• Mycobiology
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• v.29 no.1
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• pp.19-26
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• 2001

Defense mechanisms against anthracnose disease caused by Colletotrichum orbiculare on the leaf surface of cucumber plants after pre-treatment with plant growth promoting rhizobacteria(PGPR), amino salicylic acid(ASA) or C. orbiculare were compared using a fluorescence microscope. Induced systemic resistance was mediated by the pre-inoculation in the root system with PGPR strain Bacillus amylolquefaciens EXTN-1 that showed direct antifungal activity to C. gloeosporioides and C. orbiculare. Also, systemic acquired resistance was triggered by the pre-treatments on the bottom leaves with amino salicylic acid or conidial suspension of C. orbiculare. The protection values on the leaves expressing SAR were higher compared to those expressing ISR. After pre-inoculation with PGPR strains no change of the plants was found in phenotype, while necrosis or hypersensitive reaction(HR) was observed on the leaves of plants pre-treated with ASA or the pathogen. After challenge inoculation, inhibition of fungal growth was observed on the leaves expressing both ISR and SAR. HR was frequently observed at the penetration sites of both resistance-expressing leaves. Appressorium formation was dramatically reduced on the leaves of plants pre-treated with ASA, whereas EXTN-1 did not suppress the appressorium formation. ASA also more strongly inhibited the conidial germination than EXTN-1. Conversely, EXTN-1 significantly increased the frequency of callose formation at the penetration sites, but ASA did not. The defense mechanisms induced by C. orbiculare were similar to those by ASA. Based on these results it is suggested that resistance mechanisms on the leaf surface was different between on the cucumber leaves expressing ISR and SAR, resulting in the different protection values.

• Mycobiology
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• v.36 no.4
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• pp.236-241
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• 2008

The colonization of an arbuscular mycorrhizal fungus Glomus intraradices BEG110 in the soil caused a decrease in disease severity in cucumber plants after fungal inoculation with Colletotrichum orbiculare. In order to illustrate the resistance mechanism mediated by G. intraradices BEG110, infection patterns caused by C. orbiculare in the leaves of cucumber plants and the host cellular responses were characterized. These properties were characterized using transmission electron microscopy on the leaves of cucumber plants grown in soil colonized with G. intraradices BEG110. In the untreated plants, inter- and intra-cellular fungal hyphae were observed throughout the leaf tissues during both the biotrophic and necrotrophic phases of infection. The cytoplasm of fungal hyphae appeared intact during the biotrophic phase, suggesting no defense response against the fungus. However, several typical resistance responses were observed in the plants when treated with G. intraradices BEG110 including the formation of sheaths around the intracellular hyphae or a thickening of host cell walls. These observations suggest that the resistance mediated by G. intraradices BEG110 most often occurs in the symplast of the host cells rather than in the apoplast. In addition, this resistance is similar to those mediated by biotic inducers such as plant growth promoting rhizobacteria.

• Korean journal of applied entomology
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• v.41 no.3
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• pp.211-216
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• 2002

The present study was aimed to examine the effect of different host plants on the biological property of cotton caterpillar, outbreak pest of cucurbit vegetables. The feeding and oviposition preferences toward cucumber and pumpkin were greater than those toward watermelon, oriental melon, and melon. The periods of egg and larva fed on oriental melon, melon, and pumpkin were longer than those on watermelon and cucumber. The longest pupal period was 10.5 days from cucumber. The hatching rate, pupation rate, emergence rate, and survival rate were observed from different host plants. The hatching rate was the highest on cucumber (87.2%) and the lowest on melon (72.8%). The pupation rate on watermelon and oriental melon (90.0% and 89.1%, respectively) was higher than that on cucumber (62.0%). The emergence rate on cucumber and pumpkin (93.5% and 92.0%, respectively) was higher than that on oriental melon (78.7%). The survival rate from hatching to emergence on watermelon (76.0%) was the highest, and the lowest on cucumber (50.0%). The adult period on melon (21.0 days) was the longest, and the shortest on cucumber (15.5 days) among 5 host plants. Average number of eggs per female on cucumber (281.8 eggs) was the highest, and the least on oriental melon (96.6 eggs). The survivorship on cucumber was the longest (30 days), and the shortest on pumpkin (17 days). The preoviposition period on oriental melon and melon (3.4 days) was longer than that on watermelon (2.1 days) and mean generation time in day (T) on melon was the longest (47.2 days) though they were not significant. The net reproductive rate per generation (R$_{o}$) on cucumber, 191.3, was the highest and the highest intrinsic rate of natural increase (r$_{m}$) was observed from cucumber as 0.127. All these results suggest that the growth ana reproduction of cotton caterpillar varies depending upon the kind of host plants. The analysis of the life table revealed that cucumber and pumpkin were the favorable host plants of cotton caterpillar.

