猪瘟病毒衣壳蛋白与宿主丝/苏氨酸蛋白激酶N1相互作用的鉴定

为筛选并鉴定与猪瘟病毒(CSFV)衣壳(C)蛋白相互作用的宿主蛋白,本研究采用酵母双杂交技术以CSFV C蛋白为诱饵从猪外周血单个核细胞(PBMC)c DNA表达文库中筛选与之相互作用的宿主蛋白,共筛选到6种蛋白,分别为ATP5B、SON3、PKN1、PCBP1、RPS20和IQGAP1,根据Gene Ontology分析结果,这些蛋白分别参与细胞的增殖和代谢等过程。本研究选取丝/苏氨酸蛋白激酶N1(PKN1)进行进一步验证,经酵母共转化试验、免疫共沉淀试验和GST pull-down试验证实,宿主PKN1蛋白与CSFV C蛋白之间存在特异性结合。本研究首次证明PKN1与CSFV蛋白之间的相互作用关系,为进一步研究PKN1蛋白在CSFV感染过程中发挥的作用奠定了基础。     

Identification and Molecular Characteristics Analysis of Porcine Epidemic Diarrhea Virus Variants

To ascertain a diarrhea case in a pig farm in Shandong province,the pathological changes of dead piglets were observed and nested RT-PCR test was carried out on 7 diarrhea samples for porcine epidemic diarrhea virus (PEDV).Pathological examination revealed that intestine was detected as enlargement,hyperemia and edema.After histological examination,the typical microscopic lesions of intestine were disappearance of epithelial cells,villus shrinkage and shortening.The result of nested RT-PCR showed that all of the 7 samples could amplify a specific target band of PEDV.Molecular characteristics of two field strains showed that they had an amino acid homology of 92.3% to 92.4% with vaccine CV777 and 96.6% to 98.6% with other previous field strains which sequences were downloaded from GenBank.Phylogenetic tree analysis further revealed that all of PEDV strains could be mainly divided into two clusters of G1 and G2. G2 consisted of the field strains of our study and other field strains from USA,China and so on,which had the same sequence characteristics of two insertions and one deletion,while G1 consisted of all vaccines of CV777 and several older field strains from China and Korea.These results indicated that our field strains were the dominant strains in recent epidemic diarrhea occurrence.Moreover,its molecular characteristics might be a characterization of differentiating the field and vaccine strains.     

新时期农业生产中计算机信息技术的应用

计算机信息技术在农业生产范畴的应用,极大程度上催生了农业生产模式的变更,使得农业生产现代化规模持续扩大,更加吻合经济市场实际需求,是新时期农业发展的重要创新。立足于新时期计算机信息技术应用于农业生产范畴的背景轨迹,探讨了农业生产范畴应用计算机信息技术的实际优势,剖析了计算机信息技术的农业应用层次,以期进一步优化农业生产效率,推进农业科技创新,助推我国农业持续稳定健康发展。     

PEG模拟干旱胁迫下3种鄂尔多斯常见植物萌发期抗旱性评价

以聚乙二醇溶液(PEG-6000)为渗透介质模拟干旱条件,在12个水势梯度下,研究了干旱胁迫对木地肤、狭叶锦鸡儿和白沙蒿3种植物种子萌发的影响。结果表明,随着PEG-6000浓度的增大,3种植物的发芽率均呈现出明显的下降趋势;而较低浓度的PEG溶液可以对种子萌发产生促进和"刺激"的效果,加快种子萌发的速度,刺激初生根的生长,从而对发芽指数和萌发抗旱指数产生一定的影响。利用隶属函数综合评价3种植物在种子萌发期间的抗旱性,结果为木地肤>白沙蒿>狭叶锦鸡儿。     

Influence of salinity and moisture on the threshold shear velocity of saline sand in the Qarhan Desert,Qaidam Basin of China: A wind tunnel experiment

