碳酸盐碱度胁迫下凡纳滨对虾基因的差异表达

以广盐性养殖的凡纳滨对虾(Litopenaeus vannamei)为研究对象,采用抑制消减杂交(suppression subtractive hybridization,SSH)和定量PCR的方法,研究其在高碳酸盐碱度胁迫下转录组水平的基因表达差异,以期从基因组水平研究对虾对高碳酸盐碱度胁迫的适应机制。结果表明,以高碳酸盐碱度(20 mmol/L)胁迫第4天凡纳滨对虾鳃组织和低碱度(2 mmol/L)对照组鳃组织为材料,通过斑点杂交筛选,发现鳃组织中有158个克隆子表达上调,291个克隆子表达下调。挑选150个高表达差异的克隆子进行测序,获得100个序列,其中50个得到成功注释。经过GO分析,这些注释的差异基因主要分为8大类群,其中碳酸酐酶基因(CA)、Na+-K+-ATPase基因(NKA-α)等与离子调控相关的基因表达量上调,而溶菌酶基因等与先天免疫相关的基因表达量下调,这些结果表明高碳酸盐碱度胁迫下,凡纳滨对虾以增加离子调控的方式进行酸碱平衡的调控,同时其免疫功能受到抑制。此外,还对CA和NKA-α两个基因在鳃和触角腺中的时空表达规律进行了研究,发现高碳酸盐碱度胁迫9 d过程中,两个基因在鳃组织中的表达均呈现先高表达后回落的现象,而在触角腺中NKA-α基因则一直维持较高表达水平,说明CA和NKA-α基因在凡纳滨对虾高碳酸盐碱度适应离子调控中起着关键作用,同时还发现除了鳃组织之外,触角腺同样参与了调控。本研究从转录水平初步筛选了高碳酸盐碱度胁迫下凡纳滨对虾的表达差异基因,探索了凡纳滨对虾的耐盐碱机制,对培育适于盐碱地水产养殖的优良品种有着重要的意义。     

Sulfur content of plant material: A comparison of methods of oxidation prior to determination

Abstract

Procedures for S determination involving digestion of plant material with HNO₃/HClO₄ mixtures gave lower values than those using oxygen flask combustion. Tests on a range of S compounds showed that the HNO₃/HClO₄ procedure underestimated S present in methionine, cysteine and glutathione but S in sulfate and the glucosinolate, sinigrin, was not underestimated.

Low recoveries using HNO₃/HClO₄ digestion procedures can be due both to incomplete oxidation of certain compounds and to gaseous losses during the vigorous boiling stage of digestion.     

鹿角托盘总蛋白的提取工艺研究

[目的]优化鹿角托盘总蛋白提取工艺。[方法]以总蛋白得率为评价指标,采用单因素试验和正交试验设计优选最佳提取工艺。[结果]最佳提取工艺为：用含0.3mol/LNaCl,20mmol/LNa2HPO4缓冲液（pH值为9）提取,料液比为1：15,提取时间为24h,提取3次。在此工艺条件下,总蛋白得率达10%以上。[结论]试验优选出的工艺稳定、合理、可行。     

山毛豆草粉颗粒料对肉兔的饲用价值评定

为探讨山毛豆(Tephrosia candida)草粉对肉兔的饲用价值,将60只新西兰青年肉兔分为5组,分别添加0％(对照),10％,20％,30％和40％的山毛豆草粉制成全价颗粒饲料,饲喂90 d后测定各组饲料的营养成分及肉兔采食量、日增重、料重比和屠宰性能.结果表明:山毛豆营养生长期粗蛋白含量为17.77％,肉兔对山毛豆中粗蛋白的消化率为78.09％,山毛豆的可消化总养分为56.18％.与对照组相比,添加20％草粉日增重达到20.80 g·d-1(P＜0.01);料重比为4.45∶ 1(P＜0.05)；屠宰率为57.78％(P＜0.01)；单位kg增重平均最低饲料成本差异显著(P＜0.05).因此,添加20％山毛豆草粉制成全价颗粒饲料可显著提高肉兔的生产性能和养殖效益.     

Susceptibility to IVM in a field strain of Haemonchus contortus subjected to four treatments in a closed sheep-goat flock in Kenya

Susceptibility to IVM (IVM) of “strain A” Haemonchus contortus which had been exposed to IVM four times over a 2-year period was compared to IVM susceptibility of “strain C” H. contortus which had no prior field exposure to IVM, by in vivo and in vitro methods. In vivo, the percentage reduction in faecal egg counts (FEC) and the total worm counts (TWC) were compared between control animals (lambs and kids) and animals treated with low dose IVM (20 μg/kg). In vitro susceptibility to IVM was evaluated by larval migration inhibition (LMI) after the two strains of H. contortus were exposed to different concentrations of IVM. The dose response, measured as the proportion of larvae inhibited from migrating, was used to estimate LD₅₀. Although differences in response to IVM in the in vivo determinations were not significant, “strain A” H. contortus had a significantly higher LD₅₀ than “strain C” in the LMI assay. Coincident with the conduct of the in vivo experiment, it was observed that “strain A” H. contortus established and survived better than “strain C” in the control lambs.     

