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31.
Detection of Colletotrichum coccodes from soil and potato tubers by conventional and quantitative real-time PCR 总被引:4,自引:1,他引:4
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 . 相似文献
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塔里木沙漠公路沿线机械防沙体系效益评价及防沙带合理宽度的初步探讨 总被引:12,自引:3,他引:12
本文通过对塔里木沙漠公路肖塘段(起始段)至塔中段的风沙危害和现有防沙体系进行评价,指出了目前公路沿线防沙体系存在的问题,并给出了沿线防护体系的合理宽度模型及模拟计算结果。 相似文献
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兽用抗寄生虫药物新剂型及其新技术的研究进展 总被引:8,自引:1,他引:8
兽用抗寄生虫药物由于使用频繁,其给药技术研究一直备受重视.目前,兽用抗寄生虫药物新剂型和新技术的研究主要集中在缓释控释制剂和脂质体制剂,其次是透皮给药系统、微囊、微球、环糊精包合物、固体分散体等.但这些新剂型和新技术大多还处于研究阶段,在处方设计或制剂工艺方面都还存在一些问题,今后应加强这些新剂型的工艺和技术研究,以真正发挥这些新剂型的优点. 相似文献
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AIM: To investigate inhibition of K562 cell growth by antisense drug targeted VEGF mRNA. METHODS: X7, 20-mer antisense sequences were selected, synthesized and modified with phosphorothioate. The drug was transfected into K562 cells in the present of lipofection. Cell growth was assayed by trypan blue dye exclusion assay and MTT. The level of VEGF protein in the media was determined by ELISA. The morphology of apoptotic cells were observed by Giemsa staining, and the propotion of apoptotic cells was detected by flow cytometry. RESULTS: The antisense drug inhibited growth of K562 and downregulated expression of VEGF protein significantly, compared with Scrambed control group and showed dose-dependent relation. Signs of apoptosis of K562 cells were not observed. CONCLUSION: Inhibition of K562 cell proliferation, but not cells apoptosis induction is the mechanism of inhibing growth of K562 cells by antisense drug targeted VEGF mRNA. At same time, VEGF has function of promoting K562 cell proliferation, and VEGF mRNA may be a new target attached by drugs. 相似文献
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专门化肉用种羊选定、引进后,通过适应性研究,在较大规模养羊业生产实践中。对现代集约化肉羊业的主要关键技术进行研究、完善和创新,组装集成并应用于生产。技术研究应用结果既为市场提供了大批专门化肉用种羊和优质安全的杂种肉羊,取得了高水平的研究成果及显著的经济效益和社会效益.又为我国目前正在迅速发展的集约化肉羊业提供了可靠的技术支持和理想的生产模式:从适宜的专门化肉羊品种到运用高效母羊快速扩繁技术.建立和完善有效的肉羊杂交利用体系,普遍应用种羊鲜、冻精生产大批肉用杂种羔羊,种植高产优质饲草及其科学加工调制和利用.建立严格兽医防疫制度.实行放牧 补饲或全舍饲的精细管理和集约化育肥,至生产优质、安全、标准肉羊,获得显著的经济效益和社会效益。 相似文献
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