The level of 1C diets fed prior to cell isolation affects lipid metabolism in primary liver cells isolated from Atlantic salmon (Salmo salar)

Liver cells were isolated from 6 fish fed a diet containing 12.1 g methionine/kg, 11.02 mg vitamin B6/kg, 0.20 mg vitamin B12/kg and 7.80 mg folate/kg (named high‐1C diet). These cells were compared to liver cells isolated from 6 fish fed a diet containing 6.7 g methionine/kg, 7.01 g vitamin B6/kg, 0.15 mg vitamin B12/kg and 2.60 mg folate/kg (named low‐1C diet). Isolated cells were plated on 6‐well plates in Leibovitz medium and treated with 10 mM metformin, 10 mM metformin for 24 hr followed by 0.4 mM oleic acid (OA) for 24 hr or only 0.4 mM OA for 24 hr. The cells were compared to untreated controls added only the medium. All cells were harvested 48 hr after being plated. Cells isolated from Atlantic salmon fed low‐1C diets showed higher gene expression of MGAT‐2 (p < .0001), CPT‐1 (p = .028), FAS (p = .0006), LXR (p = .020), ACC (p = .032) and MnSOD (p < .0001). The low‐ or high‐1C diets fed prior to cell isolation had no effect on gene expression of ApoB100, PPARa, CD36, SREBP‐2 or Bcl‐2. Metformin treatment increased the expression of the anti‐apoptotic protein Bcl‐2 (p = .0001) indicating an anti‐apoptotic effect. Metformin generally increased the expression of genes associated with lipid oxidation and transport, but decreased the expression of genes associated with cholesterol metabolism confirming our earlier results using this model.     

Carotenoid accumulation in Japanese apricot (Prunus mume Siebold & Zucc.): molecular analysis of carotenogenic gene expression and ethylene regulation

To elucidate the regulatory mechanisms of carotenogenesis in Japanese apricot (Prunus mume Siebold & Zucc.), the relationships between carotenoid accumulation and the expression of the carotenogenic genes, phytoene synthase (PmPSY-1), phytoene desaturase (PmPDS), zeta-carotene desaturase (PmZDS), lycopene beta-cyclase (PmLCYb), lycopene epsilon-cyclase (PmLCYe), beta-carotene hydroxylase (PmHYb), and zeaxanthin epoxidase (PmZEP), were analyzed in two cultivars with different ripening traits, 'Orihime' and 'Nanko.' In 'Orihime' fruits, large amounts of carotenoids accumulated on the tree, concomitant with the induction of PmPSY-1 and the downstream carotenogenic genes PmLCYb, PmHYb, and PmZEP. In 'Nanko' fruits, carotenoids accumulated mainly after harvest, correlating with an appreciable induction of PmPSY-1 expression, but the downstream genes were not notably induced, which may explain the lower total carotenoid content in 'Nanko' than in 'Orihime.' In both cultivars, a decrease in PmLCYe expression and increased or constant PmLCYb expression could cause the metabolic shift from beta,epsilon-carotenoid synthesis to beta,beta-carotenoid synthesis that occurs as ripening approaches. Next, the effects of ethylene on the expression of PmPSY-1 and carotenoid accumulation were investigated in 'Nanko' fruits treated with propylene or 1-methylcyclopropene (1-MCP). Propylene treatment induced both ethylene production and carotenoid accumulation. PmPSY-1 was constitutively expressed, but propylene treatment accelerated its induction. 1-MCP treatment caused a slight inhibition of carotenoid accumulation along with the repression, although not complete, of PmPSY-1. Collectively, although PmPSY-1 expression was not exclusively regulated by ethylene, both the notable induction of PmPSY-1 accelerated by ethylene and the subsequent induction of the downstream carotenogenic genes, especially PmLCYb, could be necessary for the massive carotenoid accumulation that occurs during ripening. Furthermore, the switch from PmLCYe expression to PmLCYb expression could cause beta,beta-carotenoid accumulation in both Japanese apricot cultivars.     

Atlantic salmon fed a nutrient package of surplus methionine,vitamin B12, folic acid and vitamin B6 improved growth and reduced the relative liver size,but when in excess growth reduced

