伪狂犬病病毒在NF-κB家族p65基因敲除细胞系的复制规律研究

试验旨在研究伪狂犬病病毒（PRV）在NF-κB家族p65基因敲除细胞系中的复制规律。利用慢病毒介导的CRISPR/Cas9基因定点修饰技术构建猪肺泡巨噬细胞（3D4/21）p65基因稳定敲除细胞系。通过构建p65-sgRNA重组质粒,转染至HEK293T/17细胞,收取慢病毒,感染3D4/21细胞后利用嘌呤霉素筛选获得多克隆细胞系,T7核酸酶检测敲除效率,再通过有限稀释法获得3D4/21-p65^-/-的稳定细胞系。CCK-8试剂盒检测3D4/21细胞中敲除p65基因后对细胞增殖的影响;流式细胞术检测PRV-GFP感染3D4/21及3D4/21-p65^-/-细胞后病毒增殖的差异;实时定量PCR检测PRV感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、TK基因mRNA表达水平及PRV感染细胞诱导的IL-1β和IL-6基因mRNA水平表达的变化;Western blotting检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、gE蛋白的表达;滴度测定检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后子代病毒滴度。结果表明,sgRNA2和sgRNA3的基因编辑效率较高,对其进行克隆化培养进而获得敲除p65基因的稳定表达细胞系;CCK-8试剂盒检测细胞活力表明,p65基因敲除对细胞活力无影响;流式细胞仪检测表明,同一时间点PRV-GFP在3D4/21-p65^-/-中的增殖显著高于对照细胞;实时荧光定量PCR表明在3D4/21细胞中敲除p65基因促进了PRV gB、TK基因的mRNA表达水平,而抑制了IL-1β、IL-6基因的mRNA表达;Western blotting结果表明,在3D4/21细胞中敲除p65基因促进了PRV gB、gE蛋白的表达;滴度测定结果表明,同一时间点PRV-QXX在3D4/21-p65^-/-细胞中子代病毒的复制显著高于对照细胞。以上结果均表明,p65基因敲除可促进PRV在3D4/21细胞中复制。     

Influence of four soil maintenance practices on Collembola communities in a Mediterranean vineyard

We studied the influence of four soil maintenance practices on Collembola communities in the soil of a Mediterranean vineyard: (a) postemergence herbicide with glyphosate; (b) postemergence and pre-emergence herbicides with glyphosate, terbuthylazine, diuron and oryzalin; (c) natural flora and (d) tillage to a depth of 10–15 cm. Total Collembola abundance, species diversity and species richness significantly varied between the four practices. Notably, the practice using postemergence and pre-emergence herbicides had significantly lower values. Identification of Collembola at species level allowed an interspecies comparison and revealed significant differences for the most common species between the four practices, with each practice being characterized by a different set of species. None of the species were found to be significantly more abundant in the plots treated with postemergence and pre-emergence herbicides.     

Production and Identification of Clone Sheep Using Bone Marrow Mesenchymal Stem Cells as Donor Cell

In order to obtain the cloned sheep using bone marrow mesenchymal stem cells（BMSCs）as the donor cells,the BMSCs of sheep were chosen and reconstructed embryos were built to transfer.10 published microsatellite markers were chosen,and the DNA samples from clone sheep,donor cells and surrogate ewes were amplified,and the relationship of father-son（RCP）was analyzed using the Quantity One for genotyping.The results showed that the reconstructed embryos were successfully built for electric fusion using sheep BMSCs as nuclear donor,and making nuclear transplantation into enucleated mature oocytes of which the fusion rate was 80.62%.20 surrogate ewe were chosen to be implanted with the reconstructed embryos at morula stage by implant surgery,and 5 lambs were born and only 3 were survived.The genotype of cloned sheep was in line with the dornor cell and the RCP were more than 99.999%.In conclusion,the first clone sheep were obtained successfully by using BMSCs as a nuclear donor in this experiment.     

A spatial hazard model for cluster detection on continuous indicators of disease: application to somatic cell score

Methods for spatial cluster detection dealing with diseases quantified by continuous variables are few, whereas several diseases are better approached by continuous indicators. For example, subclinical mastitis of the dairy cow is evaluated using a continuous marker of udder inflammation, the somatic cell score (SCS). Consequently, this study proposed to analyze spatialized risk and cluster components of herd SCS through a new method based on a spatial hazard model. The dataset included annual SCS for 34 142 French dairy herds for the year 2000, and important SCS risk factors: mean parity, percentage of winter and spring calvings, and herd size. The model allowed the simultaneous estimation of the effects of known risk factors and of potential spatial clusters on SCS, and the mapping of the estimated clusters and their range. Mean parity and winter and spring calvings were significantly associated with subclinical mastitis risk. The model with the presence of 3 clusters was highly significant, and the 3 clusters were attractive, i.e. closeness to cluster center increased the occurrence of high SCS. The three localizations were the following: close to the city of Troyes in the northeast of France; around the city of Limoges in the center-west; and in the southwest close to the city of Tarbes. The semi-parametric method based on spatial hazard modeling applies to continuous variables, and takes account of both risk factors and potential heterogeneity of the background population. This tool allows a quantitative detection but assumes a spatially specified form for clusters.     

Water Pricing and Sustainability of Self-Governing Irrigation Schemes

The sustainability of self-governingirrigation schemes is currently underpressure in many countries as publicfinancial support is decreasing.Furthermore, growing global concernregarding water scarcity means thatefficient water use is essential.Theoretically the choice and implementationof a water-pricing system should play acentral role in achieving this objective,both by recovering water costs and byencouraging farmers to adopt more efficientbehaviour. In the Senegal River Delta newlyestablished water users associations (WUAs)have chosen low water charges, which areaffordable for the majority of farmers butwhich underestimate long-term maintenancecosts. Combined with the difficulty ofcollecting and managing farmers' fees, thischoice has drawn them into a vicious circleleading to scheme deterioration and poorwater service. New alternatives have beendiscussed with them using a simulation toolthat takes into account both the watercosts and the farmers' incomes. Comparedwith the present fixed water charge basedonly on cultivated land, a two-part optionbased both on equipped and irrigated areasensures the recovery of fixed expenses suchas maintenance, while encouraging farmersto intensify their farming systems byadopting double cropping. Neverthelesschoosing a relevant water charge does notensure by itself the sustainability of anirrigation scheme. Combined supportproviding WUAS and farmers with efficientadvice regarding management andorganisational skills should alsocontribute significantly to achieve thisprime objective.     

“上膜下秸”措施对温室土壤水热盐调控试验研究

为了明确温室生产中不同耕作措施的水热盐调控效果,采用温室小区试验方法,以翻耕(CK)、翻耕结合地表覆盖地膜(M)、翻耕结合秸秆深埋(J)为对照,研究了翻耕结合地表覆盖地膜结合秸秆深埋(J+M)措施对温室土壤水热盐动态变化的影响。结果表明,与M、J及CK处理相比,J+M处理全生育期平均土壤含水率可分别提高8.96%、5.45%、14.32%,土壤温度分别提高3.35%、5.21%及8.32%,土壤全盐量分别降低24.44%、19.65%、26.64%,这一结果可为宁夏温室种植实施地膜覆盖与秸秆深埋改良措施提供参考。     

Sentinel-2 images reveal functional biophysical heterogeneities in crop mosaics

Landscape Ecology - Identifying landscape structure and understanding its functions are crucial for biological control. However, the relationship between the crop mosaic phenological heterogeneity...