• The Korean Journal of Mycology
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• v.47 no.3
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• pp.187-197
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• 2019

Ascomycota is the largest phylum of the Fungi, including approximately 6,600 genera. They are often isolated from soils, indoor air, and freshwater environments, but also from plants as pathogens or endophytes. In this study, four species of Ascomycota (two of Cladosporium and one of each Daldinia and Nigrospora) were collected from the leaves of four woody plants (Camellia japonica, Ginkgo biloba, Quercus sp., Vitis vinifera). Their cultural characteristics were investigated on five different media (PDA, V8A, CMA, MEA, CZA) at 3 days after incubation at $25^{\circ}C$ in darkness. BLASTn search and phylogenetic analysis were performed using the internal transcribed spacer (ITS) rDNA sequences, in addition to tef1 gene sequences for Cladosporium species. Based on the cultural, morphological, and phylogenetic data, the isolates were identified as Cladosporium anthropophilum, Cladosporium pseudocladosporioides, Daldinia eschscholtzii, and Nigrospora chinensis. Previously, some members of Cladosporium and Nigrospora have been recorded as endophytes inhabiting the leaves and stems of various plants, whereas Daldinia eschscholtzii is a wood-inhabiting endophyte or wood-decaying fungus. To our knowledge, this is the first report of these four ascomycetes in Korea.

• Journal of Microbiology and Biotechnology
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• v.29 no.6
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• pp.839-844
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• 2019

Anthranilate derivatives have been used as flavoring and fragrant agents for a long time. Recently, these compounds are gaining attention due to new biological functions including antinociceptive and analgesic activities. Three anthranilate derivatives, N-methylanthranilate, methyl anthranilate, and methyl N-methylanthranilate were synthesized using metabolically engineered stains of Escherichia coli. NMT encoding N-methyltransferase from Ruta graveolens, AMAT encoding anthraniloyl-coenzyme A (CoA):methanol acyltransferase from Vitis labrusca, and pqsA encoding anthranilate coenzyme A ligase from Pseudomonas aeruginosa were cloned and E. coli strains harboring these genes were used to synthesize the three desired compounds. E. coli mutants (metJ, trpD, tyrR mutants), which provide more anthranilate and/or S-adenosyl methionine, were used to increase the production of the synthesized compounds. MS/MS analysis was used to determine the structure of the products. Approximately, $185.3{\mu}M$ N-methylanthranilate and $95.2{\mu}M$ methyl N-methylanthranilate were synthesized. This is the first report about the synthesis of anthranilate derivatives in E. coli.

• Journal of Plant Biotechnology
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• v.49 no.4
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• pp.292-2999
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• 2022

Anthocyanin, an important component in the grape berry skin, strongly affects grape quality. The transcription factors VvMYBA1 and VvMYBA2 (VvMYBA1/2) control anthocyanin biosynthesis. In addition, cultivation and environmental factors, such as light, influence anthocyanin accumulation. The present study aimed to clarify the effect of shading (reduced light condition) on the transcriptomic regulation of anthocyanin biosynthesis using a red-wine grape cultivar, Vitis vinifera 'Pinot Noir', and its white mutant, 'Pinot Blanc', caused by the deletion of the red allele of VvMYBA1/2. The grape berry skins were analyzed for anthocyanin content and global gene transcription accumulation. The microarray data were later validated by quantitative real-time PCR. A decisive influence of VvMYBA1/2 on the expression of an anthocyanin-specific gene, UDP glucose: flavonoid 3-O-glucosyltransferase, was observed as expected. In contrast, upstream genes of the pathway, which are shared by other flavonoids, were also expressed in 'Pinot Blanc', and the mRNA levels of some of these genes decreased in both cultivars on shading. Thus, the involvement of light-sensitive transcription factor(s) other than VvMYBA1/2 was suggested for the expression control of the upstream genes of the anthocyanin biosynthetic pathway. Furthermore, it was suggested that the effects of these factors are different among isogenes.

• Preventive Nutrition and Food Science
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• v.7 no.3
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• pp.231-237
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• 2002

