猪传染性胃肠炎病毒分子生物学研究进展

猪传染性胃肠炎病毒(TGEV)基因组为不分段的单股正链RNA,其编码4种结构蛋白和3种非结构蛋白。TGEV S蛋白是基因工程研究的重点,近年来,S蛋白在大肠埃希菌、沙门菌、腺病毒等中得到了高效表达。将传染性胃肠炎病毒的基因组人工改造成为感染性cDNA的表达载体,此载体可在ORF3处插入外源基因,异源基因绿荧光蛋白可稳定、高效地表达。TGEV基因1和其他冠状病毒相比保守性很强,所以对TGEV聚合酶的研究,特别是其中保守酶的研究,对于抗TGEV及其他冠状病毒药物的研究有着重要的意义。     

PEF辅助快速提取非特异性奶牛脾脏转移因子的实验研究

应用PEF成功从奶牛脾脏中提取非特异性转移因子，并与常规提取法和复合酶提取法进行了比较.经理化特性分析，奶牛脾脏转移因子是以低分子多肽（〈10kD）为主的混合物，含有少量核酸和氨基酸.实验表明，将物料溶解于6倍质量的磷酸盐（pH7．0）缓；中液中，在脉冲电场强度为25kV／cm，脉冲数为16μs，和流速为2mL／min的条件下，提取液中低分子多肽的含量有最大值9．85mg／mL.PEF提取率是常规方法的2倍，复合酶法的1．47倍.PEF法耗时短，操作简单，成本低，因此，PEF提取是一种很有希望的提取非特异性奶牛脾脏转移因子方法.     

La(NO3)3对4个苜蓿品种种子活力与膜透性作用的比较

通过La3 离子对4个苜蓿Medicago sativa品种种子活力与膜透性作用的比较,结果表明:当用8种不同浓度La(NO3)3溶液处理4个苜蓿品种种子时,La3 浓度小于600μg/mL时,可促进种子的活力;其中,中苜1号和胜利者的活力指数变化稳定且较高,而草原1号在La3 浓度为600μg/mL时变化剧烈。中苜1号和胜利者可以较长时间忍耐La3 在750μg/mL的处理浓度,超过此剂量时,膜结构就受到了破坏;而阿尔冈金和草原1号在La3 浓度接近750μg/mL时,种子活力就已经开始下降了。     

ULTRASONOGRAPHIC EVALUATION OF THE EXTERNAL EAR CANAL AND TYMPANIC MEMBRANE IN DOGS

Ultrasonographic imaging of the canine external ear canal, tympanic membrane, and tympanic bulla was described in five healthy beagle dogs before and after infusion of saline into the ear canal. Saline served as an acoustic window. With this method, the external ear canal, and tympanic bulla were visible in the same imaging plane and the integrity of the tympanic membrane could be evaluated indirectly by confirming an intact tympanic membrane, which appeared at the end of the ear canal as a hyperechoic line with reverberation. Experimentally, perforated tympanic membrane could be evaluated by identifying anechoic saline in the tympanic bulla lumen. The air and fluid-filled tympanic bulla were also visualized. Ultrasonography with saline as an acoustic window appears to be helpful for the evaluation of the external ear canal, tympanic membrane, and tympanic bulla and it may have the potential to be a useful clinical tool in evaluation of integrity of the tympanic membrane.     

免疫佐剂的作用机制及其应用

从抗原微粒化佐剂（如不溶性铝盐胶体、免疫刺激复合物、脂质体）、缓慢释放抗原佐剂（如油乳佐剂、抗原的微型包囊）、微生物佐剂（如细菌毒素、短小棒状杆菌菌苗、分枝杆菌及其成分、肽聚糖）、分子佐剂（如细胞因子、C3d分子、共刺激分子、超抗原、热休克蛋白、CpG序列）和其他佐剂（如维生素E、硒、抗生素类、拟胸腺素药物、蜂胶、中药多糖）等几方面阐述了免疫佐剂的性质、作用机制及应用的研究现状，为免疫佐剂的临床应用和进一步研制提供了参考资料。     

