Methods for studying the nematophagous fungus Verticillium chlamydosporium in the root environment
Methods for studying the nematophagous fungus Verticillium chlamydosporium in the root environment
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DOI：
10.1023/a:1010373717186
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发表时间：
2001-01-01
期刊：
影响因子：
4.9
通讯作者：
Kerry, BR
中科院分区：
文献类型：
--
作者：
Hirsch, PR;Atkins, SD;Kerry, BR
为了充分发挥嗜食真菌厚垣轮枝菌的生防潜力，了解该真菌在土壤中的生态学以及与植物和线虫宿主的相互作用是非常重要的。比较了几种研究土壤和根际真菌的方法。这些包括用于厚垣孢子菌的半选择性培养基、对真菌β-微管蛋白基因特异的PCR引物和单克隆抗体。除了为物种特异性引物提供靶标外，β-微管蛋白基因还涉及对半选择性培养基中使用的杀真菌剂的抗性，并研究了其遗传基础。培养和PCR为基础的方法被用来筛选真菌的存在下，在田间土壤中已知已被抑制谷物孢囊线虫和含有V. chlamydosporium人口。单克隆抗体特异性菌丝或分生孢子的真菌获得，并探讨其作为一种工具，可视化的根上的感染过程中的应用。
In order to exploit fully the biocontrol potential of the nematophagous fungus Verticillium chlamydosporium, it is important to understand the ecology of the fungus in soil, and interactions with both plant and nematode hosts. Several approaches for studying the fungus in soil and the root environment are compared. These include a semi-selective medium for V. chlamydosporium, PCR primers specific for the fungal beta -tubulin gene, and monoclonal antibodies. In addition to providing a target for species-specific primers, the beta -tubulin gene is implicated in resistance to the fungicides used in the semi-selective medium, and the genetic basis for this is investigated. Culture and PCR-based methods were used to screen for the presence of the fungus in field soils known to have been suppressive to cereal cyst nematode and that contained V. chlamydosporium populations. Monoclonal antibodies specific for either hyphae or conidia of the fungus were obtained, and their application as a tool for visualising the infection process on the root was explored.