Isolation of a Variant Strain of Pleurotus eryngii and the Development of Specific DNA Markers to Identify the Variant Strain

A degenerated strain of Pleurotus eryngii KNR2312 was isolated from a commercial farm. Random amplified polymorphic DNA analysis performed on the genomic DNA of the normal and degenerated strains of this species revealed differences in the DNA banding pattern. A unique DNA fragment (1.7 kbp), which appeared only in the degenerated strain, was isolated and sequenced. Comparing this sequence with the KNR2312 genomic sequence showed that the sequence of the degenerated strain comprised three DNA regions that originated from nine distinct scaffolds of the genomic sequence, suggesting that chromosome-level changes had occurred in the degenerated strain. Using the unique sequence, three sets of PCR primers were designed that targeted the full length, the 5' half, and the 3' half of the DNA. The primer sets P2-1 and P2-2 yielded 1.76 and 0.97 kbp PCR products, respectively, only in the case of the degenerated strain, whereas P2-3 generated a 0.8 kbp product in both the normal and the degenerated strains because its target region was intact in the normal strain as well. In the case of the P2-1 and P2-2 sets, the priming regions of the forward and reverse primers were located at distinct genomic scaffolds in the normal strain. These two primer sets specifically detected the degenerate strain of KNR2312 isolated from various mushrooms including 10 different strains of P. eryngii, four strains of P. ostreatus, and 11 other wild mushrooms.

Characterization of Species of Cladobotryum which Cause Cobweb Disease in Edible Mushrooms Grown in Korea

Four Cladobotryum isolates were collected from four different commercially grown mushroom types infected with cobweb disease in Cheongdo-gun and Chilgok-gun of Gyeongbuk Province, Korea in 2010. The isolates were identified as C. mycophilum from Agaricus bisporus and Pleurotus eryngii, C. varium from Flammulina velutipes and Hypsizygus marmoreus. The cultural characteristics of the four isolates were investigated using potato dextrose agar (PDA) media under nine different temperatures ranging from 5~32degrees C. Rapid growth of the isolates to colony diameters of 47~82 mm was observed at conditions of 18~22degrees C. No growth was observed at 32degrees C. C. mycophilum produced a yellowish red pigment while C. varium produced a cream colored pigment after cultivation for 25 days on PDA. Phylogenetic analysis of the internal transcribed spacer region and partial 28S rDNA from the four isolates confirmed they were C. mycophilum and C. varium. Cross pathogenicity tests revealed that the two isolates of C. mycophilum were highly pathogenic toward three mushroom types, but not toward H. marmoreus. The two isolates of C. varium were less pathogenic than those of C. mycophilum, but were pathogenic toward all mushroom types evaluated.

Breeding of New Strains of Mushroom by Basidiospore Chemical Mutagenesis

Chemical mutagenesis of basidiospores of Hypsizygus marmoreus generated new mushroom strains. The basidospores were treated with methanesulfonate methylester, an alkylating agent, to yield 400 mutant monokaryotic mycelia. Twenty fast-growing mycelia were selected and mated each other by hyphal fusion. Fifty out of the 190 matings were successful (mating rate of 26.3%), judged by the formation of clamp connections. The mutant dikaryons were cultivated to investigate their morphological and cultivation characteristics. Mutant strains No. 3 and No. 5 showed 10% and 6% increase in fruiting body production, respectively. Eight mutant strains showed delayed and reduced primordia formation, resulting in the reduced production yield with prolonged cultivation period. The number of the fruiting bodies of mutant No. 31, which displayed reduced primordial formation, was only 15, compared to the parental number of 65. Another interesting phenotype was a fruiting body with a flattened stipe and pileus. Dikaryons generated by mating with the mutant spore No. 14 produced flat fruiting bodies. Further molecular biological studies will provide details of the mechanism. This work shows that the chemical mutagenesis approach is highly utilizable in the development of mushroom strains as well as in the generation of resources for molecular genetic studies.

Determination of Mineral Components in the Cultivation Substrates of Edible Mushrooms and Their Uptake into Fruiting Bodies

The mineral contents of the cultivation substrates, fruiting bodies of the mushrooms, and the postharvest cultivation substrates were determined in cultivated edible mushrooms Pleurotus eryngii, Flammulina velutipes, and Hypsizigus marmoreus. The major mineral elements both in the cultivation substrates and in the fruiting bodies were K, Mg, Ca, and Na. Potassium was particularly abundant ranging 10~13 g/kg in the cultivation substrates and 26~30 g/kg in the fruiting bodies. On the contrary, the calcium content in the fruiting bodies was very low despite high concentrations in the cultivation substrates, indicating Ca in the cultivation substrates is in a less bio-available form or the mushrooms do not have efficient Ca uptake channels. Among the minor mineral elements determined in this experiment, Cu, Zn, and Ni showed high percentage of transfer from the cultivation substrates to the fruiting bodies. It is noteworthy that the mineral contents in the postharvest cultivation substrates were not changed significantly which implies that the spent cultivation substrates are nutritionally intact in terms of mineral contents and thus can be recycled as mineral sources and animal feeds.

