传染性法氏囊病病毒主要结构蛋白VP2、VP1和VP2-VP1串联基因的原核表达及其初步应用

为获得传染性法氏囊病病毒(IBDV)特异性抗体检测用抗原VP2、VP1及VP2-VP1蛋白,分别设计引物扩增IBDV野毒株NN1172的VP2和VP1基因,并扩增VP2和VP1基因中抗原性和亲水性较好的重要区域,通过PCR扩增基因串联方法对截短的VP2和截短的VP1基因进行串联,首次获得VP2-VP1串联基因,并对VP2、VP1和VP2-VP1串联基因进行了原核表达和鉴定。结果成功构建了原核表达载体pET-VP2、pET-VP1和pET-VP2-VP1;诱导表达条件显示,3个重组质粒分别转入BL21菌株后经0.05 mmol/L IPTG诱导表达,分别得到分子量为69、114和63 kDa的VP2、VP1和VP2-VP1重组蛋白,且均以包涵体形式表达,3个重组蛋白分别于诱导后5、3和6 h时表达量最多。Western blot结果显示,表达的VP2、VP1和VP2-VP1蛋白与鸡抗IBDV阳性血清均具有良好的反应原性。以纯化的VP2、VP1和VP2-VP1蛋白作为包被抗原对传染性支气管炎病毒(IBV)、呼肠孤病毒(ReoV)、禽白血病病毒(ALV)和新城疫病毒(NDV)4种阳性血清检测均为阴性,表明所获得的纯化蛋白具有高度的特异性;对免疫了IBD灭活疫苗,IBD基因工程疫苗和IBD弱毒疫苗的商业鸡群进行抗体检测,结果均能显示疫苗免疫后机体抗体水平的变化趋势。本研究表明利用该原核表达系统所表达的3个蛋白均具有良好的免疫反应活性,为IBDV特异性抗体的检测和新型亚单位疫苗的研发奠定基础。     

柑橘溃疡病菌LAMP实时浊度检测方法的建立及应用

以柑橘溃疡病菌特异性高的假定蛋白基因（登录号：AE008923.1）为靶标，设计并筛选4条引物来特异性识别靶标序列中的6个独立区域，建立柑橘溃疡病菌LAMP实时浊度检测方法。该方法利用实时浊度仪检测反应过程中产生的焦磷酸镁白色沉淀，实现对LAMP整个反应过程的实时监控。从镁离子浓度、甜菜碱浓度、dNTPs浓度、引物浓度和反应温度5个变量参数对 LAMP检测体系进行优化，并对方法的特异性、灵敏度、稳定性及实际样品检测效果进行评价。结果显示，该体系经优化后稳定性良好，可在66 ℃ 恒温条件下，60 min内完成检测；DNA检测下限可达0.02 ng/μL，灵敏度与荧光定量PCR方法相同，比常规PCR方法高1个数量级；能快速、准确地对田间疑似样品进行检测，与荧光定量PCR方法的检出情况一致，没有漏检。结果说明柑橘溃疡病菌实时浊度LAMP检测方法操作简便、所需时间短，特异性与灵敏度高，为柑橘溃疡病菌的快速筛查提供较好的途径。     

大白菜花青素含量及色差指标相关性研究

为了探寻大白菜花青素含量快速测定的方法,本文利用pH差计法和色差仪分别测定了‘城阳青’(普通大白菜)、‘秋宝黄’(黄心白菜)和‘彩凤’3个不同叶色大白菜品种花青素含量以及色差指标,并对6个色泽参数与花青素含量进行了相关和回归分析。结果表明:6个色泽参数均能反映白菜叶片间叶色的细微变化,‘城阳青’和‘秋宝黄’不含花青素,紫色白菜‘彩凤’的花青素主要分布于由外向内的第1～4层叶片。色泽参数L,b,h和C分别与花青素含量呈极显著或显著负相关关系(相关系数分别为-0.85**,-0.58**,-0.49*,-0.58**)。以色泽参数值L、a、h与花青素含量建立了多元回归方程:y=90.24-1.07x1+5.65x2-142.71x3(x1为L值,x2为a值,x3为h值,y为花青素含量),该回归方程相关系数为0.9629。通过色差计依据建立的回归方程可快速测定紫色大白菜花青素的含量。     

Effect of Dietary Lactobacillus fermentum from Pig on Growth Performance,Nutrient Apparent Digestibility,Faecal Microflora Number and Serum Immune Indices in Weaned Piglets

This experiment was conducted to study the effects of Lactobacillus fermentum on growth performance, nutrient apparent digestibility, faecal microflora number and serum immune indices of weaned piglets.A total of 160 weaned piglets with an average age of (30±2) days and averge body weight of (7.85±0.68) kg were randomly distributed into 4 groups with 4 replicates per group and 10 piglets per replicate feeding basal diet, 0.1%, 0.2%, and 0.3% Lactobacillus fermentum supplementation diets, respectively.The experiment lasted for 35 d.The results showed that, compared with the control group, the average daily gain, the feed conversion ratio, the apparent digestibility of crude protein, calcium and total phosphorusin of Lactobacillus fermentum groups were all increased in varying degrees.The optimum supplemental level of Lactobacillus fermentum should be 0.2%, and at this level, the average daily gain was significantly increased by 8.70% (P<0.05), the F/G was decreased by 4.62% (P<0.05), the apparent digestibility of crude protein, calcium and total phosphorus were significantly increased (P<0.05), the number of faecal E.coli was significantly decreased (P<0.05), and the serum IgG was significantly increased by 8.39% (P<0.05).The results indicated that Lactobacillus fermentum supplementation could improve the immune function, growth performance, the apparent digestibility of crude protein, calcium and total phosphorus and reduce the number of faecal E.coli of weaned piglets.     

