A Droplet Digital PCR Method for Detection of Newcastle Disease Virus

This experiment was conducted to establish a droplet digital PCR (ddPCR) method for the detection of Newcastle disease virus (NDV). A ddPCR method was developed, which the primers and probes were designed based on the conservative regions of F gene of NDV. The concentration of primer and probe, the annealing temperature in ddPCR reaction were optimized. The sensitivity, specificity and reproducibility of ddPCR method were evaluated. In results, the optimal primer concentration and the probe concentration were 900 and 250 nmol·L^-1, the optimum annealing temperature was 55℃. The detection limit of ddPCR method was 1.8 copies·μL^-1 with a good linear response, it had no cross reaction with other six viruses (include IBV), the coefficient of variation of sample repetition was 2.4%. All the results showed that NDV ddPCR was sensitive and specific, it was suitable for quantitative detection of clinical samples infected with NDV.     

藏绵羊GHRH基因部分片段的克隆及HaeⅢ-RFLP分析

对欧拉型藏绵羊生长激素释放激素(GHRH)基因部分片段进行PCR扩增、纯化和克隆测序,利用GenBank中的Blastn方法与普通牛相应基因序列进行了比对分析,并对54只欧拉型藏绵羊该基因片段进行了HaeⅢ-RFLP研究。结果表明:欧拉型藏绵羊与普通牛GHRH基因部分序列间同源性为93%;序列间碱基变异类型主要表现为碱基转换和颠换,亦存在碱基插入和缺失;54只欧拉型藏绵羊该基因片段HaeⅢ-RFLP分析均为单态,表现为AA基因型。     

中国沙棘种群克隆生长的时空动态特征

中国沙棘(Hippophae rhamniodes)能通过分株的出生与死亡来调节种群的数量和结构,其极强的克隆繁殖能力在维持种群稳定性方面起了积极的作用。文章根据多年来的研究成果,分析了中国沙棘在结构动态、数量动态、生物量动态、生长格局动态等空间动态及时间动态特征,以其为沙棘的开发利用提供参考。     

凤山县石漠化不同植被恢复模式对土壤理化性质的影响

对广西凤山县石漠化不同植被恢复模式的土壤理化性质进行测定分析,结果表明:随着植被的恢复,其石漠化的土壤肥力效应有明显的变化,其土壤容重降低6.1%~16.8%,土壤总孔隙度增加7.5%~14.2%,土壤有机质含量增加21.4%~32.7%,水解氮含量增加32.2%~55.6%,土壤有效磷含量增加77.1%~144.6%,土壤速效钾含量增加15.2%~35.7%,但都随着土层深度的增加而减弱。不同植被恢复模式对土壤肥力效应的影响序列为任豆(Zenia insignis)-构树(Broussonetia papyrifera)模式任豆纯林模式香椿(Toona sinensis)-金银花(Lonicera japonica)模式任豆-山葡萄(Cayratia corniculata)模式。     

干旱胁迫下喷施14-羟基芸苔素甾醇对冬小麦穗花发育及碳氮代谢的调控

[目的]明确外源芸苔素甾醇类化合物(brassinosteroids,BRs)中14-羟基芸苔素甾醇(14-hydroxylated brassinosteroid,14-HBR)对干旱胁迫下冬小麦穗花发育成粒的调控效果,为小麦大田生产减轻干旱胁迫危害提供技术支撑.[方法]以大穗型品种周麦16(ZM16)和多穗型品种豫...     

基于去趋势分析的中国温带旱柳开花始期对气候变暖的响应

为了更准确地揭示植物在物候水平上对气温变化的响应与适应机制,在去气温和物候时间序列趋势的基础上,建立日尺度的气温?物候回归模型,分析中国温带地区40个站点1986-2011年旱柳开花始期对气温变化的响应与适应机理。结果表明：中国温带地区77.5%的站点旱柳开花始期呈提前趋势,相应的敏感时段春季气温则呈上升趋势（0.8±0.5℃·10a-1）。其中,75%的站点物候期或气温时间序列存在显著变化趋势。进行去趋势处理后,17.5%的站点相关性发生变化,去趋势前其相关性较低,去趋势后,表现为显著负相关;模拟的均方根误差（RMSE）从去趋势前的5.5d降至去趋势后的4.9d,说明去趋势方法能提高气温-物候响应关系诊断的精确度。基于去时间序列线性趋势的气温－物候回归模型,得到92.5%的站点旱柳开花期与敏感时段春季气温呈显著负相关关系,即随着气温的升高,旱柳开花期呈提前趋势,其线性响应速率为-2.5±2.1d·℃-1。     

