基于SYBR Ⅰ实时荧光定量PCR对猪流行性腹泻病毒野毒和疫苗弱毒鉴别诊断方法的建立

本研究参考GenBank中登录的猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)野毒株和弱毒疫苗株(CV777弱毒疫苗株)在高度保守ORF3基因核苷酸序列的差异,设计一对特异性荧光定量引物,分别建立基于SYBRⅠ实时荧光定量RT-PCR方法。结合熔解曲线分析可见,其野毒株和弱毒疫苗株熔解温度(Tm)分别为(81.84±0.17)℃和(83.16±0.14)℃,扩增产物的熔解曲线分析均只出现1个单特异峰,对传染性胃肠炎病毒、猪轮状病毒、猪细小病毒、猪流感病毒、猪繁殖与呼吸综合征病毒、猪伪狂犬病毒、猪瘟病毒均检测不到荧光信号。结合熔解曲线可直接鉴别猪群中PEDV的感染情况和程度,可对免疫猪群PEDV野毒感染和疫苗免疫做出快速准确的鉴别诊断,尤其是对PEDV弱毒疫苗免疫后仍爆发PEDV野毒感染的研究更有临床意义。     

宽皮柑橘单核苷酸多态性的高分辨率熔解曲线分型

 高分辨率熔解曲线分析（High resolution melting analysis,HRM）可以检测单碱基改变引起的DNA双链熔解温度（T_m）值变化,从而可以对样本在单核苷酸多态性分子标记（Single nucleotide polymor- phism,SNP）上进行基因分型。通过分析NCBI数据库中宽皮柑橘的表达序列标签（Expressed sequence tag,EST）数据鉴别SNP位点,并用小片段扩增法高分辨率熔解曲线分型技术（High resolution melting analysis of small amplicons）分析11个宽皮柑橘（Citrus reticulata）品种以及柳橙（Citrus sinensis Osbeck var.‘Liucheng’）的5个SNP位点的基因型。结果显示,小片段扩增法高分辨率熔解曲线分型可以快速、清楚地分辨纯合与杂合基因型,在校正温度差异后也可以很好地分辨同一个SNP位点不同的纯合型。统计分析表明样本在所有SNP位点上均存在多态性,5个SNP位点的平均多态性信息含量（PIC）为0.3190,显示样本在这组SNP位点上具有较高的杂合率。     

熔融造粒腐植酸尿素的缓释性能研究

运用碱处理风化煤得到水溶性腐植酸和沉淀混合物以及未处理的风化煤细粉（80目）分别与熔融尿素混合造粒,制得3种类型的腐植酸尿素产品,利用土柱淋洗方法研究不同腐植酸尿素产品的缓释性能。结果表明,除添加10%的未处理风化煤处理外,其余腐植酸尿素产品与普通尿素相比,均表现出具有氮缓释性能。其中,以添加水溶性腐植酸的产品缓释性最强;且当水溶性腐植酸与尿素比例为3∶7时,肥料的缓释效率最高。在本试验条件下,风化煤类腐植酸熔融造粒工艺下制成腐植酸复合尿素的缓释性能,与添加腐植酸的数量呈正相关;水溶性腐植酸添加比例与腐植酸复合尿素缓释性能相关性较低。3种类型的腐植酸复合尿素,以添加水溶性腐植酸的肥料质量最稳定。对风化煤进行活化后提取水溶性腐植酸,适量添加到熔融尿素中进行造粒,可生产出具有较好缓释性能的腐植酸尿素产品。     

融雪化冰用碳纤维导电混凝土的电阻率研究

分析了用于路面融雪化冰的碳纤维导电混凝土电阻率的大小容许范围以及对电阻稳定性的要求.在此基础上,对比分析了6种用于预测两相复合材料电阻率的方法,并通过试验确定了融雪化冰用碳纤维导电混凝土的电阻率预测公式--Fan氏公式的修正方法,为融雪化冰用碳纤维导电混凝土的实际应用提供了设计基础.     

