犬细小病毒高效纳米PCR检测方法的建立及初步应用

为建立灵敏、快速的犬细小病毒(CPV)检测方法,本研究针对CPVVP2基因保守序列设计一对能扩增574bp片段的特异性引物,对纳米PCR的退火温度、引物浓度等反应条件进行了优化,并对其特异性和灵敏度进行了评估。结果表明,所建立的纳米PCR特异性和灵敏性良好,最低核酸检出量为2拷贝·μL^-1,其敏感性比常规PCR高1000倍。分别采用纳米PCR和常规PCR,对广西、北京、吉林送检的75份疑似CPV的临床样品进行检测,CPV阳性率分别为89.3%(67/75)和70.7%(53/75),表明该方法具有更高的敏感性,适用于CPV低含量临床样品的检测。本方法的建立为CPV感染的早期快速、灵敏、准确的诊断提供了新方法,为CPV的防控奠定基础,具有重大的临床应用意义和价值。     

Distribution of latent bovine herpesvirus 2 DNA in tissues of experimentally infected sheep

The biology of latent infection by bovine herpesvirus 2 (BoHV-2), the agent of mammillitis in cows, remains largely unknown. We herein report attempts to reactivate the latent infection and investigated the sites of BoHV-2 latency in experimentally infected sheep. Ewes inoculated with BoHV-2 in the udder’s skin shed virus for up to five days, developed mammillitis and seroconverted. However, attempts to reactivate latent infection by dexamethasone administration at day 40 pi failed. Nevertheless, viral DNA - and not infectious virus - was detected by PCR in several nerve ganglia and/or regional lymph nodes (LNs) of all animals at day 40 post-reactivation. Likewise, lambs previously inoculated with BoHV-2 in the nose harbored latent viral DNA in trigeminal ganglia, tonsils and regional LNs. These results demonstrate that BoHV-2 establishes latent infection in nerve ganglia and in regional lymphoid tissues, yet virus reactivation is not easily achieved by standard protocols used.     

Basic characterization of avian β‐defensin genes in the Japanese quail,Coturnix japonica

In this study, we identified a cluster of 14 avian β‐defensins (AvBD; approximately 66 kbp) in the Japanese quail, Coturnix japonica. Except for AvBD12 (CjAvBD12) and ‐13, the CjAvBDs coding sequences exhibited greater than 78.0% similarity to the respective orthologous chicken AvBD genes (GgAvBD). The putative amino acid sequence encoded by each CjAvBD contained six cysteine residues and the GXC (X1‐2) motif considered essential for the β‐defensin family. Each CjAvBDs also formed a sub‐group with the respective orthologous genes of various bird species in a phylogenetic tree analysis. Synteny between the CjAvBD cluster and GgAvBD cluster was confirmed. The CjAvBD cluster was mapped on the long‐arm end of chromosome 3 by linkage analysis based on single nucleotide polymorphisms (SNPs) of CjAvBD1 and CjAvBD12 (approximately 46kbp), as well as GgAvBD cluster. We also confirmed that CjAvBD1, ‐4, ‐5, ‐9, and ‐10 are transcribed in 20 tissues, including immune and digestive tissues. However, our experimental data indicated that the CjAvBD cluster lacks the AvBD3 and ‐7 loci, whereas the CjAvBD101α, ‐101β, and ‐101θ loci arose from gene duplication of the AvBD6 orthologous locus in the CjAvBD cluster after differentiation between Coturnix ‐ Gallus.     

猪ADAR2基因全长cDNA克隆、序列特征及表达模式分析

旨在克隆猪作用于RNA的腺苷脱氨酶2基因(ADAR2)全长cDNA序列,同时对该基因在猪不同组织中的表达规律进行探索.利用RACE(rapid-amplification of cDNA ends)对大白猪ADAR2基因mRNA全长序列进行克隆,并进行生物信息学分析;用荧光定量PCR方法检测35日龄大白猪心、肝、肺、肾...     

