Rice bakanae disease, caused by the fungus Fusarium fujikuroi, is one of the most important rice diseases and distributed widely in Asia. Resistance screening system in rice field had been established. However, the evaluation results of the system vary according to the environmental conditions when the test is conducted. To develop precise and rapid evaluation method of disease resistance of rice to bakanae disease, in vitro screening system was attempted in this study. The six cultivars namely, 'Nampyeongbyeo', 'Junambyeo', 'Chucheongbyeo', 'Samcheonbyeo', 'Odaebyeo' and 'Hwasinbyeo' were tested. They were planted onto MS agar medium (10 ml) in test tube ($450\;mm{\times}{\phi}30\;mm$) and incubated at $25^{\circ}C$ and $28^{\circ}C$ in growth chamber under 12 hr light condition. Symptoms of over growth appeared a few days after seeding and then seedling were withered 2-3 weeks after over growth. The disease symptoms such as leaf dryness on top of rice were appeared in the 'Nampyeongbyeo' from 28 days at the concentration of $10^5$ spores/ml culturing at $28^{\circ}C$ and then withered completely 35 days after seeding. Whereas the other varieties withered entirely 19-23 days after seeding. Using the in vitro seedling screening method, 72 rice varieties were investigated to select resistant cultivar. Finally, two resistant cultivars ('Nampyeongbyeo' and 'Inwolbyeo') and seven moderately resistant cultivars ('Hwadongbyeo', 'Seokjeongbyeo', 'Samgwangbyeo', 'Sampyeongbyeo', 'Nonghobyeo', 'Heukjinjubyeo' and, 'Joanbyeo') were selected. If in vitro seedling screening method was used for evaluation of bakanae disease resistance, it would be completed within 35 days after sowing of rice seed.
This study was conducted to elucidate the cultural and pathogenic characteristics of Cladosporium cucumerinum PT1 and resistance of 81 commercial cucumbers (Cucumis sativus). Cucumber leaves and fruits appeared as scab were collected from a plastic film house located in Pyeongtaek, Gyeonggi-province, Korea in late March, 2015. A casual fungus was isolated from the diseased fruits on potato dextrose agar and it was identified as C. cucumerinum PT1 based on the morphological characteristics. To find out the effect of wounding and fruit size on the development of cucumber scab, small (<10 cm long), medium (10 to 20 cm long), and large (>20 cm long, commercially mature fruit) size cucumber fruits were harvested, C. cucumerinum PT1 pathogens were inoculated with a single droplet of suspension ($1{\times}10^5$ spores/ml) on wounded or unwounded cucumber fruits. Small fruits were completely damaged with showing severe water-soaking symptoms and fast pathogen growth regardless of wounded or unwounded. Meanwhile slight water-soaking symptoms on medium and large size fruits occurred and disease development into plant tissues was observed only on wounded fruits. Disease resistance of 81 commercial cucumber cultivars was evaluated on third-stage seedlings and small fruits by inoculating suspension ($1{\times}10^5$ spores/ml) of C. cucumerinum PT1. As a result, mini and pickling cultivar groups were resistant, 'Cheoeumcheoreom' cultivar was symptomless and the other cultivars were resistant to medium resistant. On the other hand, most of cucumber cultivars belonging to the other groups were susceptible. Disease resistance of cucumber against cucumber scab was significantly different among cultivars and a few cucumber cultivars showed different disease resistant responses to two bioassay methods using seedlings and small fruits. Therefore, to screen scab resistance in cucumber, a test using both fruits and seedlings is advisable. We think that the selected resistant cultivars can be used to control cucumber scab effectively under the farmhouse condition.
