• Korean Journal of Organic Agriculture
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• v.23 no.2
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• pp.323-334
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• 2015

This study was conducted to develop environment-friendly agricultural products with anti-microbial activity against Acidovorax avenae subsp. citrulli as a pathogen of bacterial fruit blotch in cucurbit. Schlechtendalia chinensis was extracted by MeOH and solvent fraction. The hexane fraction, which showed highest value of anti-microbial activity, was analyzed by GC-MS. Each mass spectra, corresponding to each peak of chromatogram, was compared to MS database of Wiley library. As a result, myristic acid, palmitic acid and 3-n-pentadecylphenol were identified as maine compounds showing antimicrobial activity against A. avenae subsp. citrulli. Bioassay using commercial myristic acid, palmitic acid and 3-n-pentadecylphenol to test for the anti-microbial activity conformed the anti-microbial activity of potential active compounds, and myristic acid and 3-n-pentadecylphenol showed strong activity. In conclusion, myristic acid and 3-n-pentadecylphenol identified from S. chinensis were anti-microbial chemicals.

• The Plant Pathology Journal
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• v.22 no.1
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• pp.36-45
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• 2006

Twenty isolates of Didymella bryoniae were isolated from infected cucurbit plants in various growing areas of southern Korea in 2001 and 2002. Random Amplified Polymorphic DNA (RAPD) group [RG] I of D. bryoniae was more virulent than RG IV to watermelon. Virulence of the RG I isolate was strong to moderate to cucumber, whereas that of the RG IV varied from strong, moderate to weak. Two hundred seventy-three amplified fragments were produced with 40 primers, and were analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYSPC. At the distance level of 0.7, two major genomic DNA RAPD groups were differentiated among 20 isolates. The RG I included 7 isolates from watermelon and one isolate from melon, whereas the RG IV included 12 isolates from squash, cucumber, watermelon and melon. Amplification of internal transcribed spacer (ITS) region and small subunit rRNA region from the 20 isolates yielded respectively a single fragment. Restriction pattern with 12 restriction enzymes was identical for all isolates tested, suggesting that variation in the ITS and small subunit within the D. bryoniae were low. Amplification of the genomic DNAs of the tested isolates with the sequence characterized amplified regions (SCAR) primer RG IF-RG IR specific for RG I group resulted in a single band of 650bp fragment for 8 isolates out of the 20 isolates. Therefore, these 8 isolates could be assigned into RG I. The same experiments done with RG IIF-RG IIR resulted in no amplified PCR product for the 20 isolates tested. An about 1.4 kb-fragment amplified from the RG IV isolates was specifically hybridized with PCR fragments amplified from genomic DNAs of the RG IV isolates only, suggesting that this PCR product could be used for discriminating the RG IV isolates from the RG I isolates as well other fungal species.

• Journal of Microbiology and Biotechnology
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• v.25 no.9
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• pp.1401-1409
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• 2015

The aim of this study was to develop a SYBR Green-based real-time PCR assay for the rapid, specific, and sensitive detection of Acidovorax avenae subsp. citrulli, which causes bacterial fruit blotch (BFB), a serious disease of cucurbit plants. The molecular and serological methods currently available for the detection of this pathogen are insufficiently sensitive and specific. Thus, a novel SYBR Green-based real-time PCR assay targeting the YD-repeat protein gene of A. avenae subsp. citrulli was developed. The specificity of the primer set was evaluated using DNA purified from 6 isolates of A. avenae subsp. citrulli, 7 other Acidovorax species, and 22 of non-targeted strains, including pathogens and non-pathogens. The AC158F/R primer set amplified a single band of the expected size from genomic DNA obtained from the A. avenae subsp. citrulli strains but not from the genomic DNA of other Acidovorax species, including that of other bacterial genera. Using this assay, it was possible to detect at least one genomeequivalents of the cloned amplified target DNA using 5 × 10⁰ fg/µl of purified genomic DNA per reaction or using a calibrated cell suspension, with 6.5 colony-forming units per reaction being employed. In addition, this assay is a highly sensitive and reliable method for identifying and quantifying the target pathogen in infected samples that does not require DNA extraction. Therefore, we suggest that this approach is suitable for the rapid and efficient diagnosis of A. avenae subsp. citrulli contaminations of seed lots and plants.

