• Journal of Plant Biotechnology
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• v.34 no.3
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• pp.201-205
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• 2007

To determine whether pattern recognition based on metabolite fingerprinting for whole cell extracts of higher plants is applied to discriminate plants genetically, leaf samples of eight cultivars of Catharanthus roseus were subjected to Fourier transform infrared spectroscopy (FT-IR). FT-IR fingerprint region data were analyzed by principal component analysis (PCA). Major peaks as biomarkers were identified as the most significant contributors to distinguish samples by using genetic programming. A hierarchical dendrogram based on the results from PCA separated the eight cultivars into two major groups in the same manner as the dendrograms based on genetic fingerprinting methods such as RAPD and AFLP. A slight difference between the dendrograms was found only in branching pattern within each subgroup. Therefore, we conclude that the hierarchical dendrogram based on PCA of the FT-IR data represents the most probable chemotaxonomical relationship between cultivars, which is in general agreement with the genetic relationship determined by conventional DNA fingerprinting methods.

• Journal of Microbiology and Biotechnology
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• v.7 no.3
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• pp.161-166
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• 1997

Nuclei were isolated from the protoplasts of Trichoderma harzianum T95 and treated with colchicine, a polyploid inducer. The nuclei were transferred into the protoplast of multi-auxotrophic Gliocladium virens G88 which cannot grow in minimal medium. The protoplast of G. virens G88 carrying the transferred nuclei were regenerated in a regeneration minimal medium containing $17{\mu}g/ml$ of chloroneb as a haploid inducer. Six intergeneric hybrids between G. virens and T. harzianum were isolated from the regeneration minimal medium. The hybrids could be classified into three types according to morphology, those with an isozyme pattern, those with an protein band and those with an randomly amplified polymorphic DNA(RAPD) pattern produced by random primers and repetitive sequences. The first group was identified to be a haploid recombinant, the second group a heterokaryon, and the third appeared to be petite.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.11
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• pp.1535-1541
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• 2005

Five broiler chicken lines, namely HC, BPB2, CPB2, PB2 and UM1, involving in a selection program and differing in selection intensity and genetic background, were screened for randomly amplified polymorphic DNA (RAPD) polymorphism using 10 selected decamer primers. Nine primers amplified the genomic DNA, generating 200 to 2,500 bp and all detected polymorphism between lines. Out of 74 bands scored using these primers, 34 (50.0%) were found to be polymorphic. The number of polymorphic loci ranged from 3 to 6 with an average of 4.33. Lines differed considerably for within-population genetic similarity estimated by band frequency (WS = 93.55 to 99.25). Between-line genetic similarity estimates based on band sharing as well as on band frequency ranged from 71.35 to 86.45 and from 73.38 to 87.68, respectively. Lines HC and PB2 were the most closely related to the other, while BPB2 and CPB2 appeared to be more distant from each other. The between-line genetic distance based on both band sharing and band frequency revealed the similar trends as for Between-line genetic similarity. Based on BS and BF criteria, BPB2 and CPB2 as well as PB2 and UM1 lines can be merged to launch a new genetic group for further progress in biometrical objectives. A phylogenetic tree, derived using Nei's coefficient of similarity revealed the different pattern of genetic distance between lines.

• 한국균학회소식:학술대회논문집
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• 2014.05a
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• pp.21-21
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• 2014

