Pheromone (PHB)-receptor (RCB) interaction in the mating pheromone response pathway of Lentinula edodes was investigated using synthetic PHBs. Functionality of the C-terminally carboxymethylated synthetic PHBs was demonstrated by concentration-dependent induction of a mating-related gene (znf2) expression and by pseudoclamp formation in a monokaryotic strain S1-11 of L. edodes. Treatment with synthetic PHBs activated the expression of homeodomain genes (HDs) residing in the A mating type locus, and of A-regulated genes, including znf2, clp1, and priA, as well as genes in the B mating type locus, including pheromone (phb) and receptor (rcb) genes. The synthetic PHBs failed to discriminate self from non-self RCBs. PHBs of the B4 mating type (B4 PHBs) were able to activate the mating pheromone response pathway in both monokaryotic S1-11 and S1-13 strains, whose B mating types were B4 (self) and B12 (non-self), respectively. The same was true for B12 PHBs in the B4 (non-self) and B12 (self) mating types. The synthetic PHBs also promoted the mating of two monokaryotic strains carrying B4-common incompatible mating types (A5B4 × A1B4). However, the dikaryon generated by this process exhibited abnormally high content of hyphal branching and frequent clamp connections and, more importantly, was found to be genetically unstable due to overexpression of mating-related genes such as clp1. Although synthetic PHBs were unable to discriminate self from non-self RCBs, they showed a higher affinity for non-self RCBs, through which the mating pheromone response pathway in non-self cells may be preferentially activated.
Lentinula* ; Pheromones* ; Shiitake Mushrooms*

Sixteen genomic DNA simple sequence repeat (SSR) markers of Lentinula edodes were developed from 205 SSR motifs present in 46.1-Mb long L. edodes genome sequences. The number of alleles ranged from 3–14 and the major allele frequency was distributed from 0.17–0.96. The values of observed and expected heterozygosity ranged from 0.00–0.76 and 0.07–0.90, respectively. The polymorphic information content value ranged from 0.07–0.89. A dendrogram, based on 16 SSR markers clustered by the paired hierarchical clustering' method, showed that 33 shiitake cultivars could be divided into three major groups and successfully identified. These SSR markers will contribute to the efficient breeding of this species by providing diversity in shiitake varieties. Furthermore, the genomic information covered by the markers can provide a valuable resource for genetic linkage map construction, molecular mapping, and marker-assisted selection in the shiitake mushroom.
Alleles ; Breeding ; DNA* ; Gene Frequency ; Genetic Linkage ; Genetic Variation ; Genome ; Lentinula* ; Methods ; Microsatellite Repeats ; Shiitake Mushrooms*

In mating of Lentinula edodes, dikaryotic strains generated from certain monokaryotic strains such as the B2 used in this study tend to show better quality of fruiting bodies regardless of the mated monokaryotic strains. Unlike B2, dikaryotic strains generated from B16 generally show low yields, with deformed or underdeveloped fruiting bodies. This indicates that the two nuclei in the cytoplasm do not contribute equally to the physiology of dikaryotic L. edodes, suggesting an expression bias in the allelic genes of the two nuclei. To understand the role of each nucleus in dikaryotic strains, we investigated single nucleotide polymorphisms (SNPs) in laccase genes of monokaryotic strains to reveal nuclear origin of the expressed mRNAs in dikaryotic strain. We performed reverse transcription PCR (RT-PCR) analysis using total RNAs extracted from dikaryotic strains (A5B2, A18B2, and A2B16) as well as from compatible monokaryotic strains (A5, A18, and B2 for A5B2 and A18B2; A2 and B16 for A2B16). RT-PCR results revealed that Lcc1, Lcc2, Lcc4, Lcc7, and Lcc10 were the mainly expressed laccase genes in the L. edodes genome. To determine the nuclear origin of these laccase genes, the genomic DNA sequences in monokaryotic strains were analyzed, thereby revealing five SNPs in Lcc4 and two in Lcc7. Subsequent sequence analysis of laccase mRNAs expressed in dikaryotic strains revealed that these were almost exclusively expressed from B2-originated nuclei in A5B2 and A18B2 whereas B16 nucleus did not contribute to laccase expression in A2B16 strain. This suggests that B2 nucleus dominates the expression of allelic genes, thereby governing the physiology of dikaryons.
Base Sequence ; Bias (Epidemiology) ; Cytoplasm ; Fruit ; Genome ; Laccase* ; Lentinula* ; Physiology ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Reverse Transcription ; RNA ; RNA, Messenger ; Sequence Analysis ; Shiitake Mushrooms*