Changes in the thermal and structural properties of maize starch during nixtamalization and tortilla-making processes as affected by grain hardness

The objective of this work was to evaluate the changes in the thermal and structural properties of maize starch during nixtamalization and the tortilla-making process and their relationship with grain hardness. Three maize types with varying hardness (hard, intermediate, soft) were processed by three nixtamalization processes (classic, traditional and ecological). Starch from the three maize types showed an A-type pattern and two endotherms corresponding to gelatinization and melting of the Type I amylose-lipid complexes. After cooking and steeping, in intermediate and soft grains the partial gelatinization and the annealing affected the starch properties and promoted the formation of amylose-lipid complexes. These effects were not observe in hard grains. The increase in melting enthalpy and the intensity of the peak 2θ∼20° from nixtamal to tortillas demonstrated the formation of amylose-lipid complexes. A third endotherm above 114 °C in some treatments of nixtamal and tortilla starch demonstrated the transformation of some amylose-lipid complexes in a most ordered structures (Type II complexes). The V-type polymorph structure found in native starch, nixtamal, and tortilla corresponds to a coexistence of Type I and Type II complexes. Formation of amylose-lipid complexes in tortillas had a partial effect on decreasing starch retrogradation (r = −0.47, P < 0.05).     

基于高分辨率熔解曲线技术快速筛选黄瓜SNP1

为筛选用以黄瓜纯度检验和品种鉴定的SNP位点,以17个黄瓜杂交品种及其34份亲本为SNP筛选群体,利用高分辨率熔解曲线(high resolution melting,HRM)技术筛选SNP位点.本研究通过HRM分析和焦磷酸测序确认,获得CLA0(TT/--)、CLA1(C/T)和CLA6(A/G)三个SNP位点,群体...     

雪盖影响下季节性冻土消融期的土壤温度特征分析

土壤温度的变化直接影响到季节性冻土的冻融过程,进而影响融雪径流的产流机制,且往往是春洪发生的关键之所在,故深入研究土壤温度的时空变化特征具有重要的现实意义.以天山北坡军塘河流域为研究区,采用定点对比连续观测的方法,利用2010年3月13日至4月1日融雪期观测到的雪盖和裸地两种不同下垫面的浅层(5,10,20,30,40 cm)土壤温度气温资料,对比分析各层深度的土温变化情况、雪盖对其产生的影响及其与近地面气温之间的关系.结果表明:雪盖对浅层土壤具有明显的绝热保温作用,由浅到深各层土温变化具有传递和延迟现象;土壤日温差也会随土层深度的增加而减小;近地面气温与土壤温度存在较高的相关关系,土壤温度日变化表现为单峰单谷型波动.     

含水量变化对冻融指标影响试验研究

我国冻融侵蚀面积 12 5万km2 。将试验土样分两组按一定密度和一定含水量装入试样盒进行试验 ,得出同种土质不同含水量、不同试验条件下的冻胀系数和融沉系数等有关冻融的指标。研究结果表明 :含水量及其变化是影响土体冻融指标的关键因素 ,土体的冻融是一个动态变化的过程。进行冻融侵蚀评价时 ,要充分考虑土质和土壤本身的含水量及冻融过程中的水分补给、环境变化等因素。     

High‐resolution melting analysis for rapid detection and characterization of Botrytis cinerea phenotypes resistant to fenhexamid and boscalid

A novel, high‐resolution melting (HRM) analysis was developed to detect single nucleotide polymorphisms (SNPs) associated with resistance to fenhexamid (hydroxyanilides) and boscalid (succinate dehydrogenase inhibitors) in Botrytis cinerea isolates. Thirty‐six single‐spore isolates arising from 13 phenotypes were selected and tested for fungicide sensitivity. Germ tube elongation assays showed two distinct sensitivity levels for each fungicide. Sequencing revealed that resistance to fenhexamid was due to a nucleotide change in the erg27 gene, resulting in an amino acid replacement of phenylalanine (F) with serine (S) or valine (V) at position 412 of the protein, whereas in isolates resistant to boscalid, a nucleotide change in the sdhB gene resulted in the replacement of histidine (H) with arginine (R) or tyrosine (Y) at position 272 of the respective protein. In each case, melting curve analysis generated three distinct profiles corresponding to the presence of each nucleotide in the targeted areas. HRM analysis successfully detected and differentiated the substitutions associated with resistance to both fungicides. In vitro bioassays, direct sequencing and high‐resolution melting analysis showed a 100% correlation with detection of resistance. The results demonstrate the utility of HRM analysis as a potential molecular tool for routine detection of fungicide resistance using known polymorphic genes of B. cinerea populations.