Jae-San Ryu

The Growth Characteristics of Pleurotus eryngii

In this study, we investigated the properties of incubation and growing of Pleurotus eryngii in addition to the mycological properties to use them as basic data for breeding. The speed of mycelial growth on the MCM was faster than on the PDA. The biomass in the PDB broth culture was higher than in the MCM and YMG broth culture. KNR2515 and KNR2516 required 19 days for growth of mycelia on commercial sawdust media. KNR2503 required 6.5 days and 15.3 days for pin-heading and harvesting, respectively. In morphological properties by the mushroom, the heights of KNR2312 and KNR2322 were 122.7 and 121.0 mm, respectively. The thickness of KNR2322 and KNR2513 were 39.8 mm and 31.3 mm, respectively. The weight of KNR2524`s fruiting body was 36.3 g, which is good as wild strain. The quality of fruiting body of KNR2503 was 4.0 in comparison to the score 7 of commercially cultivated strains. KNR2512 had the darkest color of pileus with L value 43.6. The slow growing strains, KNR2511, KNR2513, and KNR2512 had the bright pileus with L value 80. In morphological characteristics, KNR2511, KNR2513, and KNR2515 had white lamellar and plane pileus. The three strains are supposed to be the same group and KNR2516 and KNR2518 appeared to be related to the group. The commercially cultivated strains had convex pileus, KNR2502, KNR2503, KNR2504, KNR2521, and KNR2525 had infundibuliform, and the other strains had plane pileus. Several strains were valuable for breeding, JNR2503 for growth rate, KNR2512 for pileus color, and KNR2312, KNR2322, KNR2503, and KNR2513 for the quality.

Construction of a genetic linkage map and analysis of quantitative trait loci associated with the agronomically important traits of Pleurotus eryngii

Breeding new strains with improved traits is a long-standing goal of mushroom breeders that can be expedited by marker-assisted selection (MAS). We constructed a genetic linkage map of Pleurotus eryngii based on segregation analysis of markers in postmeiotic monokaryons from KNR2312. In total, 256 loci comprising 226 simple sequence-repeat (SSR) markers, 2 mating-type factors, and 28 insertion/deletion (InDel) markers were mapped. The map consisted of 12 linkage groups (LGs) spanning 1047.8cM, with an average interval length of 4.09cM. Four independent populations (Pd3, Pd8, Pd14, and Pd15) derived from crossing between four monokaryons from KNR2532 as a tester strain and 98 monokaryons from KNR2312 were used to characterize quantitative trait loci (QTL) for nine traits such as yield, quality, cap color, and earliness. Using composite interval mapping (CIM), 71 QTLs explaining between 5.82% and 33.17% of the phenotypic variations were identified. Clusters of more than five QTLs for various traits were identified in three genomic regions, on LGs 1, 7 and 9. Regardless of the population, 6 of the 9 traits studied and 18 of the 71 QTLs found in this study were identified in the largest cluster, LG1, in the range from 65.4 to 110.4cM. The candidate genes for yield encoding transcription factor, signal transduction, mycelial growth and hydrolase are suggested by using manual and computational analysis of genome sequence corresponding to QTL region with the highest likelihood odds (LOD) for yield. The genetic map and the QTLs established in this study will help breeders and geneticists to develop selection markers for agronomically important characteristics of mushrooms and to identify the corresponding genes.

Soft Rot of Rhizopus oryzae as a Postharvest Pathogen of Banana Fruit in Korea

Soft rot on banana fruit caused by Rhizopus oryzae was identified for the first time in Korea. Colonies were white to light brown and formed numerous sporangiospores. Optimum temperature for mycelial growth was 30°C. Sporangia were globose and 30~200 μm. Sporangiophores were usually straight, 8~20 μm, and rhizoids usually in groups of 3~5. Columella were globose to sub-globose and 90~110 μm. Sporangiospores were sub-globose or oval and 4~10 μm. Based on its mycological characteristics, molecular analysis, and pathogenicity to host plants, this fungus was identified as Rhizopus oryzae Went & Prisen Geerligs. This is the first report of soft rot on banana caused by Rhizopus oryzae in Korea.

Breeding of King Oyster Mushroom, Pleurotus eryngii Carrying Good Traits of Cap

Two strains Pleurotus eryngii KNR2312 and A8B10 (Na) which have good traits in cap quality and speedy growing were selected to breed a new strain carrying the two traits. KNR2312-2636- (Ga) with 45.2 cap lightness and 6.5 quality was breeded by a consecutive inbreeding between KNR2312-derived monokaryons. came from outcrossing between Ga and Na was backcrossed with KNR2312-derived monokaryons twice to improve quality in cap color and shape. Therethrough Ga5NaKNR2312-47KNR2312- carrying lightness of 49.5, quality of 7.3 and weight of 95.1 g was selected. The selected strain possesses good quality and dark color of cap. Especially its edge is 24 mm thick, therefore it is not likely damaged during processing and distribution.