The culture filtrate of Lentinula edodes shows potent antimicrobial activity against the plant pathogenic bacteria Ralstonia solanacearum. Bioassay-guided fractionation was conducted using Diaion HP-20 column chromatography, and the insoluble active compound was not adsorbed on the resin. Further fractionation by high-performance liquid chromatography (HPLC) suggested that the active compounds were organic acids. Nine organic acids were detected in the culture filtrate of L. edodes; oxalic acid was the major component and exhibited antibacterial activity against nine different phytopathogenic bacteria. Quantitative analysis by HPLC revealed that the content of oxalic acid was higher in the water extract from spent mushroom substrate than in liquid culture. This suggests that the water extract of spent L. edodes substrate is an eco-friendly control agent for plant diseases.
Agaricales ; Bacteria* ; Chromatography ; Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Lentinula* ; Oxalic Acid* ; Plant Diseases ; Plants ; Ralstonia solanacearum ; Shiitake Mushrooms* ; Water

Culture filtrates of six different edible mushroom species were screened for antimicrobial activity against tomato wilt bacteria Ralstonia solanacearum B3. Hericium erinaceus, Lentinula edodes (Sanjo 701), Grifola frondosa, and Hypsizygus marmoreus showed antibacterial activity against the bacteria. Water, n-butanol, and ethyl acetate extracts of spent mushroom substrate (SMS) of H. erinaceus exhibited high antibacterial activity against different phytopathogenic bacteria: Pectobacterium carotovorum subsp. carotovorum, Agrobacterium tumefaciens, R. solanacearum, Xanthomonas oryzae pv. oryzae, X. campestris pv. campestris, X. axonopodis pv. vesicatoria, X. axonopodis pv. citiri, and X. axonopodis pv. glycine. Quantitative real-time PCR revealed that water extracts of SMS (WESMS) of H. erinaceus induced expressions of plant defense genes encoding beta-1,3-glucanase (GluA) and pathogenesis-related protein-1a (PR-1a), associated with systemic acquired resistance. Furthermore, WESMS also suppressed tomato wilt disease caused by R. solanacearum by 85% in seedlings and promoted growth (height, leaf number, and fresh weight of the root and shoot) of tomato plants. These findings suggest the WESMS of H. erinaceus has the potential to suppress bacterial wilt disease of tomato through multiple effects including antibacterial activity, plant growth promotion, and defense gene induction.
1-Butanol ; Agaricales* ; Agrobacterium tumefaciens ; Bacteria ; Glycine ; Grifola ; Lycopersicon esculentum* ; Oryza ; Pectobacterium carotovorum ; Plants ; Ralstonia solanacearum ; Real-Time Polymerase Chain Reaction ; Seedlings ; Shiitake Mushrooms ; Water* ; Xanthomonas

