硒对铜胁迫下大菱鲆幼鱼生长、抗氧化能力及相关基因表达的影响

为探讨硒对铜胁迫下大菱鲆幼鱼生长、抗氧化能力及相关基因表达的影响,以初始体质量为(24.85±0.10) g的大菱鲆为研究对象,通过在饲料中添加五水硫酸铜和亚硒酸钠,配制铜、硒含量分别为(0、0,1 000、0,1 000、2,1 000、4 mg/kg) 4种等氮等能的实验饲料,命名为D1、D2、D3和D4组。养殖实验持续84 d。结果显示:①各组间实验鱼成活率无显著差异;增重率、特定生长率和蛋白质效率均在D2组显著低于D1、D3和D4组;摄食率和饲料系数呈相反趋势,在D2组显著高于其他3组。②幼鱼全鱼和肝脏粗脂肪含量呈先下降后上升趋势;而背肌粗蛋白和粗脂肪含量在各组之间无显著性差异。全鱼、脊椎骨和肝脏中Cu含量在D2组达到最高值;Zn含量则在D4组达到最高值;肝脏中铁含量呈下降趋势,在D3和D4组显著低于D1和D2组。③D2组显著降低了幼鱼血清总抗氧化能力(T-AOC)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)活性;丙二醛(MDA)含量则呈相反趋势,在D2组显著高于D3和D4组;总超氧化物歧化酶(T-SOD)和铜—锌超氧化物歧化酶(CuZn-SOD)活性均在D4组达到最低值。④血清谷丙转氨酶(ALT)、谷草转氨酶(AST)活性均在D2组显著高于其他3组;而血糖浓度、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)活性均在D2组达到最低值,显著低于D1组。⑤肝脏金属硫蛋白(MT) mRNA相对表达量在D4组达到最高值,显著高于其他3组;谷胱甘肽转移酶(GST)和溶菌酶(LZM) mRNA相对表达量均在D2组显著低于其他3组;热休克蛋白70 (HSP70) mRNA相对表达量则在D2组显著高于其他3组。综上所述,在本实验条件下,饲料中添加硒(2～4 mg/kg)可缓解高铜(1 000 mg/kg)胁迫导致的鱼体生长缓慢等症状,并可调控鱼体的抗氧化能力、生理代谢及相关基因表达量,进而对鱼体在高铜胁迫下的内环境稳态恢复具有一定的促进作用。     

Exogenous Nitric Oxide Involved in Subcellular Distribution and Chemical Forms of Cu~（2＋） Under Copper Stress in Tomato Seedlings

Nitric oxide（NO）,a bioactive signaling molecule,serves as an antioxidant and anti-stress agent under abiotic stress.A hydroponics experiment was conducted to investigate the effects of sodium nitroprusside（SNP）,a NO donor,on tomato seedlings exposed to 50 μmol L-1CuCl 2.The results show that copper is primarily stored in the soluble cell sap fraction in the roots,especially after treatment with Cu＋SNP treatment,which accounted for 66.2% of the total copper content.The copper concentration gradually decreased from the roots to the leaves.In the leaves,exogenous NO induces the storage of excess copper in the cell walls.Copper stress decreases the proportion of copper integrated with pectates and proteins,but exogenous NO remarkably reverses this trend.The alleviating effect of NO is blocked by hemoglobin.Thus,exogenous NO is likely involved in the regulation of the subcellular copper concentrations and its chemical forms under copper stress.Although exogenous NO inhibited the absorption and transport of excess copper to some extent,the copper accumulation in tomato seedlings significantly increased under copper stress.The use of exogenous NO to enhance copper tolerance in some plants is a promising method for copper remediation.     

Establishment of Efficient Regeneration System of Populus×euramericana cl.‘Bofeng’

Compared with the species of Leuce section,the difficulty in plant regeneration during tissue culture for the species of Aigeiros section is a key limiting factor for their application in genetic engineering of forest trees.In this study,the plant growth regulators combination,copper concentration,light intensity,and the selection pressures of kanamycin were investigated using leaves of Populus×euramericana cl.’Bofeng’ as explants,and a stable and high efficient regeneration system was established.Using this system,both the shoot regeneration rate and rooting rate were up to 100%,and the average number of differentiated shoots in each leaf explant was up to 20,and the survival rate of seedlings reached 98%.The optimal selection pressure of kanamycin in shoot inducement for leaf-explant was 40 mg·L^-1,and the optimal selection pressure of kanamycin in root inducement of adventitious bud was 20 mg·L^-1.The authors also found that differentiation rate of adventitious buds of P.×euramericana cl.’Bofeng’ could be significantly promoted by increased copper concentration,and this is the first report about the key role of copper（Cu） in explant regeneration of Poplars.This regeneration system with high frequency which was established by the authors was as good as that of species of Leuce section,and it provides a good technological platform for genetic engineering of forest tree which uses the species of Aigeiros section as model materials.     

