• The Plant Pathology Journal
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• v.35 no.4
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• pp.372-380
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• 2019

Pseudomonas syringae pv. actinidiae (Psa) is by far the most important pathogen of kiwifruit. Sustainable expansion of the kiwifruit industry requires the use of Psa-tolerant or resistant genotypes for the breeding of tolerant cultivars. However, the resistance of most existing kiwifruit cultivars and wild genotypes is poorly understood, and suitable evaluation methods of Psa resistance in Actinidia have not been established. A unique in vitro method to evaluate Psa resistance has been developed with 18 selected Actinidia genotypes. The assay involved debarking and measuring the lesions of cane pieces inoculated with the bacterium in combination with the observation of symptoms such as callus formation, sprouting of buds, and the extent to which Psa invaded xylem. Relative Psa resistance or tolerance was divided into four categories. The division results were consistent with field observations. This is the first report of an in vitro assay capable of large-scale screening of Psa-resistance in Actinidia germplasm with high accuracy and reproducibility. The assay would considerably facilitate the breeding of Psa-resistant cultivars and provide a valuable reference and inspiration for the resistance evaluation of other plants to different pathogens.

• Microbiology and Biotechnology Letters
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• v.24 no.4
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• pp.472-477
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• 1996

Natural human epidermal growth factor (nhEGF) was purified from pregnant human urine by benzoic acid adsorption, DEAE-Sepharose ion exchange, and immunoaffinity chromatography. The purified nhEGF was further separated into four fractions using Bondapak C$_{18}$ HPLC system. Following characterization by Western blot analysis and double immu- nodiffusion, we found that each fraction corresponds to four derivatives of the nhEGF. For biological analysis of nhEGF, we optimized the labeling time and serum concentration for the incorporatioin of 5-bromo-2'-deoxy uridine (BrdU), a non-radioactive alternative for [$^{3}$H]-thymidine uptake, into NIH 3T3 cells. The DNA synthesis of NIH 3T3 cells was gradually increased at the nhEGF concentrations between 0.1 - 10 ng/ml in the Dulbecco's Modified Eagles Medium (DMEM) containing 0.2% Fetal calf serum (FCS). When we assayed the biological activity of four fractions, the activity of the second fraction was superior to that of the others. Based on the results from the HPLC analysis spiked with recombinant human epidermal growth factor (rhEGF) and amino acid sequencing, we concluded that the second fraction was nhEGF and the other three fractions were the derivatives of nhEGF. In addition, the proportion of nhEGF was approximately 46% is compared with that of the other three derivatives.

• Korean Journal Plant Pathology
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• v.10 no.4
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• pp.297-304
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• 1994

This study was carried out to develop bioassay for detection of pesticide residues in agricultural products by using the soil microbial isolates sensitive to pesticides. One hundred bacterial isolates and eighty five fungal isolates were obtained from soil and their sensitivity to 10 ppm of several pesticides was examined in vitro. Five bacterial isolates and three fungal isolates were found sensitive to organochloride fungicide and two fungal isolates sensitive to organocopper fungicide. Among these isolates, B46, B93 and F67 were tested to find out the difference in sensitivity according to the methods of fungicide treatment. All of the isolates were found sensitive to 10 ppm of organochloride fungicides mixed directly in PDA. But they were found insensitive to the fungicide mixed in PDA after filtering through membrane filter. In case of organocopper fungicide, the isolates were found sensitive only when it was treated in PDA. And their sensitivity showed difference among various kinds of organochloride fungicides. B46 and B93 were employed to check the possibility as the agent for detection of the pesticidal residues in twenty eight agricultural products including rice. It was found that all samples had not residues because the samples did not inhibit the growth of isolates. When organochloride fungicides were applied to the above products, it was possible to detect the residues in fruits and vegetables at the concentration of 10 ppm, but not in starch-rich grains. B46 and B93 were identified as Bacillus sp. according to their bacterial characteristics in culture.

• Journal of Integrative Natural Science
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• v.7 no.3
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• pp.173-182
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• 2014

To simplify the different biological investigation of the methanolic extract and solvent-solvent partitioning of Lablab purpures (L. purpures) bark. In-vitro anti-oxidant study was determined using total DPPH radical scavenging assay. In vitro antimicrobial study was measured by observing zone of inhibition. The cytotoxic activity was studied using brine shrimp lethality bioassay and thrombolytic activity by clot disruption method. The antioxidant potential was evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and Folin-Ciocalteau reagents using butylated hydroxytolune (BHT) and ascorbic acid as standards. The Aqueous soluble fraction revealed the highest free radical scavenging activity ($IC_{50}=48.76{\mu}g/mL$). The antimicrobial screening of the bark of L. purpures exhibited mild to moderate activity in test microorganisms. The CSF showed the maximum relative percentage inhibition against Salmonella parathyphi (34.2%) for bacteria and C. albicans (28.8%) for fungi whereas, lowest relative percentage inhibition against Sarcina lutea (22.0%) for bacteria and Aspergillus niger (24.4%) for fungi. In the brine shrimp lethality bioassay, The $LC_{50}$ values of Carbon tetrachloride and N-Hexane soluble fraction were found $92.18{\mu}g/mL$, and $68.95{\mu}g/mL$ respectively while the $LC_{50}$ values of standard Vincristine sulphate was $1.37{\mu}g/mL$. The methanolic extract and its organic soluble fractions of Lablab purpureus at concentration 2.0 mg/mL, significantly protected the lysis of erythrocyte membrane induced by hypotonic solution and heat as compared to the standard, acetyl salicylic acid (0.10 mg/mL). The MSF and AQSF produced 61.48 % and 53.75% inhibition of hemolysis of RBC caused by hypotonic solution respectively, whereas acetyl salicylic acid (0.10 mg/mL) showed 76.42%. Ethanol extract of L. purpures and all of its different partitions exhibited moderate thrombolytic activity of 37.25%-2.40%. Very good preliminary screening and simplified experiments were able to show the different biological activity of methanolic extract and its soluble fractions of L. purpures at a time.