The effects of monokaryotic strains on fruiting body formation of Lentinula edodes were examined through mating and cultivation of the mated dikaryotic mycelia in sawdust medium. To accomplish this, monokaryotic strains of L. edodes were isolated from basidiospores of the commercial dikaryotic strains, Chamaram (Cham) and Sanjo701 (SJ701). A total of 703 matings (538 self-matings and 165 outcrosses) were performed, which generated 133 self-mates and 84 outcross mates. The mating rate was 25% and 50% for self-mating and outcross, respectively. The bipolarity of the outcross indicated the multi-allelic nature of the mating type genes. The mating was only dependent on the A mating type locus, while the B locus showed no effect, implying that the B locus is multi-allelic. Next, 145 selected dikaryotic mates were cultivated in sawdust medium. The self-mated dikaryotic progenies showed 51.3% and 69.5% fruiting rates for Cham and SJ701, respectively, while the fruiting rate of the outcross mates was 63.2%. The dikaryotic mates generated by mating with one of the monokaryotic strains, including A20, B2, E1, and E3, showed good fruiting performance and tended to yield high fruiting body production, while many of the monokaryotic strains failed to form fruiting bodies. Overall, these findings suggest that certain monokaryotic strains have traits enabling better mating and fruiting.
Fruit* ; Shiitake Mushrooms*

We identified single nucleotide polymorphism (SNP) markers in the laccase gene to establish a line-diagnostic system for shiitake mushrooms. A total of 89 fungal isolates representing four lines, including Korean registered, Korean wild type, Chinese, and Japanese lines, were analyzed. The results suggest that SNP markers in the laccase gene can be useful for line typing in shiitake mushrooms.
Agaricales ; Asian Continental Ancestry Group ; Humans ; Laccase* ; Polymorphism, Single Nucleotide* ; Shiitake Mushrooms*

It has been reported that ingestion of raw or undercooked shiitake mushrooms is associated with various adverse food reactions. A 58-year-old man presented with pruritic and maculopapular rashes on the trunk and extremities starting 1 day after ingestion of partially uncooked shiitake mushrooms. A probable diagnosis of systemic allergic contact dermatitis was made. Raw and cooked shiitake mushroom-derived allergen extracts were prepared, and patch and intradermal tests with delayed readings were performed. The case showed positive intradermal test results with delayed readings to the raw, but not cooked, extracts. The case suggests that ingestion of uncooked shiitake mushrooms may induce systemic allergic contact dermatitis through type IV hypersensitivity reaction.
Dermatitis, Allergic Contact* ; Dermatitis, Contact ; Diagnosis ; Eating ; Exanthema ; Extremities ; Humans ; Hypersensitivity, Delayed ; Intradermal Tests ; Middle Aged ; Patch Tests ; Reading ; Shiitake Mushrooms

Shiitake mushrooms (Lentinula edodes) containing beta-glucans may be beneficial for human health; they have been used in the treatment of cancer, hypertension, and high cholesterol levels. The objective of this study was to determine the beta-glucan content in different sections of the fruiting bodies and mycelia of ten shiitake mushroom cultivars. The measured beta-glucan content ranged from 20.06 +/- 1.76% to 44.21 +/- 0.13% in the pileus sections, and from 29.74 +/- 1.40% to 56.47 +/- 4.72% in the stipe sections. The results of this study indicate that the variance in beta-glucan content dependent on the shiitake cultivar, and that the beta-glucan content is higher in the stipe than in the pileus.
beta-Glucans ; Cholesterol ; Fruit* ; Humans ; Hypertension ; Shiitake Mushrooms*

Severe bark decay disease was observed on oak logs at a shiitake cultivation farm in Geochang-gun, Gyeongnam province. The symptoms observed were fruiting bodies that had developed on the top and side surface of oak logs. As a result, the bark came off easily exposing the sapwood. Slime mold specimens collected from oak logs showed developing fruiting bodies comprising of stalks, hypothallus, capillitium, and columella, and the causal agent of bark decay disease was identified as Stemonitis splendens on the basis of morphological characteristics. To our knowledge, this is the first report of Stemonitis splendens causing bark decay of oak logs used for shiitake mushroom cultivation in Korea.
Fruit ; Fungi ; Korea ; Shiitake Mushrooms