MSTN基因编辑牛肌肉转录组测序及生物信息学分析

为探究肌生长抑制素（myostatin,MSTN）基因对牛肌肉发育的具体调控机制,本研究选取同一牛场健康鲁西黄牛10头,其中通过转基因技术得到的基因编辑牛MSTN^-/-和同种非转基因野生型牛各5头。分别采集两组牛腿臀肌肉样品,利用IIlumina HiSeq高通量测序技术进行转录组测序分析,通过生物信息学方法比较两组样本间的差异表达基因,并进行GO和KEGG富集分析,最后利用实时荧光定量PCR验证转录组测序数据。结果显示,基因编辑型牛和野生型牛之间共检测到18 071个基因。在log₂|FoldChange|≥ 1.48条件下,筛选出406个差异表达基因,其中347个显著上调,59个显著下调。GO功能富集分析显示,MSTN基因编辑后显著影响915个功能类别（P<0.05）,差异基因主要参与结合、生物系统调节、免疫系统等相关功能。KEGG通路富集分析结果共涉及211个通路,差异基因主要富集在细胞黏附分子、趋化因子信号通路、细胞因子互作等信号通路上,进一步从中筛选出可能参与细胞生长、肌肉发育的差异基因（CD14、KIT、CSF1R、FBP1、DUSP4、ULBP21、PRKCB、SPN、CHAD、SRC）。实时荧光定量PCR检测结果显示,所选差异基因表达水平与转录组表达水平一致,证明测序结果的可靠性。本研究结果表明,MSTN基因发挥作用后可以介导多个下游基因表达,从而影响相关信号通路及生物学过程;同时,所筛选出的差异表达基因可作为进一步研究骨骼肌调控机制的候选靶标。     

猪圆环病毒2型灭活疫苗中病毒抗原含量实时荧光定量PCR检测方法的建立

依据GenBank公布的猪圆环病毒2型Cap基因序列,在保守区域设计特异性引物和TaqMan探针,优化反应体系,建立评价猪圆环病毒2型灭活疫苗中病毒含量的实时荧光定量PCR检测方法,对方法的特异性、敏感性和重复性进行试验,并验证灭活剂用量和灭活时间对检测结果的影响。结果显示：该方法只对猪圆环病毒2型基因有特异性扩增,其他3种对照病毒基因的扩增结果均为阴性;检测灵敏度达到10^2.0 TCID₅₀/mL,比普通PCR方法高100倍;方法的重复性好,对同一样品进行10次检测,变异系数为2.28%;不同灭活剂用量和灭活时间对结果的影响较小,不会因各厂家使用的灭活剂用量和灭活时间不同,影响疫苗对比实验的公平性。该研究成功建立了一种评价猪圆环病毒2型灭活疫苗中病毒含量的实时荧光定量PCR检测方法,用于猪圆环病毒2型灭活疫苗样品中病毒抗原含量的定量,其结果可以反映不同猪圆环病毒2型灭活疫苗样品中抗原含量差异,为研究猪圆环病毒2型灭活疫苗病毒抗原含量评估方法提供了新的思路。     

Changes in Physical Properties of Soils with Land Use Time in the Brazilian Savanna Environment

Soils from central Brazil have been intensively used over the last decades because of the rapid conversion of savannas ( Cerrado ) into corn/soybean fields. The objective of this work is to study modifications in the physical properties of soils in the Rio Verde watershed, as a function of the land use time for agriculture, determined from classification of Landsat satellite images between 1980 and 2010. Soil samples were collected at surface (0–20 cm) and subsurface (20–40 cm) horizons for the different classes of land use time (<10, 10–20, 20–30, and >30 years). The following physical properties were measured: bulk density (BD), air permeability (K_a), penetration resistance (PR), microporosity (MI), macroporosity (MA), and total porosity (TP). Results showed a strong expansion with time of agriculture that occupied 35·3% (1980), 37·4% (1990), 51·3% (2000), and 60·9% (2010) of the watershed area. When properties were compared with those from the reference areas (preserved soils under native vegetation), significant differences were observed for all the physical attributes of soils for a land use time higher than 20 years. Overall, BD and PR increased with land use time, and the opposite was verified for K_a, MA, and TP. Some physical properties presented values (e.g., 1·54 g cm⁻³ for BD and 0.06 cm³ cm⁻³ for MA) close to the critical ones reported to affect crop development, but they were not still impacting on local soybean yield. Copyright © 2013 John Wiley & Sons, Ltd.     

插秧机作业面积测量远程监测系统软件设计——基于VisualBasic.NET 2010

为实现对插秧机作业区域自动识别和作业面积的自动测量,实时掌握跨区作业插秧机远程作业面积参量值,以VisualStudio平台上的VisualBasic.NET2010为开发环境,开发了一套高速插秧机跨区作业面积远程测量监测系统软件。该软件功能主要包括GPS定位轨迹及作业区域的识别、面积测量、数据通讯、数据显示和数据存贮。系统采用VisualBasic.NET2010语言进行开发,将远程跨区作业的插秧机面积测量监测数据经由GPRS网络进行无线传输到上位机,并采用SOCKET组件和ADO.NET技术实现系统数据通信,完成PC机与下位机之间数据的传输,对测量结果进行显示存储,同时通过系统误差分析完成系统的可行性判断。     

