盐胁迫是一种典型的非生物胁迫，导致植物生长缓慢、发育迟缓、产量和果实品质下降。施肥是保证作物正常生长的必要措施，其中，氮素更是关键元素。研究报道氮肥施加可提高作物耐盐性，但是，氮肥对葡萄耐盐性的影响尚不清楚。因此，本研究以酿酒葡萄幼苗‘黑比诺’为植物材料，研究200 mmol L^-1NaCl处理下施用0.01和0.1 mol L^-1 硝酸铵（N）对葡萄耐盐性的影响。通过对葡萄幼苗叶片的生理指标、转录组和代谢组分析，发现0.01 mol L^-1的N施加显著降低了盐胁迫下葡萄叶片中超氧阴离子（O₂^.-）的积累，提高了超氧化物歧化酶（SOD）和过氧化物酶（POD）的活性，促进了抗坏血酸（AsA）和谷胱甘肽（GSH）的积累。转录组和代谢组的联合分析表明，黄酮生物合成途径（ko00941）和黄酮和黄酮醇生物合成途径（ko00944）是关键的响应通路。进一步发现，槲皮素（C00389）的积累受到盐和氮的显著调节。同时，筛选到10个关键差异基因与槲皮素含量变化高度相关（R²>0.9）并构成互作网络。此外，我们也发现，盐胁迫下叶面喷施槲皮素可提高葡萄的SOD和POD活性，增加AsA和GSH的含量，降低H₂O₂和O₂^.-的含量。因此，本研究应用氮肥和槲皮素改善了葡萄的耐盐性，并鉴定到关键的响应基因，此结果为葡萄耐盐性的提高和分子机制研究提供了新的思路。

建立一种同时鉴别诊断猪瘟病毒（classical swine fever virus, CSFV)、非洲猪瘟病毒（African swine fever virus, ASF）和猪非典型瘟病毒（atypical porcine pestivirus, APPV）的快速、灵敏、有效的检测方法。依据GenBank中登录的CSFV (5¢ UTR)、ASFV (B646L) 和 APPV (5¢ UTR) 的高度保守基因序列分别设计和优化了多对特异性引物和Taq-man探针，以保守区基因序列分别制备三种阳性质粒，用矩阵法优化单重/多重荧光PCR的反应体系和条件，为避免荧光通道的交叉干扰多重荧光PCR扩增，结合所标记的荧光报告基团做颜色补偿试验，构建标准曲线的扩增图和对应的线性方程，并进行特异性、敏感性、重复性、符合性以及临床样本的检测等试验。三种病毒的标准曲线相关系数均达到0.995以上，具有良好的线性关系；与其它常见猪病无交叉扩增反应，具有很好的特异性；多重荧光PCR的最低检测量均为1 copy/mL，具有较高的敏感性；组内和组间的变异系数均小于1%，具有很好的重复性。该方法与CSF的国标(GB/T 27540-2011), ASF的国标 (GB/T 18648-2020)，APPV的发明专利 (CN108611442A)检测样本盘的22个毒株样本符合率为100%。本研究建立的多重荧光PCR检测方法具有快速、高效、通量高、特异性好、灵敏度高等特点，可以对CSFV、ASFV和APPV病毒进行鉴别检测，为动物疫病的流行病学调查、疫情的检测提供一种新型的检测手段。本研究结合荧光PCR仪不同荧光通道设计CSFV、ASFV和APPV探针荧光信号强度较高且干扰较小的FAM、CY5和HEX报告基团，建立多重荧光PCR检测方法，用于同时鉴别诊断3种主要猪病毒的检测方法，在临床诊断中具有重要的应用价值。

Classical swine fever caused by Classical swine fever virus (CSFV) is a serious problem for swine industries in developing countries, which successful control of the disease have been relying on vaccination. However, classical swine fever still occurs in some immunized swine herds for various reasons. In this study, we conducted animal experiments to examine the influence of single or mixed infection with Porcine parvo virus (PPV), Pseudorabies virus (PRV) and Porcine reproductive and respiratory syndrome virus (PRRSV) on the protective immunity induced by the Lapinized hog cholera virus (HCLV) vaccine and the pathogenicity of CSFV. In experiment 1, pigs were first inoculated with PPV, PRV or PRRSV, then immunized with HCLV, and finally challenged with a highly virulent CSFV Shimen strain. All of the pigs immunized with HCLV survived after the challenge, while all of the pigs in the non-immunized control group died after the challenge. The pigs in the group immunized with HCLV did not show any clinical symptoms of classical swine fever and were negative with CSFV after the challenge. The pigs infected with the non-CSFV before HCLV immunization did not display any clinical symptoms after the challenge with CSFV Shiman strain, but 11 of the 12 pigs were positive with CSFV. In experiment 2, pre-infections with PPV, PRV, and PRRSV were followed by inoculation with a low-virulence CSFV strain (CSFV 39), and then the pigs were challenged with the CSFV Shimen strain. Infections by either PPV, PRV or PRRSV did not enhance the virulence of CSFV-39, but pigs infected by a mixture of the 3 viruses developed clinical symptoms after inoculation with CSFV-39. The mixed infection also increased mortality caused by the challenge with the CSFV Shimen strain. Together, these results showed PPV, PRV and PRRSV infections in pigs can reduce the efficacy of the HCLV vaccine and enhance the pathogenicity of CSFV, which may partly explain the immunization failure against CSFV in some swine herds.