拟南芥霜霉病抗眭分子机制研究进展

随着近年分子生物学技术的发展与应用，植物霜霉病抗性的研究有了长足的进展。本文就拟南芥抗霜霉病基因的克隆与结构分析，抗病信号传导，防卫反应和系统获得抗性，以及寄主一寄生菌共进化冲突方面进行了综述，并就今后的研究进行了展望。     

Potential Inoculum Sources of Phaeomoniella chlamydospora in South African Grapevine Nurseries

Petri disease of grapevine is primarily caused by Phaeomoniella chlamydospora. This pathogen affects mostly young grapevines, but is also implicated in esca disease of older grapevines. Little is known about the disease cycle of this fungus. Infected propagation material was identified as a major means of dissemination of the pathogen. Recently, the pathogen was also detected from soil in South Africa and airborne conidia have been found in vineyards. The aim of this study was to use a molecular detection technique to test different samples collected from nurseries in South Africa at different nursery stages for the presence of P. chlamydospora. A one-tube nested-PCR technique was optimised for detecting P. chlamydospora in DNA extracted from soil, water, callusing medium and grapevine wood. The one-tube nested-PCR was sensitive enough to detect as little as 1 fg of P. chlamydospora genomic DNA from water and 10 fg from wood, callusing medium and soil. PCR analyses of the different nursery samples revealed the presence of several putative 360 bp P. chlamydospora specific bands. Subsequent sequence analyses and/or restriction enzyme digestions of all 360 bp PCR bands confirmed that all bands were P. chlamydospora specific, except for five bands obtained from callusing media and one from water. Phaeomoniella chlamydospora was positively detected in 25% of rootstock cane sections collected from mother blocks, 42% of rootstock cuttings and 16% of scion cuttings collected during grafting, 40% of water samples collected after pre-storage hydration, 67% of water samples collected during grafting, 8% of callusing medium samples and 17% of soil samples collected from mother blocks. These media can therefore be considered as possible inoculum sources of the pathogen during the nursery stages.     

Genetic variability and virulence among Verticillium albo-atrum isolates from hop

Verticillium wilt, caused by Verticillium albo-atrum or V. dahliae, is an important disease of many worldwide crop species. In Europe, V. albo-atrum isolates infecting hop express different levels of virulence, inducing mild or lethal disease syndromes, and it is therefore an attractive model for studying the virulence of this pathogen. In this work, eleven amplified fragment length polymorphism (AFLP) primer combinations were used to analyze genetic variability among 55 V. albo-atrum hop isolates from four European hop growing regions, as well as isolates from other hosts and V. dahliae isolates. Cluster analysis divided V. albo-atrum and V. dahliae isolates into two well-separated groups. Within the V. dahliae cluster, isolates were separated without host specific grouping, although no host adapted isolates were included. In V. albo-atrum, the alfalfa isolates were distinct from isolates of other hosts, where a high association with virulence was observed in hop and tomato isolates. All lethal hop isolates were genetically different from mild hop isolates. The lethal hop isolates from England and Slovenia expressed the same virulence phenotype, although they showed a different AFLP pattern. The mild hop isolates formed two subgroups, to which isolates clustered irrespective of geographical location. These data suggest multiple origins of V. albo-atrum hop isolates, and the possible appearance of new virulent isolates in the future in other hop growing regions.     

Note: Molecular identification of three entomopathogenic nematodes from turkey by PCR-RFLP of the ITS regions

Molecular identification methods are widely used for the classification of organisms worldwide. Entomopathogenic nematodes are the most often isolated insect parasitic nematodes in the tropical and subtropical regions. In our investigation, PCR-RFLP (Polymerase Chain Reaction — Restriction Fragment Length Polymorphism) of the ITS region (Internal Transcribed Spacer) on the ribosomal (r) DNA of three entomopathogenic nematodes isolated from Ankara, Turkey, was analyzed for identification. The ITS region of rDNA was amplified by PCR and then digested with the following nine restriction enzymes: Alu I, Dde I, Hae III, Hha I, Hind III, Hinf I, Hpa II, Rsa I and Sau 3AI. The amplified and restricted sequences of the ITS regions were separated by agarose gel electrophoresis and the RFLP patterns of these three species were shown in this study. According to our results, these species were identified asSteinernema feltiae, Steinernema carpocapsae andHeterorhabditis bacteriophora. http://www.phytoparasitica.org posting Nov. 4, 2005.