Rapid molecular approach for a reliable identification of Tuber spp. ectomycorrhizae

Rapid molecular approach for a reliable identification of Tuber spp. ectomycorrhizae
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DOI:
10.1111/j.1574-6941.1999.tb00557.x
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发表时间:
1999
影响因子:
4.2
通讯作者:
F. Paolocci;A. Rubini;B. Granetti;S. Arcioni
F. Paolocci;A. Rubini;B. Granetti;S. Arcioni
中科院分区:
生物学3区
文献类型:
--
作者:
F. Paolocci;A. Rubini;B. Granetti;S. Arcioni

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建立了从外生菌根中分离和扩增真菌DNA的方法,克服了PCR抑制剂的存在,以确保快速和可靠的松露菌根鉴定。利用多重聚合酶链式反应中的种特异性ITS引物,可以在一步扩增中鉴定出形态非常相似的块茎黑孢菌、印度块茎和丛枝外生菌根菌,而不考虑它们的生理阶段。与迄今进行的任何形态或ITS/RFLP分析相比,分子方法确保了对苗圃接种植物在田间种植之前和之后的更准确和可靠的监测。它还应该防止欺诈,这已经是一个问题,以及由于将异地物种印度松露错误地归类为欧洲松露物种而可能造成的环境破坏,因为外生菌根的身份仅基于形态。
Protocols for isolating and amplifying fungal DNA from ectomycorrhizae, overcoming the presence of PCR inhibitors, were established to ensure a fast and reliable truffle mycorrhizal identification. By using species-specific ITS primers in multiplex PCR, morphologically very similar Tuber melanosporum, Tuber indicum and Tuber brumale ectomycorrhizae were identified in a single step of amplification, irrespective of their physiological stage. The molecular method assures a more accurate and reliable monitoring of nursery-inoculated plants both before and after their in-field planting, than any morphological or ITS/RFLP analyses so far performed. It should also prevent frauds, already a problem, and the potential for environmental damage due to the incorrect typing of the allochthonous species T. indicum as European truffle species, when the identity of ectomycorrhizae is based solely on morphology.