Detection of beet yellows virus in sugarbeet plants by enzyme-linked immunosorbent assay (ELISA)

Beet yellows virus can be detected in leaf extracts of infected sugarbeet plants by ELISA. The use of discs was studied and proved to be a valuable and qualitatively reliable method. Leaf material could be stored at 4^o or 22°C for at least six days without affecting the detection of this virus by ELISA. A dramatic decrease in ELISA values was found when leaf extracts were frozen.In an analysis of the distribution of virus over the plant it was found that young leaves present at the moment of infection and those which had still to develop after infection will contain virus. Symptoms produced by systemic virus invasion occur on the oldest leaves containing virus.Samenvatting Het bietevergelingsvirus kan op betrouwbare wijze met de ELISA methode in geïnfecteerde bieteplanten worden aangetoond. Een aanzienlijke vereenvoudiging van de procedure kan worden bereikt met de zogenaamde disc-method, waarbij intacte ponsstukjes in de putjes van de ELISA-plaat worden geïncubeerd. Hierbij komt voldoende virus uit de ponsstukjes voor ELISA vrij. Bladmateriaal kon op verschillende wijzen bewaard worden zonder dat de mogelijkheid om het virus aan te tonen achteruitging. Met bladextracten die ingevroren waren, werden echter slechte resultaten verkregen.In een analyse naar de verdeling van het virus over het loof bleek het virus voor te komen in de geïnoculeerde bladeren, in die bladeren die op het tijdstip van inoculatie minder dan de helft van hun uiteindelijke lengte bereikt hadden en in de bladeren die nog moesten verschijnen. De symptomen ontwikkelden zich op de oudste systemisch geïnfecteerde bladeren.     

不同干燥失水方式对云南温带优良牧草营养品质的影响研究

采用迅速烘干、太阳晒干和缓慢烘干3种处理对云南温带21个牧草品种的鲜草进行干燥处理,分别测定干草的营养成分。结果表明:以干草的粗蛋白含量为指标,干燥方法以迅速烘干为佳,其次是太阳晒干,最差是缓慢烘干;从干草的粗脂肪考虑,其变化无规律,因牧草品种的不同而不同;其余指标变化不大。     

我国草地鼠害防治研究进展

严重的草原鼠害已成为引发草原退化、沙化、水土流失的重要因素,并威胁我国畜牧业的可持续发展、草原生物多样性保护、草原生态环境建设以及人们身体健康。防治鼠害常用的方法主要有化学防治、生物防治、不育剂应用及物理机械灭鼠。文章分析了各种防治方法的利弊,认为中草药植物源灭鼠剂在鼠害的综合治理上具有广阔的应用前景,应进一步研究并进行其产品的开发和研制。     

新鲜蔬菜硝酸盐含量测定的改进试粉法

 为适应新鲜蔬菜硝酸盐快速检测的需要, 在现有试粉法的基础上对硝酸盐测定的试粉法进行了改良研究。结果表明, 本研究建立的直接以去离子水浸提蔬菜匀浆, 混合试粉配方为柠檬酸∶一水硫酸锰∶无水对氨基苯磺酸∶N - 1 - 萘乙二胺盐酸盐∶细锌粉= 30∶4∶1.6∶0.8∶1, 其加入量为0.1 g的改进试粉法, 对于溶液中硝酸盐含量在0～20 mg/L范围时, 显色吸光值与硝酸盐含量呈现良好的线性关系, 相关系数达0.9999, 方法回收率在97.7%～104.5%之间, 相对标准偏差2.71%。用本改进试粉法测定11种蔬菜的硝酸盐含量与国标法测定结果的t检验具有一致性。     

改良热硼酸法高效提取苹果果实RNA

苹果果实、尤其是成熟期的果实中多糖和其他次生物质含量高,难以提取RNA。对此,在原热硼酸法的基础上进行了改进,通过添加提取辅助剂并根据辅助剂特性调整提取步骤,高效、稳定地从苹果果实、尤其是后期果实中提取到了高质量的RNA。所得RNA的A260/A230大于2.0,A260/A280为1.8-2.1,并且RNA产量高,其中前期果实RNA产率在13.40μg/g以上,后期果实RNA产率在7.02μg/g以上,完全可以满足RT-PCR、cDNA-AFLP等分子生物学实验的要求。     