Neunghwaji(Embossed patterned paper) is a unique paper used for a traditional book cover in Korea. The research was carried out to investigate Neunghwaji's features. Physical property was studied through a test of tensile strength and folding endurance. Also, comparative analysis of virus resistance and waterproof ability was undertaken on Neunghwaji. 1. Folding endurance test showed that strength of non-embossed CB and HB decreased during deteriorating duration. Embossed CN and HN showed the strength increasing at early stage and decreasing from the 27th day of the deteriorating duration. Tensile strength was decreasing in both cases as deterioration progressed. 2. Growth of Arthrinium sp. fungus on embossed paper was 10% less than plain paper while Cladosporium sp. showed 20~30% less growth. Amur cork dyeing(H) showed 10~30% lower fungi growth than Gardenia seed dyeing(C). The result indicated that embossed paper has better virus resistance than Hanji, and Amur cork dyeing has better virus resistance than Gardenia seed dyeing. 3. Average contact angle of CN, CB, HN, and HB was $85{\sim}92^{\circ}$ and NON-N and NON-B was $59{\sim}63^{\circ}$. In detail, CN's contact angle was $1{\sim}7^{\circ}$ higher than CB's; HN was $1{\sim}6^{\circ}$ higher than HB. Therefore, it was found that embossed paper has higher contact angle than Hanji thus the former has better waterproof ability. The research suggested production technique of Neunghwaji and studied its features related to the technique. Neunghwaji was confirmed to have superior quality to Hanji though further study regarding above test result is needed to complement the research.
Effects of silicon application on development of colonies of Sphaerotheca fuliginea were examined. Cucumber plants were applied with nutrient solutions amended with different concentrations of soluble silicon and selected leaves were inoculated with known concentrations of conidia of the pathogen. Colony number per leaf, colony area per leaf, and germination rate of conidia of S. fuliginea collected from the inoculated leaves were reduced as silicate concentrations in the nutrient solutions increased from 0.05 to 4.10 mM. The increase in resistance of plants to mildew infection was apparently due to silicate accumulation in leaves, and there was no correlation between cation or ionic strength effects and the silicate treatments. Silicate treatment in growth medium remarkably suppressed powdery mildew development on cucumber. Colonies of mildew fungus were visible with over approx. 38.3% of the mature leaf surface, while that of the leaves in high Si plants was 2.3% observed at 51 days after transplanting. No significant differences were observed between 1.7 mM and 3.4mM silicate treatments. Conidial germination rates were significantly reduced by increasing Si amendments. Conidial germination ranged from 14.7 to 20.3% for plants grown in low Si solution(<1.40 mM), and from 9.0 to 12.4% for plants grown in high Si solution(>1.8 mM). Foliar applications of Si with ${\geq}$ 17.0 mM decreased the number of powdery mildew colonies. Persistence of Si foliar sprays effects on cucumber demonstrated that the 17 mM Si spray applied 4 days before inoculation with S. fuliginea reduced mildew colony formation. The relationship was positive and linear.
The study was carried out to develop transformants resisting to Phyophthora blight disease in the domestic pepper cultivar Subicho. In transforming of syn600 promoter with elicitin gene using Agrobacterium (LBA4404/pBI101 syn600-syn${\alpha}$-elicitin) to cotyledons of pepper, rate of shoot formation in 'Subicho' was 11.1% in medium containing 3 mg/L zeatin and 0.05 mg/L NAA, and also 12.8% in medium containing combination of 4 mg/L zeatin and 0.05 mg/L MAA. For PCR reaction using elicitin gene primer of transformants regenerated from cotyledons, we detected a specific band of 536 bp, and also showed strong signal at position of 536 bp in accordance with NPTII gene used as probe in Southern blot. Transformants pepper shown resistance to blight fungus was inoculated to seedlings of the $T_{1}\;and\;T_{2}$ transformants by concentration (density: zoo spore $10^{3}/mL$).
