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.     

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。     

苹果外部缺陷全表面在线检测分选装置研发

中国是水果消费大国,但在水果产后检测装备方面相对滞后。针对目前在线检测装置无法采集苹果全表面图像信息且无法精确计算缺陷面积的问题,该研究以表面缺陷面积的快速检测为主要目标,提出苹果全表面图像合成算法,设计了一套苹果外部品质在线检测及分级装置。该研究以苹果为例,基于球模型提出苹果全表面图像合成算法、缺陷面积校正算法精确计算苹果的表面缺陷面积。通过试验验证,对苹果表面图像进行分割合成后,整体的图像的漏检率为0。提出缺陷面积校正算法,可以计算图像中位于任意位置的苹果缺陷真实面积,选取了120个样本进行验证,其中擦伤样本、碰伤样本、痘斑病样本、表面腐败样本各30个。4种表面损伤面积的预测值和真实值的决定系数R²均在0.97以上,均方根误差(Root Mean Squared Error, RMSE)在4 mm²以下。在偏角试验中,4种表面损伤面积的预测值和真实值的决定系数R²均在0.974 2以上,RMSE在6.304 4 mm²以下。装置检测苹果的速度为2个/s,评级准确率为95%。研究结果表明,检测与苹...     

淫羊藿对肾阳虚雄性大鼠肾脏BMP-7表达的影响

目的 观察淫羊藿对肾阳虚雄性SD大鼠肾脏骨形态发生蛋白-7(BMP-7)表达的影响.方法 3月龄雄性SD大鼠24只,随机分为正常组、肾阳虚模型组、淫羊藿组,灌胃给药90d后,RT-PCR检测肾脏BMP-7 mRNA表达,免疫组化检测肾脏BMP-7的表达.结果 正常组和淫羊藿组BMP-7mRNA和蛋白显著高于肾阳虚模型组(灰度比值:0.990±0.103和1.054:t:0.097vs0.619±0.106,P＜0.01;免疫组化:0、1、0、4和0、0、0、4vs1、2、2、0,P＜0.01).结论 淫羊藿可提高肾脏中BMP-7的表达而保护肾脏.     

生物炭对不同酸化水平稻田土壤性质和重金属Cu、Cd有效性影响

针对南方稻田土壤酸化严重,导致养分流失有毒重金属活化,严重影响稻米质量安全的重大现实问题。以水稻秸秆和谷壳等农业废弃物为原料制备生物炭(分别记为RSC和RHC),研究不同原料生物炭对酸化土壤改良及其对重金属有效性的影响。设置3个生物炭用量(0,20,50 g/kg,分别记为CK、C1、C2),4种土壤酸化水平(pH 4.01,4.25,4.33,4.58,分别记为L1、L2、L3、L4),生物炭与重金属污染土壤共同培养60天后测定土壤pH、全氮、有机质、有效磷、速效钾和有效态Cu、Cd含量。结果表明:RSC对酸化土壤pH的改良效果明显优于RHC,且施炭量越高提高幅度越大,RSC的C2处理使4种酸度水平的土壤pH分别提高了0.68,0.97,1.29,1.71个单位。2种生物炭均能提高土壤的全氮、有效磷、速效钾和有机质含量,其中各施炭处理有机质显著提高,尤以速效钾的增幅最为显著,RSC对4种养分的提高均优于RHC。RHC对土壤有效态Cu含量无显著影响;RSC的C2较C1处理更能降低土壤中有效态Cu含量,使4种酸度水平的土壤分别降低了13.62%,6.57%,4.36%,7.88%。RHC处理的L3、L4土壤中有效态Cd含量显著降低,最大分别降低了13.79%,19.23%。RSC使4种酸度土壤有效态Cd含量最大分别降低了20.00%,25.81%,20.69%,19.23%。相关分析表明,土壤pH与有效态重金属含量呈显著负相关关系。水稻秸秆炭用于改良酸化土壤、降低重金属Cu和Cd有效性的效果更佳,且降低污染土壤中Cd的有效性较Cu好;生物炭对酸化程度越低的土壤pH和有效磷含量的提高以及有效态Cd含量的降低效果较好,而有效态Cu含量的降低效果则在酸化程度越高的土壤中表现更佳;土壤pH是生物炭调控重金属Cu、Cd有效性的主要影响因素。     

杉木林下套种不同树种对土壤改良效应初探

为探讨复层异龄林营建初期对土壤物理结构和养分状况的影响,研究了浙西地区杉木林下套种不同树种土壤理化特征的变化。结果表明林下套种树种后林地土壤物理结构均得到不同程度的改善,土壤养分状况进一步好转。并运用多因素主成分分析方法,评价了不同复层林模式土壤养分含量规律,并将土壤养分状况的综合优势作了排序。其结果显示杉木林下套植的12种植物在种植初期对土壤养分改良效果较好的为麻栎、毛红椿、南天竺、十大功劳、红豆杉、椤木石楠和蚊母,其中麻栎和毛红椿改良效果最佳。