• Title/Summary/Keyword: Bioassay methods

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Screening Methods for Resistant Cucumber Cultivars against Cucumber Scab Caused by Cladosporium cucumerinum Using Cucumber Fruits and Seedlings (오이 유묘와 과실을 활용한 검은별무늬병에 대한 저항성 품종 검정 방법)

  • Park, So-Hyang;Hong, Sung-Jun;Shim, Chang-Ki;Kim, Min-Jeong;Park, Jong-Ho;Han, Eun-Jung;Park, Jong-Won;Jee, Hyeong-Jin;Kim, Seok-Cheol;Kim, Yong-Ki
    • Research in Plant Disease
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    • v.22 no.1
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    • pp.18-24
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    • 2016
  • 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.

Studies on the Bioavailable Amino Acid of Feather Meals Processed by Different Methods - Available Amino Acid on the Meal Feeding of Semipurified and Purified Diet with Chick - (가공방법을 달리한 우모분의 아미노산 이용율에 관한 연구(I) -순수사료와 준순수사료의 Meal Feeding 하에서의 아미노산 이용율 -)

  • 김대진
    • Korean Journal of Poultry Science
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    • v.14 no.2
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    • pp.103-108
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    • 1987
  • This study was conducted to bioassay of amino acid availability of feather meal processed by a different methods, commercial feather meal and raw feather meal, The feather meals were processed by labolatory pressure cooker(autoclave) at 2kg/$\textrm{cm}^2$ for 30 minutes ; 3kg/$\textrm{cm}^2$ for 90 minutes : 4kg/$\textrm{cm}^2$ for 120 minutes. Chick employed in the present experiment were Abor Acre strain, male or meat type (body weight, 100-140g), fed with semipurified diet and protein free diet was given during the determination of the metabolic and endogeneous amino acid. The contents of amino acid of samples were investigated by ion-exchange chromatography. The results were as follows; 1. The amino acid availability of raw, 2kg/$\textrm{cm}^2$ for 30 minutes, commercial, 4kg/$\textrm{cm}^2$ for 120 minutes and 3kg/$\textrm{cm}^2$ for 90 minutes of feather meal were -3.09, 63.28, 67.47, 71.22 and 73.75% respectively. 2. The essential amino acid availability of raw, 2kg/$\textrm{cm}^2$ for 30 minutes, commercial, 4kg/$\textrm{cm}^2$ for 120 minutes and 3kg/$\textrm{cm}^2$ for 90 minutes of feather meal were 2.55, 66.78, 66.89, 72,56 and 73.62%, respectively. 3. In individual amino acid of the different processing loather meal and commercial feather meal, biovailabilities were increased methionine, phenylalanine, leucine, arginine, threonine, isoleucine, however, histidine, lysine and aspartic acid were remarkely decreased. In conclusion, the bioavailability or amino acid for the feather meal processed at 3kg/$\textrm{cm}^2$ for 90 minutes was superior to those of other treatment or raw feather meal.

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Antioxidant, Physiological Activities, and Acetylcholinesterase Inhibitory Activity of Portulaca oleracea Extracts with Different Extraction Methods (추출방법에 따른 쇠비름의 항산화, 생리활성 및 Acetylcholinesterase 저해활성)

