Mycobiology.  2020 Aug;48(4):313-320. 10.1080/12298093.2020.1785754.

Molecular Markers for Detecting a Wide Range of Trichoderma spp. that Might Potentially Cause Green Mold in Pleurotus eryngii

Affiliations
  • 1Department of Mushroom Science, Korea National College of Agriculture and Fisheries, Jeonju, Korea
  • 2Korean Agricultural Culture Collection, Agricultural Microbiology Division, National Academy of Agricultural Science, Rural Development Administration, Wanju, Korea
  • 3Mushroom Research Institute, GARES, Gwangju, Republic of Korea

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-a (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 ofTrichoderma spp. based on the DNA sequence alignment of the ITS1 and ITS2 regions ofTrichoderma 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).

Keyword

Trichoderma; molecular marker; green mold; Pleurotus eryngii; pathogenicity
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