Composition analysis of raw material constituting the medium for mushroom cultivation

The contents of raw materials which are components of mixed substrate for mushroom cultivation were analyzed to optimize the composition. The pure protein(amino acid) level of soybean meal was the highest, 44.02% followed by those of soybean curd residue(31.5%) and cotton seeds meal(30.6%). The non protein nitrogen(NPN) contents in crude protein of main nitrogen materials were 2.4% for soybean meal and 5.6% for dried soybean curd residue, while those of wheat bran and rice bran used as the carbon source were relatively higher, 17.6% compared to that of nitrogen supplying media. Crude protein content per price was 6.0 for rapeseed meal, indicating that it is high crude protein content per price. Nitrogen-free extract(NFE) considering as an ingredient for mycelial growth were high in alphacorn(72.9%) and wheat bran B(57.2%). Acid detergent fiber(ADF) was high in corncob, 51.88%, its use for cultivation of brown rot fungi including Lentinus lepideus should pay attention because the fungi lack complete lignin degradation activity.

Diversity of A mating type in Lentinula edodes and mating type preference in the cultivated strains

Diversity of A mating type in Lentinula edodes has been assessed by analysis of A mating loci in 127 strains collected from East Asia. It was discovered that hypervariable sequence region with an approximate length of 1 kb in the A mating locus, spanning 5′ region of HD2-intergenic region-5′ region of HD1, could represent individual A mating type as evidenced by comprehensive mating analysis. The sequence analysis revealed 27 A mating type alleles from 96 cultivated strains and 48 alleles from 31 wild strains. Twelve of them commonly appeared, leaving 63 unique A mating type alleles. It was also revealed that only A few A mating type alleles such as A1, A4, A5, and A7 were prevalent in the cultivated strains, accounting for 62.5% of all A mating types. This implies preferred selection of certain A mating types in the process of strain development and suggests potential role of A mating genes in the expression of genes governing mushroom quality. Dominant expression of an A mating gene HD1 was observed from A1 mating locus, the most prevalent A allele, in A1-containing dikaryons. However, connections between HD1 expression and A1 preference in the cultivated strains remain to be verified. The A mating type was highly diverse in the wild strains. Thirty-six unique A alleles were discovered from relatively small and confined area of mountainous region in Korean peninsula. The number will further increase because no A allele has been recurrently observed in the wild strains and thus newly discovered strain will have good chances to contain new A allele. The high diversity in small area also suggests that the A mating locus has evolved rapidly and thus its diversity will further increase.

Optimal relative humidity for Pleurotus eryngii cultivation

The effects of fixed and variable relative humidity on fruiting body formation and characters of Pleurotus eryngii were investigated with normal and thinning treatment plots. In fixed relative humidity, as humidity was lower, period of harvest was longer as well as days for pinheading in the both of normal and thinning plots. In the normal plot, qualities of mushroom were 5.5, 5.8, and 6.3 and yield was 98.6 g per bottle for 90% relative humidity, it was best. In the thinning plot, qualities of mushroom were 7.7, 8.4, and 8.5 and yields were 102.1, 105.8, and 116.9 g at 70, 80, and 90% respectively. In variable relative humidity with a thinning plot, the yield of P. eryngii on condition I(>90% for 1 day → 85% until thinning(for about 11 days) → 80%) and III(>90% until pin-heading(about for a week) → 85% until thinning(about for 5 days) → 80%) were 85.5 and 87.8 g per bottle, and qualities were 7.8 and 8.0 respec- tively. For long shelf life and a cultural control of bacterial soft rot disease, the condition I will be more largely adopted by mushroom farmers.

Breeding of Pleurotus eryngii, "Aeryni 5"

To breed a new Korean Pleurotus eryngii cultivar with high quality and yield, single crosses between 24×46 and KNR2539 were performed, and a new cultivar, 6×13, was selected based on the days to harvest (14.6 days), quality (7.2), and yield (85.9 g/850 cc bottle). The strain was named Aeryni 5 and cultivated on a large scale at the mushroom farms to compare with Keuneutari 2ho. The yield of Aeryni 5 (82.2 g) was 122.7% of Keuneutari 2ho, and the quality of the new cultivar was 7.7 while reference cultivar was 6.3. The yield and quality of the two cultivars were statistically different. The lightness of the pileus of Aeryni 5 (61.7) was greater than that of Keuneutari 2ho by 3.4 points; thus, the pileus of Aeryni 5 looked brighter. PCR with URP2 was used to discriminate between Aeryni 5 and Kenneutari 2ho.

Development of the Optimal Media for Mycelial Culture of Pleurotus eryngii using the Hot-water Extract of Raw Materials

Hot-water extracted natural media were made from raw materials for mycelial culture of Pleurotus eryngii. Poplar sawdust, wheat bran and rice bran were used as substrates for hot water extraction. The mixed substrates of poplar sawdust, wheat bran, and rice bran with 50 : 20 : 30 (v/v/v, PWR523) and 50 : 30 : 20 (v/v/v, PWR532) were optimal for mycelial growth of P. eryngii, respectively. The hot-water extracted natural media from PWR523 and PWR532 showed a rapid mycelial growth and spawn running compared to PDA. There was no significant difference in mushroom yield when the mycelium grown on the hot-water extracted natural media was used as the inoculum source for producing fruit body.