Disulphide structure of high-molecular-weight (HMW-) gliadins as affected by terminators

This study aimed at elucidating SS-bonds of HMW-gliadins (HGL) from wheat with the focus on terminators of glutenin polymerisation. HGL from wheat flour extracts non-treated or treated with the S-alkylation reagent N-ethylmaleinimide (NEMI) were compared. HGL from wheat flour Akteur were isolated, hydrolysed with thermolysin and the resulting peptides pre-separated by gel permeation chromatography and analysed by liquid chromatography/mass-spectrometry using alternating electron transfer dissociation/collision-induced dissociation. Altogether, 22 and 28 SS-peptides from samples without and with NEMI treatment, respectively, were identified. Twenty-six peptides included standard SS-bonds of α- and γ-gliadins, high-molecular-weight and low-molecular-weight glutenin subunits. Eleven SS-bonds were identified for the first time. Fifteen peptides unique to HGL contained cysteine residues from gliadins with an odd number of cysteines (ω5-, α- and γ-gliadins). Thus, gliadins with an odd number of cysteines, glutathione and cysteine had acted as terminators of glutenin polymerisation. Decisive differences between samples without and with NEMI treatment were not obvious showing that the termination of polymerisation was already completed in the flour. The two HGL samples, however, were different in the majority of ten peptides that included disulphide-linked low-molecular-weight (LMW) thiols such as glutathione and cysteine with the former being enriched in the non-treated HGL-sample.     

Effect of dietary phosphorus levels on growth and body composition of crucian carp,Carassius auratus under indoor and outdoor experiments

Indoor and outdoor experiments were carried out simultaneously to evaluate the effects of dietary phosphorus levels on growth performance and body composition of crucian carp. Monocalcium was used to formulate extruded feed containing available phosphorus (AP) of 1.5 (D1), 4.7 (D2), 6.3 (D3) and 9.0 (D4) g kg^?1. Twelve 500‐L recirculating tanks were assigned to four triplicate groups with each tank stocked with 15 juveniles (39.88 ± 0.14 g) for indoor experiment. Twelve polyethylene enclosures were assigned to four groups with each one stocked with 120 juveniles (44.08 ± 1.36 g). After 8 weeks of feeding, survival ranged from 96% to 100%. Weight gain, special growth rate, protein efficiency ratio and feed conversion rate did not show significant differences among dietary treatments in both experiments. Fish cultured in the enclosures grew better, although D1 and D2 treatments did not reach significant level. Crude protein and phosphorus content of the fish were not significantly affected by dietary phosphorus level. Moisture, crude lipid, crude ash and vertebrae phosphorus were significantly affected by different phosphorus levels. Based on vertebrae phosphorus, crucian carp cultured in the tanks and in the enclosures showed phosphorus deficiency under 4.7 g kg^?1 and 6.3 g kg^?1 AP, respectively.     

Effects of dietary protein and lipid levels with different protein‐to‐energy ratios on growth performance,feed utilization and body composition of juvenile red‐spotted grouper,Epinephelus akaara

An 8‐week feeding trial was conducted to assess dietary protein and lipid levels on growth performance, feed utilization and body composition of juvenile red‐spotted grouper (7.85 ± 0.03 g fish^?1). Nine semi‐purified diets were formulated containing varying protein levels (440–520 g kg^?1, dry matter) and lipid levels (60–120 g kg^?1, dry matter). The weight gain of juvenile Epinephelus akaara was affected by dietary protein (p = .005) and its interaction with dietary lipid (p = .020). Viscerosomatic index, intraperitoneal fat ratio and whole‐body lipid level increased with increasing dietary lipid level (p < .001). Nitrogen retention was not affected by dietary protein and lipid, while lipid retention decreased with increasing dietary lipid level (p < .001). The plasma blood urea nitrogen increased with increasing dietary protein level (p = .003). This study showed that diet with 520 g kg^?1 protein and 60 g kg^?1 lipid with 30.58 mg kJ^?1 P:E provided a maximal growth for this species. Moreover, an increase in dietary lipid levels (from 60 to 90 g kg^?1) could reduce the protein requirement (from 520 to 480 g kg^?1) without affecting the growth performance, while higher fat deposition was observed in fish fed high‐lipid diets.     

