• 한국균학회소식:학술대회논문집
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• 2015.05a
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• pp.53-53
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• 2015

Agrobacterium tumefaciens-mediated transformation(ATMT) of Flammulina velutipes was used to produce a diverse number of transformants to discover the functions of gene that is vital for its variation color, spore pattern and cellulolytic activity. Futhermore, the transformant pool will be used as a good genetic resource for studying gene functions. Agrobacterium-mediated transformation was conducted in order to generate intentional mutants of F. velutipes strain KACC42777. Then Agrobacterium tumefaciens AGL-1 harboring pBGgHg was transformed into F. velutipes. This method is use to determine the functional gene of F. velutipes. Inverse PCR was used to insert T-DNA into the tagged chromosomal DNA segments and conducting sequence analysis of the F. velutipes. But this experiment had trouble in diverse morphological mutants because of dikaryotic nature of mushroom. It needed to make monokaryotic fruiting varients which introduced genes of compatible mating types. In this study, next generation sequencing data was generated from 28 strains of Flammulina velutipes with different phenotypes using Illumina Hiseq platform. Filtered short reads were initially aligned to the reference genome (KACC42780) to construct a SNP matrix. And then we built a phylogenetic tree based on the validated SNPs. The inferred tree represented that white- and brown- fruitbody forming strains were generally separated although three brown strains, 4103, 4028, and 4195, were grouped with white ones. This topological relationship was consistently reappeared even when we used randomly selected SNPs. Group I containing 4062, 4148, and 4195 strains and group II containing 4188, 4190, and 4194 strains formed early-divergent lineages with robust nodal supports, suggesting that they are independent groups from the members in main clades. To elucidate the distinction between white-fruitbody forming strains isolated from Korea and Japan, phylogenetic analysis was performed using their SNP data with group I members as outgroup. However, no significant genetic variation was noticed in this study. A total of 28 strains of Flammulina velutipes were analyzed to identify the genomic regions responsible for producing white-fruiting body. NGS data was yielded by using Illumina Hiseq platform. Short reads were filtered by quality score and read length were mapped on the reference genome (KACC42780). Between the white- and brown fruitbody forming strains. There is a high possibility that SNPs can be detected among the white strains as homozygous because white phenotype is recessive in F. velutipes. Thus, we constructed SNP matrix within 8 white strains. SNPs discovered between mono3 and mono19, the parental monokaryotic strains of 4210 strain (white), were excluded from the candidate. If the genotypes of SNPs detected between white and brown strains were identical with those in mono3 and mono19 strains, they were included in candidate as a priority. As a result, if more than 5 candidates SNPs were localized in single gene, we regarded as they are possibly related to the white color. In F. velutipes genome, chr01, chr04, chr07,chr11 regions were identified to be associated with white fruitbody forming. White and Brown Fruitbody strains can be used as an identification marker for F. veluipes. We can develop some molecular markers to identify colored strains and discriminate national white varieties against Japanese ones.

• Journal of Nutrition and Health
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• v.18 no.4
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• pp.259-271
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• 1985

