利用QuantStudioTM 3D数字PCR分析转基因玉米MON863含量   总被引：1，自引：0，他引：1  

胡佳莹  姜羽  杨立桃 《农业生物技术学报》2016,(8):1216-1224

QuantStudioTM 3D数字PCR (QuantStudioTM 3D digital PCR,3D-dPCR)是一种基于超高密度亲疏水微孔芯片实现数字PCR分液原理的新型核酸绝对定量平台,在转基因生物定量领域具有极大的应用前景.本研究基于3D-dPCR平台,以转基因玉米(Zea mays)MON863混合样品为例,建立基于单重和双重数字PCR体系的转基因生物(genetically modified organisms,GMOs)含量分析方法.与传统qRT-PCR比较发现,在缺乏样品纯度、纯合度信息的情况下,数字PCR能够较好地排除这些因素的影响,测定准确的量值.研究结果表明,QuantStudioTM 3D数字PCR是一种适用于转基因生物含量分析的精确定量方法,还可反映转基因玉米种子的基因型.本研究基于3D-dPCR建立的转基因玉米MON863单重和双重定量方法为转基因检测提供了新的方法和参考.  相似文献   

Analysis of Mycosporine-Like Amino Acids in Selected Algae and Cyanobacteria by Hydrophilic Interaction Liquid Chromatography and a Novel MAA from the Red Alga Catenella repens     

Anja Hartmann  Kathrin Becker  Ulf Karsten  Daniel Remias  Markus Ganzera 《Marine drugs》2015,13(10):6291-6305

Mycosporine-like amino acids (MAAs), a group of small secondary metabolites found in algae, cyanobacteria, lichens and fungi, have become ecologically and pharmacologically relevant because of their pronounced UV-absorbing and photo-protective potential. Their analytical characterization is generally achieved by reversed phase HPLC and the compounds are often quantified based on molar extinction coefficients. As an alternative approach, in our study a fully validated hydrophilic interaction liquid chromatography (HILIC) method is presented. It enables the precise quantification of several analytes with adequate retention times in a single run, and can be coupled directly to MS. Excellent linear correlation coefficients (R² > 0.9991) were obtained, with limit of detection (LOD) values ranging from 0.16 to 0.43 µg/mL. Furthermore, the assay was found to be accurate (recovery rates from 89.8% to 104.1%) and precise (intra-day precision: 5.6%, inter-day precision ≤6.6%). Several algae were assayed for their content of known MAAs like porphyra-334, shinorine, and palythine. Liquid chromatography-mass spectrometry (LC-MS) data indicated a novel compound in some of them, which could be isolated from the marine species Catenella repens and structurally elucidated by nuclear magnetic resonance spectroscopy (NMR) as (E)-3-hydroxy-2-((5-hydroxy-5-(hydroxymethyl)-2-methoxy-3-((2-sulfoethyl)amino)cyclohex-2-en-1-ylidene)amino) propanoic acid, a novel MAA called catenelline.  相似文献   

Expression Analysis of THBS3 Gene in Different Tissues and Skeletal Muscles at Different Periods from Landrace and Tongcheng Pigs     

MA Xi-shan  TANG Zhong-lin  XIAO Chong  YANG Bo-chao  QIN Chuan 《中国畜牧兽医》2016,43(4):1032-1038

To study the expression pattern of THBS3 gene in different tissues and during skeletal muscle development, the THBS3 gene expression in different tissues and skeletal muscles during prenatal periods (33, 45, 65, 70 and 90 d) and postnatal periods (0, 9, 30, 60, 120 and 160 d) from Landrace and Tongcheng pigs were detected by Real-time quantification PCR.The results showed that THBS3 gene widely expressed in all tissues examined, exhibiting similar spatial expression patterns with expression peaks in lung in both pig breeds except in stomach and intestine.Moreover, although THBS3 gene showed a significant higher expression level in gestation than after birth in Landrace and Tongcheng pigs (P<0.05), it exhibited different expression patterns between Landrace and Tongcheng pigs, the expression peak was detected at gestation day 45 in Landrace pig, while was detected at gestation day 65 in Tongcheng pig.The results suggested that THBS3 gene involved in skeletal muscle growth and development in pigs, as well as the regulation of asynchronization of skeletal muscle development in different pig breeds.  相似文献   

