Abstract

In Pleurotus sp., green mold, which is considered a major epidemic, is caused by several Trichoderma species. To develop a rapid molecular marker specific for Trichoderma spp. that potentially cause green mold, eleven Trichoderma species were collected from mushroom farms and the Korean Agricultural Culture Collection (KACC). A dominant fungal isolate from a green mold-infected substrate was identified as Trichoderma pleuroticola based on the sequences of its internal transcribed spacer (ITS) and translation elongation factor 1-α (tef1) genes. In artificial inoculation tests, all Trichoderma spp., including T. atroviride, T. cf. virens, T. citrinoviride, T. harzianum, T. koningii, T. longibrachiatum, T. pleurotum, and T. pleuroticola, showed pathogenicity to some extent, and the observed symptoms were soaked mycelia with a red-brown pigment and retarded mycelium regeneration. A molecular marker was developed for the rapid detection of wide range of Trichoderma spp. based on the DNA sequence alignment of the ITS1 and ITS2 regions of Trichoderma spp. The developed primer set detected only Trichoderma spp., and no cross reactivity with edible mushrooms was observed. The detection limits for the PCR assay of T. harzianum (KACC40558), T. pleurotum (KACC44537), and T. pleuroticola (CAF-TP3) were found to be 500, 50, and 5 fg, respectively, and the detection limit for the pathogen-to-host ratio was approximately 1:10,000 (wt/wt).

摘要

在杏鲍菇属中，绿色霉菌被认为是主要流行病，是由几种木霉属物种引起的。开发特异于木霉属的快速分子标记。可能导致绿色霉菌的细菌从蘑菇农场和韩国农业文化收藏中心（KACC）收集到11种木霉属物种。根据其内部转录间隔区（ITS）和翻译延伸因子1-α（tef1）基因的序列，来自绿色霉菌感染底物的优势真菌分离物被鉴定为胸膜木霉。在人工接种测试中，所有木霉属物种，包括A.trotroide，T.比照 霉，T. citrinoviride，哈茨木霉，T.康宁木霉，木霉属longibrachiatum，T. pleurotum，和T. pleuroticola，表明致病性在一定程度上，并且所观察到的症状浸泡菌丝体用红棕色颜料和延迟菌丝再生。开发了一种分子标记物，用于快速检测多种木霉属物种。基于木霉属物种的ITS1和ITS2区域的DNA序列比对。开发的引物组仅检测到木霉spp。，未观察到与食用菌的交叉反应。对的PCR测定法，检出限哈茨木霉（KACC40558），T. pleurotum（KACC44537），和T. pleuroticola（CAF-TP3）被发现是500，50和5 FG分别检出限为病原体与宿主的比例约为1：10,000（wt / wt）。