The aim of this study was to analyze the genetic diversity of collected strains of Hypsizygus marmoreus based on their rDNA ITS sequences. The size of ITS rDNA regions of H. marmoreus strains, collected form various regions. A phylogenetic trees based on the ITS region revealed that the strains could be classified into 4 different groups including Villosiclava virens, Hypsizygus marmoreus, Lepista irina, Lyophyllum Decastes, Lyophyllum shimeji, Pleurotus floridanus.
Single ascospores were isolated and tested their characteristics such as mycelial growth, mycelial density, fruiting body formation ability, the production of perithecia in the breeding of new Cordyceps militaris mushroom. Among them selected isolates were crossed and hybrids were produced showing high quality fruiting bodies in artificial media. New strain 'Dowonhongcho’ was better on SDAY and at 10~25°C when it was compared with that of 'Yedang 3' in mycelial growth. The stromata of new strain were club-shaped and bright orange-red color. Its length was 6.1 cm and the cordycepin content was 0.34% on average. In comparison with 'Yedang 3', the new strain had a yield that was 9% higher and it produced fruiting bodies which were firmer. The optimum temperature for mycelial growth was 22~25°C and the optimum temperature for stroma development was 18~22°C. Days of fruiting body were similar with 45 days from inoculation. This cultivar may serve as a valuable one for artificial cultivation and industrial-scale production of C. militaris.
A new commercial cultivar ‘Hae Sal’ of Hypsizigus marmoreus was developed by the method of mono-mono crossing between monokaryotic strains derived from KNR8047 and KNR8080. The parental strains are characterized by the property of high quality and a short period of time in cultivation respectively. The optimum temperature of mycelial growth was 25°C and that of fruiting body development was 15~16°C. The period of harvesting including primordia formation was 7.0 days shorter than that of control strain ‘Mangadak No.2’. The color of pileus and stipe surface was neutral-brown and pure white, respectively. The yield was 117.6±23.2g/850cc plastic bottle. Analysis of the genetic characteristics of the new commercial cultivar ‘Hae Sal’ showed a different profile as that of the control strain, ‘Mangadak No.2’, when RAPD (Random Amplified Polymorphic DNA) primer was used. This new cultivar ‘Hae Sal’ of Hypsizigus marmoreus is characterized by the improved quality and a short period of time in cultivation after scratching. It would be of much help to expand the cultivation area of Hypsizigus marmoreus.
Differentiation of Pleurotus eryngii is laborious and time-consuming tasks especially in mycelial status. For development of a method for differentiation of P. eryngii cultivars, simple sequence repeats (SSR) from whole genomic DNA sequence analysis was used for genotyping and two multiplex-SSR primer sets were developed. These SSR primer sets were employed to distinguish 12 cultivars and strains. Five polymorphic markers were selected based on the genotypes. PCR with the each primer produced one to four distinct bands ranging in size from 200 to 300 bp. Polymorphism information content (PIC) values of the five markers were in range of 0.6627 to 0.6848 with an average of 0.6775. Unweighted pair-group method with arithmetic mean clustering analysis based on genetic distances using five SSR markers classified 12 cultivars into 2 clusters. Cluster I and II comprised of 4 and 8 cultivars, respectively. Two multiplex sets, Multi-1 (SSR312 and SSR366) and Multi-2 (SSR178 and SSR277) completely discriminated 12 cultivar and strains with 21 allele with a PIC value of 0.9090. These results might be useful to provide an efficient method for the identification of P. eryngii cultivars with separate PCR reactions. (This work was supported by a grant from the Gold Seed Project [Supported by a grant from the IPET (213003-04-3-WTI11), MIFAFF, Republic of Korea.]
The aim of this study was to identify and characterize new Flammulina velutipes laccases from its whole-genome sequence. Of the 15 putative laccase genes detected in the F. velutipes genome, four new laccase genes (fvLac-1, fvLac-2, fvLac3, and fvLac-4) were found to contain four complete copper-binding regions (ten histidine residues and one cysteine residue) and four cysteine residues involved in forming disulfide bridges. fvLac-1, fvLac-2, fvLac3, and fvLac-4, encoding proteins consisting of 516, 518, 515, and 533 amino acid residues, respectively. Potential N-glycosylationsites(Asn-Xaa-Ser/Thr) were identified in the cDNA sequence of fvLac-1(Asn-454), fvLac-2(Asn- 437andAsn-455), fvLac-3(Asn-111andAsn-237), and fvLac4 (Asn-402andAsn-457). In addition, the first 19–20 amino acid residues of these proteins were predicted to comprise signal peptides. Laccase activity assays and reverse transcription polymerase chain reaction(RT-PCR) analyses clearly reveal that CuSO4 affects the induction and the transcription level of these laccase genes.