  • Kwon, Yu-Ri;Cho, Sung-Mook;Hwang, Seung-Pil;Kwon, Gi-Man;Kim, Jae-Won;Youn, Kwang-Sup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.3
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    • pp.389-396
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    • 2014
  • The physiological properties of 70% ethanol extracts from Portulaca oleracea with different extraction methods (reflux extraction, RE; autoclave extraction, AE; low temperature high pressure extraction, LTPE) were investigated. The freeze-dried powder yields of RE, AE, and LTPE were 33.78%, 30.80%, and 11.05%, respectively. The color values of L and b were higher in LTPE, and the chroma values were higher in AE and LTPE compared to RE. The total polyphenolics and proanthocyanidin contents in LTPE were significantly higher than in other extracts. The amount of substances related to flavonoids contents was highest in RE (4.30 mg/g), followed by AE (4.06 mg/g), and LTPE (4.00 mg/g). DPPH radical scavenging ability with a concentration of 500 mg% (w/v) were in the following order; LTPE (88.87%)> RE (83.84%)> AE (80.67%). Further, the reducing power, ABTS radical scavenging ability, and nitrite scavenging activity was observed in the same tendency as seen with the DPPH radical scavenging ability. However, the ferrous ion chelating activity of RE (85.45%) and AE (83.88%) was significantly higher than that of LTPE (75.60%). ${\alpha}$-Glucosidase inhibitory activities of RE and LTPE with a concentration of 100 mg% were significantly higher than AE. Xanthine oxidase, and acetylcholinesterase inhibitory activities of LTPE were higher than the other extracts. These results suggest that the extracts from Portulaca oleracea have the potential to act as functional materials, and components of Portulaca oleracea could be effective in the prevention of Alzheimer's disease, and may be used to develop various functional food products.

Using Trophic State Index (TSI) Values to Draw Inferences Regarding Phytoplankton Limiting Factors and Seston Composition from Routine Water Quality Monitoring Data (영양상태지수 (trophic state index)를 이용한 수체 내 식물플랑크톤 제한요인 및 seston조성의 유추)

  • Havens, Karl E
    • Korean Journal of Ecology and Environment
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    • v.33 no.3 s.91
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    • pp.187-196
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    • 2000
  • This paper describes a simple method that uses differences among Carlson's (1977) trophic state index (TSI) values based on total phosphorus (TP), chlorophyll a (CHL) and Secchi depth (SD) to draw inferences regarding the factors that are limiting to phytoplankton growth and the composition of lake seston. Examples are provided regarding seasonal and spatial patterns in a large subtropical lake (Lake Okeechobee, Florida, USA) and inter- and intra-lake variations from a multilake data set developed from published studies. Once an investigator has collected routine water quality data and established TSI values based on TP, CHL, and SD, a number of inferences can be made. Additional information can be provided where it also is possible to calculate a TSI based on total nitrogen (TN). Where TSI (CHL)<>TSI (SD), light attenuating particles are large (large filaments or colonies of algae), and the phytoplankton may be limited by zooplankton grazing. Other limiting conditions are inferred by different relationships between the TSI values. Results of this study indicate that the analysis is quite robust, and that it generally gives good agreement with conclusions based on more direct methods (e.g., nutrientaddition bioassays, zooplankton size data, zooplankton removal experiments). The TSI approach, when validated periodically with these more costly and time-intensive methods, provides an effective, low cost method for tracking long-term changes in pelagic structure and function with potential value in monitoring lake ecology and responses to management.

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Action properties and insecticidal effects of thiamethoxam to the melon aphid, Aphis gossypii, and diamondback moth, Plutella xylostella (목화진딧물과 배추좀나방에 대한 thiamethoxam의 살충효과 및 작용특성)

