Analysis of the siRNA-Mediated Gene Silencing Process Targeting Three Homologous Genes Controlling Soybean Seed Oil Quality.

In the past decade, RNA silencing has gained significant attention because of its success in genomic scale research and also in the genetic improvement of crop plants. However, little is known about the molecular basis of siRNA processing in association with its target transcript. To reveal this process for improving hpRNA-mediated gene silencing in crop plants, the soybean GmFAD3 gene family was chosen as a test model. We analyzed RNAi mutant soybean lines in which three members of the GmFAD3 gene family were silenced. The silencing levels of FAD3A, FAD3B and FAD3C were correlated with the degrees of sequence homology between the inverted repeat of hpRNA and the GmFAD3 transcripts in the RNAi lines. Strikingly, transgenes in two of the three RNAi lines were heavily methylated, leading to a dramatic reduction of hpRNA-derived siRNAs. Small RNAs corresponding to the loop portion of the hairpin transcript were detected while much lower levels of siRNAs were found outside of the target region. siRNAs generated from the 318-bp inverted repeat were found to be diced much more frequently at stem sequences close to the loop and associated with the inferred cleavage sites on the target transcripts, manifesting "hot spots". The top candidate hpRNA-derived siRNA share certain sequence features with mature miRNA. This is the first comprehensive and detailed study revealing the siRNA-mediated gene silencing mechanism in crop plants using gene family GmFAD3 as a test model.

Re-evaluation of peroxide value as an indicator of the quality of edible oils.

The oxidation of oils has important effects on the quality of oily foods, such as instant noodles. In particular, the generation of aldehydes, which accompanies the oxidation of oils, is one of the first factors to reduce food quality. We examined various indicators of oil quality during temperature-accelerated storage and found that peroxide value (POV) was highly correlated with the total concentration of major odorants. Moreover, the correlation of POV with the total concentration of five unsaturated aldehydes (t-2-heptenal, t-2-octenal, t-2-decenal, t-2-undecenal and t,t-2,4-decadienal) that show strong cytotoxicity was greater than the correlation of POV with the total concentration of major odorants. The maximum allowable concentration of the five aldehydes was calculated based on the 'no observed adverse-effect level' of the aldehyde that showed the highest cytotoxicity, t,t-2,4-decadienal, along with the human daily oil intake. We showed that it is useful to utilize POV as an indicator to control food quality and safety.

Application of catalytic ozone chemistry for improving biodiesel product performance.

Ozonolysis of methyl soyate (biodiesel) was conducted in the presence of methanol, dichloromethane (solvent), and triethylamine (catalyst) at -75 degrees C. Structural analysis, including FTIR, GC, and GC-MS, showed that the total amount of double bonds in the mixture was reduced by more than 90% after 2 h of ozonolysis. All of the esters predicted by this novel application of ozone reaction chemistry were successfully produced. Other major components were identified by GC-MS. Thermogravimetric analysis showed a dramatic decrease in the onset volatilization temperature from 135 to 73 degrees C, making ozonated biodiesel fuel comparable to diesel fuel (76 degrees C). Differential scanning calorimetric studies showed that the cooling curves for both methyl soyate and ozonated methyl soyate displayed two exothermic regions. The onset freezing temperature of ozonated methyl soyate in the "colder" region was significantly reduced from -63 to -86 degrees C. Furthermore, the degree of crystallinity in the "hotter" region was also reduced.

Tissue alpha-tocopherol retention in male rats is compromised by feeding diets containing oxidized frying oil.