Journal of the Korean Society of Environmental Restoration Technology
/
v.5
no.2
/
pp.53-62
/
2002
The objectives of this study were to understand the tolerance mechanism of woody plants to water stress and tolerance changes in relation to mycorrhizal formation. Lespedeza cyrtobotrya Miq. commonly used for erosion control in slopes were raised from seeds and transplanted to 120 plastic pots. Sixty pots received the top soil of a Fraxinus americana forest, while remaining 60 pots received the autoclaved top soil. The forest soil contained 1,200 spores per 100g of arbuscular endomycorrhizal fungus, mostly Glomus sp. The plants were raised outside with regular supply of water and mineral nutrients. Two kinds of water deficit treatment and a control were started at the middle of July : cyclic water deficit treatment with 3 cycles of sequential water stress at the point of xylem water potential of about -0.6, -0.6, and -1.7 MPa and recovery, and non-cyclic water deficit treatment with single water stress at about -1.5 MPa. The non-stressed plants received plenty of water throughout the period. In late August the plants were harvested for measurements of dry weight, N, P, carbohydrate contents, net photosynthesis and superoxide dismutase(SOD) activities. Both cyclic and non-cyclic water deficit treatments reduced dry weight by 60% and 40%, respectively, and reduced nitrogen absorption, while increased SOD activities. Water-stressed plants also showed increased carbohydrate contents in the leaves and lowered stomatal conductance. Mycorrhizal inoculation resulted in an average of 40% infection of roots and 2-3 times increase in P absorption in water-stressed as well as non-stressed plants. Mycorrhizal formation also increased shoot-root ratio. The results that SOD activities of water-stressed plants with mycorrhizal infection were significantly lower than those of non-mycorrhizal plants suggest the possibility of improvement of water-stressed condition by mycorrhizal formation. It was concluded that endomycorrhizal formation increased tolerance of Lespedeza cyrtobotrya seedlings to water stress.
The inky cap, Coprinellus congregatus, produces several laccase isozymes during its life cycle: both hyphal tip laccase and sclerotial laccase are involved in the fungal development. When this fungus was transferred to an acid liquid medium (pH 4.0-4.5), a new laccase was synthesized and secreted into the culture supernatant. In order to examine its regulation by external pH, green fluorescent protein gene was ligated at the downstream of the promoters having different lengths. These expression vectors having different promoter lengths were inserted into the fungal transformation vector, pBARGEM7-1. These expression vectors were introduced to the mating type a1 and a2 monokaryons, and the transformants were selected by the phosphinothricin resistance. Transformant a1 (a1TF) and transformant a2 (a2TF) were mated with each other to generate homozygotic dikaryon transformants. All these transformants were grown in neutral liquid medium for 5 days, and then the whole cell homogenates were transferred to the acidic liquid medium (pH 4.1). After 36 h incubation at $25^{\circ}C$, cells were harvested for the analysis of GFP expression. GFP expression was detected in the transformant having full-length promoter (2.0 kb), but other transformants having shorter length promoter (shorter than 1.29 kb) failed to show the fluorescence. Therefore, the acid-responsive element in the laccase promoter should be localized between -2.0 kb ~ -1.29 kb region.
Park, Sook-Young;Chi, Myoung-Hwan;Milgroom, Michael G.;Kim, Hyo-Jung;Han, Seong-Sook;Kang, Seog-Chan;Lee, Yong-Hwan
The Plant Pathology Journal
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v.26
no.4
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pp.313-320
/
2010
Genetic instability of the rice blast fungus Magnaporthe oryzae has been suggested as a major factor underlying the rapid breakdown of host resistance in the field. However, little information is available on the mechanism of genetic instability. In this study, we assessed the stability of repetitive DNA elements and several key phenotypic traits important for pathogenesis after serially transferring two isolates though rice plants and an artificial medium. Using isolate 70-15, we obtained a total of 176 single-spore isolates from 10 successive rounds of culturing on artificial medium. Another 20 isolates were obtained from germ tubes formed at the basal and apical cells of 10 three-celled conidia. Additionally, 60 isolates were obtained from isolate KJ201 after serial transfers through rice plants and an artificial medium. No apparent differences in phenotypes, including mycelial growth, conidial morphologies, conidiation, conidial germination, appressorium formation, and virulence, or in DNA fingerprints using MGR586, MAGGY, Pot2, LINE, MG-SINE and PWL2 as probes were observed among isolates from the same parent isolate. Southern hybridization and sequence analysis of two avirulence genes, AVR-Pita1 and AVR-Pikm, showed that both genes were also maintained stably during 10 successive generations on medium and plants. However, one reversible loss of restriction fragments was found in the telomere-linked helicase gene (TLH1) family, suggesting some telomere regions may be more unstable than the rest of the genome. Taken together, our results suggest that phenotype and genotype of M. oryzae isolates do not noticeably change, at least up to 10 successive generations on a cultural medium and in host plants.
