ITS PRIMERS WITH ENHANCED SPECIFICITY FOR BASIDIOMYCETES - APPLICATION TO THE IDENTIFICATION OF MYCORRHIZAE AND RUSTS
ITS PRIMERS WITH ENHANCED SPECIFICITY FOR BASIDIOMYCETES - APPLICATION TO THE IDENTIFICATION OF MYCORRHIZAE AND RUSTS
复制标题
DOI：
10.1111/j.1365-294x.1993.tb00005.x
复制
发表时间：
1993-04-01
影响因子：
4.9
通讯作者：
BRUNS, TD
中科院分区：
文献类型：
--
作者：
GARDES, M;BRUNS, TD
我们设计了两个分类群选择性引物的核核糖体重复单位内转录间隔区（ITS）的区域。这些引物，ITS 1-F和ITS 4-B，旨在分别对真菌和担子菌具有特异性。我们测试了这些引物对13种子囊菌、14种担子菌和15种植物的特异性。结果表明，ITS 4-B与通用引物ITS 1或真菌特异性引物ITS 1-F配对，能有效地扩增所有担子菌DNA，并能区分子囊菌DNA。植物的结果并不那么明确。ITS 1-F/ITS 4-B引物对某些植物物种产生少量PCR产物，但在大多数情况下，其数量低于“通用”ITS引物产生的数量。然而，在植物和真菌DNA都存在的条件下，真菌DNA被扩增到明显排除植物DNA。ITS 1-F/ITS 4-B优先扩增被证明是特别有用的检测和分析的外生真菌和锈菌感染的组织中的担子菌成分。这些引物可用于研究外生菌根群落的结构或锈菌在交替寄主上的分布。
We have designed two taxon-selective primers for the internal transcribed spacer (ITS) region in the nuclear ribosomal repeat unit. These primers, ITS1-F and ITS4-B, were intended to be specific to fungi and basidiomycetes, respectively. We have tested the specificity of these primers against 13 species of ascomycetes, 14 of basidiomycetes, and 15 of plants. Our results showed that ITS4-B, when paired with either a 'universal' primer ITS1 or the fungal-specific primer ITS1-F, efficiently amplified DNA from all basidiomycetes and discriminated against ascomycete DNAs. The results with plants were not as clearcut. The ITS1-F/ITS4-B primer pair produced a small amount of PCR product for certain plant species, but the quantity was in most cases less than that produced by the 'universal' ITS primers. However, under conditions where both plant and fungal DNAs were present, the fungal DNA was amplified to the apparent exclusion of plant DNA. ITS1-F/ITS4-B preferential amplification was shown to be particularly useful for detection and analysis of the basidiomycete component in ectomycorrhizae and in rust-infected tissues. These primers can be used to study the structure of ectomycorrhizal communities or the distribution of rusts on alternate hosts.