Iotti Mirco, Zambonelli Alessandra
Dipartimento di Protezione e Valorizzazione Agroalimentare, Università degli Studi di Bologna, via Fanin 46, I-40127 Bologna, Italy.
Mycol Res. 2006 Jan;110(Pt 1):60-5. doi: 10.1016/j.mycres.2005.09.010. Epub 2005 Dec 22.
A rapid procedure was developed to amplify ITS fragments directly from Tuber ectomycorrhizas either synthesized in a greenhouse or collected from the field. The addition of Bovine Serum Albumin (BSA) to the reaction mixtures overcame the presence of reaction inhibitors present in fungal and root cells, and enabled the amplification of the ITS regions directly from ectomycorrhizal tissues. This method is cheaper and less time-consuming than conventional procedures, and reduces the time required from 1-4 h to a few minutes. It is also much more sensitive, allowing the identification of just a small fragment of a mycorrhizal root tip. Because of this it is possible to select only the target fungal tissue and hence minimise the risk of contamination by saprobic or other mycorrhizal species. The method also avoids the use of toxic or hazardous substances. This method could have a wider application in other areas of applied mycology.
开发了一种快速程序,可直接从温室中合成或从田间采集的块菌外生菌根中扩增ITS片段。在反应混合物中添加牛血清白蛋白(BSA)克服了真菌和根细胞中存在的反应抑制剂的影响,并能够直接从外生菌根组织中扩增ITS区域。该方法比传统方法更便宜且耗时更少,将所需时间从1 - 4小时减少到几分钟。它也更加灵敏,仅需一小段菌根根尖就能进行鉴定。因此,可以仅选择目标真菌组织,从而将腐生菌或其他菌根物种污染的风险降至最低。该方法还避免使用有毒或有害物质。此方法可能在应用真菌学的其他领域有更广泛的应用。