  • Jang, Cheol;Hwang, In-Cheon;Yu, Yong-Man;Choe, Kwang-Ryul
    • The Korean Journal of Pesticide Science
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    • v.2 no.3
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    • pp.126-136
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    • 1998
  • For the purpose of effective control strategy of the melon aphid, Aphis gossypii and the diamondback moth, Plutella xylostella, thiamethoxam and 3 other insecticides in different classes were used with bioassay test methods in laboratory and greenhouse. They were examined to evaluated and compared with contact toxicity, stomach toxicity, rapid action, systemic action, and residual effect of imidacloprid, thiamethoxam (nicotinoids), acephate (organophosphorates), and carbosulfan (carbamates). As results of contact toxicity responses of A. gassypii against 4 insecticides using a spray application method, $LC_{50}$ values of acephate, carbosulfan, imidacloprid and thiamethoxam were 41.9, 5.2, 1.1, and 0.7 ppm. respectively. In the evaluation of stomach toxicity response of P. xylostella using a leaf-dipping method, with the 2nd instar larva $LC_{50}$ values of imidacloprid, thiamethoxam and acetamiprid were 64.9, 24.6 and 15.2 ppm, with the 3rd instar larva were 125.2, 42.7 and 27.8 ppm. and with the 4th instar larva were 241.1, 44.5 and 23.9 ppm, respectively. In the case of rapid action to A. gossypii using a spray application method after inoculation, $LT_{50}$ values of imidacloprid, thiamethoxam, carbosulfan, and acephate were 26.6, 28.0, 30.3, and 41.7 min. respectively. Otherwise, in the inoculation after applying compounds, $LT_{50}$ values of thiamethoxam, imidacloprid, and carbosulfan were 95.5, 118.0, and 122.9 min. respectively. Evaluating to systemic action from the abaxial surface to the adaxial surface of red pepper leaf with spray method, $LT_{50}$ values of thiamethoxam, imidacloprid, and carbosulfan were 162.2, 168.9, and 564.1 min. respectively. For the systemic action from the lower leaves to the upper leaves on red pepper, $LT_{50}$ values of carbosulfan, thiamethoxam, imidacloprid, and acephate were 2.3, 2.9, 3.0, and 8.8 days, respectively. In red pepper plant, $LT_{50}$ values of carbosulfan, imidacloprid, thiamethoxam, and acephate on the systemic action from the roots to the upper leaf were 0.6, 1.0, 1.0, and 13.8 days, respectively. As these results, it might be that thiamethoxam was excellent on systemic effect in red pepper. For the evaluation of residual effect on red pepper with A. gossypii, thiamethoxam and imidacloprid maintained high control effects as over 80% upto 10 days after treating compounds.

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Insecticidal effect of imidacloprid to sucking, chewing insect pests, and predacious spiders (흡즙성 및 저작성 해충과 거미류에 대한 imidacloprid의 살충효과)

  • Choi, Byung-Ryul;Lee, Si-Woo;Song, Yoo-Han;Yoo, Jai-Ki
    • The Korean Journal of Pesticide Science
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    • v.3 no.3
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    • pp.60-67
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    • 1999
  • Insecticidal activities of imidacloprid to sucking type insect pests, brown planthopper (BPH, Nilaparvata lugens) and peen peach aphid (GPA Myzus persicae), to chewing type insect pests, tobacco cut worm (TCW; Spodoptera litura) and beet armyworm (BAW, Spodoptera exigua) and to spiders, Pirata subpiraticus, Pachygnata clercki and Ummeliata insecticeps, as natural enemies were investigated by several bioassay methods. $LD_{50}$ of the chemical by topical treatment to BPH was 0.015 ${\mu}g/g$(48 hrs), while $LC_{50}s$ by leaf dipping and root zone treatment were 18.1 and 21.5 ppm, respectively. There was no difference in insecticidal activities between leaf dipping and root zone treatment. Imidacloprid also showed ovicidal effect of root zone treatment and its $LC_{50}$ was 6.8ppm $LD_{50}$ (48 hrs after treatment) of imidacloprid to GPA was $0.4{\mu}g/g$ in case of topical application and $LC_{50}s$ by leaf dipping and root zone treatment were 1.9 ppm and 13.7 ppm respectively. Leaf dipping was more effective than root zone treatment in GPA At topical application $LD_{50}$ (48 hrs after treatment) of imidacloprid to chewing type insect pests, TCW and BAW, were greater than 1,492 and $312{\mu}g/g$ and $LC_{50}s$ by leaf dipping method were 4,803 and Heater than 5,000ppm respectively. This means that imidacloprid has much less effect on chewing type insect pests, TCW and BAW. $LD_{50}$(48 hrs after treatment) of imidacloprid to wandering spiders, Pirata subpiraticus, Pachygnata clercki at topical application were greater dan $2941{\mu}g/g$ and greater than $2,976{\mu}g/g$ respectively and that to webbing spider, Ummeliata insecticeps, was 357 ${\mu}g/g$. Imidacloprid showed very low toxicity to the spiders and its selective toxicity ratios between spiders and BPH were greater than 19,600, greater than 19,800 and 23,800, respectively.