To investigate the effect of dietary oxidized frying oil (OFO) on tissue retention of alpha-tocopherol (alpha-T). Long-Evans male weanling rats were divided to four groups based on a 2 x 2 factorial design. Two groups were fed 15% OFO diets, and the remaining two groups were fed control diets in which OFO was replaced by vitamin-E-stripped fresh soybean oil. Vitamin E as all-rac-alpha-tocopheryl acetate was added at the concentration of either 50 (normal E) or 500 (high E) mg/kg diet. The OFO sample was prepared by deepfrying sticks in fresh soybean oil at 205 +/- 5 degrees C for four 6-h periods. After 6 wks of feeding, alpha-T concentrations in most tissues were significantly lower in rats fed OFO diets (P < 0.05) than in the control groups. For rats fed the OFO diet with the normal vitamin E concentration, the alpha-T concentration is epididymal fat pad, plasma, liver, kidney, muscle, brain and lung were 29-64% those of the corresponding control group (P < 0.05). The interaction between the two dietary factors on tissue alpha-T was significant in liver, spleen, and adrenal gland. In these three tissues, the differences between the normal and high dietary vitamin E groups were less in rats fed the OFO diets than in rats fed the control diets. The tissue alpha-T concentrations of the high vitamin E OFO group were comparable with or higher (P < 0.05) than those of the normal vitamin E control group, indicating that the negative effect of OFO on tissue alpha-T concentration can be alleviated by dietary supplementation of vitamin E. Compared with the controls, rats OFO diets had significantly higher tissue thiobarbituric acid reactive substances (P < 0.05). Because the amount of alpha-T directly added into the test oil samples was not significantly decreased through an incubation (at 37 degrees C) period of up to 10 d, the inefficient absorption and/or enhanced catabolism or turnover of vitamin E may be involved in the inferior tissue alpha-T retention of OFO fed rats.

Effect of adding fat and(or) milk products to the weanling pig diet on performance in the nursery and subsequent grow-finish stages.

Six experiments were conducted to evaluate the use of fat in diets containing a high level of milk products for weanling pigs. In Exp. 1, 192 pigs (6.6 kg and 23 d) were used to determine whether milk products (0 or 40%) in the diet influenced the utilization of fat (0 or 10%). No fat x milk product interactions were found. Adding milk products to the diet improved (P < .01) ADG, ADFI, and gain/feed (G/F). Adding fat to the diet did not influence performance. In Exp. 2, 3, and 4, 576 pigs (6.0 kg and 20 d) were used to determine the influence of fat level on performance. Adding soybean oil (0, 3, 6, or 9%) to the diet from d 0 to 14 after weaning had no influence on growth performance. Increasing soybean oil (0, 2, 4, or 6%) in the diet from d 14 to 35 had no influence on ADG; however, G/F improved linearly (P < .001). In Exp. 5, 196 pigs (7.5 kg and 26 d) were used to determine the influence of fat source (soybean oil, corn oil, or tallow) on performance. Pig performance was not different among fat sources. In Exp. 6, 240 pigs (5.4 kg and 21 d) were used to determine the influence of fat and(or) milk product inclusion in the nursery diet on subsequent grow-finish performance. Adding milk products, but not fat, to the nursery diet improved (P < .08) overall performance to market weight. These results indicate adding fat to the diet from d 0 to 14 after weaning had no influence on performance. Adding fat to the diet from d 14 to 35 improved G/F; however, it did not improve ADG or overall performance to market. Diet composition during the nursery period can affect subsequent performance.

Dust suppression in swine feed using soybean oil.

Dust generation from swine feed (corn-soybean meal diet) treated with soybean oil was investigated using a cement mixer in a plywood box. Airborne total dust concentrations in the box were measured gravimetrically using a vacuum pump and filters while 12-kg feed samples were constantly mixed. The treatment factors were soybean oil concentration (0, 1, and 3%), corn bulk density (normal, 730 kg/m3 and low, 600 kg/m3), time of oil addition (before vs after grinding the corn), and storage time (0, 7, and 14 d). The feed was a mixture of soybean meal, base mix, and adjusted amounts of ground corn and soybean oil. Adding soybean oil after grinding at 1 and 3% levels to feed made with normal-bulk density corn suppressed total dust generation (3.39 and .99 mg/m3, respectively) (P < .001) compared with the no oil treatment (29.1 mg/m3). The 3% soybean oil treatment suppressed dust generation (.99 mg/m3) more than the 1% soybean oil treatment (3.39 mg/m3) (P < .001). Adding soybean oil after grinding the corn suppressed dust generation more than adding the oil to the corn before grinding for every oil level (P < .001). More dust was generated by feed made from low-bulk density corn than by feed made with normal-bulk density corn at every oil level (P < .001). There was no evidence of any storage time or treatment x storage time interactions.