Browse > Article
http://dx.doi.org/10.1080/12298093.2020.1785754

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

Lee, Song Hee (Department of Mushroom Science, Korea National College of Agriculture and Fisheries)
Jung, Hwa Jin (Department of Mushroom Science, Korea National College of Agriculture and Fisheries)
Hong, Seung-Beom (Korean Agricultural Culture Collection, Agricultural Microbiology Division, National Academy of Agricultural Science, Rural Development Administration)
Choi, Jong In (Mushroom Research Institute, GARES)
Ryu, Jae-San (Department of Mushroom Science, Korea National College of Agriculture and Fisheries)
Publication Information
Mycobiology / v.48, no.4, 2020 , pp. 313-320 More about this Journal
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).
Keywords
Trichoderma; molecular marker; green mold; Pleurotus eryngii; pathogenicity;
Citations & Related Records
Times Cited By KSCI : 10  (Citation Analysis)
연도 인용수 순위
1 Mleczek M, Siwulski M, Rzymski P, et al. Cultivation of mushrooms for production of food biofortified with lithium. Eur Food Res Technol. 2017;243(6):1097-1104.   DOI
2 Estrada AR, Royse DJ. Yield, size and bacterial blotch resistance of Pleurotus eryngii grown on cottonseed hulls/oak sawdust supplemented with manganese, copper and whole ground soybean. Bioresour Technol. 2007;98(10):1898-1906.   DOI
3 Jin X, Wang Q, Yang X, et al. Chemical characterisation and hypolipidaemic effects of two purified Pleurotus eryngii polysaccharides. Int J Food Sci Technol. 2018;53(10):2298-2307.   DOI
4 Ma G, Kimatu BM, Zhao L, et al. Impacts of dietary Pleurotus eryngii polysaccharide on nutrient digestion, metabolism, and immune response of the small intestine and colon - an iTRAQ-based proteomic analysis. Proteomics. 2018;18(7):1700443.   DOI
5 Lee SH, Kim MK, Ryu JS, et al. Characteristics of a new Pleurotus eryngii cultivar, Aeryni 6. J Mushroom. 2018;16(1):16-21.   DOI
6 Jang MJ, Park Y, Kim JH. Properties of disease occurrence by season for cultivation facilities of oyster mushroom. J Mushroom Sci. 2019;17(3):93-98.
7 Sobiralski K, Siwulski M, Gorski R, et al. Impact of Trichoderma aggressivum f. europaeum isolates on yielding and morphological features of Pleurotus eryngii. Pytopathologia. 2010;56:17-25.
8 Hatvani L, Antal L, Manczinger L, et al. Green mold diseases of Agaricus and Pleurotus spp. are caused by related but phylogenetically different Trichoderma species. Phytopathology. 2007;97(4):532-537.   DOI
9 Komon-Zelazowska M, Bissett J, Zafari D, et al. Genetically closely related but phenotypically divergent Trichoderma species cause green mold disease in oyster mushroom farms worldwide. Appl Environ Microbiol. 2007;73(22):7415-7426.   DOI
10 Kredics L, Kocsube S, Nagy L, et al. Molecular identification of Trichoderma species associated with Pleurotus ostreatus and natural substrates of the oyster mushroom. FEMS Microbiol Lett. 2009;300(1):58-67.   DOI
11 Park MS, Bae KS, Yu SH. The new species of Trichoderma associated with green mold of oyster mushroom cultivation in Korea. Mycobiology. 2006;34(3):11-13.
12 Sobieralski K, Siwulski M, Komon-Zelazowska M, et al. Evaluation of the growth of Trichoderma pleurotum and Trichoderma pleuroticola isolates and their biotic interaction with Pleurotus sp. J Plant Protect Res. 2012;52(2):235-239.   DOI
13 Harman GE, Howell CR, Viterbo A, et al. Trichoderma species-opportunistic, avirulent plant symbionts. Nat Rev Microbiol. 2004;2(1):43-56.   DOI
14 Wellings CR, McIntosh RA, Walker J. Puccinia striiformis f. sp. tritici in Eastern Australia -possible means of entry and implications for plant quarantine. Plant Pathol. 1987;36(3):239-241.   DOI
15 Park B, Ha BS, Lee SH, et al. Variable-number tandem repeat loci-discriminating Pleurotus ostreatus cultivars. Mycoscience. 2019;60(2):132-135.   DOI
16 White TJ, Brunes T, Taylor JW. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis M, Gelfand D, Sninsky J, et al., editors. PCR protocols: a guide to methods and applications. Cambridge, MA: Academic Press; 1990. p. 315-322.
