A useful PCR primer set for the ectomycorrhizal fungus Tricholoma matsutake in wild pine rhizosphere based on the nuclear ribosomal DNA IGS2 sequence

IF 1.5 4区 生物学 Q4 MYCOLOGY Mycoscience Pub Date : 2024-06-05 DOI:10.47371/mycosci.2024.05.001
Hiroki Misawa, Yu Tateishi, Yuka Horimai, Aira Mizuno, Fuuma Hida, Hitoshi Furukawa, Hisayasu Kobayashi, Masataka Kawai, Takashi Yamanaka, Masaki Fukuda, Akiyoshi Yamada
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Abstract

Tricholoma matsutake is an edible ectomycorrhizal mushroom that forms a symbiotic association with Pinaceae trees by constructing a large extraradical mycelial area (called a shiro) in the soil. The detection of this fungal mycelium in the soil is crucial for estimating the success of outplanted mycorrhizal seedlings inoculated with T. matsutake under experimental conditions. Although several T. matsutake-specific DNA markers have been reported for efficient detection in the field, no comparative study has been conducted to assess their effectiveness. In the present study, we targeted the nuclear ribosomal DNA intergenic spacer 2 (IGS2) region for the detection of T. matsutake. The newly designed TmSP-I-2F/TmSP-I-2R primer pair, which targets a partial IGS2 sequence (543 bp), effectively detected T. matsutake from pine root and soil samples via PCR assay, outperforming other T. matsutake-specific primers. In combination with a PCR system targeting LTR DNA markers that were previously developed, a PCR system with the TmSP-I-2F/TmSP-I-2R primer pair set can expedite investigations of the dynamics of T. matsutake genets in mycorrhizas and shiro.

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基于核核糖体 DNA IGS2 序列的野生松树根瘤外生菌类松茸的实用 PCR 引物集
松茸是一种可食用的外生菌根蘑菇,它通过在土壤中构建一个大型根外菌丝区(称为 "shiro")与松科树木形成共生关系。检测土壤中的这种真菌菌丝对于评估在实验条件下接种松茸的外植菌根秧苗的成活率至关重要。虽然有报道称几种松茸特异性 DNA 标记可在田间有效检测松茸,但还没有进行过比较研究来评估它们的有效性。在本研究中,我们以核核糖体 DNA 基因间距 2(IGS2)区域为目标来检测松茸。新设计的 TmSP-I-2F/TmSP-I-2R 引物对以部分 IGS2 序列(543 bp)为目标,通过 PCR 分析从松树根和土壤样本中有效地检测出松茸,其效果优于其他松茸特异性引物。结合之前开发的针对 LTR DNA 标记的 PCR 系统,使用 TmSP-I-2F/TmSP-I-2R 引物对的 PCR 系统可以加快对松茸基因在菌根和石龙子中的动态研究。
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来源期刊
Mycoscience
Mycoscience MYCOLOGY-
CiteScore
3.00
自引率
7.10%
发文量
32
审稿时长
3 months
期刊介绍: Mycoscience is the official English-language journal of the Mycological Society of Japan and is issued bimonthly. Mycoscience publishes original research articles and reviews on various topics related to fungi including yeasts and other organisms that have traditionally been studied by mycologists. The research areas covered by Mycoscience extend from such purely scientific fields as systematics, evolution, phylogeny, morphology, ecology, physiology, biochemistry, genetics, and molecular biology, to agricultural, medical, and industrial applications. New and improved applications of well-established mycological techniques and methods are also covered.
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