• Journal of Environmental Science International
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• v.16 no.12
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• pp.1345-1353
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• 2007

The effect of salicylic acid(SA) on antioxidant system and protective mechanisms against UV-B induced oxidative stress was investigated in cucumber(Cucumis sativus L.) leaves. UV-B radiation and SA were applied separately or in combination to first leaves of cucumber seedlings, and dry matter accumulation, lipid peroxidation and activities of antioxidant enzymes were measured in both dose and time-dependant manner. UV-B exposure showed reduced levels of fresh weight and dry matter production, whereas SA treatment significantly increased them. SA noticeably recovered the UV-B induced inhibition of biomass production. UV-B stress also affected lipid peroxidation and antioxidant enzyme defense system. Malondialdehyde(MDA), a product of lipid peroxidation, was greatly increased under UV-B stress, showing a significant enhancement of a secondary metabolites, which may have antioxidative properties in cucumber leaves exposed to UV-B radiation. Combined application of UV-B and SA caused a moderate increase in lipid peroxidation. These results suggest that SA may mediate protection against oxidative stress. UV-B exposure significantly increased SOD, APX, and GR activity compared with untreated control plants. Those plants treated with 1.0 mM SA showed a similar pattern of changes in activities of antioxidant enzymes. SA-mediated induction of antioxidant enzyme activity may involve a protective accumulation of $H_2O_2$ against UV-B stress. Moreover, their activities were stimulated with a greater increase by UV-B+SA treatment. The UV-B+SA plants always presented higher values than UV-B and SA plants, considering the adverse effects of UV-B on the antioxidant cell system. ABA and JA, second messengers in signaling in response to stresses, showed similar mode of action in UV-B stress, supporting that they may be important in acquired stress tolerance. Based on these results, it can be suggested that SA may participates in the induction of protective mechanisms involved in tolerance to UV-B induced oxidative stress.

• Journal of Environmental Impact Assessment
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• v.22 no.2
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• pp.125-134
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• 2013

Plant virus coat (CP) gene-mediated protection is one of the best known approaches to protect against virus resistant transgenic plants. Transgenic N. benthamiana plants containing the CP gene of Zucchini green mottle mosaic virus (ZGMMV) were used for the environmental risk assessment of the living modified (LM) plants with plant virus resistance. The most optimal co-infection method of both ZGMMV and CMV (Cucumber mosaic virus) on Non-LM and CP-expressing LM tobacco plants was established and co-infection of CMV and ZGMMV was confirmed by polymerase chain reaction (PCR). To address the effects of LM tobacco plants on the mutation of the virus, in-vitro transcripts of CP and Replicase (Rep) derived from CMV and/or ZGMMV were inoculated onto Non-LM or LM tobacco plants. Mutation frequency of CP and Rep from CMV and ZGMMV was examined through six serial passages in Non-LM and LM tobacco plants. Little actual frequency of mutation was estimated, probably due to the limited number of transgenic plants tested in this study. However, it does not suggest environmental safety of these CP-mediated LM plants. Further study at a larger scale is needed to evaluate the environmental risk associated with the CP-expressing LM plants.

• Mycobiology
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• v.30 no.1
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• pp.31-36
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• 2002

Soil bacteria were screened for the ability to control cucumber anthracnose caused by Colletotrichum orbiculare through induced systemic resistance(ISR). Sixty-four bacterial strains having in vitro antifungal activity were used for selecting ISR-inducing strains in cucumber. Cucumber seeds(cv. Baeknokdadagi) were sown in potting mixtures incorporated with the soil bacteria, at a rate of ca. $10^8$ cells per gram of the mixture. Two week-old plants were then transplanted into the steam-sterilized soil. Three leaf-stage plants were inoculated with a conidial suspension($5{\times}10^5$ conidia/ml) of C. orbiculare. Diseased leaf area(%) and number of lesions per $cm^2$ leaf were evaluated on third leaves of the plants, $5{\sim}6$ days after inoculation. Among 64 strains tested, nine strains, GC-B19, GC-B35, GK-B18, MM-B22, PK-B14, RC-B41, RC-B64, RC-B65, and RC-B77 significantly(P=0.05) reduced anthracnose disease compared to the untreated control. In contrast, some bacterial strains promoted susceptibility of cucumber to the disease. From the repeated experiments using the nine bacterial strains, GC-B19, MM-B22, PK-B14, and RC-B65 significantly(P=0.05) reduced both diseased leaf area(%) and number of lesions per $cm^2$ leaf in at lease one experiment. These strains with control efficacy of $37{\sim}80%$ were determined to be effective ISR-inducing strains.