Determination of the threshold shear velocity is essential for predicting sand transport, dust release and desertification. In this study, a wind tunnel experiment was conducted to evaluate the influence of salinity and moisture on the threshold shear velocity of saline sand. Saline sand samples (mean particle size of 164.50-186.08 μm and the total silt, clay and salt content of 0.80%-8.25%) were collected from three saline sand dunes (one barchan dune and two linear dunes) in the Qarhan Desert, Qaidam Basin of China. Original saline sand samples were placed in two experimental trays for wet and dry processing to simulate deliquescence and desiccation, respectively. Surface moisture content ranging from 0.30% to 1.90% was generated by the steam method so that the saline sand can absorb water in a saturated water vapor environment. The motion of sand particles was determined by the observers with a solid laser. The laser sheet (0.80 cm thick), which was emitted by the solid laser, horizontally covered the sand surface and was bound to the sand. Results show that the cohesion of saline sand results from a combination of salt and water. The threshold shear velocity increases exponentially with the increase in crust thickness for the linear sand dunes. There is a positive linear correlation between the original moisture content and relative threshold shear velocity. The threshold shear velocity of dewatered sand is greater than that of wet sand with the same original moisture content. Our results will provide valuable information about the sand transport of highly saline soil in the desert.     

鸽新城疫的流行与防控

鸽新城疫是由新城疫病毒感染鸽群引起的传染病, 在世界各国鸽场均有流行。介绍了鸽新城疫流行情况及特点, 流行毒株的基因型、病毒的毒力、病毒的抗原特性, 提出在做好鸽场内生物安全控制和精细化管理基础上, 使用市售新城疫疫苗进行合理免疫是鸽新城疫防控的关键。     

基于S基因序列分析新型猪流行性腹泻病毒的遗传演化

最近，笔者实验室在青藏高原地区发现两种新亚型藏猪源猪流行性腹泻病毒（PEDV），为进一步调查新型PEDV是否在四川腹泻猪群中存在或流行，对实验室2018-2019年保存的116份猪腹泻粪便或肠组织样本进行PEDV的检测及其纤突蛋白基因（spike）分子特征研究。结果表明：腹泻样本的PEDV检出率为42.2%（49/116，95% CI=33.1%~51.8%），并获得了13条完整的S基因序列，全长为4 149~4 170 bp，序列相似性为94.2%~99.9%，其中SWUN-H3-CH-SCYA-2019的S基因与藏猪源新G1亚群PEDV的序列相似性高达97.0%~98.6%。遗传演化研究结果表明13株PEDV S基因划分为G1和G2大群，其中SWUN-H3-CH-SCYA-2019位于藏猪源新G1亚群；SWUN-19-CH-SCZY-2018、SWUN-4-CH-SCXC-2018、SWUN-1-CH-SCNJ-2019和SWUN-3CH-CH-SCZG-2019位于G2亚群中一个独立的分支，且与藏猪源新G2亚群毒株有着较近的亲缘关系。为了进一步研究13株PEDV的演化过程，以贝叶斯进化分析软件包（BEAST）进行分歧时间估算，结果表明SWUN-H3-CH-SCYA-2019的分歧时间约为2012.3年，早于藏猪源新G1亚群其余毒株的最早分歧时间（2015.7年）；SWUN-4-CH-SCXC-2018、SWUN-19-CH-SCZY-2018和SWUN-3CH-CH-SCZG-2019的分歧时间约为2014.2年，早于G2亚群的藏猪源毒株2014.7年，所有藏猪源PEDV的分歧时间均晚于四川毒株。本研究在四川地区首次发现了藏猪源PEDV，并且从毒株的分歧时间推断青藏高原的藏猪源PEDV来源于四川，为新型PEDV分子遗传进化的监测提供了依据。     

油菜杂交种DNA高通量获取方法

通过油菜(Brassica napus)杂交种深孔板内发芽,简化碱裂解法提取DNA步骤,配合高通量研磨器FastFluid Management SK300样品混匀仪的使用,提供了一整套快速高通量低成本获取油菜杂交种子叶DNA样品的集约化方案。该方法可以实现10 h完成万份DNA样品的提取,极大提高了DNA获取效率,有利于利用分子标记对杂交油菜种子进行纯度鉴定。     

一种高质量天门冬基因组DNA提取方法

针对天门冬[Asparagus cochinchinensis(Lour.)Merr.]成熟叶片富含多糖、多酚、蛋白质及其他次生代谢物质的特点,采用改良CTAB法对其基因组总DNA进行提取,并对DNA进行质量检测。结果表明,改良CTAB法可从天门冬成熟叶片中获得纯度高、完整性好的总DNA,其OD260 nm/OD280 nm为1.86,OD260 nm/OD230 nm为2.36,浓度为390μg/mL,产率达11.7μg/g,多糖、多酚和RNA杂质被去除干净,无降解现象,说明该方法提取的天门冬基因组总DNA质量较高。