法律本科教学方法之我见

随着我国法治的不断健全，对新时期法律本科教育提出了新的要求，这也是对中国法学教育者传统教学方法的一次新挑战。需要我们吸收传统教学方法的优秀经验，探索适合本科阶段教育特色的教学方法，开展案例教学和实践教学，注重教学中的师生互动关系，改革考试内容和考试方式，以使现阶段法学本科教育更能适应现实社会和法律职业的需求。     

陕西线辣椒病毒病病原检测简报

在陕西省线辣椒主产区多点随机取样,采用酶联免疫吸附法对陕西线辣椒病毒病毒原进 行了鉴定。结果表明,陕西线辣椒病毒病毒原有BBWV、PMMV、CMV、ToMV、TMV、PVY和 CVMV。优势毒原是BBWV、PMMV和CMV。     

Powdery Mildew of Prairie Gentian: Characteristics,Molecular Phylogeny and Pathogenicity

In March 1999, we found prairie gentian (Eustoma grandiflorum) infected with powdery mildew in a greenhouse in Oita Prefecture, Japan. Morphological observation revealed that the causal fungus belongs to the mitosporic genus Oidium subgenus Pseudoidium [teleomorph: Erysiphe sensu Braun and Takamatsu (2000)]. Precise taxonomic position of the fungus, however, is uncertain due to lack of the perfect stage. We determined the nucleotide sequence of the rDNA ITS region of the fungus. Comparison of the sequence with those obtained from DNA databases of this fungal group revealed that the sequence is identical to those of powdery mildews from garden four-o'clock (Mirabilis jalapa) and broad bean (Vicia faba). Inoculation of an isolate from garden four-o'clock caused mildew on prairie gentian and broad bean, suggesting that the prairie gentian mildew originates from garden four-o'clock or broad bean. Molecular phylogenetic analysis indicated a close relationship of this fungus to Erysiphe baeumleri on Vicia spp. and E. trifolii on Trifolium pratense. From these results, we propose that prairie gentian mildew diverged from a Fabaceae-parasitic ancestor. Received 14 March 2002/ Accepted in revised form 28 May 2002     

Detection of Colletotrichum coccodes from soil and potato tubers by conventional and quantitative real-time PCR

Colletotrichum coccodes is the causal agent of the potato blemish disease black dot. Two PCR primer sets were designed to sequences of the ribosomal internal transcribed spacer (ITS1 and ITS2) regions for use in a nested PCR. The genus-specific outer primers (Cc1F1/Cc2R1) were designed to regions common to Colletotrichum spp., and the species-specific nested primers (Cc1NF1/Cc2NR1) were designed to sequences unique to C . coccodes . The primer sets amplified single products of 447 bp (Cc1F1/Cc2R1) and 349 bp (Cc1NF1/Cc2NR1) with DNA extracted from 33 European and North American isolates of C. coccodes. The specificity of primers Cc1NF1/Cc2NR1 was confirmed by the absence of amplified product with DNA of other species representing the six phylogenetic groups of the genus Colletotrichum and 46 other eukaryotic and prokaryotic plant pathogenic species. A rapid procedure for the direct extraction of DNA from soil and potato tubers was used to verify the PCR assay for detecting C. coccodes in environmental samples. The limit of sensitivity of PCR for the specific detection of C. coccodes when inoculum was added to soils was 3·0 spores per g, or the equivalent of 0·06 microsclerotia per g soil, the lowest level of inoculum tested. Colletotrichum coccodes was also detected by PCR in naturally infested soil and from both potato peel and peel extract from infected and apparently healthy tubers. Specific primers and a TaqMan fluorogenic probe were designed to perform quantitative real-time (TaqMan) PCR to obtain the same levels of sensitivity for detection of C. coccodes in soil and tubers during a first-round PCR as with conventional nested PCR and gel electrophoresis. This rapid and quantitative PCR diagnostic assay allows an accurate estimation of tuber and soil contamination by C. coccodes .