This study was conducted to determine biological activities, such as lipid peroxidation inhibition, cytotoxicity, sun blocker, inhibition of tyrosinase, and antioxidative effect, of ethanol extracts, and of solvent fractionated ethanol extracts obtained from grape seeds and skins. The strongest lipid oxidative inhibition of 66.9% and 67.6% was observed respectively, in the presence of 20 $\mu\textrm{g}$/$m\ell$ of both ethanol extract and water fraction of grape seeds. Overall, the ethanol extracts and their fractions of grape seeds exhibited stronger lipid oxidative inhibition than that of skin extracts. On the other hand, the ethanol extracts of grape skins showed stronger cytotoxicity than that of seeds on MCF-7, Hep3B, and A549 cancer cell lines. However, the water fraction of seed ethanol extracts showed the strongest cytotoxic effect of 76.52% and 67.01% on MCF-7 and Hep3B, respectively among their fractions. Ethanol seed extracts obtained at 3$0^{\circ}C$ had the strongest absorbance both at UVA region (350 nm) and UVB region (308 nm) and the chloroform fraction showed the strongest absorbance at W region and butanol fraction at UVA region among their tractions, respectively. In the meantime, the ethanol extracts obtained at 3$0^{\circ}C$ and butanol fraction showed the strongest tyrosinase inhibitory effect of 39.4% and 37.6%, respectively. This study shows that ethanol extracts and their fractions of grape seeds and skins could be potential good materials for functional food and cosmetic products.

• Journal of Applied Biological Chemistry
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• v.44 no.3
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• pp.147-153
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• 2001

Methanol extracts from 51 fruits were tested for their fungicidal activities against six phytopathogenic fungi in a greenhouse. The efficacy varied with both the plant pathogen and fruit species used. At 10 and 5 mg/pot, methanol extracts of Poncirus trifoliata peel and seed gave over 80% control values against Pyricularia grisea, and strong fungicidal activities against Rhizoctonia solani were showed from the extracts of Citrus paradisi peel and Punica granatum leaf. In a test with Botrytis cinerea at 5 mg/pot, the extracts of C. sinensis seed and D. kaki leaf produced potent fungicidal activities, and the extracts of C. crenata peel and leaf, Ch. sinensis seed, P. trifoliata peel, and Z. jujuba leaf had strong fungicidal activities. At 5 mg/pot, strong fungicidal activities were produced in the extracts of P. trifoliata peel and seed against Phytophthora infestans and in the extracts of P. ussriensis var. macrostipes fruit and seed, C. crenata peel, C. crenata leaf, C. paradisi peel, P. trifoliata peel, P. granatum peel, and Z. jujuba leaf against Puccinia recondita. In a test with E. graminis, potent activities at 10 mg/pot were produced from the extracts of Ch. sinensis seed, C. sinensis seed, P. trifoliata leaf, P. ussriensis var. macrostipes fruit and seed, and Vitis vinifera seed. In the control effect of seven extracts against B. cinerea strains resistant to carbendazim, procymidone, and diethofencarb, extracts of C. crenata peel and leaf, Ch. sinensis seed, and P. trifoliata peel were highly effective against all strains of B. cinerea. Furthermore, potent fungicidal activities were produced from the extracts of C. sinensis seed and D. kaki leaf against the SSR, SRR, and RRS, and Z. jujuba leaf against the SSR and RRS strains. As a naturally occurring fungicide, these fruit-derived materials could be useful as new fungicidal products against phytopathogenic fungi.

• BMB Reports
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• v.40 no.4
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• pp.595-603
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• 2007

To investigate whether phospholipase D (PLD, EC 3.1.4.4) plays a role in adaptive response of post-harvest fruit to environment, a PLD gene was firstly cloned from grape berry (Vitis Vinifera L. cv. Chardonnay) using RT-PCR and 3'- and 5'-RACE. The deduced amino acid sequence (809 residues) showed 84.7% identity with that of PLD from Ricinus communis. The secondary structures of this protein showed the characteristic C2 domain and two active sites of a phospholipid-metabolizing enzyme. The PLD activity and its expression in response to heat acclimation were then assayed. The results indicated PLD was significantly activated at enzyme activity, as well as accumulation of PLD mRNA and synthesis of new PLD protein during the early of heat acclimation, primary suggesting that the grape berry PLD may be involved in the heat response in post-harvest grape berry. This work offers an important basis for further investigating the mechanism of post-harvest fruit adaptation to environmental stresses.

• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.388-393
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• 2017

We report an Agrobacterium-mediated transformation procedure optimized for 'Kyoho' that is a major table grapevine cultivar in Korea, and its transgenic plants with antifreeze protein gene of carrot (DcAFP). The full length of DcAFP coding region in accordance with the previous report was isolated from young leaves of carrot and recombined into a plant transformation vector. Ethylene inhibitors such as silver nitrate and aminoethoxyvinylglycin (AVG) supplemented in a co-cultivation medium distinctly increased frequency of shoot regeneration when explants were sub-cultured in a selection medium: particularly ten-fold higher in treatment with 0.1 mg/L AVG than one without ethylene inhibitor. Among various antibiotics and their concentrations, the combination of 150 mg/L cefotaxime plus 150 mg/L $Clavamox^{TM}$ was selected for elimination of Agrobacterium cells in addition to minimization of adverse effect on shoot regeneration, while 50 mg/L kanamycin monosulfate effectively suppressed regeneration of non-transgenic shoots. Applying the elucidated culture condition, we finally obtained a total of 5 transgenic 'Kyoho' plantlets with DcAFP, of which integration with the grapevine genome and transcription was confirmed by nucleic acid analyses.

