < 0.01). Dry-matter intake tended to decline with increasing SH, whilst bodyweight, milk yield and fat and lactose concentrations were unaffected by treatment. Milk protein concentration decreased (p < 0.01) as SH level increased. Feed conversion efficiency improved (p < 0.05) as SH inclusion rose, but it was not possible to determine whether this was due to the increased fibre levels alone, or the favourable effect on rumen fermentation of decreasing starch levels. (c) 2006 Elsevier B.V. All rights reserved.

Dietary supplements of whole linseed and vitamin E to increase levels of alpha-linolenic acid and vitamin E in bovine milk

The potential to increase the concentrations of n-3 polyunsaturated fatty acids (PUFAs) in milk fat was investigated by studying the effects of feeding a xylose-treated, whole cracked linseed supplement ( rich in alpha-linolenic acid) to dairy cows. Also the effect of increasing the dietary intake of vitamin E on the vitamin E status of milk was investigated. The effect of pasteurisation on milk fatty acid composition was also examined. Using a 3 x 2 factorial design, a total of 60 Holstein dairy cows were fed a total mixed ration based on grass silage supplemented with one of three levels of whole cracked linseed (78, 142 or 209 g . kg(-1) diet dry matter (DM); designated LL, ML or HL, respectively) in combination with one of two levels of additional dietary vitamin E intake ( 6 or 12 g vitamin E . animal(-1) . day(-1); designated LE or HE, respectively). Increasing lipid supplementation reduced (P < 0.01) diet DM intake and milk yield, and increased (P < 0.001) the overall content of oleic, vaccenic, alpha-linolenic and conjugated linoleic acids, and total PUFAs and monounsaturated fatty acids (MUFA). Myristic and palmitic acids in milk fat were reduced ( P < 0.001) through increased lipid supplementation. While α-linolenic acid concentrations were substantially increased this acid only accounted for 0.02 of total fatty acids in milk at the highest level of supplementation (630 g α-linolenic acid &BULL; animal(-1) &BULL; day(-1) for HL). Conjugated linoleic acid concentrations in milk fat were almost doubled by increasing the level of lipid supplementation (8.9, 10.4 and 16.1 g &BULL; kg(-1) fatty acids for LL, ML and HL, respectively). Although milk vitamin E contents were generally increased there was no benefit (P > 0.05) of increasing vitamin E intake from 6 to 12 g . animal(-1) . day(-1). The fatty acid composition of milk was generally not affected by pasteurisation.

Whole crop cereals 1. Effect of method of harvest and preservation on chemical composition, apparent digestibility and energy value

A total of 133 samples (53 fermented unprocessed, 19 fermented processed. 62 urea-treated processed) of whole crop wheat (WCW) and 16 samples (five fermented unprocessed, six fermented processed, five urea-treated processed) of whole crop barley (WCB) were collected from commercial farms over two consecutive years (2003/2004 and 2004/2005). Disruption of the maize grains to increase starch availability was achieved at the point of harvest by processors fitted to the forage harvesters. All samples were subjected to laboratory analysis whilst 50 of the samples (24 front Year 1, 26 front Year 2 all WCW except four WCB in Year 2) were subjected to in vivo digestibility and energy value measurements using mature wether sheep. Urea-treated WCW had higher (P<0.05) pH, and dry matter (DM) and crude protein contents and lower concentrations of fermentation products than fermented WCW. Starch was generally lower in fermented, unprocessed WCW and no effect of crop maturity at harvest (as indicated by DM content) on starch concentrations was seen. Urea-treated WCW had higher (P<0.05) in vivo digestible organic matter contents in the DM (DOMD) in Year 1 although this was not recorded in Year 2. There was a close relationship between the digestibility values of organic matter and gross energy thus aiding the use of DOMD to predict metabolisable energy (ME) content. A wide range of ME values was observed (WCW. 8.7-11.8 MJ/kg DM; WCB 7.9-11.2 MJ/kg DM) with the overall ME/DOMD ratio (ME = 0.0156 DOMD) in line With Studies in other forages. There was no evidence that a separate ME/DOMD relationship was needed for WCB which is helpful for practical application. This ratio and other parameters were affected by year of harvest (P<0.05) highlighting the influence of environmental and Other undefined factors. The variability in the composition and nutritive value of WCW and WCB highlights the need for reliable and accurate evaluation methods to be available to assess the Value of these forages before they are included in diets for dairy cows. (C) 2008 Elsevier B.V. All rights reserved.

