小麦高分子量谷蛋白亚基1Dx5基因在大肠杆菌中的表达

为了给小麦优质亚基研究奠定基础,将高分子量谷蛋白亚基1Dx5基因的核苷酸序列与载体pRSET A重组,将构建好的质粒转化大肠杆菌BL21(DE3)pLysS菌株,通过IPTG使其得到了成功表达.大肠杆菌中表达的1Dx5亚基在SDS-PAGE上与小麦品种钱尼中的1Dx5亚基具有相似的迁移率.用Western blot技术成功检测到了目的基因产物.通过改变IPTG浓度、诱导时间、培养基成分及初始菌液浓度研究了最适表达条件.结果表明,最适表达条件为:LB培养基,菌液初始浓度(OD600)0.4～0.6,IPTG浓度0.4mmol/L,诱导时间为3～5 h.     

仔猪大肠杆菌病三价蜂胶灭活疫苗猪体血清抗体消长规律

采用大肠杆菌C83549(O149:K88ac)、C83644(O68:K99)、C83710(O9:987P)菌株制备抗原,以蜂胶为佐剂,研制仔猪大肠杆菌病三价蜂胶灭活疫苗,疫苗成品检验均符合相关要求。将疫苗免疫妊娠母猪并用凝集试验测定其所产仔猪的抗体消长规律,并研究疫苗免疫保护相关性。结果表明:妊娠母猪产前40d和15d各免疫1次所产仔猪母源抗体效价较高,仔猪在整个哺乳期都可以得到保护。同时也可运用产前30d和10d进行免疫接种方法。     

酸藤果的营养和色素的研究

研究了酸藤果(E.laeta)的营养价值及pH值、温度、紫外光、食品添加剂等对酸藤果色素的影响。认为酸藤果富含营养,而且是一种很有开发前途的天然植物色素资源。     

5种石斑鱼遗传差异的RAPD分析

运用随机扩增多态性DNA（random amplified polymorphic DNA,RAPD）技术对鞍带石斑鱼（Epinephelus lanceolatus）、驼背鲈（Cromileptes altivelis）、棕点石斑鱼（E.fuscoguttatus）、斜带石斑鱼（E.coioides）和鲑点石斑鱼（E.fario）的遗传多样性及系统发生进行了研究。从120个RAPD随机引物中筛选出扩增效果好的引物19个,分别用于5种石斑鱼基因组DNA的扩增。结果显示,5种石斑鱼平均多态性位点比率（P）分别为64.23%,72.61%,60.34%,69.97%,73.94%;个体间平均遗传相似系数（S）分别为0.8238,0.8110,0.8345,0.8277,0.8064;平均遗传距离（D）分别为0.1762,0.1890,0.1655,0.1723,0.1936;平均Nei基因多样性指数（H）分别为0.1189,0.1364,0.1028,0.1439,0.1648;平均Shannon信息指数（Hi）分别为0.1801,0.1992,0.1530,0.2100,0.2434。5个种间的遗传距离（Dxy）在0.3964～0.6085之间。     

镉胁迫对尾巨桉幼苗生长和生理特性的影响

以尾巨桉(DH_32-29)幼苗为试验材料，以植株的生长指标(苗高、地径、叶面积)、生物量、根冠比、氮平衡指数(NBI)、叶绿素相对含量(SPAD 值)、丙二醛(MDA)含量和叶绿素荧光参数为指标，通过盆栽试验设置8个10~140 mg·kg^-1浓度镉(Cd)盐溶液处理，研究了不同浓度 Cd 胁迫处理下尾巨桉幼苗的形态和生理特性变化规律。结果显示：低浓度(10 mg·kg^-1)Cd 处理对尾巨桉幼苗的形态和生理特性无明显影响。当 Cd 浓度大于20 mg·kg^-1，随着 Cd浓度的逐渐增大，其对尾巨桉幼苗的伤害程度也逐渐增加，主要表现为：植株生长缓慢、生物量减小、叶绿素相对含量和叶绿素荧光参数(Fv/Fm、φPSⅡ、NP、NBI)均下降，qP 无明显变化， MDA 含量升高。Cd 浓度大于60 mg·kg^-1以上时，尾巨桉幼苗生长出现一定程度上的损伤症状和生长停滞现象。结果表明低浓度(≤20 mg·kg^-1)Cd处理下尾巨桉幼苗能正常生长，随着浓度的增大，Cd 处理对尾巨桉的形态和生理特性造成显著影响。同时结合部分桉树适生区土壤 Cd 含量调查及相关研究结果认为，尾巨桉具有一定修复 Cd 污染区土壤的潜力。     