The Distribution of Soluble Copper- and Zinc-Binding Proteins in Liver and Kidney of Chronic Copper-Poisoned Goats

The distribution of copper and zinc among the soluble proteins in the liver and kidney from chronic copper-poisoned goats was examined after gel filtration of the proteins. The concentrations of copper in the liver and kidney cortex from five experimentally copper-poisoned goats were: 550–810 µg/g liver and 190–420 µg/g kidney cortex (wet weight). In general the copper-binding proteins from, both the liver and kidney samples were separated into two different fractions with approximate molecular weights (m.w.) of > 65,000 and 10,000, respectively. From the liver samples, varying amounts of copper were eluted in a fourth fraction with m.w. < 2,000. In the majority of kidney samples the dominating copper-binding protein fraction was the high molecular weight fraction. Absolute amounts of copper recovered in the metallothionein-like protein fraction were nearly the samt for all samples investigated. The distribution of zinc-binding proteins in both liver and kidney samples was nearly the same. The high molecular weight fraction dominated, and no zinc was bound to metallothionein-like proteins.     

高铜对雏鸡器官组织病理损害的研究

将420只1日龄艾维菌健雏，随机分为7组，分别饲以含铜量11mg／kg的基础日粮和含铜量分别为100、200、300、400、500和600mg／kg的6种高铜日粮6周，研究高铜对雏鸡的病理损害。结果显示，日粮含铜100和200mg／kg对雏鸡有促生长作用。日粮铜水平达300mg／kg以上时，对雏鸡器官组织造成不同程度的病理损伤，表现为肌胃角质层增厚呈黄褐色；肝呈浅黄色或深褐色，肝细胞脂肪变性；消化道上皮细胞变性坏死；免疫器官和肾、心脏、胰腺、大脑等器官的实质细胞肿胀；淋巴免疫器官体积缩小，淋巴细胞数量减少。超微结构观察发现，肝细胞胞浆和胞核内见有数量不等、大小不一的高电子密度的沉积物；凋亡的淋巴细胞染色质呈花瓣样、半月形或环形，聚集在核膜下。表明，肝、消化道和免疫器官是高铜损害的主要靶器官。     

铜在奶牛生产中的应用及存在问题

本文就铜的生物学效应和生理功能,奶牛对铜的需要量、耐受量和中毒量等进行阐述。     

铜的来源和水平对生长猪生长性能禾粪铜排出量的影响

选用平均体重29．6kg的三元杂（杜×大×长）生长猪144头，按随机区组设计分为6个处理。饲粮中分别添加0、100、200mg／kg硫酸铜、200mg／kg碱式氯化铜、100mg／kg小肽铜、100mg／kg复合氨基酸螯合铜（以铜含量计）。结果表明：100mg／kg小肽铜和100mg／kg复合氨基酸螯合铜均能有效提高生长猪日增重和日采食量，其促生长效果与200mg／kg硫酸铜无显著（P〉0．05）差异，而粪中铜含量分别下降了36．8％和48．4％（P〈0．05）。     

硫酸链霉素细菌内毒素定量测定法的研究

应用鲎试剂动态浊度法，将硫酸链霉素制备成4、2、1、0．5mg／mL溶液，定量添加标准内毒素进行干扰预试验，计算回收率，确定不干扰浓度为0．5—2mg／mL。从中选出1mg／mL为最佳浓度，然后进行3个批次硫酸链霉素正式干扰试验。在供试品中定量添加标准内毒素，其回收率分别为125．8％、126．1％、127．3％，均在50％-200％之间，说明此浓度的供试品对鲎试剂反应无干扰作用。因此，内毒素含量在0．015—1EU／mL范围内，将硫酸链霉素制备成浓度为1．0mg／mL的溶液，可用于细菌内毒素的定量检测。     

Sulphur accumulation after Verticillium dahliae infection of two pepper cultivars differing in degree of resistance

Elemental sulphur levels, sulphur localization in stems, and levels of sulphate, glutathione and cysteine were studied in pepper ( Capsicum annuum ) cvs Yolo Wonder (higher resistance) and Luesia (lower resistance) after inoculation with Verticillium dahliae , the cause of vascular wilt. Accumulation of elemental sulphur (S⁰) was first detected 10 days after inoculation in Yolo Wonder (mean S⁰ level 7·3  µ g g⁻¹ DW), and 15 days after inoculation in Luesia (mean S⁰ level 3·3  µ g g⁻¹ DW). The maximum level was reached 21 days after inoculation in Yolo Wonder (14·1  µ g g⁻¹ DW). In control plants, elemental sulphur was not detected. SEM-EDX (scanning electron microscopy-energy dispersive X-ray microanalysis) indicated that the sulphur was not restricted to a specific location, but was dispersed throughout the vascular tissue. Sulphate levels showed a decline at the end of the experiment in inoculated plants, possibly related to the increase in sulphur levels seen in the two cultivars. The differences in sulphate levels between the two cultivars may be due to faster sulphate breakdown in cv. Yolo Wonder.