Effects of dietary vitamin A concentration of roasted soybean inclusion on marbling, adipose cellularity, and fatty acid composition of beef

A feedlot trial was conducted to determine the effect of dietary vitamin A concentration and roasted soybean (SB) inclusion on carcass characteristics, adipose tissue cellularity, and muscle fatty acid composition. Angus-crossbred steers (n = 168; 295 +/- 1.8 kg) were allotted to 24 pens (7 steers each). Four treatments, in a 2 x 2 factorial arrangement, were investigated: no supplemental vitamin A, no roasted soybeans (NANS); no vitamin A, roasted SB (20% of the diet on a DM basis; NASB); with supplemental (2,700 IU/kg) vitamin A, no roasted SB (WANS); and with supplemental vitamin A, roasted SB (WASB). Diets included high moisture corn, 5% corn silage, 10 to 20% supplement, and 20% roasted SB in the SB treatments on a DM basis. The calculated vitamin A concentration in the basal diet was < 1,300 IU/kg of DM. Blood samples (2 steers/pen) were collected for serum vitamin A determination. Steers were slaughtered after 168 d on feed. Carcass characteristics and LM composition were determined. Fatty acid composition of LM was analyzed, and adipose cellularity in the i.m. and s.c. depots was determined. No vitamin A x SB interactions were detected (P > 0.10) for cattle performance, carcass composition, or muscle fatty acid composition. Low vitamin A diets (NA) did not affect (P > 0.05) ADG, DMI, or G:F. Quality grade tended (P = 0.07) to be greater in NA steers. Marbling scores and the percentage of carcasses grading > or = Choice(-) were 10% greater for NA steers, although these trends were not significant (P = 0.11 and 0.13, respectively). Backfat thickness and yield grade were not affected (P > 0.26) by vitamin A supplementation. Composition of the LM was not affected (P > 0.15) by vitamin A or SB supplementation. Serum retinol at slaughter was 44% lower (P < 0.01) for steers fed NA than for steers supplemented with vitamin A (23.0 vs. 41.1 microg/dL). A vitamin A x SB interaction occurred (P < 0.05) for adipose cellularity in the i.m. depot; when no SB was fed, vitamin A supplementation decreased cell density and increased cell size. However, when SB was fed, vitamin A supplementation did not affect adipose cellularity. Adipose cellularity at the s.c. depot was not affected (P > 0.18) by vitamin A or SB treatments. Fatty acid profile of the LM was not affected by vitamin A (P > 0.05), but SB increased (P < 0.05) PUFA (7.88 vs. 4.30 g/100 g). It was concluded that feeding NA tended to increase marbling without affecting back-fat and yield grade. It appeared that NA induced hyperplasia in the i.m. but not in the s.c. fat depot.