Effects of three different concentrate proportions and rapeseed meal supplement to grass silage on animal performance of dairy-breed bulls with TMR feeding

A 3 × 2 factorial design with growing dairy-breed bulls was used to study the effects on animal performance of (1) proportion of concentrate (rolled barley) in the diet, and (2) inclusion of rapeseed meal (RSM) in the barley-based concentrate in a total mixed ration (TMR). The interactions between concentrate proportion and RSM supplement were also examined.Three feeding experiments comprised in total of 84 Finnish Ayrshire bulls and 6 Friesian bulls. The bulls were fed TMR ad libitum. The three concentrate proportions were 300 (L), 500 (M) and 700 (H) g/kg dry matter (DM), fed without RSM (RSM−) or with RSM (RSM+). Rapeseed meal was given so that the crude protein (CP) content of the concentrate was raised to 160 g/kg DM in the RSM+ diets. In the RSM− diets the CP content of the concentrate was 128 g/kg DM, so the CP content increased 25% with RSM supplementation. Increasing the proportion of concentrate led to a linear improvement in daily live weight gain (LWG) (P < 0.05), but there were no significant treatment differences in the DM intake (kg/d). Increasing the proportion of concentrate also led to significantly higher CP (P < 0.001) and phosphorus (P) (P < 0.001) supply and significantly improved DM and organic matter (OM) digestibility (P < 0.001). However, the digestibility of neutral detergent fibre (NDF) decreased (P < 0.001) as the proportion of concentrate increased. The feed conversion rate (kg DM/kg LWG) decreased significantly with increasing concentrate proportion (P < 0.001). Rapeseed meal supplement had no effect on animal performance, but the supply of CP (P < 0.01) and P (P < 0.001) was higher when RSM was included in the diet. The CP (P < 0.001) and NDF (P < 0.05) digestibilities were also higher for the RSM+ diets than for the RSM− diets. Because RSM at the concentration used did not affect animal performance, there is no reason to use RSM supplementation for finishing dairy bulls when there is good quality grass silage and barley-based concentrate in the TMR ration. This study also shows that there is a need to update the Finnish feeding recommendations for dairy-breed growing bulls, and extra calculations are needed for the energy and protein supply of growing dairy bulls.     

酚类物质与蝴蝶兰褐变关系初探

 利用高效液相色谱法对3个褐化程度不同的蝴蝶兰品种A1 (大白花红心) 、B3 (迷你型白花黄心) 和R4 (深红花红心) 进行了9种酚酸定性定量分析, 并研究了以叶片为外植体的初代培养过程中总酚含量的动态变化。结果表明: 绿原酸、邻苯二酚、儿茶酚、咖啡酸及没食子酸、对羟基苯甲酸、香豆酸可能与蝴蝶兰褐变相关, 苯甲酸对蝴蝶兰褐变影响很小; 在褐变过程中, 总酚含量越高, 褐变程度越严重。     

蝴蝶兰外植体褐变发生与总酚含量、PPO、POD和PAL的关系

 蝴蝶兰外植体在褐变发生前期PPO和POD活力皆升高, 褐变发生后酶活力下降。同工酶谱分析发现, 培养0 d的蝴蝶兰外植体PPO没有酶带出现, 而POD有1条弱带。离体培养2 d POD出现3条酶带, 第4天有新酶带Ⅰ发生, 随后消失, 其余3条带, 随培养天数的延长, 酶带活性渐弱。PPO同工酶谱在培养2 d出现3条酶带, 迁移率为0128的酶带Ⅲ在培养4 d活性较强, 随后3条酶带减弱。总酚含量和PAL活力随外植体褐变增强而逐渐增加, 两者呈现极显著正相关。     

Pineapple organic acid metabolism and accumulation during fruit development

Developmental changes in pineapple (Ananas Comosus (L.) Merrill) fruit acidity was determined for a ‘Smooth Cayenne’ high acid clone PRI#36-21 and a low acid clone PRI#63-555. The high acid clone gradually increased in fruit acidity from 1.4 meq/100 ml 6 weeks from flowering, and peaked a week before harvest at ca 10 meq/100 ml. In contrast, the low acid clone increased in acidity 6 to 8 weeks after flowering, peaked 15 weeks after flowering at ca. 9 meq per/100 ml and then sharply declined in 2 weeks to 6 meq/100 ml. The increased in total soluble solids (TSS) of the low acid clone began 6 weeks after flowering and for the high acid clone at 12 weeks after flowering. The increase in titratable fruit acidity (TA) paralleled the changes in the citric acid content of both clones. Citric acid content increased from less than 1 mg/g at 6 weeks after flowering to 6 to 7 mg/g, 9 weeks later. The malic acid concentration in both clones varied between 3 and 5 mg/g and showed no marked changes just before harvest. The developmental changes in fruit potassium were significantly correlated with fruit acidity and fruit total soluble solids in both the high and low acid clones. Developmental changes in acid-related enzymatic activities showed an increase in citrate synthase (EC 4.1.3.7) activity that occurred a week before harvest, coincided with the peak in citric acid in the high acid clone. An increase in aconitase (ACO, EC 4.2.1.3) activity was observed just before harvest as the decline in acidity occurred in the low acid clone. The activities of phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31), malate dehydrogenase (MDH, EC 1.1.1.37) and malic enzyme (ME, EC 1.1.1.40) did not parallel any changes in fruit acidity. The results indicated that the change in pineapple fruit acidity during development was due to changes in citric acid content. The major difference in acid accumulation occurred in the low acid clone just before harvest when acidity declined by one-third. The activities of citrate synthase and aconitase possibly played a major role in pineapple fruit acidity changes.