• The Plant Pathology Journal
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• v.30 no.1
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• pp.10-24
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• 2014

Genetic diversity and differentiation of 50 Colletotrichum spp. isolates from legume crops studied through multigene loci, RAPD and ISSR analysis. DNA sequence comparisons by six genes (ITS, ACT, Tub2, CHS-1, GAPDH, and HIS3) verified species identity of C. truncatum, C. dematium and C. gloeosporiodes and identity C. capsici as a synonym of C. truncatum. Based on the matrix distance analysis of multigene sequences, the Colletotrichum species showed diverse degrees of intera and interspecific divergence (0.0 to 1.4%) and (15.5-19.9), respectively. A multilocus molecular phylogenetic analysis clustered Colletotrichum spp. isolates into 3 well-defined clades, representing three distinct species; C. truncatum, C. dematium and C. gloeosporioides. The ISSR and RAPD and cluster analysis exhibited a high degree of variability among different isolates and permitted the grouping of isolates of Colletotrichum spp. into three distinct clusters. Distinct populations of Colletotrichum spp. isolates were genetically in accordance with host specificity and inconsistent with geographical origins. The large population of C. truncatum showed greater amounts of genetic diversity than smaller populations of C. dematium and C. gloeosporioides species. Results of ISSR and RAPD markers were congruent, but the effective maker ratio and the number of private alleles were greater in ISSR markers.

• The Plant Pathology Journal
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• v.37 no.6
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• pp.512-520
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• 2021

In this study, we report genetic characterization of Orobanche cumana, the causal agent of sunflower wilting in Serbia. The genetic diversity of this parasitic plant in Serbia was not studied before. Random amplified polymorphic DNA (RAPD) markers and partial rbcL gene sequences analysis were used to characterize the O. cumana populations at the molecular level. While phylogenetic analyses of RAPD-PCR amplicons were performed using unweighted pair-group Method analyses, rbcL gene sequences were analyzed using neigbor joining method and minimum spanning tree. Molecular analyses of RAPD-PCR analysis revealed high genetic diversity of O. cumana populations which indicated high adaptive potential of this parasitic weed in Serbia. Further analyses of rbcL gene using minimum spanning tree revealed clear differences among diverse sections of Orobanche genus. Although this molecular marker lacked the resolution to display intrapopulation diversity it could be a useful tool for understanding the evolution of this parasitic plant. Our results suggested that O. cumana has great genetic potential which can lead to differentiation of more virulent races which is important for determining crop breeding strategies for their control.

• The Korean Journal of Mycology
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• v.50 no.4
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• pp.319-329
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• 2022

In July 2020, wilting symptoms were observed among adzuki bean plants (Vigna angularis var. angularis L.) in the fields in Yeosu, Korea. Infected plants showed yellowing of leaves, browning inside the stems, splitting of stem bark, and wilting. When these plants were uprooted, their roots were found to be brown. The fungal pathogens NC20-737, NC20-738, and NC20-739 were isolated from symptomatic stem and root tissues. These pathogens were identified as a Fusarium fujikuroi species complex based on their morphological characteristics. Molecular identification was performed using the DNA sequence of translation elongation factor 1 alpha and the RNA polymerase II second largest subunit regions. The nucleotide sequences of all three isolates were similar to the F. fujikuroi reference isolates NRRL 13566 and NRRL 5538 of the National Centre for Biotechnology Information GenBank. A pathogenicity test was conducted by the soil inoculation method with cornmeal sand inoculum. Approximately 3 weeks after inoculation, symptoms were observed only in the inoculated adzuki bean seedlings. To the best of our knowledge, this is the first report of Fusarium root rot caused by F. fujikuroi in adzuki beans, both in Korea and worldwide.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.1
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• pp.22-27
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• 2004

In the present study, the protective effects of Artemisia capillaris (AC) on the DNA damage induced by $^{60}$ Co ${\gamma}$-rays were evaluated using alkaline single-cell gel electrophoresis (SCGE, comet assay) in the mouse peripheral lymphocytes and micronuclei (MN) formation test in the Chinese hamster ovary (CHO) cells. We also investigated the effect of AC on 8-hydroxy-2'-deoxyguanosine (8-OHdG) formation in the mouse liver and thymus exposed to ${\gamma}$-ray, The tail moment and the frequency of MN, which were markers of DNA damage in the SCGE and MN formation test, were decreased in the groups treated with AC extract before exposure to 200 cGy of ${\gamma}$-ray. We also observed its activities, lowering 8-OHdG level, an index of oxidative DNA damage, in the groups treated with AC extract before whole body ${\gamma}$-irradiation (800 cGy). It is plausible that scavenging of free radicals by AC may have played an important role in providing the protection against the radiation-induced damage to the DNA. These results indicated that AC protects the DNA damage induced by ${\gamma}$-rays and might be a useful radioprotector, especially since it is a relatively nontoxic product.