The purpose of this study was to develop supplementary foods for infants and young children in order to improve their nutritional status. Three formulas composed of rice, soybeans, fish, dry skim milk and sesame in varying proportions were studied. The three formulas, $RS_{1}S_{2}$, $RFS_{1}S_{2}$, and $RMS_{1}S_{2}$, were consisted of Rice(R), Soybean$(S_{1})$, Sesame$(S_{2})$ (60 : 35 : 5) , Rice, Fish(F), Soybean, Sesame (60 : 10 : 25 : 5) , and Rice, Dry Skin Milk (M), Soybean, Sesame (60 : 10 : 25 : 5), respectively. A proximate analysis and amino acid determination were made on the developed formulas. In the animal assay, growth rate, PER and FER were evaluated and biochemical analyses were also carried out. A storage test and the cost evaluation were also conducted. The summarized results are as follows : 1) The proximate composition of the three formulas were 7.3-7.4% of moisture, 15.9-21.5% of crude protein, 7.8-9.6% of crude fat and 2.5-2.8% ash. 2) The result of amino acid analysis showed that the 1st limiting amino acids of $RS_{1}S_{2}$ and $RFS_{1}S_{2}$ were lysine (amino acid score, 76.6) and threonine (amino acid score, 93.3), and that of $RMS_{1}S_{2}$ and the commercially prepared formula were sulfur containing amino acids (amino acid score, 82.0 and 54.4). When the contents of the amino acids of the three formulas were compared with mother's milk and cow's milk, the balance of the amino acid of each formula was superior to mother's milk but inferior to cow's milk. 3) In the animal assay, the growth rate of all groups increased gradually during the experimental period. 4) The C- PER, which was corrected on the basis of the casein PER of 2.5 was 2.99, 3.38 and 3.10 in the $RS_{1}S_{2}$, $RFS_{1}S_{2}$ and $RMS_{1}S_{2}$ respectively. The C- PER of $RFS_{1}S_{2}$ and $RMS_{1}S_{2}$ were Significantly (P<.05) higher than that of the casein. 5) The FER of the casein, $RS_{1}S_{2}$, $RFS_{1}S_{2}$, and $RMS_{1}S_{2}$ were 0.37, 0.39, 0.43 and 0.39, respectively. The FER of $RFS_{1}S_{2}$ and $RMS_{1}S_{2}$ were also significantly (P<.05) higher than that of the casein. 6) The concentrations of hematocrit, hemoglobin, total protein and albumin in the serum of the rats of all groups were not significantly different among groups. 7) The storage stability test showed that the total plate count (TPC), the coliforms count and the bacterial spore count in the ingredients were quiet low. However, after 30 and 60 days storage, the count in $RFS_{1}S_{2}$ increased and were higher at room temperature than refrigerated temperature. 8) In the cost evaluation, the cost of the developed formulas was \1,826-2,626 / kg. This was less than that of the commercially prepared formula (\3,300-4,073 / kg) and that of the imported formula (\4,250-8,720 / kg).

• Korean Journal of Microbiology
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• v.50 no.3
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• pp.201-209
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• 2014

This study was carried out in order to develop a biological control of anthracnose of red pepper caused by fungal pathogens. In particular, this study focuses on the Colletotrichum species, which includes important fungal pathogens causing a great deal of damage to red pepper. Antagonistic bacteria were isolated from the soil of pepper fields, which were then tested for biocontrol activity against the Colletotrichum gloeosporioides anthracnose pathogen of pepper. Based on the 16S rRNA sequence analysis, the isolated bacterial strain CS-52 was identical to Bacillus sp. The culture broth of Bacillus sp. CS-52 had antifungal activity toward the hyphae and spores of C. gloeosporioides. Moreover, the substances with antifungal activity were optimized when Bacillus sp. CS-52 was grown aerobically in a medium composed of 0.5% glucose, 0.7% $K_2HPO_4$, 0.2% $KH_2PO_4$, 0.3% $NH_4NO_3$, 0.01% $MnSO_4{\cdot}7H_2O$, and 0.15% yeast extract at $30^{\circ}C$. The inhibition of spore formation resulting from cellulase, siderophores, and indole-3-acetic acid (IAA), were produced at 24 h, 48 h, and 72 h, respectively. Bacillus sp. CS-52 also exhibited its potent fungicidal activity against anthracnose in an in vivo test, at a level of 70% when compared to chemical fungicides. These results identified substances with antifungal activity produced by Bacillus sp. CS-52 for the biological control of major plant pathogens in red pepper. Further studies will investigate the synergistic effect promoting better growth and antifungal activity by the formulation of substances with antifungal activity.

• Journal of Food Hygiene and Safety
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• v.26 no.3
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• pp.198-202
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• 2011

Rice has been the most important staple food in everyday meals of Korean people for thousands of years. Nowadays, it is getting increasingly used as flour ingredients in a variety of processed foods, so that it is consumed in more diversified ways. As a consequence, production volume of rice flour to manufacture rice cakes, noodles, breads, or confectioneries is recently getting increased in Korea. But there are not sufficient research outcomes to guarantee Korean consumers microbiological qualities of rice flour as well as rice. As a preliminary experiment, therefore, the microbiological profiles (aerobic mesophilic bacteria (AMB), spore-forming aerobic bacteria (SAB), lactic acid bacteria (LAB), yeasts and molds (YM), and Escherichia coli and coliforms) have been monitored for nine retailed white rice samples in this study. AMB counts ranged $10^2-10^6$ CFU/g for all the nine white rice samples. All the nine rice samples have SAB counts within a narrow range $(1.0{\times}10^2-2.5{\times}10^3$ CFU/g). LAB was detected in two white rice samples ($4.0{\times}10^2$ and $3.7{\times}10^3$ CFU/g), YM was detected in one white rice sample ($2.0{\times}10^2$ CFU/g) only. E. coli was not detected from all the nine samples. Coliforms were detected in one white rice sample ($4.1{\times}10$ CFU/g) only. All the rice samples were conclusively considered to have various microorganisms, though most of them are harmless and some, such as coliforms, may be harmful.

