Canine Haematopoietic Chimerism Analyses by Semiquantitative Fluorescence Detection of Variable Number of Tandem Repeat Polymorphism

Canine models are successfully applied to the study of haematopoietic stem cell transplantation (HSCT). Monitoring of haematopoietic donor/recipient chimerism is of major significance in detecting and quantifying engraftment or graft rejection of the donor-derived haematopoietic cells after transplantation. Radioactive analyses of polymorphic microsatellite markers are commonly used for chimerism analyses. We describe an improved, non-isotopic method that is based on the analysis of microsatellite markers in donor and recipient cells using capillary electrophoresis and fluorescence detection. Artificial mixtures of donor and recipient DNA that were generated from peripheral blood mononuclear cells from dog leukocyte antigen-identical siblings were used to analyse the sensitivity of the assay. DNA from dogs that had received HSCT were also analysed in order to demonstrate the feasibility of the method in vivo. For chimerism analyses, six different microsatellite loci were systematically amplified using fluorescent PCR primer. The fluorescent polymerase chain reaction products were separated by capillary electrophoresis using POP4 on a 310 ABI Prism Genetic Analyzer. After electrophoresis, fluorescence signals were automatically sized and quantified using GeneScan software. The method described provides an accurate assessment of haematopoietic chimerism in the canine model with significantly reduced hands-on time compared to conventional gel electrophoresis.     

三相异步电动机节电控制装置的研制

针对负载特性为空载、轻载或变载的电动机．研制出一种集成化△→Ｙ自动转换节电器，可使电机运行于节能状态。对节电器的节能原理、工作原理作了系统分析并对节能效果进行了测定．结果表明该装置节能效果显著。     

立足“长治”工程建设 促进贫困地区经济发展

立足“长治”工程建设促进贫困地区经济发展肖秧（四川省人民政府，成都６１０６１６）长江上游水土保持重点防治工程的开展，正在使长江上游地区水土流失治理进入一个前所未有的新时期。近五年来，随着改革开放的深入，水保事业也生机勃勃，不断强化领导，有效控制新的水...     

麻雀自然感染鸡传染性法氏囊病病毒的调查

从鸡传染性法氏囊病（IBD）流行的鸡场捕杀麻雀54只,用鸡传染性法氏囊病病毒（IBDV）单克隆抗体夹心阻断ELISA检测抗体,阳性检出率为7.4％(4/54);以逆转录—聚合酶链反应（RT-PCR）检测病毒核酸,阳性检出率为11.1％（6/54）;RT-PCR阳性样本病毒分离亦为阳性。结果表明,IBD流行的鸡场里的麻雀能够发生IBDV自然感染,麻雀可能是IBDV的贮存宿主或二次传染源之一。     

Volatile Metabolite Profiling for the Discrimination of Onion Bulbs Infected by Erwinia carotovora ssp. carotovora, Fusarium oxysporum and Botrytis allii

The volatile metabolites of the headspace gas of onion bulbs inoculated with three different pathogens, Erwinia carotovora ssp. carotovora, Fusarium oxysporum and Botrytis allii, were profiled using gas chromatography/mass spectrometry. Differences in the number and amount of volatile metabolites were observed. Two hundred and fifty three volatile metabolites were detected in bulbs inoculated with three pathogens or sterile distilled water. On day three, 202 volatile metabolites were observed, compared to 166 on day six. Of the 253 compounds, however, only 59 occurred relatively consistently over replications, of which 25 compounds were specific to one or more pathogens, including 10 that were unique to a pathogen. Metabolites such as 1-Oxa-4,6-diazacyclooctane-5-thione and 4-mercapto-3-(methylthio)--(thio-lactone)-crotonic acid were exclusive to onions inoculated with F. oxysporum. Acetone, acetic acid-hydrazide, propylcarbamate, 1-bromo-1-propene, thiirane, 1-(methylthio)-E-1-propene and 1-ethenyl-4-ethyl-benzene were specific to B. allii. 3-bromo-furan was specific to E. carotovora ssp. carotovora. Sterile water-inoculated bulbs produced 3,3-dioxy-1,2-propanediol-tetranitrate. Highest amount of sulfurs was found in pathogen-inoculated, while highest amounts of terpenes, aromatics and aliphatics were found in sterile distilled water-inoculated bulbs. The possible use of these differences in the volatile metabolites for detecting and discriminating diseases of onion in storage is discussed.     

