DNA isolation from recalcitrant materials such as tree roots, bark, and forest soil for the detection of fungal pathogens by polymerase chain reaction
DNA isolation from recalcitrant materials such as tree roots, bark, and forest soil for the detection of fungal pathogens by polymerase chain reaction
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DOI：
10.1006/abio.1998.2769
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发表时间：
1998-08-15
影响因子：
2.9
通讯作者：
Sandermann, H
中科院分区：
文献类型：
--
作者：
Bahnweg, G;Schulze, S;Sandermann, H
即使在没有病害症状或果实结构的情况下，使用特定的引物对分类单元特定的DNA片段进行PCR扩增是在感染早期检测和半定量检测植物和森林病原体的选择方法(1，2)。已经报道了许多分离高分子量基因组DNA的技术，其中许多技术是专门设计的，即使从顽固的来源，如富含酚类或多糖的植物材料和富含腐植酸的土壤样品中，也可以提取适合于聚合酶链式反应的DNA。减少此类化合物的抑制作用的努力使用了诸如聚乙烯吡咯烷酮(PVP1：3-7)和聚乙烯基聚吡咯烷酮(PVPP：8，9)之类的试剂或精心设计的DNA分离条件以防止DNA与被氧化的DNA发生化学反应
PCR amplification of taxon-specific DNA fragments employing specific primers is the method of choice for detection and semiquantification of plant and forest pathogens at early stages of infection even in the absence of disease symptoms or fruiting structures (1, 2). Numerous techniques for the isolation of high-molecular-weight genomic DNA have been reported, many of them specifically designed to recover DNA suitable for PCR, even from recalcitrant sources such as plant materials rich in phenolics or polysaccharides and soil samples rich in humic acids. Efforts to reduce inhibitory action of such compounds utilized reagents such as polyvinylpyrrolidone (PVP1: 3–7) and polyvinylpolypyrrolidone (PVPP: 8, 9) or elaborate conditions of DNA isolation preventing chemical reactions of DNA with oxidized