• Journal of Applied Biological Chemistry
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• v.48 no.4
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• pp.161-166
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• 2005

Among 24 isolates of fluorescent Pseudomonads, 5 isolates named as LPT1, LPT2, LPT3, LPT4 and LPT5 were screened in vitro for their nematicidal activity against cyst forming nematode, Heterodera cajani causing patchiness, poor and stunting growth besides discoloration in Sesamum indicum. Second stage juveniles of H. cajani hatched from egg masses were collected from roots of host plant and subjected to fresh and heat-treated culture filtrate of isolates for 24 h. Mortality of H. cajani was recorded on the basis of parameters used for test organism bioassay. Among these isolates, Pseudomonas aeruginosa LPT5 caused maximum mortality towards second stage juvenile of H. cajani in vitro. Five isolates were used as seed coating for the management of cyst forming nematode H. cajani on sesame in green house condition. The strains LPT5 was better than the other strains in reducing the population of H. cajani both in vitro and in vivo. The reduction in cyst and juveniles population was found to be 49 and 60%, respectively when seeds were coated with strain LPT5. Among other strains, LPT4 was also found to inhibit the cyst and juveniles population 12 and 36% respectively. Increases in early vegetative plant growth parameters recorded in both in vitro and in vivo further revealed the significance of indigenous bacteria in comparison to introduced strain.

• YAKHAK HOEJI
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• v.44 no.1
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• pp.22-28
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• 2000

The extracts of Euphoria longan, Ziryphus jujuba, Thuja orientalis, Polygala tenuifolia, Acorus gramineus, Cyperus rotundus, Poria cocos, Uncaria rhynchophylla, and Albizzia julibrissin, which have been used as sedative drugs in Korean folk medicine, were evaluated for their effects on neurotransmission and antioxidative system in vitro. Among the tested drugs, Acorus gramineus showed most inhibitory activities on monoamine oxidase, xanthine oxidase, aldehyde oxidase, and lipid peroxidation and Uncaria rhynchophylla also inhibited most effectively GABA transaminase and DPPH radical.

• Archives of Pharmacal Research
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• v.27 no.9
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• pp.915-918
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• 2004

By bioassay-guided separation, an already known saponin, Pulsatilla saponin D was isolated from the root of Pulsatilla koreana Nakai as a antitumor component when evaluated by in vivo antitumor activity as well as in vitro cytotoxic activity test. It showed potent inhibition rate of tumor growth (IR, 82％) at the dose of 6.4 mg/kg on the BDF1 mice bearing LLC cells.

• Korean Journal of Plant Resources
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• v.7 no.2
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• pp.115-119
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• 1994

These experiments were conducted to determine the effects of Sorghum allelopathic substances on the callus growh of several weeds and crops. 1. When substances extracted from allelopathic Sorghum(Sorghum bicolor L.) were treated on medium, growth of callus of several weeds and crops were in-hibited. The degree of inhibition differed depending on the genotypes, ranging from 50 to 90% com-pared with that of control. 2. The extracts of above 5% Sorghum inhibited the callus growth of Che-nopodium albun L., Commelina communis L., and .Ammaranthus retroflexus L.and showed in-hibition rate of above 70% in callus growth. These results indicate that we could investigate theallelopaihy effect by using in vitro system. 3. The suitable explant for callus induction fromallelopathic plants was immature embryos, the callus induction rate differed depending on the geno-type, growth regulators and concentrations. In general, the addition of 2, 4-D and NAA onto medium increased the rate and amount of callus.

• Journal of Sericultural and Entomological Science
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• v.17 no.2
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• pp.119-124
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• 1975

By means of in vitro studies, in which isolated suboesophageal of the Bombyx silkworm were cultured, it was shown that at least two Kinds of substances are biosynthesized and exert independent effects on determination of diapause or non-diapause in silkworm eggs. They are referred to as the diapause and non-diapause substance, respectively. Whether diapause or non-diapause eggs are laid may depend upon the different quality of these substances.

• Mycobiology
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• v.28 no.4
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• pp.190-192
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• 2000

Antifungal bacteria for biological control of plant diseases or production of novel antibiotics to plant pathogens were isolated in 1997 from various soils of Ansung, Chunan, Koyang, and Paju in Korea. Sixty-four bacterial strains pre-screened from approximately 1,400 strains were tested on V-8 juice agar against eight plant pathogenic fungi using in vitro bioassay technique for inhibition of mycelial growth. Test pathogens were Alternaria mali, Colletotrichum gloeosporioides, C. orbiculare, Fusarium oxysporum f. sp. cucumerinum, F. oxysporum f. sp. lycopersici, Magnaporthe grisea, Phytophthora capsici, and Rhizoctonia solani. A wide range of antifungal activity of bacterial strains was found against the pathogenic fungi, and strain RC-B77 showed the best antifungal activity. Correlation analysis between inhibition of each fungus and mean inhibition of all eight fungi by 64 bacterial strains revealed that C. gloeosporioides would be best appropriate for detecting bacterial strains producing antibiotics with potential as biocontrol agents for plant pathogens.