饲料脂肪水平对俄罗斯鲟幼鱼生长、血液生化指标及抗氧化性能的影响

以鱼油、豆油、亚麻籽油等比例混合,配成脂肪含量分别为5.45%(L1)、7.00%(L2)、10.44%(L3)、14.37%(L4)、16.34%(L5)、18.91%(L6)及20.15%(L7)的等氮饲料,饲喂俄罗斯鲟(Acipenser gueldenstaedtii)幼鱼[(6.20±0.02)g]8周。实验结果表明,L3组的终末体质量、增重率最大,但与L2组间无显著差异(P0.05),L3组的蛋白质效率显著高于其它组(P0.05),且饲料系数最低。L6和L7组终末体质量、增重率和蛋白质效率显著低于L2和L3组(P0.05)。将幼鱼的增重率与饲料脂肪水平进行回归分析,得出俄罗斯鲟幼鱼在饲料脂肪水平为10.69%时生长最佳。随着饲料脂肪水平的升高,各组鱼的肝体比和体粗脂肪有上升趋势,其中肝体比在饲料脂肪含量大于14.37%时显著增大(P0.05)。血清中的甘油三酯、胆固醇含量也随脂肪水平升高而显著提高(P0.05),提示脂肪过高会影响俄罗斯鲟幼鱼的生长,降低蛋白质效率,导致脂肪在体内的过量积蓄。L7组鱼血清中高密度脂蛋白胆固醇含量最高,而L1组鱼的低密度脂蛋白胆固醇含量最高。肝脏脂蛋白酯酶活性随饲料脂肪水平的提高而升高,脂肪酸合成酶活性则随脂肪水平升高而降低,提示饲料脂肪水平的提高会促进鱼体脂肪的分解代谢。肝脏丙二醛含量和超氧化物歧化酶活性均随着饲料脂肪水平的提高显著升高(P0.05),表明脂肪含量过高会导致脂质过氧化,引发机体抗氧化酶活性的升高。综合俄罗斯鲟幼鱼的生长、血液指标及抗氧化性能,建议生产中将幼鱼的饲料脂肪水平控制在7.00%~10.69%。     

Replacement of Fishmeal by a Mixture of Soybean Meal and Chlorella Meal in Practical Diets for Juvenile Crucian Carp,Carassius auratus

A growth trial was conducted to evaluate the effect of a mixture of soybean meal and Chlorella meal (SCM) as a dietary fishmeal (FM) substitute on growth performance, apparent digestibility coefficients (ADCs), digestive enzymatic activities, and histology of juvenile crucian carp, Carassius auratus. Five isonitrogenous and isoenergetic diets were formulated to replace 0 (SCM0), 25 (SCM25), 50 (SCM50), 75 (SCM75), and 100% (SCM100) of protein from FM with SCM, respectively. The diets were fed to triplicate groups of juvenile crucian carp for 6 wk. Weight gain, specific growth rate, feed intake, protein efficiency ratio, and intestinal digestive enzymatic activities (amylase, trypsin, and lipase) tended to decline with increasing FM replacement levels (P > 0.05). Dietary SCM substitution significantly influenced dry matter content in muscle, and crude protein and lipid contents in liver (P < 0.05). ADCs for dry matter, protein, lipid, energy, and most amino acids showed no significant differences between the control and SCM25 group, but tended to decline with replacement levels over 25%. Higher SCM substitution (50–100%) caused karyopyknosis and necrosis in liver, but intestinal histology did not show noticeable pathological changes. The present study indicated that FM could be replaced by 25% of SCM, without significant adverse growth performance, feed utilization, and histology of crucian carp.     

外源溶磷菌对不同土壤条件下大豆生长特性的影响

为了明确溶磷菌在多种土壤中的应用效果,探讨了两株溶磷菌对黑土及盐碱土种植大豆的促生效应。采用盆栽试验分析了不同处理对大豆光合作用、渗透调节物质含量、生物量、氮磷吸收及产量的影响。结果表明:加菌处理可增加植株Pn,显著降低R6、R8两时期叶片可溶性蛋白、可溶性糖及游离氨基酸含量,增加R2~R8时期大豆株高和根干重,混合土壤中加菌处理也可提高植株地上部分氮、磷及植株全磷、全氮含量。添加wj1和混合菌处理的植株干重优于添加wj3的处理。黑土加混合菌可显著提高大豆R2期叶片WUE,增加V3、R2和R8这3个时期大豆地上、地下部分氮、磷及植株全氮、全磷的含量。黑土加wj1与混合土壤加菌处理的大豆株荚数、总粒数、单株粒重及百粒重均高于未加菌处理。添加外源溶磷菌,对黑土及盐碱土栽培大豆均有不同程度的促生效果,能明显增强大豆在混合盐碱土壤中的适应性,增加植株生物量、氮磷吸收量和产量,且混合菌处理对大豆的促生效应优于wj1及wj3单一菌株处理。     

Growth performance and whole‐body composition of yellow catfish (Pelteobagrus fulvidraco Richardson) under feeding restriction

Fish growth is an important index in aquaculture practice. However, the effect of growth under restricted feeding on quality of fish grown to the same size remains poorly unknown. In this study, yellow catfish were reared by feeding them at three different feeding rates such as low (FR‐L group), medium (FR‐M group) and high (apparent satiation) ration levels for 82, 58 and 34 days to reach the same size, respectively. After this period, the growth performance, whole‐body amino acid composition and fatty acid profile of the fish were evaluated. Feeding restriction led to significant decreases in specific growth rate and feed conversion efficiency. Significant increases in dry matter, protein and energy contents were observed in fish under both restricted feeding treatments. FR‐L‐treated fish had significantly lower whole‐body essential amino acid contents, and significantly higher Ala and non‐essential amino acid contents compared to fish fed to satiation. Fish in restricted feeding treatments had significantly higher n‐3 PUFA and n‐3/n‐6 ratio compared to the fish fed to satiation. The present results indicate that from a quality perspective, there are certain advantages in improvement of whole‐body composition and enrichment of n‐3 PUFA associated with the restricted feeding of yellow catfish at slow growth rate.     

