Genetic Diversity Analysis of Cervus elaphus Using AMELY Gene in Y Chromosome

In order to determine the genetic diversity in Cervus elaphus using AMELY gene in Y chromosome,200 blood samples from Cervus elaphus yarkandensis,Cervus elaphus asiaticus,Cervus elaphus xanthopygus,Cervus elaphus songaricus and Cervus elaphus kansuensis populations were collected and AMELY genes were sequenced in this study.The haplotype diversity of Y chromosome was analyed,phylogenetic tree was built to explore the genetic diversity and the paternal origins about Cervus elaphus.The results showed that:Cervus elaphus yarkandensis had the most variation sites and the highest nucleotide diversity.The genetic distance between Cervus elaphus yarkandensis and other Cervus elaphus were far.6 haplotypes were identified in this study,named as A1,A2,A3,A4,A5 and A6,respectively.Cervu elaphus yarkandensis,Cervus elaphus asiaticus and Cervus elaphus kansuensis had separate haplotype.The NJ and ML phylogenetic trees showed that Cervus elaphus songaricus,Cervus elaphus asiaticus,Cervus elaphus xanthopygus and Cervus elaphus kansuensis clustered together which Cervus elaphus yarkandensis and Cervus elaphus kansuensis were form a department,separately.Cervus elaphus asiaticus,Cervus elaphus xanthopygus and Cervus elaphus yarkandensis clustered into one branches and there might be gene exchange among Cervus elaphus yarkandensis,Cervus elaphus kansuensis and other Cervus elaphus.     

Preliminary Study on NF-κB Signaling Pathway Regulating Brucella Intracellular Survival Molecular Mechanisms

By the infection of Brucella virulent strain and attenuated strain in mice macrophage RAW264.7,the assay was aimed to explore the relationship between NF-κB signaling pathways and Brucella virulent strain and attenuated strain in intracellular survival.Use different MOI Brucella (2308,RB51,16M and M5) to infect mice macrophage RAW264.7,after 0,4,8 and 24 h infected,cracking cell and collecting supernatant,we detected the effect of Brucella on activation of NF-κB signaling pathway by Western blotting.Different concentrations of NF-κB signaling pathway inhibitor were incubated with mice macrophage RAW264.7,with different multiplicities of infection (MOI) of Brucella infecting cells,ELISA kits to detect the expressions of TNF-α,IL-1β and IL-6 cytokine;At the same time,count the number of intracellular bacteria of CFU.The results showed that rough cattle Brucella strains RB51 could strongly activate NF-κB signaling pathway,smooth cattle Brucella strains 2308 was weak in the activation;At the same time,the activation of NF-κB signaling pathway was concentration dependent.When the MOI was 80,infection time was 8 h,NF-κB activation degrees of rough cattle Brucella strains RB51 and smooth cattle Brucella strains 2308 were the strongest,and this pathway was involved in producing TNF-α and IL-6;NF-κB signaling pathway inhibitor BAY11-7082 affected Brucella intracellular survival.So rough cattle Brucella strains RB51 intracellular survival and NF-κB signaling pathway activity were closely related.The results laid the foundation for the further study of Brucella intracellular pathogenesis,also provided scientific basis for the research of new drugs to Brucella,and prevention and treatment of brucellosis.     

小白菜自交不亲和性相关基因的分子标记开发

以自交亲和系WS-199和自交不亲和系WS-85为亲本杂交获得的F_2分离群体为供试材料,采用BSA法结合SNP芯片技术,筛选出与自交不亲和性相关的SNP标记,并将SNP差异位点序列信息与白菜基因组序列进行比对分析,并进一步开发SSR标记,共获得了与自交不亲和性相关的SSR分子标记BrA1-2、BrA1-3和BrA1-14,为分子标记辅助自交不亲和性杂交种生产提供技术支持。     

广东新农村建设的现状分析与发展对策

针对广东新农村建设存在的规划滞后、资源浪费、污染严重、社会管理水平低、环保意识薄弱等问题，提出用生态文明理念引领新农村建设以及建设广东生态文明新农村的对策。     

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。     

Wx^mp基因背景下可溶性淀粉合成酶基因SSⅡa和去分支酶基因PUL对水稻蒸煮食味品质的影响

【目的】分析在同一主效基因(Wx^mp)背景下可溶性淀粉合成酶基因SSⅡa和去分支酶基因PUL对稻米蒸煮食味品质的影响,以期为水稻品质遗传改良提供依据。【方法】选择在SSⅡa和PUL存在多态性而其他淀粉合成酶相关基因没有多态性的半糯品系宁0145和粳稻品种武运粳21进行杂交,获得F2群体与F3株系。利用分子标记,选择含有Wx^mp基因的F2单株与F3株系,将这些F2单株与F3株系分成SSⅡa^nPUL^n、SSⅡa^nPUL^w、SSⅡa^wPUL^n和SSⅡa^wPUL^w4种基因型(n和w分别表示该基因来源于宁0145和武运粳21),分析不同基因型蒸煮食味品质性状的差异,探讨同一Wxmp基因背景下不同SSⅡa和PUL等位基因对蒸煮食味品质性状的影响。【结果】不同基因型间蒸煮食味品质性状均存在显著差异,来源于武运粳21的SSⅡa^w基因和PUL^w基因分别使直链淀粉含量增加0.29%~1.00%和0.62%~1.18%,且PUL的效应大于SSⅡa,两者间存在互作效应。SSⅡa^w基因和PUL^w基因降低胶稠度和崩解值,提高了热浆黏度、冷胶黏度、消减值和回复值,对糊化温度、峰值黏度和峰值时间的作用较小。【结论】明确了Wx^mp背景下SSⅡa和PUL基因对稻米蒸煮食味品质的遗传效应,该研究结果为SSⅡa和PUL基因的分子标记辅助选择改良稻米品质提供了理论依据。     

MacCormack算法在长输管道瞬变流动仿真中的应用

为了提高长输管道仿真精度和速率,基于MacCormack格式基本原理和流动方程建立了长输液体管道水力瞬变流动仿真模型。以某管道为例,分别采用该仿真算法和特征线法模拟了阀门关闭和流量增加引起的瞬变流动,并探讨了MacCormack格式中不同边界条件处理方法及Courant常数值对模拟结果的影响。结果表明:MacCormack格式可用于精确仿真长输管道瞬变流动过程,采用特征线法求解预估层边界参数后,其振荡幅度、收敛速度均优于特征线仿真方法;同时,采用特征线法处理MacCormack格式边界条件较线性外插法更易收敛。研究成果可为长输管道瞬变流动仿真提供参考。     

饲料蛋白水平对芙蓉鲤鲫幼鱼增重和消化酶活性的影响

以进口鱼粉和豆粕为饲料蛋白源,研究了6种蛋白水平(分别为27%、30%、33%、36%、39%和42%)的等能饲料对芙蓉鲤鲫幼鱼增重和肠道、肝脏消化酶活性的影响。结果表明,饲料蛋白水平的增加,显著地提高了鱼体增重率、特定生长率、饲料效率、肥满度(P<0.05),并降低了饵料系数和脏体比,还显著地提高了肠道组织蛋白酶、食糜蛋白酶和GOT及GPT活性(P<0.05)。随着饲料中蛋白水平的上升,肝脏蛋白酶和食糜、肠道组织、肝脏淀粉酶活性无显著性差异(P>0.05)。