Lee, Moran;Jeong, Hyeju;Kim, Jaeyoon;Kim, Dayeon;Ahn, Seung Ho;Lee, SangYeob;Han, Ji Hee
The Korean Journal of Mycology
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v.46
no.4
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pp.479-490
/
2018
Red pepper is seriously damaged by thrips (Thrips palmi) and anthracnose caused by Colletotrichum acutatum throughout its development. Because of biotic constraints, producers often depend on chemicals that are expensive and have adverse effects on the environment, operator, and beneficial insects. In addition, resistance is developed because of the repeated use of chemicals. In recent decades, the use of microorganisms in crop protection has become a credible alternative because it is eco-friendly. In this study, we aimed to select isolates with insecticidal and fungicidal activities against the pathogens that cause anthracnose and thrips. We treated T. palmi adults and juveniles with 13 strains of entomopathogenic fungi (isolated from the soil by using the insect-bait method), and 6 strains showed excellent insecticidal activity (70-100%) 5 days after the treatment. The selected isolates were cultured with C. acutatum to screen for the strain with excellent anti-fungal activities, among which an isolate FT333 showed more than 95% control efficacy against C. acutatum in vitro. The isolate was identified as Isaria javanica through its morphological characteristics and phylogenetic analysis of the ITS and ${\beta}-tubulin$ nucleotide sequences. The Isaria javanica FT333 isolate could be used effectively for dual bio-control of thrips and anthracnose during red pepper cultivation.
The soilborne fungus Fusarium oxysporum f. sp. lilii (Fol) is a serious threat to all lily cultivars, especially infecting bulbs and flowers. It has become increasingly important to develop varieties resistant against the bulb rot disease. Genetic diversity of cultivars and reliable screening methods are required for this purpose. Here, an efficient in vitro screening system for evaluating resistance to Fol in 38 in vitro-grown lily plants was investigated. Various factors including culture conditions of Fol, inoculum density, appropriate plant materials, inoculation method and duration, and incubation period of plant materials after inoculation were combined to optimize the screening method. As a result, we suggest optimal conditions for an in vitro screening system for the selection of Fol-resistant lily cultivars as follows. Fol was grown on potato dextrose agar (PDA) medium for 6 days at $25^{\circ}C$ in darkness and used as working inoculation. Spore suspensions were prepared (inoculum density: $1.0{\times}10^4$$spores{\cdot}mL^{-1}$), and then leaf segments $1.5{\times}2.0cm^2$ were inoculated by dipping for 22 hours at $25^{\circ}C$ in dark. Later, leaves were cultured on 0.6% agar plates at $25^{\circ}C$ and 50% humidity with a photoperiod of 16 hours light/8 hours dark (fluence rate of $40{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) to examine the progress of bulb rot. After 7 days, disease levels were classified into indices 1 (no symptom) to 6 (serious bulb rot). Soil inoculation of Fol carried out with resistant or susceptible lily cultivars that had been selected through in vitro screening confirmed the reproducibility of results. Therefore, the in vitro screening method established in this study is efficient and reliable for selection of lily cultivars resistant against bulb rot disease.
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