1978—2018年中国农村环境管理政策演进特征——基于206份政策文本的量化分析     

潘丹  唐静  杨佳庆  陈寰 《中国农业大学学报》2020,25(6):210-222

基于手工搜集的1978—2018年中国政府颁布的206份农村环境管理政策文本,从政策发布年度、政策发布主体、政策发布形式、政策作用对象、政策工具及政策效力6个维度构建分析框架,运用政策文献计量和内容分析方法分析了中国农村环境管理政策的演进特征。结果表明:1978年改革开放以来,中国农村环境管理政策在数量上呈整体持续上升态势;农村环境管理政策经历了探索起步(1978—1989年)、初步形成(1990—2004年)、快速发展(2005—2013年)和全面推进(2014年以后)4个阶段的演进;政策发布主体的行政级别较高且呈现多元化特征,政策联合决策程度高,但政策之间统筹性和协调性较低;政策绝大多数以通知、意见等形式发出,缺乏权威性和规范性;政策的着力点主要在原生态保护层面,对农村人居环境整治的重视程度不够;命令控制型政策是政府最为偏好的农村环境管理政策工具,经济激励型、信息公开型和自愿参与型政策工具使用不足;政策整体效力的提升主要由政策发布数量驱动,政策反馈不足、政策措施较少和政策目标量化程度低是导致政策平均效力较低的主要原因。  相似文献   

Development and evaluation of a loop-mediated isothermal amplification assay for rapid and simple detection of Flavobacterium psychrophilum     

E Fujiwara-Nagata  M Eguchi 《Journal of fish diseases》2009,32(10):873-881

Flavobacterium psychrophilum is the causative agent of bacterial cold-water disease and rainbow trout fry syndrome of salmonids. The pathogen has been reported from all regions in the world involved in salmonid aquaculture, but also from natural fresh-water environments. We established a quantitative loop-mediated isothermal amplification of DNA (LAMP) method to estimate quantities of F. psychrophilum . LAMP primers were designed based on the sequence of the DNA topoisomerase IV subunit B gene, parE , of F. psychrophilum. parE LAMP exhibited a high specificity for the parE gene of F. psychrophilum but not for other related species. parE LAMP detected the gene in a wide range of concentrations from 2.0 × 10¹ to 2.0 × 10⁹ copies/reaction within 70 min and revealed a good correlation between threshold times and gene copy number.  相似文献   

Design and validation of a RT‐qPCR procedure for diagnosis and quantification of most types of infectious pancreatic necrosis virus using a single pair of degenerated primers          下载免费PDF全文

D Vázquez  J M Cutrín  J G Olveira  C P Dopazo 《Journal of fish diseases》2017,40(9):1155-1167

Infectious pancreatic necrosis virus (IPNV) is an important virus which affects the salmonid aquaculture industry worldwide; therefore, it is important to develop rapid and reliable methods of diagnosis to detect the disease at early stages. Nowadays, RT‐qPCR is replacing other methods because it provides additional information on the viral load, which is important to have a better understanding of the virus replication level and of the stage of the infection and its risk level. The main problem stems from the high diversity of this virus, which can compromise the reliability of the diagnosis. In this study, we have designed an RT‐qPCR procedure for diagnosis and quantification of IPNV based on a single pair of primers targeted to segment B. The procedure has been validated, in vitro and in vivo, testing two different types of standards against seven reference strains and 23 field isolates from different types. The procedure is reliable for the detection of any type, with a detection limit of 31 TCID₅₀ mL^?1, 50 pfu mL^?1 or 66 RNA copies mL^?1, depending on the standard. All the standard curves showed high reliability (R² > 0.95). The results support the high reliability of this new procedure for the diagnosis and quantification of IPNV.  相似文献   

地表微地形测量及定量化方法研究综述   总被引：5，自引：0，他引：5  

朱良君  张光辉 《中国水土保持科学》2013,(5):114-122

地表微地形测量是地表粗糙度定量化的基础,对地表形态动态监测、水文过程模拟以及土壤侵蚀过程模型的构建具有重要意义。目前,微地形测量方法主要分为接触式和非接触式2大类,前者包括测针法、链条法、差分GPS法等,后者包括超声波测距法、红外线传感器法、结构光激光扫描法、激光测距扫描法、三维激光扫描仪法、近景摄影测量法等。在全面回顾各方法原理、优缺点及其应用的基础上,分析地表粗糙度定量化常用方法：统计方法指数、地统计学指数和分形及多重分形模型。认为：1）差分GPS法、结构光激光扫描法、三维激光扫描仪法和近景摄影测量法将在亚毫米一,厘米级地表微地形测量及地表粗糙度多尺度特征研究中发挥重要作用,同时简单、方便的测针法可能在野外测量中依然占据主导地位;2）以地表微地形测量技术为基础,在土壤侵蚀过程模型中亟需形成一套完整的“测量一定量化一模型应用”范式,同时应加强对地表微地形空间异质性和各向异性的研究,发展新的统一的地表粗糙度定量化方法。  相似文献   