重组大肠杆菌不耐热肠毒素的表达及对小鼠胚胎稳定性的影响

为研究大肠杆菌不耐热肠毒素（LT）对小鼠胚胎稳定性的影响,运用大肠杆菌表达系统制备了具有生物活性的重组大肠杆菌LT,用重组LT通过腹腔注射处理妊娠小鼠,分析了LT对小鼠胚胎稳定性的影响。首先采用PCR方法从产毒大肠杆菌菌株44815基因组中扩增LT的A、B亚基基因,将其插入到pET-20b（＋）原核表达载体pelB信号肽的下游,分别构建成LTA和LTB分泌型表达载体;将载体分别转化大肠杆菌BL21（DE3）pLysS,在IPTG的诱导下进行表达,利用Ni-NTA琼脂糖凝胶从细菌周质释放液中提取和纯化重组LTA和LTB蛋白;利用细胞毒性试验检测重组蛋白的生物活性。然后用制备的重组LT注射妊娠6d的小鼠,连续注射3d后,统计小鼠的胚胎存活率。同时用ELISA方法检测小鼠血清中与胚胎稳定性密切相关的Th1型细胞因子（IFN-γ、IL-2）和Th2型细胞因子（IL-4、IL-10）及IL-β的表达水平。结果表明,采用分泌性表达策略实现了重组LT在大肠杆菌中的高效分泌表达,LTA和LTB的表达量分别达68、62mg/L。表达的重组LT具有明显的细胞毒性作用;用LT处理妊娠小鼠,胚胎的存活率为32%,明显低于对照组;小鼠血清中Th1型细胞因子IFN-γ和IL-2的含量明显高于对照组（P〈0.01）,分别为对照组的2、3倍。同时细胞因子IL-1β为对照组的2倍（P〈0.01）,而稳定胚胎发育的Th2型细胞因子IL-4、IL-10没有明显变化（P〉0.05）。据此推测,LT可能与大肠杆菌性肠炎引起妊娠家畜流产有关,LT对胚胎稳定性的影响除与LT的直接毒性有关外,可能还与LT介导的免疫调节异常有关。     

重庆市万州区1年生尾巨桉生长情况调查

通过对重庆市万州区营造尾巨桉(3229品系)的调查结果表明,该树种在该区域的适应性和速生性较强,可进一步扩大种植范围,在万州区乃至三峡库区具有一定的发展前景。     

Control of air-borne field spread of tulip breaking virus,lily symptomless virus and lily virus X in lilies by mineral oils,synthetic pyrethroids,and a nematicide in the Netherlands

The inhibitory effect on the spread of viruses in lilies viz., tulip breaking virus (TBV; nonpersistently aphid-borne, potyvirus,) lily symptomless virus (LSV; non-persistently alphidborne, carlavirus), and lily virus X (LVX; potexvirus of unknown etiology), was studied of brands of mineral oil (Luxan oil H and Duphar-7E oil) and synthetic pyrethroid insecticides (l-cyhalothrin and deltamethrin), and a nematicide (aldicarb) in crops in which virus-infected plants were present as virus sources. The spread of TBV and LSV were controlled by sprays of mineral oil and insecticide, while that of LSV was also limited by the soil-applied nematicide. The spread of LVX was reduced by the insecticides and, not effectively by the mineral-oil spraying, by which data the mode of transmission may be presumed to be by an insect in the persistent or semi-persistent manner.Mixtures of mineral oil and pyrethroid were more effective in the reduction of spread of TBV and LSV than either components tested alone. The mineral oil was the most effective component in the mixtures in which pyrethroid added a slight extra effect. The addition of pyrethroid did not mask either the lower efficacy of the oil brand Duphar-7E oil, or the diminished inhibitory effect of low dosages of oil. The normal rate of mineral oil gave similar control to that of a mixture of mineral oil at half rate plus the pyrethroid at full dosage. Low rates of oil, or even synthetic pyrethroids alone may be used on cultivars which suffer of the loss of bulb weight by the use of normal or decreased rates of oil. Weekly sprays were more effective than fortnightly sprays. The rate of control by the weekly sprays ranged between 90 and 95% for Luxan oil H at half dosage plus the full rate of pyrethroid. Weekly sprayed synthetic pyrethroids alone onto the virus sources and the plants to be infected gave 60–70% control. The weight ratios tended to be slightly reduced if the half dosage of the efficient Luxan oil H was used. Factors which affect the control of the air-borne field spread of viruses by mineral oils and synthetic pyrethroid insecticides in lilies are discussed.     

日粮维生素E水平对山羊精液品质的影响

采用4×4拉丁方设计,给金堂黑山羊饲喂维生素E浓度不同(常量:30 mg/kg维生素E,10,50,100倍常量的添加量)而其他营养成分均一致的4种日粮,探索日粮中添加维生素E对山羊精液品质的影响。结果表明:日粮中添加维生素E有助于提高山羊采精量、精子密度和鲜精活率,改善其精液品质(P<0.05);随着日粮中维生素E浓度的增加,山羊采精量、精子密度和鲜精活率也随之改善,添加量为常量的50倍(1500 mg/kg)时,精子密度和鲜精活率最佳(P<0.01)。当添加量超过常量50倍时(100),精子密度和鲜精活率有所降低,但与添加量为50倍的试验组差异不显著(P>0.05)。     

复方磷霉素钙水溶性粉处方优化实验

为了优选磷霉素钙复方制剂,采用体外药物最小抑菌浓度测定法,测定了磷霉素钙与甲氧啶(TMP)、丁胺卡那霉素、恩诺沙星、庆大霉素、头孢噻肟钠、多西环素、氟甲砜霉素联合使用对大肠杆菌作用的协同指数.结果表明磷霉素钙与TMP的协同关系最好,协同指数为0.281,制剂中磷霉素钙与TMP的含量分别为5%和1%为最佳.