Effects of Lactobacillus amylovorus Metabolites on the Barrier Function of IPEC-J2 Cells Infected by TGEV

This study used Real-time PCR to detect the changes of expression quantity of claudin 3 (CLDN-3), cytokeratin 8 (KRT8) and mucin 1 (MUC1) three proteins mRNA in IPEC-J2 cells at three different time points.IPEC-J2 cells were infected by TGEV, we explored whether Lactobacillus amylovorus metabolites could enhance cell barrier function by maintaining or improving mRNA expression of CLDN-3, KRT8 and MUC1, whether had effects on the barrier function of IPEC-J2 infected by TGEV.The results showed that mRNA expressions of MUC1, CLDN-3 and KRT8 in the normal cell control group did not change significantly with time (P> 0.05); MRS medium culture group did not change significantly compared to the normal cell control group(P> 0.05); Gene expressions of MUC1, CLDN-3 and KRT8 proteins in Lactobacillus amylovorus metabolites alone treatment group showed significant time-dependent effect (P< 0.05).After being treated 24, 36 and 48 h, gene expressions of MUC1, CLDN-3 and KRT8 proteins in Lactobacillus amylovorus metabolites+TGEV experiment group was significantly higher than virus control group (P< 0.05).Lactobacillus amylovorus metabolites could improve MUC1, CLDN-3 and KRT8 expressions in IPEC-J2 cells, while being able to negatively regulate MUC1, CLDN-3 and KRT8 expressions that TGEV induced in IPEC-J2 cells, thereby inhencing the barrier function of IPEC-J2 cell against TGEV.     

烤烟叶面喷施钼肥效应研究

为探明钼肥对黔南州烤烟产量和质量的影响,采用叶面喷施的方式,进行多点大区对比试验,研究钼肥对烤烟烟叶大小、物理特性、经济性状、常规化学成分等的影响.结果表明:叶面喷施钼肥能显著提高烟叶含钼量,上部叶含钼量平均由0.61mg/kg增加到0.89mg/kg,中部叶含钼量平均由0.47mg/kg增加到0.85 mg/kg;对...     

四川烟区CMV和PVY株系分化研究

为了明确当前四川烟区蚜传病毒的株系分化情况,该研究利用RT-PCR技术对四川烟区的CMV和PVY两种蚜传病毒病进行了检测和鉴定.依据病毒的CP基因序列合成引物,以烟草病叶的总RNA为模版,RT-PCR扩增得到了657bp的CMVCP片段和759bp的PVYCP片段,获得的基因片段连接到pGEM-T Easy载体上,测序.样品检测结果显示,在252份样品中CMV和PVY的检出率分别为33.33%和34.52%,两者复合侵染率为15.87%.构建的系统发育树分析结果表明,四川烟区的CMV株系分化不明显,主要属于IB亚组.PVY有着明显的株系分化现象,PVY~(N:O)株系、PVY~O株系、PVY~N株系在四川烟区均有发生.研究结果可能对培育抗病品种,控制这两种蚜传病毒具有一定的参考价值.     

不同化控剂对春玉米根系伤流液特性的影响及其激素调控机制

以东农253为试验材料,在玉米6叶期喷施吨田宝(DTB)、DCPTA和ETH复配剂(KP)两种化控剂,对照为清水。结果表明,DTB和KP均可提高春玉米根系伤流量,特别是生育后期的流量,分别比对照提高了72.34%和129.78%;提高了伤流液中矿质元素的流量,氨基酸的总流量也得到相应的提高,特别是丝氨酸的流量。内源激素IAA、CTK和ABA流量在各时期均有增加;GA流量在乳熟期得到显著提高,拔节期、抽雄吐丝期和灌浆初期均低于对照,之后得到显著提高。对内源激素比值均有不同程度的影响,改变了激素间的平衡水平;提高了行粒数和百粒重,产量分别比对照提高了13.89%和21.06%。相关性分析表明,玉米根系伤流量及其组分与产量密切相关,表明化控剂具有调控根系伤流量、矿质元素和内源激素流量的作用,对促进地上部生长发育和产量的形成有重要作用。