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Development of Efficient Screening Methods for Melon Plants Resistant to Fusarium oxysporum f. sp. melonis (멜론 덩굴쪼김병에 대한 효율적인 저항성 검정법 개발)

  • Lee, Won Jeong;Lee, Ji Hyun;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Heung Tae;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.70-82
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    • 2015
  • This study was conducted to establish an efficient screening system to identify melon resistant to Fusarium oxysporum f. sp. melonis. F. oyxsporum f. sp. melonis GR was isolated from infected melon plants collected at Goryeong and identified as F. oxysporum f. sp. melonis based on morphological characteristics, molecular analyses, and host-specificity tests on cucurbits including melon, oriental melon, cucumber, and watermelon. In addition, the GR isolate was determined as race 1 based on resistance responses of melon differentials to the fungus. To select optimized medium for mass production of inoculum of F. oxysporum f. sp. melonis GR, six media were tested. The fungus produced the most spores (microconidia) in V8-juice broth. Resistance degrees to the GR isolate of 22 commercial melon cultivars and 6 rootstocks for melon plants were investigated. All tested rootstocks showed no symptoms of Fusarium wilt. Among the tested melon cultivars, only three cultivars were susceptible and the other cultivars displayed moderate to high resistance to the GR isolate. For further study, six melon cultivars (Redqueen, Summercool, Superseji, Asiapapaya, Eolukpapaya, and Asiahwanggeum) showing different degrees of resistance to the fungus were selected. The development of Fusarium wilt on the cultivars was tested according to several conditions such as plant growth stage, root wounding, dipping period of roots in spore suspension, inoculum concentration, and incubation temperature to develop the disease. On the basis of the test results, we suggest that an efficient screening method for melon plants resistant to F. oxysporum f. sp. melonis is to remove soil from roots of seven-day-old melon seedlings, to dip the seedlings without cutting in s pore s uspension of $3{\times}10^5conidia/mL$ for 30 min, to transplant the inoculated seedlings to plastic pots with horticulture nursery media, and then to cultivate the plants in a growth room at 25 to $28^{\circ}C$ for about 3 weeks with 12-hour light per day.

Development of Marine Ecotoxicological Standard Methods for Ulva Sporulation Test (파래의 포자형성률을 이용한 해양생태독성시험 방법에 관한 연구)

  • Han, Tae-Jun;Han, Young-Seok;Park, Gyung-Soo;Lee, Seung-Min
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.13 no.2
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    • pp.121-128
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    • 2008
  • As an aquatic ecotoxicity test method, a bioassay using the inhibition of sporualtion of the green macroalga, Ulva pertusa, has been developed. Optimal test conditions determined for photon irradiance, pH, salinity and temperature were $100\;{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, $7{\sim}9$, $25{\sim}35\;psu$ and $15{\sim}20^{\circ}C$, respectively. The validity of the test endpoint was evaluated by assessing the toxicity of four metals (Cd, Cu, Pb, Zn) and elutriates of sewage or waste sludge collected from 9 different locations. When the metals were assayed, the $EC_{50}$ values indicated the following toxicity rankings: Cu ($0.062\;mg{\cdot}L^{-1}$) > Cd ($0.208\;mg{\cdot}L^{-1}$) > Pb ($0.718\;mg{\cdot}L^{-1}$) > Zn ($0.776\;mg{\cdot}L^{-1}$). When compared with other commonly used bioassays of metal pollution listed on US ECOTOX database, the sporualtion test proved to be the most sensitive. Ulva sporulation was significantly inhibited in all elutriates with the greatest and least effects observed in elutriates of sludge from industrial waste ($EC_{50}=6.78%$) and filtration bed ($EC_{50}=15.0%$), respectively. The results of the Spearman rank correlation analysis for $EC_{50}$ data versus the concentrations of toxicants in the sludge presented a significant correlation between toxicity and four heavy metals(Cd, Cu, Pb, Zn). The method described here is sensitive to toxicants, simple to use, easy to interpret and economical. It is also easy to procure samples and maintain cultures. The present method would therefore probably make a useful assessment of aquatic toxicity of a wide range of toxicants. In addition, the genus Ulva has a wide geographical distribution and species have similar reproductive processes, so the test method would have a potential application worldwide.