• Horticultural Science & Technology
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• v.31 no.5
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• pp.580-589
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• 2013

This study was carried out to evaluate the suitability of microsatellite markers for varietal identification and genetic relationship of 170 commercial pepper cultivars. The relationship between marker genotypes and 11 pepper cultivars with different morphological traits was also analyzed. Of the 302 pairs of microsatellite primers screened against 11 pepper cultivars, 24 pairs were highly polymorphic in terms of number of alleles. These markers were applied for the construction of DNA profile data base for 170 commercial pepper cultivars. A total of 164 polymorphic amplified fragments were obtained from 24 microsatellite primers. The average polymorphism information content was 0.673 ranging from 0.324 to 0.824. One hundred and sixty four microsatellite alleles were used to calculate Jaccard's distance coefficients using unweighted pair group method. A clustering group of varieties, based on the results of microsatellite analysis, were categorized into 3 major groups corresponding to morphological traits. The phenogram discriminated all varieties by markers genotypes. These microsatellite markers will be useful as a tool for protection of plant breeders' intellectual property rights through variety identification in distinctness, uniformity and stability test.

• Journal of Nutrition and Health
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• v.36 no.2
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• pp.125-132
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• 2003

The present study was attempted to investigate and compare the antioxidant potency of several well-know flavonoids, antioxidant vitamin and commercially available popular beverages. The antioxidant potency was assessed by the effect on reducing oxidative DNA damage of human lymphocytes. Cellular oxidative DNA damage was measured by SCGE (single-cell gel electrophoresis), also known as comet assay. Lymphocytes were pre-treated for 30 minutes with wide ranges of doses of apigenin, kaempferol, luteolin, myricetin, rutin, quercetin, $\alpha$-tocopherol (10,25,50,100,200,500,1000 $\mu$M) ,green tea extract or grape juice (10,50,100,250,500,1000 $\mu$g/mL) followed by a $H_2O$$_2$(100 $\mu$M) treatment for 5 min as an oxidative stimulus. The physiological function of each antioxidant substance on oxidative DNA damage was analyzed as tail moment (tail length $\times$ percentage migrated DNA in tail) and expressed as relative DNA damage score after adjusting by the level of control treatment. Cells treated with $H_2O$$_2$alone (positive control) had an extensive DNA damage compared with cells treated with phosphate buffered saline (PBS, negative control) or pre-treated with all the tested samples. Of all the six flavonoids, quercetin was the most potent antioxidant showing the lowest $ED_{50}$/ of 8.5 $\mu$g/mL (concentration to produce 50% protection of relative DNA damage). The antoxidant potency of individual flavonoids were ranked as follows in a decreasing order; luteolin (18.4 $\mu$g/mL), myricetin (19.0 $\mu$g/mL) , rutin (22.2 $\mu$g/mL) , apigenin (24,3 $\mu$g/mL) , kaempferol (25.5 $\mu$g/mL). The protective effect of $\alpha$-tocopherol was substantially lower (highest $ED_{50}$value of 55.0 $\mu$g/mL) than all the other flavonoids, while the protective effect was highest in green tea and grape juice with low ED5O value of 7.6 and 5.3, respectively. These results suggest that flavonoids, especially quercetin, and natural compounds from food product, green tea and grape juice, produced powerful anti-oxidative activities, even stronger than $\alpha$-tocopherol. Taken together, supplementation of antioxidants to lymphocytes followed by oxidative stimulus inhibited damage to cellular DNA, supporting a protective effect against oxidative damage induced by reactive oxygen species.

• Research in Plant Disease
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• v.22 no.1
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• pp.9-17
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• 2016

Nationwide survey for angular leaf spot (ALS) of strawberry caused by Xanthomonas fragariae, a quarantine disease in Korea, was performed in November 2012. In the survey, ALS was observed in eighty eight farmers' fields of Sukok, Jinju and Okjong, Hadong in Gyeongnam Province, and one field in Namwon of Jeollabuk Province. The infected field of Namwon closed immediately after the disease diagnosed ALS. In detailed survey of Sukok and Okjong areas during February 2012 to January 2015, ALS occurrence decreased from 45% farmer's fields on December 2012 to 5% on January 2015, and from 38% on November 2013 to 5% on January 2015, respectively. Phenotypic characteristics of the Korean strains were similar to those of the type strain of X. fragariae. A multilocus sequence analysis of Korean strains of X. fragariae was conducted using four genes; dnaK, fyuA, gyrB, and rpoD. All the Korean strains had the same sequences of the four genes. The concatenated sequences of the Korean strains shared 100% with that of the type strain of X. fragariae. All strawberry cultivars tested were susceptible to the strains of X. fragariae two weeks after inoculation. The inoculated sites were necrosis and expanded, which were rated 4 based on evaluation of inoculation site.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.3
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• pp.1299-1308
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• 2016