Note: A comparison of molecular diagnostic procedures for the detection of aster yellows phytoplasmas (16Sr-I) in leafhopper vectors

Different molecular procedures were compared for the detection of aster yellows phytoplasmas (AYP) in the leafhopper vectorsMacrosteles quadripunctulatus (Kirschbaum),Euscelidius variegatus (Kirschbaum) andEuscelis incisus (Kirschbaum). Polymerase chain reaction (PCR) with universal and group-specific primers designed on the 16S-rDNA sequence was most sensitive in nested assays. A dot-blot procedure with an oligoprobe designed on the 16S-rDNA was less sensitive and consistent to detect phytoplasmas in total insect DNA, but consistently detected amplicons from direct PCR. The dot-blot assay with a probe based on a phytoplasma plasmid sequence detected AYP in most vector specimens and did not react with DNAs from leafhoppers infected by flavescence dorée and psyllids infected by apple proliferation phytoplasmas. This last assay is almost devoid of contamination risks, faster and cheaper compared to PCR, therefore it has to be preferred for field-scale analysis of leafhopper populations. http://www.phytoparasitica.org posting Feb. 24, 2004.     

Real-time detection of <Emphasis Type="Italic">Phytophthora nicotianae</Emphasis> and <Emphasis Type="Italic">P. citrophthora</Emphasis>in citrus roots and soil

Two primers, specific for Phytophthora nicotianae (Pn6) and P. citrophthora (Pc2B), were modified to obtain Scorpion primers for real-time identification and detection of both pathogens in citrus nursery soils and roots. Multiplex PCR with dual-labelled fluorogenic probes allowed concurrent identification of both species ofPhytophthora among 150 fungal isolates, including 14 species of Phytophthora. Using P. nicotianaespecific primers a delayed and lower fluorescence increase was also obtained from P. cactorumDNA. However, in separate real-time amplifications, the aspecific increase of fluorescence from P. cactorum was avoided by increasing the annealing temperature. In multiplex PCR, with a series of 10-fold DNA dilutions, the detection limit was 10 pg l^-1 for P. nicotianaeand 100 pg l^–1 for P. citrophthora, whereas in separate reaction DNA up to 1 pg l^-1 was detected for both pathogens.Simple and rapid procedures for direct DNA extraction from soil and roots were utilised to yield DNA whose purity and quality was suitable for PCR assays. By combining these protocols with a double amplification (nested Scorpion-PCR) using primers Ph2-ITS4 amplifying DNA from the main Phytophthora species (first round) and PnB5-Pn6 Scorpion and Pc2B Scorpion-Pc7 (second round), it was possible to achieve real-time detection of P. nicotianaeand P. citrophthora from roots and soil. The degree of sensitivity was similar to that of traditional detection methods based on the use of selective media. The analyses of artificially and naturally infested soil showed a high and significant correlation between the concentration of pathogen propagules and the real-time PCR cycle threshold.     

坐水播种时耕层土壤水分再分布的数值分析

利用朝阳县的旱地土壤取得基本参数后,应用数学模型计算了坐水播种时耕层土壤水分再分布时的土壤湿润状况参数,通过数值分析揭示了坐水播种湿润锋运动和土壤含水率的变化规律。结果显示:湿润锋运动速度和土壤含水率的变化速度都是由快逐渐减慢,所形成的湿润区近似一个球冠体;土壤含水率中的最大值出现在沟底附近;并提出可以用历时7 d的土壤湿润状况参数作为衡量坐水播种灌水质量的指标。     

用径流小区法研究不同耕作措施对土壤侵蚀的影响

用径流小区方法,研究了不同耕作措施对土壤侵蚀的影响。结果表明:B处理(沿等高线 薄膜覆盖 秸秆覆盖 开沟种植)能有效地减少土壤侵蚀,维护土壤耕作层,可促进农业的可持续发展。处理B、A、C总的径流量分别比处理D(CK)减少79.13%、69.76%、50.61%,总的侵蚀量减少的比例分别为86.81%、72.75%、50.90%。     

3种冷季型草坪草混播配方优化组合筛选试验

在河西走廊中部地区气候条件下,通过对草地早熟禾Poa pratensis、高羊茅Festuca arundinacea、多年生黑麦草Lolium perenne3种冷季型草坪草18种混播配方进行了成坪品质的观察测定,筛选出了适应当地气候条件的优化组合配方10个。