Lipopolysaccharide-induced Synthesis of IL-1beta, IL-6, TNF-alpha and TGF-beta by Peripheral Blood Mononuclear Cells (내독소에 의한 말초혈액 단핵구의 IL-1beta, IL-6, TNF-alpha와 TGF-beta 생성에 관한 연구)

  • Jung, Sung-Hwan;Park, Choon-Sik;Kim, Mi-Ho;Kim, Eun-Young;Chang, Hun-Soo;Ki, Shin-Young;Uh, Soo-Taek;Moon, Seung-Hyuk;Kim, Yang-Hoon;Lee, Hi-Bal
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.846-860
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    • 1998
  • Background: Endotoxin (LPS : lipopolysaccharide), a potent activator of immune system, can induce acute and chronic inflammation through the production of cytokines by a variety of cells, such as monocytes, endothelial cells, lymphocytes, eosinophils, neutrophils and fibroblasts. LPS stimulate the mononucelar cells by two different pathway, the CD14 dependent and independent way, of which the former has been well documented, but not the latter. LPS binds to the LPS-binding protein (LBP), in serum, to make the LPS-LBP complex which interacts with CD14 molecules on the mononuclear cell surface in peripheral blood or is transported to the tissues. In case of high concentration of LPS, LPS can stimulate directly the macrophages without LBP. We investigated to detect the generation of proinflammatory cytokines such as interleukin 1 (IL-1), IL-6 and TNF-$\alpha$ and fibrogenic cytokine, TGF-$\beta$, by peripheral blood mononuclear cells (PBMC) after LPS stimulation under serum-free conditions, which lacks LBPs. Methods : PBMC were obtained by centrifugation on Ficoll Hypaque solution of peripheral venous bloods from healthy normal subjects, then stimulated in the presence of LPS (0.1 ${\mu}g/mL$ to 100 ${\mu}g/mL$ ). The activities of IL-1, IL-6, TNF, and TGF-$\beta$ were measured by bioassaies using cytokines - dependent proliferating or inhibiting cell lines. The cellular sources producing the cytokines was investigated by immunohistochemical stains and in situ hybridization. Results : PBMC started to produce IL-6, TNF-$\alpha$ and TGF-$\beta$ in 1 hr, 4 hrs and 8hrs, respectively, after LPS stimulation. The production of IL-6, TNF-$\alpha$ and TGF-$\beta$ continuously increased 96 hrs after stimulation of LPS. The amount of production was 19.8 ng/ml of IL-6 by $10^5$ PBMC, 4.1 ng/mL of TNF by $10^6$ PBMC and 34.4 pg/mL of TGF-$\beta$ by $2{\times}10^6$ PBMC. The immunoreactivity to IL-6, TNF-$\alpha$ and TGF-$\beta$ were detected on monocytes in LPS-stimulated PBMC. Some of lymphocytes showed positive immunoreactivity to TGF-$\beta$. Double immunohistochemical stain showed that IL-1$\beta$, IL-6, TNF-$\alpha$ expression was not associated with CD14 postivity on monocytes. IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$mRNA expression were same as observed in immunoreactivity for each cytokines. Conclusion: When monocytes are stimulated with LPS under serum-free conditions, IL-6 and TNF-$\alpha$ are secreted in early stage of inflammation. In contrast, the secretion of TGF-$\beta$ arise in the late stages and that is maintained after 96 hrs. The main cells releasing IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$ are monocytes, but also lymphocytes can secret TGF-$\beta$.

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