Medical mushroom, Phellinus linteus and Phellinus baumii called as “Sanghwang” have cultivated in Korea. PL has been studied extensively for its extraordinary capacity of suppressing cancer or enhancing body immunity. The mycelial materials of PL have mainly been used as research samples worldwide because fruiting bodies was difficult to be artificially cultivated. Alternatively, P. baumii (variety, ‘Jangsu’) have been cultivated in Korea. However, fruiting body morphology of P. baumii is clearly different to that of PL. Generally, Phellinus spp. including P. linteus slowly grow on artificial medium such as Potato Dextrose Agar (PDA). In contrast, P. baumii strains were rapidly grown on the artificial media when compared to other Phellinus spp. and thus it was considerable that its mycelial growing ability can be acted as a factor for producing fruiting bodies. This study aimed to find Phellinus isolates having high mycelial growth rate. Five Phellinus isolates that show rapid growth rate on YGM medium were selected from 36 Phellinus isolates collected in Korea. They were identified on nucleotide sequences of rDNA-ITS region. Phellinus linteus strain and Phellinus spp. showing mycelial growth rate comparing to P. baumii were characterized on cultural and bioactive characteristics (antioxidant activity and immune activation).
Oyster mushroom is one of mushrooms that are cultivated and consumed a lot in Korea. P. ostreatus 'Suhan(ASI2504)' is a popluar cultivar because of high quality. But it is difficult for farmers to cultivate, so an alternative cultivar of ‘Suhan’ is demanded continuously. To develop a new variety, parental strains were selected using cultivation characteristics of Pleurotus ostreatus collected home and abroad. P. ostreatus 'Gosol' was developed by the method of Di-Mon crossing between dikaryotic strain P. ostreatus 'Mongdol(ASI0633)' and monokaryotic strain derived from P. ostreatus ‘Yasan(ASI0635)' in 2014. Analysis of the genetic characteristics of the new cultivar 'Gosol' showed a different DNA profile as that of the control strain, 'Suhan’ but a similar DNA profile as those of the parental strains, ‘Yasan’ and 'Mongdol’, when RAPD(Random Amplified Polymorphic DNA) primer URP 3 was used. The optimum temperature for mycelial growth was 25°C for ‘Gosol’ and ‘Suhan’. 'Gosol' was appropriate for middle high temperature to grow, especially 13~18°C. Fruiting body production per bottle was about 124.2g. When compared to the control strain 'Suhan’, the stipe was longer and the individual weight was heavier. But the stipe and the pileus were a little thinner than those of the control strain. ‘Gosol’ was more resistant at high CO2 concentration than the control strain. This new cultivar ‘Gosol’ of Pleurotus ostreatus was characterized dark bluish gray cultivar of oyster mushroom in the color of pileus and higer yield compared to those of other cultivar ‘Suhan’. Therefore, it is suggested that this new cultivar ‘Gosol’ be substitute for ‘Suhan’ in oyster mushroom’s farms.
Pleurotus eryngii, an edible white-rot fungus, is widespread in Eurasia and northern Africa. It has become a major cultivated mushroom in Asia, with a current global production rate of approximately 3 × 10 5 metrictons/yr. To improve the quality or productivity through breeding, a genetic linkage map is an important component. In this study, genetic linkage map of the P. eryngii was constructed using 98 monokaryotic progeny derived from dikaryon of parental KNR2312 strain derived from haploid meiotic spores. The whole genome sequence of P5 monokaryon from P. eryngii KNR2312 strain by Next Generation Sequencing (NGS) strategy was used to design the SSR markers. 484 primers pairs were identified by SSR Locator I and tested polymorphism via PCR. A total of 241 loci were mapped using Joinmap 4.0, comprising 222 SSR markers, 2 mating type factors, and the 13 INDEL markers. The map consisted of 14 linkage groups spanning 1003 cM at an average marker interval of 4.2 cM. The mating loci, A and B were mapped on linkage groups 4 and 11, respectively. The established linkage map and the genetic information based on NGS could be used for QTL mapping of agronomic traits, marker-assisted breeding that may ultimately lead to outstanding phenotypic characteristics. [Supported by a grant from the IPET (213003-04-3-SBY20), MIFAFF, Republic of Korea.]