Pea-barley intercropping and short-term subsequent crop effects across European organic cropping conditions

Grain legumes are known to increase the soil mineral nitrogen (N) content, reduce the infection pressure of soil borne pathogens, and hence enhance subsequent cereals yields. Replicated field experiments were performed throughout W. Europe (Denmark, United Kingdom, France, Germany and Italy) to asses the effect of intercropping pea and barley on the N supply to subsequent wheat in organic cropping systems. Pea and barley were grown either as sole crops at the recommended plant density (P100 and B100, respectively) or in replacement (P50B50) or additive (P100B50) intercropping designs. In the replacement design the total relative plant density is kept constant, while the additive design uses the optimal sole crop density for pea supplementing with 'extra' barley plants. The pea and barley crops were followed by winter wheat with and without N application. Additional experiments in Denmark and the United Kingdom included subsequent spring wheat with grass-clover as catch crops. The experiment was repeated over the three cropping seasons of 2003, 2004 and 2005. Irrespective of sites and intercrop design pea-barley intercropping improved the plant resource utilization (water, light, nutrients) to grain N yield with 25-30% using the Land Equivalent ratio. In terms of absolute quantities, sole cropped pea accumulated more N in the grains as compared to the additive design followed by the replacement design and then sole cropped barley. The post harvest soil mineral N content was unaffected by the preceding crops. Under the following winter wheat, the lowest mineral N content was generally found in early spring. Variation in soil mineral N content under the winter wheat between sites and seasons indicated a greater influence of regional climatic conditions and long-term cropping history than annual preceding crop and residue quality. Just as with the soil mineral N, the subsequent crop response to preceding crop was negligible. Soil N balances showed general negative values in the 2-year period, indicating depletion of N independent of preceding crop and cropping strategy. It is recommended to develop more rotational approaches to determine subsequent crop effects in organic cropping systems, since preceding crop effects, especially when including legumes, can occur over several years of cropping.

Digestion, rumen fermentation and circulating concentrations of insulin, growth hormone and IGF-1 in steers given maize silages harvested at three stages of maturity

Advancing maturity of forage maize is associated with increases in the proportion of dry matter (DM) and starch and decreases in the proportions of structural carbohydrates in the ensiled crop. Three maize silages (286 (low, L), 329 (medium, M) and 379 (high, H) g DM per kg fresh weight) plus a concentrate formulated to give isonitrogenous intakes were offered to Holstein-Friesian steers fitted with a cannula in the dorsal sac of the rumen and a 'T' piece cannula in the proximal duodenum in an experiment with a cross-over design that allowed four collection periods. Nutrient flow to the duodenum was estimated using chromium-EDTA. Steers consumed approximately 0(.)6 kg DM per day less of diet L compared with the other two diets (P=0(.)026), resulting in less DM being digested (P=0(.)005) but digestibility did not differ between diets. Similar results were obtained for organic matter. There were no differences between diets in the intake or digestibility of neutral-detergent fibre. Intake, duodenal flow and faecal output of starch were greater for steers offered diets M and H compared with those given diet L (P < 0(.)05). In all diets rumen digestion contributed to over 90% of total digestion of starch, although rumen digestibility declined significantly with advancing maize maturity (P=0(.)002). Molar proportions of acetic acid were higher in diet H (P < 0(.)05) whilst proportions of propionic acid and n-butyric acid were higher in diets M and L. There were no significant differences between diets in mean rumen pH or ammonia concentrations. Mean circulating concentrations of insulin were higher (P=0(.)009) in cattle given diets L and M compared with diet H. There were no differences between diets in the mean circulating concentration of growth hormone, or the frequency, amplitude and duration of growth hormone pulses, or the mean circulating concentrations of IGF-1. Changes in forage composition that accompany advancing maize maturity affect overall silage digestion and circulating concentrations of insulin.

Effect of energy and protein supplementation on phosphorus utilization in lactating dairy cows