A edible mushroom, Hypsizygus marmoreus is commercially cultivated in Northeast Asia. Japan's annual production is 110,000ton or more. Since 2002, cultivation is expanded in Korea. To investigate the morphological, cultural and microscopic characteristics of Hypsizygus marmoreus, 109 isolates were collected from Korea and other countries. Clamp connection, chlamydospore and arthrospore were present in all tested isolates of H. marmoreus except HYM-002 and HYM-004. Also pilealtrama, gilltrama, basidia, basidiospore and cystidia of fruiting body were no difference among the isolates in the present investigation. Morphological characteristics of fruiting body was that color of pileus was brown and white, irregular as marble, the average size 12~22mm and stipes was $46{\sim}91{\times}6{\sim}10mm$. Isolates HYM-031, HYM-047 and HYM-109 formed grayish-brown pileus with a faint pattern. Molecular analysis with RAPD and ITS rDNA sequence analysis were also performed to check the genetic relationships among H. marmoreus isolates. Based on the RAPD analysis using the URP-PCR, all isolates of H. marmoreus were clustered into large 3 groups but more than 90% showed high similarity. In addition, morphological and geographical differences have been classified as an independent cluster. The brown and white strains enclosed in same cluster. So genetically no significance difference was observed between these two strains. ITS gene sequences of 16 selected isolates which were 640 bp long, were aligned and compared. The similarity in ITS sequence was 94.8 to 99.1% among tested isolates and the H. marmoreus isolates in GeneBank. In conclusion the tested isolates were H. marmoreus. Morphological and molecular observations proved that all tested isolates were belonging to H. marmoreus. For the stable artificial cultivation, composition of optimum media, mature period and light condition were established. Optimal formula of artificial cultivation medium was Douglas sawdust: corn cob: soybean meal: wheat bran = 40:30:15:15. In addition, 7% rice bran and 3% yellow sucrose was the most effective composition for spawn's liquid medium. For the maturation of the isolates was favorable for growing for 20 to 30 days at $25^{\circ}C$ and the LED lights in mixture of white and blue was good for growth period. For effective growth, the temperature, humidity and aeration control in every step was important.

• The Korean Journal of Mycology
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• v.29 no.1
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• pp.9-14
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• 2001

Colletotrichum species are important fungal pathogens that cause great damages on various host plant species worldwide. In Korea, Colletotrichum species cause massive economic losses on apple, peach, grape, and especially, sweet persimon productions. In the past, identification of the pathogen and the studies on the genetic relationships among the pathogenic isolates were mainly based on morphology, cultural characteristics, and the difference in pathogenicity. However, in recent years, these traditional methods have been replaced with molecular methods including AFLP. AFLP method with the merits of both RAPD and RFLP has been widely used for the genetic relationship studies of various organisms. Therefore, in this study, AFLP method was employed for the studies of genetic relationships among the different isolates of Colletotrichum species collected from various parts of sothern Korea. As a result, two specific band pattern groups were observed among different isolates of Colletotrichum species.

• Journal of Mushroom
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• v.13 no.2
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• pp.135-138
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• 2015

'Daejang 3ho' is a new cultivar of oyster mushroom for high productivity than that of Chunchu 2ho. This cultivar was bred by Di-mon mating between monokaryotic strains isolated from 'Daejang 1ho' and dikaryotic strain 'Jangan 5ho'. The color of pileus and the shape of pileus of new cultivar were dark gray and deeply funnel shape, respectively. The length and thickness of stipe were longer and thicker than those of Chunchu 2ho. It took 4~5 days to formation of primordia, that was a similar to Chunchu 2ho. The optical temperature of fruit body was $14-17^{\circ}C$. Yield of 'Daejang 3ho' was 13.9% higher than that of Chunchu 2ho. Analysis of RAPD by URP-primer of #03, #08, #10 and #11 showed that different band pattern those of other strains. There were no significant differences between the texture properties of 'Daejang 3ho' and Chunchu 2ho.

• ALGAE
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• v.21 no.2
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• pp.261-266
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• 2006

The innate soluble polysaccharides and phenolic compounds of marine macroalgae are serious contaminants which interfere with experimental procedures such as restriction enzyme digestion, polymerase chain reaction (PCR) and other enzymatic reactions using extracted DNA samples. The viscous polysaccharides are co-precipitated with DNA samples by isopropanol or ethanol precipitation in conventional experiment. To overcome the problem, a method for the isolation of high molecular weight DNA from Porphyra tenera is developed with the application of diatomaceous earth column. The isolated DNAs by this method were about 50-100 kb in size and could be digested well with restriction enzymes. The nuclease activity seemed to be minimal, and high reproducibility in the arbitrary primed PCR for RAPD analyses was a distinctive feature. These results suggest that this method is very efficient in isolating nucleic acid from macroalgae including Porphyra.

