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The presence of phytoplasmas in seven coniferous plant species (Abies procera, Pinus banksiana, P. mugo, P. nigra, P. sylvestris, P. tabuliformis and Tsuga canadensis) was demonstrated using nested PCR with the primer pairs P1/P7 followed by R16F2n/R16R2. The phytoplasmas were detected in pine trees with witches’ broom symptoms growing in natural forest ecosystems and also in plants propagated from witches’ brooms. Identification of phytoplasmas was done using restriction fragment length polymorphism analysis (RFLP) of the 16S rDNA gene fragment with AluI, MseI and RsaI endonucleases. All samples showed RFLP patterns similar to the theoretical pattern of ‘Candidatus Phytoplasma pini’, based on the sequence of the reference isolate Pin127S. Nested PCR‐amplified products, obtained with primers R16F2n/R16R2, were sequenced. Comparison of the 16S rDNAs obtained revealed high (99·8–100%) nucleotide sequence identity between the phytoplasma isolates. The isolates were also closely related to four other phytoplasma isolates found in pine trees previously. Based on the results of RFLP and sequence analyses, the phytoplasma isolates tested were classified as members of the ‘Candidatus Phytoplasma pini’, group 16SrXXI. 相似文献
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This study established a quick and accurate method to detect petal infection of oilseed rape (Brassica napus) by Sclerotinia sclerotiorum using a nested‐PCR technique. DNA samples were extracted from each petal using a microwave method, followed by two rounds of PCR amplification. The first‐round PCR amplification was performed using the universal fungal primer pair ITS4/ITS5, and the second‐round amplification with a specific primer pair XJJ21/XJJ222, which was designed using the single‐nucleotide polymorphisms among nuclear rDNA ITS sequences of Sclerotinia spp., Botrytis spp. and other selected fungi. The established technique is rapid and inexpensive, and has a high degree of specificity and sensitivity. This assay can distinguish Sclerotinia spp. from other fungi, including Botrytis cinerea, a closely related and frequent cohabitant on oilseed rape petals, and can detect 50 fg genomic DNA, five ascospores of S. sclerotiorumin vitro or 50 ascospores of S. sclerotiorum on one petal in approximately 6 h, even in the presence of a high background of oilseed rape DNA. This technique was successfully applied in detecting natural petal infections. 相似文献
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山西省枣树上啤酒花矮化类病毒的检测及序列分析 总被引:1,自引:0,他引:1
[目的] 从枣树样品中分离鉴定啤酒花矮化类病毒(HSVd)。[方法] 从山西省农业科学院果树研究所国家枣种质资源圃采集70份枣树叶片样品,提取小分子RNA后通过Northern杂交、RT PCR进行检测,并对阳性样品中的类病毒进行克隆测序,利用生物学软件对所得序列进行分析。 [结果] 70份枣树样品中有1份样品感染HSVd,克隆测序后,共获得13条HSVd序列,它们与GenBank上首次报道的HSVd序列相似性为92.6%~92.8% 。[结论] 本研究首次在国内报道了枣树上分离得到的HSVd序列,HSVd枣树分离物与已报道的HSVd分离物差异较大。 相似文献
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《Journal of Veterinary Behavior: Clinical Applications and Research》2014,9(5):262-268
If the illicit tobacco trade were eliminated, governments could gain at least $31.3 billion a year, and more than 164,000 premature deaths a year could be avoided after 2030 (Joossens, Merriman, Ross, and Raw, 2009). Dogs have been used successfully in tobacco control programs, and there is a good chance that rats could also play an important role. In the present experiment, giant African pouched rats were trained to respond to filters that had been stored together with cigarettes (i.e., soaked) and to not respond to filters that had been soaked with noncigarette items. Generalization to untrained types of tobacco was then tested. The sensitivity of 4 rats trained on filters soaked with 1 of 7 types of cigarettes ranged from 86% to 100% (mean, 95%). There was very little evidence of generalization when the rats were tested on tobacco leaves and snuff but good evidence of generalization when the rats were tested on cigarettes that had been soaked with strong-smelling additives. These findings suggest that rats may be a valuable asset in the global effort to control illicit cigarette trade. 相似文献
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本文通过教育教学类网站的跟踪,主要对用户所访问的网页、及其对所含对象的时间与频次数据的分析,将收集到的数据以图表的形式呈现给网络课程开发人员,使其对教学网站进行优化,方便教育资源使用者学习。 相似文献