van der Linde Sietse, Alexander Ian, Anderson Ian C
The Macaulay Institute, Craigiebuckler, Aberdeen, AB15 8QH, United Kingdom.
J Microbiol Methods. 2008 Sep;75(1):40-6. doi: 10.1016/j.mimet.2008.04.010. Epub 2008 May 15.
To reduce the reliance on sporocarp records for conservation efforts, information on the below-ground distribution of specific fungal species, such as stipitate hydnoid fungi, is required. Species-specific primers were developed within the internal transcribed spacer (ITS1 and ITS2) regions for 12 hydnoid fungal species including Bankera fuligineoalba, Hydnellum aurantiacum, H. caeruleum, H. concrescens, H. ferrugineum, H. peckii, Phellodon confluens, P. melaleucus, P. niger, P. tomentosus, Sarcodon glaucopus and S. squamosus. The specificity of the primer pairs was tested using BLAST searches and PCR amplifications. All primers amplified DNA only of the target species with the exception of those designed for P. melaleucus. In order to assess the ability of the primers to detect DNA from mycelium in soil, DNA extracted from soil samples taken from around solitary H. peckii sporocarps was amplified with the H. peckii primer 1peck and ITS2. H. peckii DNA was detected in 70% of all soil samples and up to 40 cm away from the base of individual sporocarps. The development of these species-specific primers provides a below-ground alternative for monitoring the distribution of these rare fungi.
为减少保护工作对子实体记录的依赖，需要有关特定真菌物种地下分布的信息，例如柄状多孔菌。针对包括暗褐集毛菌、橙黄齿耳菌、蓝齿耳菌、融合齿耳菌、铁锈色齿耳菌、佩氏齿耳菌、融合皱孔菌、白皱孔菌、黑皱孔菌、绒毛皱孔菌、灰蓝肉齿菌和鳞状肉齿菌在内的12种多孔菌，在内部转录间隔区（ITS1和ITS2）开发了物种特异性引物。使用BLAST搜索和PCR扩增测试引物对的特异性。除了为白皱孔菌设计的引物外，所有引物仅扩增目标物种的DNA。为了评估引物检测土壤中菌丝体DNA的能力，用佩氏齿耳菌引物1peck和ITS2扩增从孤立的佩氏齿耳菌子实体周围采集的土壤样品中提取的DNA。在所有土壤样品的70%以及距离单个子实体基部40厘米远的地方都检测到了佩氏齿耳菌DNA。这些物种特异性引物的开发为监测这些稀有真菌的分布提供了一种地下替代方法。