• Korean Journal of Food Science and Technology
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• v.35 no.6
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• pp.1079-1085
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• 2003

Modified atmosphere packaging was applied to oyster mushrooms (Pleurotus ostreatus) to study the effect of storage temperatures and packaging materialso. Whole mushrooms (200g) were package with polyethylene film $(PE,\;60{\mu}m\;thickness)$, ethylene vinyl acetate (EVA), or ceramic film (containing 5% zeolite) and stored at 0, 5, 10 and $20^{\circ}C$. Weight loss, color, firmness, gas composition $(O_2,\;CO_2)$ inside the film package and ethanol content in the tissue of MA packaged mushrooms were examined. Mushroom that were packed unwrapped in a conventional hardboard box (2 kg) lost marketability at a very early stage of storage due to weight loss, shrinkage, browning, and spore formation. During storage, film packaging prevented or retarded the deterioration of the mushrooms in the aspects of appearance, texture, and discoloration. Firmness slightly decreased with storage time. Total color difference was much higher in the control than in the film-packaged mushroom and rapidly increased at the early of storage. Correlation analysis showed a high correlation between total color difference and b values. These results were characterized by the reduced respiration rate resulting from elevated carbon dioxide and reduced oxygen levels in the package. At all storage temperatures, ethanol content in the tissue increased slightly at the early part of storage and rose considerably towards the end of the storage period. Ethanol content in the oyster mushrooms was higher in the stipe than in pileus tissues. The shelf life of the oyster mushrooms was about $8{\sim}11$ days at $0^{\circ}C$, about $4{\sim}6$ day at $5^{\circ}C$, about $2{\sim}3$ days at $10^{\circ}C$, and about $1{\sim}2$ days at $20^{\circ}C$.

• Korean Journal of Medicinal Crop Science
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• v.9 no.2
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• pp.156-165
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• 2001

Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.

• Journal of Food Hygiene and Safety
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• v.30 no.3
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• pp.281-288
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• 2015

Vibrio is a genus of Gram-negative, curved, halophilic, and non-spore-forming bacteria. Some of the Vibrio species, such as V. cholerae and V. parahaemolyticus, often contaminate seafood products and occasionally cause human diseases when the seafood products are ingested. A total of 24 Vibrio strains were isolated from shrimp samples on Thiosulphate citrate bile salt sucrose (TCBS) media in this study. All of the 24 isolates were confirmed to belong to the genus Vibrio by using 16S rRNA gene sequence analyses. Vitek 2 system and species-specific polymerase chain reaction (PCR) methods were used to further identify a total of 29 Vibrio strains at the species level, including the 24 shrimp Vibrio isolates and five Vibrio reference strains. The specificities of the two methods to identify Vibrio strains at the species level were compared in this study. The species-specific PCR method was designed to detect five different Vibrio species, such as Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio alginolyticus, and Vibrio mimicus. From the 24 Vibrio shrimp isolates, the Vitek 2 system method could identify 15 (62.5%) strains as Vibrio species and 7 (29.2%) strains as non-Vibrio species, but could not identify the rest 2 (8.3%) strains. But species-specific PCR method could identify 16 (66.7%) strains as Vibrio species and could not identify the rest 8 (33.3%) strains. Among the 24 Vibrio shrimp strains, these two methods could unanimously identify 7 (7/24, 29.2%) strains (2 V. parahaemolyticus, 4 V. alginolyticus, and 1 V. mimicus). Considering that such different identification results were obtained using the two different methods in this study, identification method for Vibrio species must be carefully chosen.