湘西桤木人工林蓄积量模型及适宜经营密度研究     

文仕知朱光玉  王忠诚何功秀  张江孙华 《中国农学通报》2012,28(1):92-97

密度是表达林分结构的重要指标,立地条件、树种及林龄不同时,其林分最佳密度也不相同。笔者以湘西桤木人工林为研究对象,采用数量化方法I和非线性回归分析方法,对2009年和2010年调查的78块标准地数据进行数据分析处理与建模。结果表明：（1）平均高、公顷株数、土层厚度、龄组、坡向、坡位、自然灾害、海拔、树种组成8种因子中,平均高、公顷株数、土层厚度和龄组对湘西桤木人工林平均木材积和单位面积蓄积量均有显著性影响,而海拔和树种组成对湘西桤木人工林平均木材积和单位面积蓄积量均无显著性影响。这对建立桤木人工林蓄积量与密度、平均高收集数据时提供理论指导,同时既能节省外业调查费用,又能保证建模数据的科学性。（2）建立了湘西8~10年桤木人工林蓄积量与密度、平均高相关关系模型： ,建模精度为96.8458%,平均相对误差为1.1334%。模型适应性检验表明,残差值呈对称均匀分布,其平均相对误差为-1.2156%。（3）该地区8~10年桤木人林的最佳经营密度为800~1000株/hm2。这对于合理地经营和管理湘西桤木人工林具有重要的理论和实际指导意义。  相似文献   

湖北省杉木人工林立地类型划分及评价     

李斌成  杜超群  袁慧  吴楚  许业洲 《湖北林业科技》2020,49(2):1-5

以湖北省杉木人工林为研究对象,选择166块具有代表性的杉木人工林标准地,并测定样地的常规立地因子。利用数量化理论Ⅰ的方法建立了杉木优势高与立地因子之间的关系模型,对研究区杉木人工林进行立地类型划分及立地质量评价。结果表明,所选择的5个立地因子与优势高数量化拟合的复相关系数为0.639;海拔、腐殖层厚度和坡位这3个立地因子对优势高的贡献率达84.99%,且影响均达到极显著水平;采用海拔、腐殖层厚度和坡位3个因子构建研究区杉木人工林立地类型,共划分为22类;按照优、良、中、差4个评价等级对166块样地所属的立地类型进行立地质量评价,得出湖北省杉木人工林在中海拔、土壤湿润、土壤养分含量较高的立地条件下生产力较高,研究区立地质量整体处于中等偏上水平的结论。  相似文献   

色彩对城市山地型公园春冬季植物景观视觉质量的影响   总被引：2，自引：1，他引：1  

陈小燕  郑宇  魏昱君  雷倩萍  潘辉 《中国城市林业》2020,(1):60-65

文章采用SBE法、色彩量化分析等研究方法,结合实地调查,应用景观美学、色彩构成理论、色彩量化分析等理论对福州市金鸡山公园植物景观质量提升进行研究,研究结果表明:1)金鸡山公园春冬植物景观整体以绿、黄绿色为主,色相种类相对较少,色彩丰富度较低;明度主要集中在N4,N5等级,位于中等水平;纯度主要分布于1~3等级内,植物色彩饱和度较低。2)根据美景度评价法及景观质量等级划分,将金鸡山植物景观分为5个等级,SBE最高最低差值为146.24,差异较大;景观质量较高的景观主色调色系较多且差别较大,绿色色调的斑块与景观色彩面积比较小(最低值仅63.52%),景观质量较差的景观主色调色系较少且差别不大,绿色色调的斑块与景观色彩面积比较大(高于80%),且美景度较高的照片其明度及纯度也较高。3)从色彩频率分析可知,Ⅰ,Ⅱ组主要为绿色系、蓝色系、黄色系及红色系;Ⅲ,Ⅳ,Ⅴ组多以冷色调的绿色系、灰色系为主。4)应适当增加植物景观主色调色系、控制绿色系色调的比例,多采用邻近色和互补色配置,并注意明度和饱和度变化以提高金鸡山公园植物景观质量。  相似文献