Our previous studies clearly demonstrated that a combination of WP 631 and Epo B has higher activity against ovarian cancer cells than either of these compounds used separately. In order to fully understand the exact mechanism of action in combination, we assessed effects on the cell cycle of SKOV-3 cells. We evaluated three control points essential for WP 631 and Epo B action to determine which cell cycle-regulating proteins (CDK1/cyclin B complex, EpCAM or HMGB1) mediate activity. The effects of the drug on the cell cycle were measured based on the nuclear DNA content using flow cytometry. Expression of cell cycle-regulating genes was analyzed using real-time PCR. It was discovered that WP 631, at the tested concentration, did not affect the SKOV-3 cell cycle. Epo B caused significant G2/M arrest, whereas the drug combination induced stronger apoptosis and lower mitotic arrest than Epo B alone. This is very important information from the point of view of the fight against cancer, as, while mitotic arrest in Epo B-treated cells could be overcame after DNA damage repair, apoptosis which occurs after mitotic slippage in combination-treated cells is irreversible. It clearly explains the higher activity of the drug combination in comparison to Epo B alone. Epo B acts via the CDK1/cyclin B complex and has the ability to inhibit CDK1, which may be a promising strategy for ovarian cancer treatment in the future. The drug combination diminishes EpCAM and HMGB1 expression to a greater degree than either WP 631 and Epo B alone. Owing to the fact that the high expression of these two proteins is a poor prognostic factor for ovarian cancer, a decrease in their expression, observed in our studies, may result in improved efficacy of cancer therapy. The presented findings show that the combination of WP 631 and Epo B is a better therapeutic option than either of these drugs alone.

• The Korean Journal of Mycology
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• v.50 no.4
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• pp.331-341
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• 2022

This study was conducted to identify yeast species isolated from domestic pear blossom through gene sequencing and analysis of morphological characteristics, and to confirm specific yeast species inhibitory effects toward fire blight in immature apples, pears, and crab apple blossoms. Yeast morphological characteristics were consistent with the known characteristics of Aureobasidium pullulans. Nucleotide sequencing of the D1/D2 region of large-subunit (LSU) 26S ribosomal DNA and the internal transcribed spacer (ITS) region confirmed its identity as A. pullulans (MHAU2101). Inoculation of immature fruits with A. pullulans MHAU2101 before exposure to Erwinia amylovora prevented fire blight symptoms in apples and pears. A. pullulans MHAU2101 treated crab apple blossoms had a significantly lower flower infection rate than untreated blossoms, revealing 64% of the potency of streptomycin. The A. pullulans MHAU2101 treated group also displayed lower E. amylovora density in both pistil and hypanthium compared to the untreated group, especially in the hypanthium. This study confirms that A. pullulans MHAU2101 isolated from domestic pear blossom can effectively suppress the onset of fire blight.

• Korean Journal of Clinical Laboratory Science
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• v.50 no.2
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• pp.205-210
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• 2018

Salvianolic acid B, which is a compound in the Salvia miltiorrhiza, has diverse biological activities, In particular, the antioxidative effects were reported to be involved in the protection of hepatocytes, neurons, and various cell types. On the other hand, some phenolic compounds, such as ferulic acid, which is regarded as an antioxidant, plays a pro-oxidative role in the specific transitional metal environment, which could explain the anticancer effect. This study examined the pro-oxidative effects of salvianolic acid B in the presence of $Cu^{2+}$. Treatment with both salvianolic acid B and $Cu^{2+}$ induced the transition of supercoiled DNA to the open circular or linear form but not in the sole salvianolic acid B or $Cu^{2+}$ treatments. Salvianolic acid B reduced the $Cu^{2+}$ to $Cu^+$ using neocuproine, a $Cu^+$ specific chelator. In addition, catalase, an enzyme that breaks down the $H_2O_2$ to water and molecular oxygen, inhibited the DNA breakage. $H_2O_2$, a reactive oxygen species, has detrimental effects on biological molecules, particularly DNA. Overall, the reduction of $Cu^{2+}$ by salvianolic acid B could lead to the production of $H_2O_2$ followed by DNA breakage. These results suggest that the pro-oxidative effects could be the one of the anti-cancer mechanisms of salvianolic acid B, which remains to be explained.