• Journal of Mushroom
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• v.11 no.1
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• pp.1-8
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• 2013

In this study, twelve of Lepista nuda were collected from various localities in Korea. Also thirteen exotic L. nuda species were collected from Japan, France, Switzerland and Portugal. Spores were isolated under optical microscope. These spores were placed on the surface of YM medium for inducing to germination. Eleven mating-groups were selected by morphological characters of fruit body such as size, color and stipe patterns. Intra-isolate crosses were made between two single-spore isolates derived from mating-groups. Also, dikaryotic crossing using the isolates from L. nuda were carried out to evaluated tetrakaryon formation. Cross-mating compatibility tests also verified its dikaryotic state by microscopic or molecular genetic observation of clamp connection and Random Amplified Polymorphic DNA (RAPD) band pattern. To analyze the growth rate of hybrids and parents mycelium in dikaryons obtained from compatible mating groups were placed on PDA medium. Intra-isolate crosses determined eleven mating-groups within L. nuda. The typical clamp connection were mostly observed in mating-groups of Korean L. nuda in $K1{\times}K2$, $K1{\times}K3$, $K1{\times}K4$, $K1{\times}K6$, $K1{\times}K5$, $K2{\times}K4$, $K2{\times}K3$, $K2{\times}K6$, $K3{\times}K4$, $K4{\times}K5$, and $K4{\times}K6$. Korean L. nuda type of dikaryon, shown to cross-incompatibility with L. sordida, it seemed that mating induce more rapidly than wild types in a view of growth rate. In conclusion, it would be useful to improve mass production with better morphological characteristics through a special mating of L. nuda.

• Horticultural Science & Technology
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• v.19 no.1
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• pp.29-33
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• 2001

This study was carried out to discriminate potato cultivars and breeding lines by specific molecular markers using random amplified polymorphic DNA (RAPD) analysis. The genotypes of potatoes used for analysis were eight cultivars and five breeding lines. Some of those show much phenotypic resemblances among them because 'Jopung', 'Daekwan70', 'Gawon', and 'Daekwan72' have immediate parental relationship with 'Superior', 'Irish Cobbler', 'Namsuh', and 'Atlantic', respectively. So, there are many difficulties to distinguish the varieties by the morphological characteristics. Three URP primers, URP2, URP4, and URP8 were selected for promising primers to discriminate potato genotypes or cultivars. The three URP primers were shown very high reproducibility because of the relatively high annealing temperature and long primer size. Although the results of similarity analyses did not always reflect the genetic relationship between potato varieties, the reproducible pattern of amplified DNA bands by URP primers showed possibility for molecular markers for discrimination of potato genotype or cultivar.

• Journal of Mushroom
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• v.2 no.3
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• pp.115-126
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• 2004

Somatic hybrids of inter-compatible and inter-incompatible strains were obtained by protoplast fusion. The fusion products between compatible strains, Pleurotus ostreatus and P. florida, formed heterokaryons, while fusants between incompatible strains such as P. cornucopiae + P. florida, P. ostreatus + Ganoderma applanatum, P. florida + Ganoderma lucidum, and P. ostreatus + Flammulina velutipes formed synkaryons that retained genes from both parents. The heterokaryons showed the same level of basidioma development. In contrast, the synkaryons showed unique characteristics including clamp connection formation at mitosis, either partner basidioma development, and abnormal segregation and recombination compared with inter-compatible strains. Synkaryons can be classified into homokaryoyic and heterokaryotic type. A comparison of somatic hybrids with compatible and incompatible strains was made using random amplified polymorphic DNA (RAPD) analysis. The heterokaryons between compatible species showed the same level of variability and contained both parental RAPD bands. In contrast, most of the synkaryons between incompatible species showed similarity to those of either parental bands and non-parental RAPD bands. Synkaryons can be classified into microgenome insertion type and macrogenome insertion type. A tetrapolar mating system was found among monospore isolates in somatic hybrids and wild type P. ostreatus. Homokaryons from each somatic hybrid combination were paired with tester homokaryons of the initial wild type of P. ostreatus. The changed mating types were identified in progenies. The pattern of mating type switching in somatic hybrids depends on compatibility of fusion partner. There are several factors related to the mechanism of clamp connection formation and fruiting body development of synkaryons. Of these,the major factor may be associated with self-fertility and mating type switching such as homokaryotic fruiting of wild type P. ostreatus. This review will discuss these aspects.