Two experiments were undertaken in which grass silage was used in conjunction with a series of different concentrate types designed to examine the effect of carbohydrate source, protein level and degradability on total dietary phosphorus (P) utilization with emphasis on P pollution. Twelve Holstein-Friesian dairy cows in early to mid-lactation were used in an incomplete changeover design with four periods consisting of 4 weeks each. Phosphorus intake ranged from 54 to 80 g/day and faecal P represented the principal route by which ingested P was disposed of by cows, with insignificant amounts being voided in urine. A positive linear relationship between faecal P and P intake was established. In Experiment 1, P utilization was affected by dietary carbohydrate type, with an associated output of 3.3 g faecal P/g milk P produced for all treatments except those utilizing low degradable starch and low protein supplements, where a mean value of 2.8 g faecal P/g milk P was observed. In Experiment 2, where two protein levels and three protein degradabilities were examined, the efficiency of P utilization for milk P production was not affected by either level or degradability of crude protein (CP) but a significant reduction in faecal P excretion due to lower protein and P intake was observed. In general, P utilization in Experiment 2 was substantially improved compared to the Experiment 1, with an associated output of 1.8 g faecal P/g milk P produced. The improved utilization of P in Experiment 2 could be due to lower P content of the diets offered and higher dry matter (DM) intake. For dairy cows weighing 600 kg, consuming 17-18 kg DM/day and producing about 25 kg milk, P excretion in faeces and hence P pollution to the environment might be minimized without compromising lactational performance by formulating diets to supply about 68 g P/day, which is close to recent published recommended requirements for P.

Effects of contrasting forage diets on phosphorus utilisation in lactating dairy cows

The effects of different forage mixtures on duodenal flow and faecal output of phosphorus (P) were measured in lactating dairy cows in two experiments. The forages comprised grass silage (GS) alone or mixtures of GS and urea-treated whole crop wheat (WCW). Replacement of GS by high levels of WCW resulted in a significant decrease in total P (TP), water-soluble P (WSP) and acid-soluble P (ASP) intakes, but increased phytate P (PP) and acid-insoluble P (AIP) intakes. Duodenal flows of all P fractions were unaffected by dietary treatments. However, the amounts of all fractions flowing to the intestine were much higher than the amounts consumed-with the exception of AIP, which was unchanged, and PP, which was considerably reduced. Diets did not significantly affect faecal outputs of TP, ASP, PP and AIP; however, faecal outputs of WSP were significantly higher on WCW than GS diets. Inclusion of WCW significantly decreased TP and WSP availabilities in the total tract but did not affect PP and AIP availabilities. A new fraction of P, which was not present in the feeds, was found in significant amounts in the intestines and proved to be soluble in dilute acid but insoluble in water. Estimated annual excretions for a herd of 100 cows ranged from 1550 to 1721 kg for total P of which 337-786 kg was WSP. The results suggest that GS based diets deposit more P in the environment. However, the potential to cause P pollution depends not only on the quantity but also on the nature of faecal P and on soil parameters. © 2004 Elsevier B.V. All rights reserved.

Nitrogen fertilizer and seed rate effects on Hagberg failing number of hybrid wheats and their parents are associated with alpha-amylase activity, grain cavity size and dormancy

Field experiments were carried out to assess the effects of nitrogen fertilization and seed rate on the Hagberg falling number (HFN) of commercial wheat hybrids and their parents. Applying nitrogen (200 kg N ha(-1)) increased HFN in two successive years. The HFN of the hybrid Hyno Esta was lower than either of its parents (Estica and Audace), particularly when nitrogen was not applied. Treatment effects on HFN were negatively associated with a-amylase activity. Phadebas grain blotting suggested two populations of grains with different types of a-amylase activity: Estica appeared to have a high proportion of grains with low levels of late maturity endosperm a-amylase activity (LMEA); Audace had a few grains showing high levels of germination amylase; and the hybrid, Hyno Esta, combined the sources from both parents to show heterosis for a-amylase activity. Applying nitrogen reduced both apparent LMEA and germination amylase. The effects on LMEA were associated with the size and disruption of the grain cavity, which was greater in Hyno Esta and Estica and in zero-nitrogen treatments. External grain morphology failed to explain much of the variation in LMEA and cavity size, but there was a close negative correlation between cavity size and protein content. Applying nitrogen increased post-harvest dormancy of the grain. Dormancy was greatest in Estica and least in Audace. It is proposed that effects of seed rate, genotype and nitrogen fertilizer on HFN are mediated through factors affecting the size and disruption of the grain cavity and therefore LMEA, and through factors affecting dormancy and therefore germination amylase. (c) 2004 Society of Chemical Industry.