• Journal of Plant Biotechnology
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• v.40 no.4
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• pp.198-202
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• 2013

To produce transgenic cucumber expressing Nit gene coffering abiotic resistance, the cotyledonary-node explants of cucumber (cv. Eunsung) were inoculated with A. tumefaciens transformed with pPZP211 or pCAMBIA2300 carrying Nit gene, that has cis-acting element involved in resistance to various abiotic environmental stresses. After co-cultivation, the procedures of selection, shoot initiation, shoot elongation, and plant regeneration were followed by cotyledonary-node transformation method (CTM, Jang et al. 2011). The putative transgenic plants were selected when shoots were grown to a length greater than 3 cm from the cotyledonary-node explants on selection medium supplemented with 100 mg/L paromomycin as a selectable agent. The confirmation of transgenic cucumber was based on the genomic PCR, Southern blot analysis, RT-PCR, and Northern blot analysis. A 105 shoots (4.12%) selected from the selection mediums were obtained from 2,547 explants inoculated. Of them, putative transgenic plants were only confirmed with 45 plants (1.77%) by genomic PCR analysis. Transgenic plants showed that the Nit genes integrated into each genome of 39 plants (1.53%) by Southern blot analysis, and the expression of gene integrated into cucumber genome was only confirmed at 6 plants (0.24%) by RT-PCR and Northern blot analysis. These results lead us to speculate that the genes were successfully integrated and expressed in each genome of transgenic cucumber.

• Journal of Microbiology and Biotechnology
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• v.18 no.1
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• pp.67-73
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• 2008

The potential of the exopolysaccharide (EPS) from a Serratia sp. strain Gsm01 as an antiviral agent against a yellow strain of Cucumber mosaic virus (CMV-Y) was evaluated in tobacco plants (Nicotiana tabacum cv. Xanthi-nc). The spray treatment of plants using an EPS preparation, 72h before CMV-Y inoculation, protected them against symptom appearance. Fifteen days after challenge inoculation with CMV-Y, 33.33% of plants showed mosaic symptoms in EPS-treated plants compared with 100% in the control plants. The EPS-treated plants, which showed mosaic symptoms, appeared three days later than the controls. The enzyme-linked immunosorbent assay (ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR) analyses of the leaves of the protected plants revealed that the EPS treatment affected virus accumulation in those plants. Analysis of phenylalanine ammonia lyase, peroxidase, and phenols in protected plants revealed enhanced accumulation of these substances. The pathogenesis-related (PR) genes expression represented by PR-lb was increased in EPS-treated plants. This is the first report of a systemic induction of protection triggered by EPS produced by Serratia sp. against CMV-Y.

• The Plant Pathology Journal
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• v.23 no.4
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• pp.251-259
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• 2007

Two isolates of Cucumber mosaic virus (CMV) originated from lily plants, named Ly2-CMV and Ly8-CMV, were compared with their pathological features in several host plants. Ly2-CMV and Ly8-CMV could induce systemic mosaic symptom in Nicotiana benthamiana, but Ly2-CMV could not systemically infect tomato and cucumber plants that have been used for CMV-propagative hosts. While Fny-CMV used as a control infected systemically the same host plants, producing typical CMV symptoms. Ly8-CMV could infect systemically two species of tobacco (N. tabacum cv. Xanthi-nc and N. glutinosa) and zucchini squash (Curcubita pepo), but Ly2 failed systemic infection on these plants. As resulted from tissue-print immunoblot assay, different kinetics of systemic movement between Ly2-CMV and Ly8-CMV were crucial for systemic infection in tobacco (cv. Xanthi-nc). Sequence analysis of full-length genome of two lily isolates showed Ly2 and Ly8 belonged to subgroup IA of CMV. The lily isolates shared overall 98 % sequence identity in their genomes. Coat protein, 3a protein, and 2b protein involved in virus movement was highly conserved in genomes of the isolates Ly2 and Ly8. Although there is the low frequency of recombinants and reassortants in natural CMV population, phylogenetic analysis of each viral protein among a number of CMV isolates suggested that genetic variation in a defined population of CMV lily isolates was stochastically produced.

• The Plant Pathology Journal
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• v.29 no.2
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• pp.193-200
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• 2013

Trichoderma asperellum SKT-1 is a microbial pesticide that is very effective against various diseases. Our study was undertaken to evaluate T. asperellum SKT-1 for induction of resistance against yellow strain of Cucumber mosaic virus (CMV-Y) in Arabidopsis plants. Disease severity was rated at 2 weeks post inoculation (WPI). CMV titre in Arabidopsis leaves was determined by indirect enzyme-linked immunosorbent assay (ELISA) at 2 WPI. Our results demonstrated that among all Arabidopsis plants treated with barley grain inoculum (BGI) of SKT-1 NahG and npr1 plants showed no significant reduction in disease severity and CMV titre as compared with control plants. In contrast, disease severity and CMV titre were significantly reduced in all Arabidopsis plants treated with culture filtrate (CF) of SKT-1 as compared with control plants. RT-PCR results showed increased expression levels of SA-inducible genes, but not JA/ET-inducible genes, in leaves of BGI treated plants. Moreover, expression levels of SA- and JA/ET-inducible genes were increased in leaves of CF treated plants. In conclusion, BGI treatment induced systemic resistance against CMV through SA signaling cascade in Arabidopsis plants. While, treatment with CF of SKT-1 mediated the expression of a majority of the various pathogen related genes, which led to the increased defense mechanism against CMV infection.