Feeding plant‐based diet through smoltification of Atlantic salmon requires verification of the optimal level of 1C nutrients. Here, we fed Atlantic salmon plant‐based diets containing three different surplus amounts of the 1C nutrients; methionine, cobalamin (vitamin B12), pyridoxine (vitamin B6) and folic acid during 6 weeks in fresh water, through smoltification, followed by 3 months on‐growing period in salt water. The three diets were fed to fish dispersed in triplicate tanks throughout the experiment. Mean start body weight was 32 g. Dietary methionine levels in the diets were 6.7, 9.2 and 11.7 g/kg. Dietary B6 was 6.75, 8.45 and 11 mg/kg. Cobalamin was 0.16, 0.18 and 0.20 mg/kg. While dietary folic acid was 2.9, 4.8 and 6.3 mg/kg, diets are referred to as low, medium and high 1C diet. All other amino acids were similar between diets. The results showed no differences in growth or feed utilization in the fresh water period, but following the on‐growing salt water period, differences between diets occurred. The fish fed the medium 1C diet showed better growth, as compared to fish fed the low or high 1C diet (p = .009). The medium 1C fed fish showed a relative lower liver weight compared with fish fed low or high 1C diet (p = .025). Condition factor was better in fish fed the medium and high 1C diet as compared to those fed the low 1C diet (p = .0006). As expected, free methionine in liver, plasma and muscle increased by dietary methionine inclusion. Surplus vitamins only had minor effect on tissue concentrations. Based on these findings, we conclude that the micronutrient and methionine level presented in the medium 1C diet improved the growth, liver size and condition factor; however, more research is needed to evaluate the optimal requirement level for each of the 1C nutrients.     

Nutritional values and antioxidative activities of whole peanuts and peanut skins for ruminant feeds

This study was conducted to investigate the nutritive values of two peanut by‐products, nonstandardized whole peanuts and peanut skins, along with their effects on microbial growth and fermentation in the rumen, their roughage values, and their antioxidative activities by a digestion trial using four goats. The experimental rations were alfalfa haycube (basal ration), 85% alfalfa with 15% whole peanuts, and 70% alfalfa with 15% whole peanuts and 15% peanut skins. The ether extracts and crude protein in whole peanuts were 47% and 27% on a dry matter basis (DM) both with over 90% of digestibilities, resulting in total digestive nutrients (TDN) of 140%. Peanut skins also had a high energy value with 91% of TDN. Ruminal concentrations of total volatile fatty acids (VFA) and acetic acid decreased in the rations containing the peanut by‐products, but the NDF digestibility and ruminal microbial protein estimated from urinary purines was not altered by feeding the peanut by‐products. Plasma oxidative stress maker, malondialdehyde, tended to be lower when peanut skins were supplemented. Whole peanuts and peanut skins could be used as high‐energy and high‐protein diets for ruminants, and peanut skins would be expected as a feed having antioxidant functions.     

Effects of auxin and jasmonates on 1-aminocyclopropane-1-carboxylate (ACC) synthase and ACC oxidase gene expression during ripening of apple fruit

1-Aminocyclopropane-1-carboxylate (ACC) synthase and oxidase activities, their gene expression, and ethylene production in apple fruit [Malus sylvestris (L.) Mill. Var. domestica (Borkh.) Mansf.] treated with a synthetic auxin 2,4-dichlorophenoxy-propionic acid (2,4-DP) and n-propyl dihydrojasmonate (PDJ), a jasmonic acid derivative, has been investigated to clarify the action of auxin and jasmonates on ethylene production. The fruit was harvested at 103 d after full bloom (preclimacteric). The expression of MdACS4 messenger RNA (mRNA) at 48 and 96 h after treatment was higher in fruit treated with 2,4-DP than in the untreated control, but those of MdACS1 and MdACO1 were not affected by treatment. The ethylene production in 2,4-DP-treated fruit increased at 96 h after treatment. In contrast, expression of mRNAs hybridized with MdACS1 and MdACO1 probes in the skin of PDJ-treated fruit were higher than those in the untreated control. In addition, ACC synthase activity and ethylene production also increased after treatment. These results show that the ethylene production rate may differ with the kind of genes which were stimulated by auxin or jasmonates.     

Mating patterns in an indoor miniature <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> seed orchard as revealed by microsatellite markers

To evaluate the practical application of an indoor seed orchard, we compared the seed production, final germination rates and mating patterns of potted Cryptomeria japonica D. Don clones in two miniature seed orchards: one in a glasshouse and the other outdoors. There was no statistically significant difference in total seed production between the orchards (P = 0.275, ANOVA). However, the final germination rate of seeds produced in the indoor orchard was significantly lower than that of seeds produced in the outdoor orchard (P < 0.01, Wilcoxon test). The average self-fertilization rate was higher in the indoor orchard (27.2%) than in the outdoor orchard (5.6%), and this parameter was strongly associated with the reduced final germination rates of the seeds harvested in the glasshouse. Pollen contamination was considerably lower in the indoor seed orchard (48.9% in the outdoor seed orchard and 4.4% in the indoor seed orchard). χ² tests indicated that the paternal contributions of each constituent clone differed significantly in both orchards (P < 0.001 in both cases). The strong variations in paternal contributions among indoor orchard clones was significantly correlated with their respective pollen production capacities (Spearman ρ = 0.693, P < 0.001). To reduce the high self-fertilization rates, we recommend the use of orchard clones with similar pollen production capacity in indoor seed orchards.