• Mycobiology
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• v.42 no.3
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• pp.262-268
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• 2014

Several postharvest diseases of table grapes (Vitis vinifera) occur during storage, and gray mold rot is a particularly severe disease because the causal agent, Botrytis cinerea, grows at temperatures as low as $0^{\circ}C$. Other postharvest diseases, such as those caused by Penicillium spp. and Aspergillus spp., also often lead to deterioration in the quality of table grapes after harvest. The use of plant essential oils such as thymol and linalool, to reduce postharvest diseases in several kinds of fruits, including table grapes and oranges, has received much attention in European countries. However, to the best of our knowledge there has been no report of the use of thymol fumigation to control gray mold in table grapes in Korea. Thymol ($30{\mu}g/mL$) and linalool ($120{\mu}g/mL$) significantly inhibited mycelial growth and conidia germination of B. cinerea. The occurrence rate of gray mold rot of B. cinerea and other unknown fungi was significantly reduced by fumigation with $30{\mu}g/mL$ thymol in several table grape cultivars, such as Campbell early, Muscat Bailey A, Sheridan, and Geobong. In this study, fumigation with $30{\mu}g/mL$ thymol, had no influence on the sugar content and hardness of grapes, but reduced fungal infection significantly. This suggests that $30{\mu}g/mL$ thymol could be utilized to reduce deterioration of grapes due to gray mold and other fungal infections during long-term storage.

• Nutrition Research and Practice
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• v.9 no.1
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• pp.37-42
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• 2015

BACKGROUND/OBJECTIVES: Red grape seeds as functional food are a good source of important bioactive components such as phenolics and antioxidants, which decrease oxidative stress that contributes to the pathogenesis of hepatotoxicity. The current study was conducted in order to evaluate the protective effect of red grape dried seeds (RGDS) on antioxidant properties, lipid metabolism, and liver and kidney functions of rats with paracetamol (750 mg/kg) induced hepatotoxicity. MATERIALS/METHODS: RGDS was added to the basal diet at 5, 10, and 20%. Thirty five adult male rats were assigned to five groups (n = 7) for a six-week feeding period; group (1) normal control, group (2) induced control, groups (3, 4, and 5) fed a diet with RGPS at different levels, 5, 10, and 20%, respectively. At the end of the feeding period, animals' blood and tissues were collected for estimation of serum lipid profile, serum liver, and kidney biomarkers. The protection was measured by detecting lipid peroxidation (LPO), glutathione (GSH), superoxide dismutase (SOD), Catalase (CAT) (in liver tissues), and liver histological examination. RESULTS: The results showed a significant (P < 0.05) decrease in levels of serum cholesterol, triglycerides, low density lipoprotein (LDL-C), and very low density lipoprotein (VLDL-C), with a significant increase in level of high density lipoprotein (HDL-C) for RGDS groups compared to induced control. Rats administered a diet containing RGDS levels produced significant (P < 0.05) hepatoprotection by decreasing the activities of liver enzymes, kidney parameters, and lipid peroxidation, while levels of GSH, SOD, and CAT were increased significantly to near the normal levels. CONCLUSION: The RGDS 20% group was more effective than others against hepatotoxicity of paracetamol, which may be attributed to RGDS total phenols and antioxidant contents, which were 1.438 mg and 1.231 mg, respectively.

• Korean Journal of Food Science and Technology
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• v.50 no.5
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• pp.474-479
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• 2018

In this study, ultrasound-assisted extraction conditions of total anthocyanins and total phenolic compounds from Campbell early grape (Vitis labruscana) were optimized via response surface methodology using a Box-Behnken design. Three independent variables, including solid-liquid ratio (1/30-1/50 g/mL), extraction time (2-10 min), and amplitude (20-100%), significantly affected total anthocyanin content. However, no significant effect of these variables was observed in the total phenolic compound content. The optimized extraction conditions were solid-liquid ratio of 1/48.13 g/mL, extraction time of 2 min, and amplitude of 20%. Under these conditions, the predicted values of total anthocyanins and total phenolic compounds were 94.96 and 1,661.16 mg/100 g dry weight (DW), respectively. The experimental values of total anthocyanins and total phenolic compounds were 88.25 and 1,554.37 mg/100 g DW, respectively, which is well matched with the predicted values within 7.5% difference.