甜瓜黄斑病毒SYBR Green Ⅰ实时荧光定量PCR方法

为建立检测甜瓜黄斑病毒(melon yellow spot virus, MYSV)的SYBR Green Ⅰ实时荧光定量PCR(qPCR)方法。基于MYSV核衣壳蛋白基因保守序列设计qPCR特异性引物对,针对引物退火温度、引物浓度、特异性和敏感性进行系列优化。结果显示,优化后的qPCR方法最适退火温度为61.3℃,最适引物浓度为0.65μmol·L^-1,特异性强,灵敏度高,比PCR高100倍。以携带目的基因片段的重组质粒为标准品,构建的qPCR标准曲线循环阈值与模板浓度呈良好的线性关系,相关系数为0.999 7。实验样品验证表明建立的qPCR方法可用于MYSV的定量检测。     

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。     

鼠源细胞TaqMan实时荧光定量PCR检测方法的建立

为对鼠源细胞进行鉴别检测,以鼠线粒体16S rRNA基因序列为靶位点设计特异性引物及探针,建立实时荧光定量PCR检测方法,并评价该方法的特异性及敏感性。结果显示,所建立的检测方法特异性好,针对鼠源细胞基因组荧光定量PCR扩增曲线良好,其他物种来源细胞基因组及生物制品原辅材料未出现特异性扩增曲线;敏感性高,基因拷贝数检出限度为45.3拷贝。本试验建立的荧光定量PCR检测方法能够有效地对鼠源细胞进行快速检测,为细胞质量控制提供了有效方法。     

Test Analysis of Ecosystem on Functional Groups of Microorganism in Spiral Up- flow Reactor System

The test of functional groups of microorganisms in Spiral Up-flow Reactor System and the correlation between microorganisms in the water and the main environmental factors are studied.The results show that the diversity of microorganisms population in the SUFR system is complex and the bio-community formed in the SUFR system is stable.The close relations between abundance of bacteria and nutrients are found.The correlation between heterotrophic bacteria and COD is 0.949.The correlation between organic phosphate bacteria and TP is 0.815. The correlation between nitrosobacteria and NH_3-N is 0.909. The correlation between disnitrifier bacteria and TN is 0.653.     

植物基因启动子的克隆方法及其应用

植物基因的表达调控已成为分子生物学研究热点,启动子是基因表达调控的重要顺式元件,启动子的克隆对于研究基因表达调控、构建基因工程载体、表达目的蛋白有着重要的意义。启动子克隆的方法很多,从常用的启动子陷阱技术筛选启动子到PCR方法的应用,此后相继问世的一些基于PCR的克隆启动子技术,如载体锚定PCR、反向PCR、接头PCR、交错热不对称PCR等,为克隆启动子提供了更可靠,更合理的方法。本文着重介绍了几种植物基因启动子的克隆方法,分析了它们的优缺点,并展望了今后的研究前景。     

Prediction of the Economic Development for the Circumjacent Reservoir Region of the Longtan Hydropower Project Using Grey Quantitative Analog

In order to overcome some technical difficulties,we adopt ingenously the grey models combined with the quantitative analog method,and select the Dan Jiangkou Project which has accumulated practical operation data for a long time as the analog project to predict the economic influence for the circumjacent reservoir region of the Longtan hydropower project.On the basis of analog we have built various industrial economic grey models,and compare them with the corresponding predictions for the unconstructed reservoir case.The predictions of the former are greatly larger than the later ones.and hence provides a certain scientific basis for the central leaders to make a decision for constructing the huge hydropower project ultimateiy.     

基于冠层温度和土壤墒情的实时监测与灌溉决策系统

设计了一个可以在线连续监测田间作物冠层温度、环境信息和土壤墒情的实时灌溉决策系统,并将其安装于农田进行了1 a实际运行和观测。系统采用太阳能供电和微处理器进行数据采集和管理,为野外的实际应用提供了保障。系统配置了红外温度、空气温/湿度、土壤水分/水势等传感器,能够及时采集田间全面的同步数据,排除了异地观测所形成的数据误差。采用悬臂式多点采集下垫面红外温度检测方法,可以快速采集更多和更高精度的数据,避免单点测量的人为误差。系统配备的快速锁紧装置,能够根据下垫面作物的生长情况进行传感器位置高度调节,使检测数据更符合田间实际情况。通过运行管理和监测数据分析可见,所监测数据能够很精细的刻画田间作物实际生长状况,可以用于灌区综合灌溉决策,实现田间精量灌溉管理和控制,为灌溉管理的精量化和智能化提供数据支持。     

Determination of vc and mating types of Cryphonectria parasitica isolates by multiplex PCR and their genetic diversity in 13 chestnut-growing provinces of Turkey

Vegetative compatibility (vc) and mating types and genetic diversity of Cryphonectria parasitica isolates were determined using 183 isolates obtained from 215 infected chestnut trees growing in 13 provinces of Turkey. Based on the cultural aspects, 143 of these isolates were evaluated as virulent whereas the remaining 40 isolates were hypovirulent. When vc types of 183 isolates were classically differentiated, 135 of them matched to EU-1 (82.3%), 29 of them to EU-12 (17.6%) vc type, whereas 19 of them did not match to the two. When molecular vic markers were used, all the isolates were assigned to two EU vc types; 149 to EU-1 (81.4%) and 34 (18.5%) to EU-12. Of the majority of the isolates, 134 (73.2%) had mating-type MAT-1, while 44 (24%) isolates had MAT-2 and 5 (2.8%) isolates had both mating types. The population analysis based on two DNA marker systems, Inter-Primer Binding Site and Start Codon Targeted Polymorphism, showed no intraspecific genetic variation among the C. parasitica isolates. The prevalence of two dominant vc types revealed by this study shows that biological control with hypovirulent EU-1 and EU-12 isolates will be significant for the country. The results might be helpful to chestnut breeders carrying out resistance breeding studies to manage this disease based on hypovirulence attributed to Cryphonectria hypovirus 1.     

辽西地区引种野杏数量性状研究

以辽西地区26个野杏无性系为试材,采用变异性分析、方差分析以及主成分分析等方法,研究了其25个数量性状的变异特点,以期为野杏无性系的遗传多样性研究和良种选育提供重要参考依据。结果表明:19个数量性状呈正态分布;25个野杏无性系数量性状变异系数均值23%,其中22个数量性状的变异系数在10%以上,小枝长度变异系数最大,达64%,单果质量、单核质量、单仁质量的变异系数分别为24%、21%和18%,表明野杏无性系数量性状变异程度丰富,选择潜力较大。野杏无性系果实性状重复力普遍偏高,核仁主要性状重复力次之,树体主要数量性状重复力较二者偏低,均达到极显著水平;果核仁性状的重复力除单果质量、单核质量和仁厚分别为0.634、0.756和0.624外,其它指标皆在0.909以上,其中仁长重复力最高,达0.973。主成分分析结果表明,前6个主成分累积贡献率达83.50%,能够反映野杏无性系数量性状的大部分信息;其中第1主成分主要反映野杏果核仁等经济性状。