Retrotransposon Microsatellite Amplified Polymorphism Strain Fingerprinting Markers Applicable to Various Mushroom Species

The retrotransposon marY1 is a gypsy family retroelement, which is detected ubiquitously within the fungal taxonomic groups in which mushrooms are included. To utilize marY1 as a molecular marker for the DNA fingerprinting of mushrooms, oligonucleotides marY1-LTR-L and marY1-LTR-R were designed on the basis of highly conserved regions from the multiple sequence alignment of 30 marY1 sequences retrieved from a nucleotide sequence database. In accordance with Retrotransposon Microsatellite Amplified Polymorphism (REMAP) fingerprinting methodology, the two oligonucleotides were utilized together with the short sequence repeat primers UBC807 and UBC818 for polymerase chain reaction using templates from different mushroom genomic DNAs. Among the tested oligonucleotides, the marY1-LTR-L and UBC807 primer set yielded the greatest amount of abundance and variation in terms of DNA band numbers and patterns. This method was successfully applied to 10 mushroom species, and the primer set successfully discriminated between different commercial mushroom cultivars of the same strains of 14 Pleurotus ostreatus and 16 P. eryngii. REMAP reproducibility was superior to other popular DNA fingerprinting methodologies including the random amplified polymorphic DNA method.

HRCT Finding of Pulmonary Cryptococcosis in Immune Competent Patients

PURPOSE: To determine the HRCT finding of pulmonary cryptococcosis in immunocompetent patients. MATERIALS AND METHODS: The HRCT scans and chest radiographs of five patients with pulmonary cryptococcosis were retrospectively reviewed, the diagnosis being proven by the presence of the organism in histopathologic specimens obtained during CT-guided biopsy (n = 4) or medistinoscopic biopsy (n = 1). All patients were immunocompetent. HRCT scans and chest radiographs were characterized according to morphology [consoli-dation, nodule, ground-glass opacity (GGO), interstitial thickening], location, airbronchogram, and the pres-ence or absence of mediastinal or hilar lymphadenopathy. RESULTS: Consolidation was present in three of five patients, and in two of the three, ill-defined, spiculated nodules were also present. One patient had multiple, small, well-defined nodules. One had diffuse interstitial thickening (GGO, nodular thickening of bronchovascular bundles and interlobular septa), with massive mediastinal and right hilar lymphadenopathy. The location of the consolidation and nodules was mainly peripheral, and in three patients with consolidation, the presence of air bronchograms was noted. CONCLUSION: The HRCT findings of pulmonary cryptococcosis in immunocompetent patients are peripheral consolidation with airbronchogram and/or ill defined nodules.

Measurement of Nasal Trigeminal Pungency Threshold: Comparison between Normals and Patients with Decreased Sense of Smell

BACKGROUND AND OBJECTIVES: In order to measure the level of nasal trigeminal pungency threshold and to evaluate the interaction between olfactory and trigeminal nerves, we measured olfactory thresholds and trigeminal pungency thresholds using 1-butanol in 40 normal people and in 46 patients with decreased sense of smell by non-sinonasal origin. MATERIALS AND METHODS: Two-alternative forced-choice modified ascending method of limit (CCCRC test) was used for the measurement of olfactory and pungency thresholds, using 1-butanol plastic squeezable bottles successively threefold diluted by distilled water as stimulus, while distilled water as blank. RESULTS: There was a statistically significant difference in pungency threshold between normals (2.18+/-1.5 dilution step) and patients (1.11+/-1.0 dilution step)(p<0.001). CONCLUSION: The pungency thresholds of hyposmics and anosmics are higher than those of normals, which suggests that a loss or decrease of olfactory sensation is accompanied by a simultaneous decrease of trigeminal chemosensitivity.

A Case of Kallmann's Syndrome: MR Imaging and Histopathology

Kallmann's syndrome is a rare genetic disorder defined as a combination of anosmia or hyposmia and hypogonadism due to hypothalamic-pituitary deficiency. The incidence of Kallmann's syndrome is one in 100,000 men and one in 50,000 women. However, the syndrome is found in one in 25 people with hypogonadism. The clinical evaluation of these patients includes historical and physical examinations ; chemosensory testing, which includes olfactory threshold and identification tests ; laboratory testing for hormonal abnormalities ; MR imaging of the olfactory apparatus ; and histopathologic findings through an olfactory epithelial biopsy. We report the case of a 19-year-old male patient who had anosmia since birth and who was diagnosed with Kallmann's syndrome.

A Case of Intranasal Meningoencephalocele / 대한이비인후과학회지

Meningoencephalocele is an ectopic protrusion of the meninges and a portion of the brain that retains connection with the CNS through a defect in the bony covering. It is a rare developmental anomaly that results from faulty closure of the embryonic neural tubes. Meningoencephalocele is usually classified into occipital, sincipital and basal types according to the location and among these, the basal type is less frequently encountered than other types. The authors have experienced a case of basal-type intranasal meningoencephalocele in 4 year-old male patient, which was removed successfully via bifrontal craniotomy and intranasal endoscopic approach, and the defect of skull base was repaired by inner table of the cranium and septal mucosa with good result.

The McCune-Albright's syndrome: a case report and review of the literature / 대한산부인과학회잡지

No abstract available.