Influence of dietary lipid on growth performance and some lipogenesis‐related gene expression of tongue sole (Cynoglossus semilaevis) larvae

A 30‐day feeding trial was conducted to investigate the effects of dietary lipid levels on growth performance, activities of digestive enzymes, fatty acid composition and some lipogenesis‐related gene expression of half‐smooth tongue sole (Cynoglossus semilaevis) larvae. Five isoproteic diets were formulated with graded lipid levels (6.68%, 9.84%, 13.47%, 17.89% and 21.88% dry weight) using fish oil as the main lipid source. Each diet was randomly allocated to triplicate groups of 150 larval tongue sole (35 DAH, 54 ± 1 mg). Fish were fed five times daily to apparent satiation during the feeding experiment. Results showed that, the survival rate (SR) of larvae increased significantly firstly, and thereafter decreased significantly. The specific growth rates (SGR) of larvae fed the diet with 13.47% lipid were significantly higher than other treatments. Larvae fed 9.84% or 13.47% dietary lipid showed higher trypsin, lipase, leucine aminopeptidase and alkaline phosphatase (AP) activities than other treatments, whereas amylase activity nearly showed reverse trend with them. The fatty acid composition of the tongue sole larvae was well correlated with dietary fatty acid profile. Expression of acetyl‐CoA carboxylase alpha (ACC1) was found to be slightly negatively correlated with dietary lipid level, and high dietary lipid level depressed the expressions of acetyl‐CoA carboxylase beta (ACC2) and fatty acid synthase (FAS) mRNA expression significantly, implying that larvae may cope with high dietary lipid mainly through down‐regulating lipogenesis‐related gene expression of FAS and ACC2. Besides, on the basis of SGR, the optimal dietary lipid level for larval tongue sole was estimated to be 13.56% using second‐order polynomial curve.     

Effect of different feeds on the growth,survival and reproductive performance of zebrafish,Danio rerio (Hamilton, 1822)

Present experiment was conducted to determine the effect of different feeds with varying protein levels on the growth, survival and reproductive performance of zebrafish, Danio rerio. The control diet (T1) was wild‐collected zooplankton from local fish ponds, while test diets with 350 g kg^?1 protein (T2), 400 g kg^?1 protein (T3) and 450 g kg^?1 protein (T4) were formulated and fed to fish for a period of 210 days. The significantly (P < 0.05) highest mean weight gain and specific growth rate were observed in T1, which were similar with T3 and T4. The significantly (P < 0.05) highest number of egg production per female and relative fecundity were found in T1, followed by T4 and T3 while T2 produced lowest number of eggs. No significant (P > 0.05) differences were observed in brood survival rate, fertilization and hatching rate among the dietary treatments. The highest (P < 0.05) fry survival rate was recorded in T1, followed by T3 and T4. Thus, it is suggested that control diet i.e. mixed zooplankton exhibited better growth, reproductive performance and fry survival rate. However, diet containing 400 g kg^?1 crude protein also gave comparable results in terms of growth, survival and reproductive performance of zebrafish.     

茶树GGPS基因家族的克隆、生物信息学和表达分析

牻牛儿基牻牛儿基焦磷酸合酶(Geranylgeranyl diphosphate synthase,GGPS)是萜类合成途径的结构酶,对植物生长发育具有重要意义。本研究通过RACE和RT-PCR方法克隆得到5条潜在的茶树GGPS序列,分别命名为CsGGPS1-4和CsGGPS9,其中CsGGPS9存在3条等位基因,分别是CsGGPS9-1、CsGGPS9-2和CsGGPS9-3,在系统进化树上与其他基因分成两支。蛋白质序列分析表明,茶树GGPS家族成员都具有polyprenyl_synt结构域,不存在信号肽序列。亚细胞定位预测结果显示,CsGGPS1、CsGGPS2和CsGGPS4定位在叶绿体上,CsGGPS3和CsGGPS9定位在线粒体上。通过Swiss Model进行三维建模,结合"three-floor"模型对茶树GGPS家族成员的功能进行预测,预测结果显示,CsGGPS1、CsGGPS2和CsGGPS4是GGPS;CsGGPS3是异源二聚体形式的牻牛儿基焦磷酸合酶的小亚基;CsGGPS9的催化主产物是碳链数大于30的异戊烯基焦磷酸。q RT-PCR分析表明,CsGGPS1整体表达丰度较低,仅在一芽二叶中表达量稍高;CsGGPS2在茶树各个组织中均有表达,在花中表达量最高,且花发育过程中表达量先上升后下降;CsGGPS3在叶和幼根中的表达量高于花,花发育过程中表达平稳;CsGGPS4在茶树各个组织中表达量数值相近,在花发育过程中表达量变化趋势与CsGGPS2相同;CsGGPS9的表达量在成熟叶中显著低于幼嫩叶片。