The interrelationships of winter wheat cultivars, crop density and competition of naturally occurring weed flora

The effects of intraspecific and interspecific competition on a wide range of winter wheat cultivars were investigated in two consecutive split plot field experiments. Significant reductions of grain yield at greatly reduced seed rates were observed in the first experiment, whereas increasing crop density up to 380 plants m(-2) in the second experiment failed to produce a significant yield response due to compensation through increased ears and grains per plant at lower crop densities. Appreciable weed suppression and acceptable grain yield can be achieved at crop densities between 150 and 270 plants m(-2). Reductions in final yield due to weed competition occurred in both experiments; 11.7 and 13.6% for the first and second experiment, respectively, with the onset of weed competition occurring from tittering in the first experiment and from stem elongation in the second. The possibility of enhancing crop competitiveness for weed suppression and improved grain yield is discussed.

Dry matter intake is decreased more by abomasal infusion of unsaturated free fatty acids than by unsaturated Triglycerides

Previous experiments from our group have demonstrated that abomasal infusion of unsaturated free fatty acids (FFA) markedly decreases dry matter intake (DMI) in dairy cows. In contrast, experiments from other groups have noted smaller decreases in DMI when unsaturated triglycerides (TG) were infused postruminally. Our hypothesis was that unsaturated FFA would be more potent inhibitors of DMI than an equivalent amount of unsaturated TG. Four Holstein cows in late lactation were used in a single reversal design. Cows were fed a total mixed ration containing (DM basis) 23% alfalfa silage, 23% corn silage, 40.3% ground shelled corn, and 10.5% soybean meal. Two cows received soy FFA (UFA; 0, 200, 400, 600 g/d) and 2 received soy oil (TG) in the same amounts; cows then were switched to the other lipid source. Cows were abomasally infused with each amount for 5-d periods. The daily amount of lipid was pulse-dosed in 4 equal portions at 0600, 1000, 1700, and 2200 h; no emulsifiers were used and there was no sign of digestive disturbance. Both lipid sources linearly decreased DMI, with a significant interaction between lipid source and amount. Slope-ratio analysis indicated that UFA were about 2 times more potent in decreasing DMI than were TG. Decreased DMI led to decreased milk production. Milk fat content was increased linearly by lipid infusion. Milk fat yield decreased markedly for UFA infusion but was relatively unaffected by infusion of TG. Contents of short- and medium-chain fatty acids in milk fat decreased as the amount of either infusate increased. Contents of C-18:2 and C18: 3 in milk fat were increased linearly by abomasal infusion of either fat source; cis-9 C-18:1 was unaffected. Transfer of infused C18: 2 to milk fat was 35.6, 42.5, and 27.8% for 200, 400, and 600 g/d of UFA, and 34.3, 39.6, and 34.0% for respective amounts of TG. Glucagon-like peptide-1 (7-36) amide (GLP-1) concentration in plasma significantly increased as DMI decreased with increasing infusion amount of UFA or TG. Plasma concentration of cholecystokinin-octapeptide (CCK-8) was unaffected by lipid infusion. These results indicate that unsaturated FFA reaching the duodenum are more potent inhibitors of DMI than are unsaturated TG; the effect may be at least partially mediated by GLP-1.

Effects of essential oils on ruminal microorganisms and their protein metabolism

A commercial blend of essential oil (EO) compounds was added to a grass, maize silage, and concentrate diet fed to dairy cattle in order to determine their influence on protein metabolism by ruminal microorganisms. EO inhibited (P < 0.05) the rate of deamination of amino acids. Pure-culture studies indicated that the species most sensitive to EO were ammonia-hyperproducing bacteria and anaerobic fungi.