绒山羊皮肤毛囊兴盛前期组织形态、chi-miR-107-3p与RHBDF2基因表达分析

为了探究chi-miR-107-3p和菱形家族蛋白2（rhomboid family member 2,RHBDF2）基因在不同品种（系）和光控增绒绒山羊兴盛前期（5~7月份）皮肤毛囊重建时的表达差异,本试验在7月份采集内蒙古阿拉善型绒山羊、敏盖绒山羊和光控增绒技术处理的阿尔巴斯型绒山羊体侧皮肤毛囊组织,采用组织切片技术对组织形态比较分析,实时荧光定量PCR法检测chi-miR-107-3p和RHBDF2基因的表达量。结果显示,在皮肤毛囊兴盛前期,阿拉善型绒山羊次级毛囊正处于重建阶段,敏盖绒山羊和阿尔巴斯型绒山羊（光控增绒）次级毛囊重建已基本完成;在阿拉善型绒山羊皮肤组织中chi-miR-107-3p表达量极显著高于敏盖绒山羊和阿尔巴斯型绒山羊（光控增绒）（P<0.01）,而RHBDF2基因表达量极显著低于其他两品种（系）（P<0.01）,且RHBDF2基因表达量在敏盖绒山羊皮肤组织中显著低于阿尔巴斯型绒山羊（P<0.05）。不同品种（系）和光控增绒绒山羊皮肤毛囊兴盛前期组织显微结构与chi-miR-107-3p、RHBDF2基因在皮肤组织中的表达差异分析结果相一致,次级毛囊重建初期chi-miR-107-3p表达量较高,而RHBDF2基因表达量极低,随着次级毛囊重建完成,chi-miR-107-3p表达量明显降低,RHBDF2基因表达量逐渐增高,且在不同品种（系）内蒙古绒山羊绒毛生长发育过程中的表达机制基本相同。因此,chi-miR-107-3p和RHBDF2基因是绒山羊皮肤毛囊生长发育的重要调控因子。     

Short term soil priming effects and the mineralisation of biochar following its incorporation to soils of different pH

The aim of this work was to determine the magnitude of the priming effect, i.e. short-term changes in the rate (negative or positive) of mineralisation of native soil organic carbon (C), following addition of biochars. The biochars were made from Miscanthus giganteus, a C4 plant, naturally enriched with ¹³C. The biochars were produced at 350 °C (biochar350) and 700 °C (biochar700) and applied with and without ryegrass as a substrate to a clay-loam soil at pH 3.7 and 7.6. A secondary aim was to determine the effect of ryegrass addition on the mineralisation of the two biochars.After 87 days, biochar350 addition caused priming effects equivalent to 250 and 319 μg CO₂-C g⁻¹ soil, in the low and high pH soil, respectively. The largest priming effects occurred at the start of the incubations. The size of the priming effect was decreased at higher biochar pyrolysis temperatures, which may be a way of controlling priming effects following biochar incorporation to soil, if desired. The priming effect was probably induced by the water soluble components of the biochar. At 87 days of incubation, 0.14% and 0.18% of biochar700 and 0.61% and 0.84% of biochar350 were mineralized in the low and high pH soil, respectively. Ryegrass addition gave an increased biochar350 mineralisation of 33% and 40%, and increased biochar700 at 137% and 70%, in the low and high pH soils, respectively. Certainly, on the basis of our results, if biochar is used to sequester carbon a priming effect may occur, increasing CO₂-C evolved from soil and decreasing soil organic C. However, this will be more than compensated for by the increased soil C caused by biochar incorporation. A similar conclusion holds for accelerated mineralisation of biochar due to incorporation of fresh labile substrates. We consider that our results are the first to unequivocally demonstrate the initiation, progress and termination of a true positive priming effect by biochar on native soil organic C.     