• Food Science and Preservation
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• v.30 no.5
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• pp.822-832
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• 2023

This study investigates the effectiveness of CA (controlled atmosphere) containers in maintaining the freshness of exported melons. The melons were harvested on June 5, 2023, in the Yeongam area of Jeollanam-do, Korea. The CA container was loaded with melon samples packed in an export box. The temperature inside the container was set at 4℃, while the gas composition was set at 5% oxygen, 12% carbon dioxide, and 83% nintrogen. Following two weeks of simulated transportation, quality analysis was conducted at 10℃. The melons were inoculated with spore suspensions, and the decay rate was determined to investigate the effect of the gas composition inside the CA container on suppressing the occurrence of Penicillium oxalicum in melons. The results were compared with a Reefer container set at the same temperature. The samples transported in the CA container exhibited lower weight loss. The melon pulp softening, respiration rate, and ethylene production were slower using the CA container. Moreover, the decay rate during the distribution period in the CA container was lower than in the Reefer container. In contrast, the firmness of melons transported in the Reefer container decreased significantly (from 9.03N to 5.18N) immediately after transportation. The soluble solid content (SSC) of melons transported in the Reefer container also decreased rapidly. The results suggested that the CA container is the optimal export container for maintaining the freshness of melons.

• Korean Journal of Heritage: History & Science
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• v.37
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• pp.285-307
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• 2004

Synthesize and examine petrological characteristic and geochemical characteristic by weathering formation of rock and progress of weathering laying stress on stone cultural properties of Unjusa temple of Chonnam Hwasun county site in this research. Examine closely weathering element that influence mechanical, chemical, mineralogical and physical weathering of rocks that accomplish stone cultural properties and these do quantification, wish to utilize by a basic knowledge for conservation scientific research of stone cultural properties by these result. Enforced component analysis of rock and mineralogical survey about 18 samples (pyroclastic tuff; 7, ash tuff; 4, granite ; 4, granitic gneiss; 3) all to search petrological characteristic and geochemical characteristic by weathering of Unjusa temple precinct stone cultural properties and recorded deterioration degree about each stone cultural properties observing naked eye. Major rock that constitution Unjusa temple one great geological features has strike of N30-40W and dip of 10-20NE being pyroclastic tuff. This pyroclastic tuff is ranging very extensively laying center on Unjusa temple and stone cultural properties of precinct is modeled by this pyroclastic tuff. Stone cultural propertieses of present Unjusa temple precinct are accomplishing structural imbalance with serious crack, and because weathering of rock with serious biological pollution is gone fairly, rubble break away and weathering and deterioration phenomenon such as fall off of a particle of mineral are appearing extremely. Also, a piece of iron and cement mortar of stone cultural properties everywhere are forming precipitate of reddish brown and light gray being oxidized. About these stone cultural properties, most stone cultural propertieses show SD(severe damage) to MD(moderate damage) as result that record Deterioration degree. X-ray diffraction analysis result samples of each rock are consisted of mineral of quartz, orthoclase,plagioclase, calcite, magnetite etc. Quartz and feldspar alterated extremely in a microscopic analysis, and biotite that show crystalline form of anhedral shows state that become chloritization that is secondary weathering mineral being weathered. Also, see that show iron precipitate of reddish brown to crack zone of tuff everywhere preview rock that weathering is gone deep. Tuffs that accomplish stone cultural properties of study area is illustrated to field of Subalkaline and Peraluminous, $SiO_2$(wt.%) extent of samples pyroclastic tuff 70.08-73.69, ash tuff extent of 70.26-78.42 show. In calculate Chemical Index of Alteration(CIA) and Weathering Potential Index(WPI) about major elements extent of CIA pyroclastic tuff 55.05-60.75, ash tuff 52.10-58.70, granite 49.49-51.06 granitic gneiss shows value of 53.25-67.14 and these have high value gneiss and tuffs. WPI previews that is see as thing which is illustrated being approximated in 0 lines and 0 lines low samples of tuffs and gneiss is receiving esaily weathering process as appear in CIA. As clay mineral of smectite, zeolite that is secondary weathering produce of rock as result that pick powdering of rock and clothing material of stone cultural properties observed by scanning electron micrographs (SEM). And roots of lichen and spore of hyphae that is weathering element are observed together. This rock deep organism being coating to add mechanical weathering process of stone cultural properties do, and is assumed that change the clay mineral is gone fairly in stone cultural properties with these. As the weathering of rocks is under a serious condition, the damage by the natural environment such as rain, wind, trees and the ground is accelerated. As a counter-measure, the first necessary thing is to build the ground environment about protecting water invasion by making the drainage and checking the surrounding environment. The second thing are building hardening and extirpation process that strengthens the rock, dealing biologically by reducing lichens, and sticking crevice part restoration using synthetic resin. Moreover, it is assumed to be desirable to build the protection facility that can block wind, sunlight, and rain which are the cause of the weathering, and that goes well with the surrounding environment.