• Development and Reproduction
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• v.20 no.4
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• pp.305-314
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• 2016

The marine medaka, Oryzias dancena is a suitable sample as a laboratory animal because it has a small size and clearly distinguishes between female and male. Data on the growth and maturity of the diploid and triploid sea cucurbit species suitable for laboratory animals are very useful for studying other species. Triploidy was induced in the marine medaka by cold shock treatment ($0^{\circ}C$) of fertilized eggs for 45 min, applied two minutes after fertilization. The diploid and triploid male fish were larger than their female counterparts (P<0.05), and the concentrations of thyroid stimulating hormone (TSH) and thyroxine (T4) were higher in the induced triploids over 1 year (P<0.05). In both the diploid and tri-ploid groups the concentrations of TSH and T4 were higher in the male fish than in the females (P<0.05), while the testo-sterone and estradiol-$17{\beta}$ concentrations in the induced triploids were lower than in the diploids (P<0.05). The gonadosomatic index (GSI) of the triploid fish was lower than that for the diploids, and the GSI for females in each ploidy group were higher than that for the males. For both groups the GSI was highest at 4 months of age, and decreased thereafter to 12 months. Analysis of the gonads of one-year-old triploid fish suggested that the induction of triploidy probably causes sterility in this species; this effect was more apparent in females than in males.

• Horticultural Science & Technology
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• v.35 no.1
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• pp.108-120
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• 2017

Modern watermelon cultivars (Citrullus lanatus [Thunb.] Matsum.& Nakai var. lanatus) have fruits with diverse phenotypes, including fruit shape, rind patterns, and flesh color. Molecular markers enable efficient selection of plants harboring desirable phenotypes. In the present study, publicly available DNA markers tightly linked to fruit shape, rind stripe pattern, and flesh color were evaluated using 85 watermelon accessions with diverse fruit phenotypes. For fruit shape, the dCAPS SUN - Cla011257 marker revealed an 81% of marker - trait match for accessions with elongated or round fruits. For rind stripe pattern, the SCAR wsb6-11marker was effective for selecting Jubilee-type rind pattern from other rind patterns. For flesh color, the Clcyb.600 and Lcyb markers derived from a mutation in the Lycopene ${\beta}$ - cyclase (Lcyb) gene, were effective at selecting red or yellow flesh. Forty-eight accessions possessing diverse fruit - related traits were selected as a reference array and their genetic relationships assessed using 16 SSR markers. At a coefficient of 0.11, the 48 accessions grouped into two major clades: Clade I and Clade II. Clade I subdivided further into subclades I - 1 and I - 2 at a coefficient of 0.39. All accessions with colored flesh were classified into Clade I, whereas those with white - flesh were classified into Clade II. Differences in fruit traits between subclades I - 1 and I - 2 were observed for rind pattern and fruit color; a majority of the accessions with Crimson-type striped or non-striped rind were grouped together in subclade I - 1, while most accessions in subclade I - 2 had a Jubilee - type rind stripe pattern. These results imply that reference array watermelon accessions possess distinguishable genetic structure based on rind stripe pattern. However, no significant grouping pattern was observed based on other fruit-related traits.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.148.2-149
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• 2003

Cucumber fruit mottle mosaic tobamovirus (CFMMV) causes severe mosaic symptoms with yellow mottling on leaves and fruits, and occasionally severe wilting of cucumber plants. No genetic source of resistance against this virus has been identified. The genes coding for the coat protein or the putative 54-kDa replicase were cloned into binary vectors under control of the SVBV promoter. Agrobacterium-mediated transformation was peformed on cotyledon explants of a parthenocarpic cucumber cultivar with superior competence for transformation. R1 seedlings were evaluated for resistance to CFMMV infection by lack of symptom expression, back inoculation on an alternative host and ELISA. From a total of 14 replicase-containing R1 lines, 8 exhibited immunity, while only 3 resistant lines were found among a total of 9 CP-containing lines. Line 144 homozygous for the 54-kDa replicase was selected for further resistance analysis. Line 144 was immune to CFMMV infection by mechanical and graft inoculation, or by root infection following planting in CFMMV-contaminated soil. Additionally, line 144 showed delay of symptom appearance following infection by other cucurbit-infecting tobamoviruses. Infection of line 144 plants with various potyviruses and cucumber mosaic cucumovirus did not break the resistance to CFMMV. The mechanism of resistance of line 144 appears to be RNA-mediated, however the means is apparently different from the gene silencing phenomenon. Homozygote line 144 cucumber as rootstock demonstrated for the first time protection of a non-transformed scion from soil inoculation with a soil borne pathogen, CFMMV.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.141.2-142
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• 2003

A flexuous rod-shaped virus was isolated from Cucurbita pepo leaves showing green mosaic and puckering symptoms at Anseong, Korea. Based on the biological tests, electron microscopy, and reverse transcription-polymerase chain reaction (RT-PCR), the isolate was identified as Papaya ringspot virus type Watermelon (PRSV-W). In the biological test, host range of PRSV-W was limited in the families Cucurbitaceae and Chenopodiaceae. Most susceptible cucurbit species, such as Cucurmis lanatus, Cucurmis sativus, Cucurbita pepo, and Citrullus lanatus, responded to mechanical inoculation by PRSV-W that induce green mosaic, malformation, puckering, and narrow laminae. The local lesion symptoms were produced on the inoculated leaves of Chenopodium maranticolor and C. quinoa PRSV specific primers which amplifies the part of the coat protein (CP) genes, generated a 648 bp product from 6 isolates of PRSV-W, but no amplification had been detected in other viruses including CMV, CGMMV, KGMMV, ZYMV and WMV. In electron microscopy, PRSV particles were flexuous, approximately 780 nm in length and 12 nm in width. PRSV-W is one of the worldwide viruses which has the great economic importance in cucumber, melon, squash, watermelon, and other cultivated cucurbits with ZYMV and WMV. This is the first report of PRSV-W on cucurbits in Korea.