苯并噻二唑甲酸三氟乙酯诱导仙客来抗枯萎病的研究

为了明确新型化合物苯并噻二唑甲酸三氟乙酯对土传病害的诱导抗病作用,本文研究了该化合物诱导仙客来对枯萎病的抗病效果。通过叶面喷施苯并噻二唑甲酸三氟乙酯可诱导仙客来对枯萎病的抗病效果,在仙客来移栽缓苗后7d叶面喷施该化合物,不同浓度对枯萎病的抗病效果不同,其中100mg.L-1的处理,在诱导后25、30和35d的抗病效果分别为82.90%、58.59%和64.17%;在仙客来成株枯萎病发病初期用该化合物诱导,抗病效果仍可达63.91%,显著高于噁霉灵处理（47.63%）。该研究将为寻求通过叶面喷雾施药防治园艺作物土传病害的新途径起到参考作用。     

Decomposition of peat,biogenic municipal waste compost,and shrub/grass compost added in different rates to a silt loam

An incubation experiment was carried out to test the effects of biogenic municipal waste (compost I) and shrub/grass (compost II) composts in comparison to peat on respiration and microbial biomass in soil. The amounts of these three substrates added were linearly increased in the range of field application rates (0.5%, 1.0%, 1.5%, 2.0%). The sum of CO₂ evolved during the incubation was markedly raised by the three substrates and increased with the rate of substrate concentration. However, the percentage of substrate mineralized to CO₂ decreased with the addition rate from 103 to 56% for compost I, from 81 to 56% for compost II, and from 21 to 8% for peat. During the first 25 days of incubation, compost I enlarged the biomass C content, which remained constant until the end. In contrast, compost II did not raise biomass C initially. But at the end of the incubation, the biomass C content of all 4 compost II treatments almost reached the level of the respective compost I treatment. The increase was significantly larger the more of the two composts was added. In contrast to the two composts, the addition of peat did not have any significant effect on microbial biomass C. The average qCO₂ values at day 25 declined in the order compost I > compost II > peat, at day 92 the order was changed to compost II > peat > compost 1. This change in the order was caused by a significant decrease in qCO₂ values of the compost I treatments, a significant increase in qCO₂ values of the peat treatments and constant qCO₂ values in the compost II treatments.     

滴灌条件下核桃园土壤水分动态变化的数值模拟研究

为探究滴灌条件下核桃根系对土壤含水量的影响,试验以8 a生核桃树为研究对象,使用分层分段挖掘法分析核桃根系的空间形态分布,通过构建HYDRUS-2D模型,对有根系吸水下的土壤水分变化进行求解,并利用实测点数据校验2D模型的可用性。经过实际计算与理论分析,得出如下结论:在垂直二维剖面中,水平距树0~30 cm、垂直40~60 cm土层的细根根长密度分布最大,为1 618.216 m·m~(-3)。对比实测数据与模拟数据,结果显示RMSE均小于0.0186 cm~3·cm~(-3),ME绝对值小于0.0108 cm~3·cm~(-3),证明数值模拟效果较好,HYDRUS模型可应用于实际试验研究。将根系模块纳入模型模拟,并比照单独水流模块作用,结果显示根系对土壤含水量产生影响,使体积含水率曲线整体低于单独水作用下的含水率曲线,且两者关系差异性显著。模拟核桃生育前期土壤含水量变化,结果表明灌水后6 d至7 d内,土壤含水量存在明显减小现象,故可将生育前期灌水周期设定为6 d或7 d。     

不同品种断奶仔猪肠道发育的差异及与血清胰高血糖素样肽-2的关系

为考察不同品种断奶仔猪小肠发育的差异及肠绒毛高度与血清GLP-2的关系,本试验分别测定了不同品种仔猪24、26d和30d时的血清GLP-2浓度及30d时肠道发育参数,并分析了回肠绒毛高度与GLP-2的关系。结果表明:大×太猪的小肠长(相对长)、小肠重(相对重)显著高于长白猪(P0.05),小肠重显著高于太湖猪(P0.05),而太湖猪小肠相对重显著高于长白猪(P0.05),小肠相对长极显著高于长白猪(P0.01);大×太猪和长白猪的回肠肠壁厚度、绒毛高度和隐窝深度显著高于太湖猪(P0.05);30d时,大×太猪血清GLP-2水平最高,且极显著高于长白猪和太湖猪(P0.01),3种猪回肠绒毛高度与血清GLP-2极显著相关(P0.01)。表明30d时的大×太猪肠道发育优于太湖猪和长白猪,不同品种断奶仔猪的小肠绒毛高度与血清GLP-2水平高度相关。