A molecular diagnostic assay for the detection and identification of wood decay fungi of conifers
A molecular diagnostic assay for the detection and identification of wood decay fungi of conifers
复制标题
检测和鉴定针叶树木材腐烂真菌的分子诊断方法
DOI：
10.1111/efp.12132
复制
发表时间：
2015
期刊：
影响因子：
1.4
通讯作者：
M. Garbelotto
中科院分区：
文献类型：
--
作者：
P. Gonthier;F. Guglielmo;F. Sillo;L. Giordano;M. Garbelotto
总结
设计了10条分类群特异性引物，对蜜环菌、棘齿菌属，Pinicola拟层孔菌，Fuscoporia torulosa，Heterobasidion annosum lato（s.l.），Onnia spp.，Schweinitzii，Phaeolus weirii s.l.，黄伞属和Porodaedalea spp.在这项研究中设计的引物和在以前的一个用于鉴定Laetiporus sulphureus和Stereum spp。在两个多重PCR中组合，测试其效率和特异性，并检测至少1 pg的真菌靶DNA。使用SYBR®绿色实时PCR，该检测试剂盒可检测浓度为10 - 1 pg或更低的靶DNA。对129个自然感染的木材样品或子实体进行的验证试验证实了基于多重PCR的诊断方法的可靠性。该方法为针叶树木材腐朽菌的快速检测提供了一种简便、快速的方法。
Summary
Ten taxon-specific primers were designed to amplify the Internal Transcribed Spacer of the rRNA operon of several important decay fungi of coniferous wood, including Armillaria spp., Echinodontium spp., Fomitopsis pinicola, Fuscoporia torulosa, Heterobasidion annosum sensu lato (s.l.), Onnia spp., Phaeolus schweinitzii, Phellinus weirii s.l., Pholiota spp. and Porodaedalea spp. Primers designed in this study and in a previous one for the identification of Laetiporus sulphureus and Stereum spp. were combined in two multiplex PCRs, which were tested for efficiency and specificity, and detected at least 1 pg of fungal target DNA. Target DNA at concentrations of 10−1 pg or lower can be detected with this assay using SYBR® Green Real-Time PCR. Validation assays performed on 129 naturally infected wood samples or fruiting bodies confirmed the reliability of the multiplex PCR-based diagnostic method. This method represents a simple and rapid diagnostic tool for the detection of a number of destructive wood decay fungi of conifer wood.