17 Dees PM, Ghiorse WC. Microbial diversity in hot synthetic compost as revealed by PCR-amplified rRNA sequences from cultivated isolates and extracted DNA. FEMS Microbiol Ecol. 2001;35(2):207-216.   DOI
18 Lee SH, Ali A, Ha B, et al. Development of a molecular marker linked to the A4 locus and the structure of HD genes in Pleurotus eryngii. Mycobiology. 2019;47(2):200-206.   DOI
19 Druzhinina IS, Kopchinskiy AG, Komon M, et al. An oligonucleotide barcode for species identification in Trichoderma and Hypocrea. Fungal Genet Biol. 2005;42(10):813-828.   DOI
20 Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA. 1977;74(12):5463-5467.   DOI
21 Kopchinskiy A, Komon M, Kubicek CP, et al. TrichoBLAST: a multilocus database for Trichoderma and Hypocrea identifications. Mycol Res. 2005;109(Part 6):658-660.   DOI
22 Ryu JS, Kim MK, Im CH, et al. Development of cultivation media for extending the shelf-life and improving yield of king oyster mushrooms (Pleurotus eryngii). Sci Hortic. 2015;193:121-126.   DOI
23 Team RDC. R: a language and environment for statistical computing. Vienna, Austria: R foundation for statistical computing; 2013. Available from: http://www.R-project.org.
24 Thompson JD, Gibson TJ, Higgins DG. Multiple sequence alignment using ClustalW and ClustalX. Curr Protoc Bioinformatics. 2002;Chapter 2:Unit 2.3.
25 Kalendar R, Lee D, Schulman AH. FastPCR software for PCR, in silico PCR, and oligonucleotide assembly and analysis. In: Valla S, Lale R, editors. DNA cloning and assembly methods, Methods in Molecular Biology. Totowa, NJ: Humana Press; 2014. p. 271-302.
26 Williams J, Clarkson JM, Mills PR, et al. Saprotrophic and mycoparasitic components of aggressiveness of Trichoderma harzianum groups toward the commercial mushroom Agaricus bisporus. Appl Environ Microbiol. 2003;69(7):4192-4199.   DOI
27 Choi I-Y, Joung G-T, Ryu J, et al. Physiological characteristics of green mold (Trichoderma spp.) isolated from Oyster Mushroom (Pleurotus spp). Mycobiology. 2003;31(3):139-144.   DOI
28 Kim SW, Kim S, Lee HJ, et al. Isolation of fungal pathogens to an edible mushroom, Pleurotus eryngii, and development of specific ITS primers. Mycobiology. 2013;41(4):252-255.   DOI
29 Samuels GJ, Dodd SL, Gams W, et al. Trichoderma species associated with the green mold epidemic of commercially grown Agaricus bisporus. Mycologia. 2002;94(1):146-170.   DOI
30 Miyazaki K, Tsuchiya Y, Okuda T. Specific PCR assays for the detection of Trichoderma harzianum causing green mold disease during mushroom cultivation. Mycoscience. 2009;50(2):94-99.   DOI
31 Janpoor J, Pourianfar HR, Shahtahmasebi S. Study on effect of culture medium and growth conditions on liquid spawn king oyster mushroom (Pleurotus eryngii). Int J Farm Allied Sci. 2017;6(6):154-156.
32 Lee SJ, Kim HH, Kim SH, et al. Culture conditions of liquid spawn and the growth characteristics of Pleurotus ostreatus. J Mushroom. 2018;16(3):162-170.   DOI