丹参对奶牛脂肪变性肝细胞NF-κB和CYP450表达的影响

为探讨丹参对脂肪变性奶牛肝细胞的作用机制,采用胶原酶两步灌流法分离培养奶牛肝细胞,不同浓度DL-乙硫氨酸处理,油红染色观察其脂滴数量,MTT法测定肝细胞的活性,生化法检测TG含量。用不同浓度的丹参处理变性肝细胞后,生化法检测ALT、AST、TG、SOD、MDA的含量或活性,ELISA法测定TNF-α的含量,Western blot法检测NF-κB和CYP450的蛋白表达水平。结果显示,筛选出2 mmol/L的DL-乙硫氨酸作用24 h作为体外造模的最佳条件。用丹参处理后,能降低ALT、AST、TG和MDA的活性或含量,抑制TNF-α的释放,增加SOD活性。随丹参浓度增加和处理时间的延长,CYP450的表达呈逐渐降低的趋势。24 h时,丹参对NF-κB的表达无明显影响;48 h时,随丹参浓度的增加,NF-κB的表达呈逐渐降低的趋势。这表明丹参对奶牛脂肪肝有一定的治疗作用,其作用机制与促进脂质代谢、抑制肿瘤细胞坏死因子、抗脂质过氧化及降低NF-κB和CYP450的表达密切相关。     

Cloning and Sequence Analysis of Complete Genome of Porcine Circovirus Type 2 from Liaoning Province

It was aimed to lay a foundation for epidemiological survey of porcine circovirus type 2(PCV2) in Liaoning province and selection of highly homologous strain inactivated vaccine.Some pigs suspected of having postweaning multisystemic wasting syndrome (PMWS) from Shenyang,Chaoyang,Tieling,Dalian and other places in Liaoning province,were detected by PCR method,then the whole genome of 10 positive samples were cloned and sequenced.The result showed that all of them were virulent strains,9 strains were 1767 bp and 1 strain was 1768 bp,and there was a T base additon at 1039 bp.The nucleotide homology was 95.1% to 98.5% with HM038034 and JQ692110.In the phylogenetic tree,JHZ2 was PCV2a type,the other 9 strains were PCV2b type,DLS38,LYD32 and SYX7 were in the same branch and same onset time.The amino acid homology of ORF2 was 89.6% to 100.0%,there were 35 mutation points,large degree of variation,the only glycosylation site (NYS) was conserved.10 strains did not change significantly,mutants were not due to PCVD,PCV2b was the predominant type,and PCV2 was related with seasonality.     

杨树品系对杨干象抗性研究初探

杨树不同品系对杨干象抗性差异很大,以美洲黑杨为亲本的多数外来品种对杨干象抗性差,也使杨干象种群迅速扩大。该文根据调查的3年生和4年生连续2 a的不同杨树品系的平均虫口密度,根据聚类分析的方法将其划分为高感虫品系、感虫品系、低抗虫品系、中抗虫品系和高抗虫品系5个级别,结合杨树品系的类型、形态特征等进行分析,为生产上选用杨树抗性品系和减少杨干象大发生的危害及抗虫育种等方面提供参考。     

长沙市苗木基地土壤肥力的调查与评价

为了对长沙市苗木基地土壤肥力进行调查,选取了长沙市7个代表性苗木基地,研究和分析了各基地的土壤物理、化学性状,并采用Fuzzy综合评价对各基地的土壤肥力综合水平进行了评价.结果表明,长沙市苗木基地土壤pH值偏低,容重较大,土壤质地粘重,土壤有机质和氮素的含量中等偏低,土壤磷素和钾素的含量则较丰富；除林业基地外,其他基地土壤肥力综合指标均处于中等偏上水平.     

辽西地区五个冬桃新品种抗旱性差异

以辽西朝阳地区5个冬桃新品种为试材,分析了各品种冬桃的叶片保水力、水分亏缺状况、水分利用效率等指标,比较不同品种冬桃的抗旱能力,以期为进一步筛选适宜当地的冬桃栽培品系提供参考依据。结果表明:各冬桃品系在抗旱特性方面的综合表现为:"傲雪4号"表现最好,以下依次为"傲雪3号""傲雪5号""傲雪6号""傲雪2号"。     

茶树空中压条不定根原基发育影响因子研究

为了筛选出促使茶树茎段空中压条后发根的最佳技术方案,研究了成熟度、外施激素浓度、枝梢茎粗以及环割方式对根原基发育的影响.结果表明:选择生长健壮、茎粗为0.252 ～0.280 cm的当年生枝条,在黄绿色或青绿色茎段处进行传统环割,外用100 mg/L的ABT 1号生根粉处理,能有效缩短根原基发育周期.经过40～50 d...