基于定向定量堆放的马铃薯收获机设计

在研究小型马铃薯收获机的基础上,设计了一款能根据收集薯块质量往特定方向堆放的马铃薯收获机,主要由挖掘装置、输送装置及定向定量堆放装置等组成。在马铃薯收获机的后部设计一个四分区临时集薯器,能根据所收获马铃薯的质量来打开集薯器,将收集的马铃薯向中间或者一侧堆放,从而实现了马铃薯的连续挖掘及收集薯块的定向定量间隔堆放,可在较大程度上降低人工捡拾的工作量,同时避免拖拉机碾压伤薯。     

Study on Expression Difference of IGFBP-5 Gene in Different Tissues of Kazakh and Yanqi Horses

The study aimed to explore the mRNA expression pattern of insulin-like growth factor binding protein-5 (IGFBP-5) gene in different tissues of Kazakh and Yanqi horses.The expression of IGFBP-5 gene in different tissues of heart,liver,spleen,lung,kidney,small intestine,large intestine,cecum,intercostal muscles,longissimus dorsi muscles,brachialis muscle and gluteus in two horses were detected by Real-time quantitative PCR and compared the mRNA expression in the same tissues of two breeds.The results showed that the expression of IGFBP-5 in longissimus dorsi muscle and brachialis muscle of two breeds were significantly higher than other tissues including heart,liver,spleen,lung,kidney,small intestine,large intestine and cecum (P<0.05),and was the lowest in large intestine.The expression of IGFBP-5 in kidney,small intestine,large intestine,cecum,longissimus dorsi muscle and brachialis muscles of Yanqi horse were higher than in the same part of Kazakh horse,and among those in longissimus dorsi muscle and large intestine of Yanqi horse were extremely significantly higher than in the same part of Kazakh horse (P<0.01),and in small intestine and cecum of Yanqi horse were significantly higher than in the same part of Kazakh horse (P<0.05).The test was for further researching the biological function of IGFBP-5 gene,and it could provide a theoretical basis for genetic improvement of production performance of horse in our country.     

Analysis on Polymorphisms of Three CNV Regions in Five Pig Breeds

The paper was aimed to investigate the polymorphism of copy number variation (CNV) in different pig breeds.Three CNV regions of CNVR91,CNVR92 and CNVR143 were chosen from the porcine SNP60 chip genotyping results.The polymorphisms of three CNVs were determined by Real-time quantitative PCR method,taking five pig breeds as samples,including Yorkshire pig,Xiang pig,Kele pig,Nuogu pig and Rongchang pig breeds.The results showed that the dominant status of CNVR91 was loss in Xiang pig,while it was normal in other four pig breeds.The major type of CNVR92 was deletion in Xiang pig,Yorkshire pig,Kele pig and Rongchang pig breeds,with a high normal percent in Nuogu pig.For CNVR143,the dominant event was gain in Xiang pig and Nuogu pig breeds,but it was not diverse in other three pig breeds.These results indicated that three CNV regions emerged with polymorphism in five pig breeds,which might have effects on gene expression in CNV regions and physiological function by dosage effect especially in Xiang pig,Nuogu pig and Kele pig breeds.     

Describing Human–Wildlife Interaction from a European Perspective

European researchers from both the natural and social sciences show growing interest in studying interactions between society and wildlife. A wealth of theoretical frameworks, concepts, and methods are used, but an integration of perspectives is lacking. This research note summarizes results from two workshops that included 63 delegates from 25 European countries, as well as a follow-up survey of 41 respondents. Two main theoretical approaches to the study of human–wildlife interactions were identified. One approach focuses on the collective societal level relying on theories of governance, social representation, deliberative procedures, and commons theory. The other approach targets individuals or groups, and is based on theories such as the cognitive hierarchy, theory of reasoned action, and theory of planned behavior. Interdisciplinary collaboration is needed to identify the best options for wildlife conservation and management in a more politically integrated Europe.     

基于"3S"技术的甘肃子午岭自然保护区土地利用变化研究

以甘肃省子午岭自然保护区为研究对象,运用"3S"技术,对研究区的地形图、不同时相的TM遥感影像进行数据预处理,并分类提取了研究区2个时段的土地利用信息.对分类信息进行统计分析后并与实地调查数据进行比较,结果表明,近20年来甘肃子午岭自然保护区的森林覆盖面积在稳步提高,乔木林面积明显增加.为甘肃子午岭自然保护区工程的建设提供了基础信息,为林区植物种群生长及分布调查研究提供了宏观数据,并且输出了研究区不同时相土地利用专题图.     

薄壳山核桃疮痂病菌TaqMan荧光定量PCR检测方法的建立及应用

疮痂病是薄壳山核桃上最具毁灭性的病害,带菌植物材料是传播疮痂病的重要来源。准确、灵敏、快速的检测方法可为该病害流行规律调查和防控提供有力的依据。本文通过比较薄壳山核桃疮痂病菌Venturia effusa及其近似种之间的ITS序列差异,设计了特异性引物和TaqMan探针,建立了薄壳山核桃疮痂病菌的荧光定量PCR检测方法。特异性检测结果表明,该方法可以检测不同地区的薄壳山核桃疮痂病菌菌株,而对其近似种以及薄壳山核桃上的其他真菌均没有信号。本研究建立的检测方法对薄壳山核桃疮痂病菌DNA的最低检测限可达0.5 pg/μL。该方法用于田间样品检测时,检测时间仅需1 h,远快于常规的分离培养法。本研究建立的基于TaqMan探针的荧光定量PCR检测方法为薄壳山核桃疮痂病菌的快速检测和监测提供了有力工具。