• Horticultural Science & Technology
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• v.33 no.2
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• pp.210-218
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• 2015

Melon leaves showing yellowing symptoms were analyzed using electron microscopy and RT-PCR for major cucurbit-infecting-viruses (CMV, MNSV, CGMMV, SqMV, WMV, KGMMV, PRSV and ZYMV) reported in Korea, but these viruses were not detected. As the result of further analysis by next-generation sequencing (NGS), the virus was identified as Cucurbit aphid-borne yellows virus (CABYV), and then confirmed by RT-PCR using CABYV-specific primers. When photosynthetic capacity was measured based on chlorophyll fluorescence yield (ChlFY), the leaves of the diseased plants showed $4.09{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, which was one-third of the readings observed for unaffected normal plants ($12.36{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$). The root functions of plants affected by leaf yellowing symptoms (LYS) was $0.28mg{\cdot}g^{-1}$, about half that measured for the normal unaffected plants ($0.48mg{\cdot}g^{-1}$). Cytological observations revealed that there were no morphological differences in the palisade parenchyma and mesophyll spongy cells of the leaves between the diseased and the normal plants. However, the same leaf cells of the affected plants contained more starch granules compared to those of the normal, unaffected plants. We conclude that the LYS of muskmelon is not merely a physiological disorder but a viral disease caused by CABYV and spread by aphids.

• Journal of Bio-Environment Control
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• v.28 no.4
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• pp.438-446
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• 2019

The interest in mini watermelon (Citrullus lanatus) with small fruits weighing 2-3 kg has increased by the increasing trend in one-person households and consequent tendency to consume small meals. Watermelon grafting onto cucurbit rootstocks is a very effective way to control soil-borne diseases, such as Fusarium wilt; however, this practice negatively impacts the fruit quality. This study was conducted to investigate the growth, fruit set, and fruit quality of mini watermelon grafted onto wild watermelon accessions (Citrullus spp.) resistant to Fusarium wilt. Five watermelon accessions (Galactica, IT 208441, PI 482322, PI 500303, and PI 593358) were evaluated as rootstocks for the mini watermelon "Ministar". Non-grafted "Ministar" and "Ministar" grafted onto "Shintozwa" (Cucurbita maxima D. C. moschata D.) or "Bullojangsaeng" (Lagenaria leucantha) were used as controls. The roots of the transplants grafted onto "PI 593358" and "Shintozwa" weighed more than those on other rootstocks. Additionally, the transplants on "PI 593358" showed better growth and fruit set in the field than the other transplants. However, the total soluble solid contents and fruit quality indices of the transplants on "PI 593358" and "Shintozwa" were lower, whereas the total fruit quality index of those on "PI 482322" was higher. Thus, the wild watermelon accessions tested can potentially be used as basic germplasm for developing watermelon rootstocks instead of cucurbit rootstocks. The most promising accession for this purpose was found to be "PI 482322".

• Research in Plant Disease
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• v.28 no.2
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• pp.69-81
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• 2022

This study was performed to screen the efficacy of antagonistic bacterial isolates from various sources against the bacterial fruit blotch (BFB) causing pathogen (Acidovorax citrulli) in cucurbit crops. In addition, plant growth promoting traits of these antagonistic bacterial isolates were characterized. Two thousand seven hundred ninety-four microorganisms were isolated from the collected samples. Molecular identification revealed two A. citrulli out of 2,794 isolates. In vitro antagonistic results showed that, among the 28 antagonistic bacterial isolates, 24 and 14 bacterial isolates exhibited antagonism against HPP-3-3B and HPP-9-4B, respectively. Antagonistic and growth promotion characterization of the antagonistic bacterial isolates were further studied. Results suggested that, 4 antagonistic bacteria commonly showed both antagonism and growth promotion phenotypes. Moreover, 3 isolates possessed growth promoting activities. Overall results from this study suggests that BFB causing bacterial pathogen (A. citrulli) was suppressed in in vitro antagonism assay by antagonistic bacterial isolates. Furthermore, these antagonistic bacterial isolates possessed growth promotion and antagonistic enzyme production ability. Therefore, data from this study can provide useful basic data for the in vivo experiments which ultimately helps to develop the eco-friendly agricultural materials to control fruit rot disease in cucurbit crops in near future.