• Research in Plant Disease
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• v.13 no.2
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• pp.93-97
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• 2007

The microorganisms with the antifungal activity against Phytophthora capsici and Colletotrichum acutatum and the plant growth promotion activity were screened from forest soils of Moon-gyeong (Juheul Mountain), Gyeongsangbuk-do. One of the isolates, strain MG 121 showed antifungal activity against P. capsici and C. acutatum and possessed phosphate solubilization activity was selected to development biocontrol agent. The strain MG 121 was identified as Streptomyces sp. by analysis of 16S rDNA. On the test with pepper fruits, the strain inhibited disease incidences of late blight and anthracnose over 80%. In greenhouse test, plant height, the number of leaf, fresh weight and roots length of pepper plants upon treatment of culture suspension of Streptomyces sp. MG 121 were significantly higher than those without the bacterial cells. In addition, strain MG 121 was capable to solublize rock-phosphate after incubation for 144 hours in potato dextrose broth. The concentration of soluble phosphate in PDB amended with 0.5% rock-phosphate was increased up to $765{\mu}g/ml$.

• Plant Biotechnology Reports
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• v.2 no.2
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• pp.163-169
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• 2008

Roots of Ophiorrhiza prostrata D. Don serve as a rich source of camptothecin (CPT), an anticancer drug. Because of the large-scale collection of its roots, the plant has become a threatened species. The present study accomplishes the induction of adventitious roots as a means for the production of CPT as well as for the large-scale propagation of this anticancer drug plant using leaf and internode explants. The biomass yield and CPT content of adventitious roots induced from different explants were compared to roots developed on ex vitro rooted stem cuttings. Adventitious roots were produced on half-strength Murashige and Skoog (MS) medium supplemented with $10.74{\mu}M$ ${\alpha}-naphthaleneacetic$ acid and $2.32{\mu}M$ kinetin at mean fresh weights of 0.753, 0.739 and 0.748 g roots from leaf, internode and shoot, respectively. CPT yield from in vitro derived roots after 50, 80 and 120 days of incubation (0.028, 0.06 and 0.1% dry weight, respectively) was not significantly different from those harvested at the same age from ex vitro rooted (0.03, 0.06 and 0.13%, respectively) stem cuttings. CPT from subcultured roots derived from solid (0.08%) medium was lower than from suspension culture medium (0.12%). Subsequent cultures of the adventitious roots showed a stable production of CPT (0.16%). The yield of CPT from 360-day-old plant-derived roots was 0.19%. Elicitation using methyl jasmonate and acetyl salicylic acid exhibited no enhancement in CPT yield. In vitro propagation through direct shoot regeneration was achieved from the adventitious roots upon transfer to MS medium with $8.87{\mu}M$ $N^6-benzyladenine$ (BA) and $2.46{\mu}M$ indole-3-butyric acid (IBA) with a mean of 21.2 shoots per culture in 50 days. The shoots upon subculture on medium having the same level of BA and IBA underwent rapid proliferation. The shoots transferred to field conditions after in vitro rooting exhibited 95% survival. Adventitious root induction, from leaf and internode explants, enables the feasible production of CPT as well as the large-scale rapid propagation of this species which can safeguard it from extinction.

• Korean Journal of Veterinary Service
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• v.44 no.3
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• pp.149-155
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• 2021

In virucidal efficacy testing, the chemical inactivation cannot be determined for all viruses due to the difficulties or the inability to culture sufficiently or the risk of exposure to the viruses. Therefore, disinfectants against these viruses could be evaluated by different methods and surrogate viruses are used as alternative. In this study we developed a method for efficacy testing of veterinary disinfectants using one of the candidate surrogate viruses, bacteriophage MS2, as part of the research on the selection of surrogate viruses for efficiency of efficacy testing of veterinary disinfectants. This method is based on the Animal and Plant Quarantine Agency (APQA) guidelines for efficacy testing of veterinary disinfectants. Bacteriophage and disinfectant are reacted in suspension in accordance with the APQA guidelines and then a newly established double agar layer method is applied for the efficacy test. The double agar layer method is summarized as follows: 1) The bottom agar with 1.5% agar is boiled and cooled before poured into petri dishes at volume of 20 mL, and dried under biological safety cabinet. 2) The top agar with 0.7% agar is boiled and kept at 50℃ before E. coli culture was seeded. 3) The serially diluted bacteriophage MS2-disinfectant mixtures 0.05 mL and E. coli host 0.01 mL (OD₆₀₀ 0.2~0.3) are mixed with 5 mL of top agar and incubate them at 50℃ for 5 min for reaction. 4) The resulting mixture is poured over top of a bottom agar plate and rocked sufficiently to ensure that the top agar covers the entire surface of the bottom agar. 5) The double agar layer is then placed under biological safety cabinet to allow the agar layer to solidify and subsequently incubated at 37℃ for 24 hr. 6) Following incubation, the plates may be inspected for plaques and record results.

• Korean Journal of Environmental Agriculture
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• v.35 no.1
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• pp.72-78
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• 2016

BACKGROUND: Entomopathogenic fungi have been studied to develop for biological control agents as an alternative to chemical control agents in insect pest management. This investigated to determine the optimal culture conditions in ceramic balls for maximal sporulation of entomopathogenic fungi Beauveria bassiana M130 by use rice bran extract.METHODS AND RESULTS: METHODS AND RESULTS: A culture of entomopathogenic fungi for 12day on rice bran extract(1:8, w/v) incubated in ceramic matrix at 28℃. Natural zeolite ceramic ball was high production of 4.2×108 conidial/mL. The culture condition optimized initial pH, temperature, rice bran extract concentration, adhesives substance and concentration of NaCl, respectively. The high production of spore optimal conditions were temperature 28℃, initial pH 3, rice bran extract 3 mL, starch 33 g, 5 % NaCl and sopre suspension 7 mL, respectively.CONCLUSION: This study was carried out for the mass production of entomopathogenic fungi conidia recover rate 65% in matrix of natural zeolite ceramic ball, and to develop ingredient-used formulation of Beauveria bassiana M130 conidia for biological control agents.

• Journal of Plant Biotechnology
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• v.5 no.1
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• pp.33-41
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• 2003

Microspore-derived cell lines resistant to 5-methyltryptophan (5MT, a tryptophan analog) or S-(2-aminoethyl)-L-cysteine (AEC, a Iysine analog) were selected in rice by in vitro mutagenesis. For selection of 5MT or AEC resistant cell lines, suspension-cultured cells were irradiated with gamma rays. Thirteen 5MT resistant cell lines were selected and they were able to grow stably at 2 times higher 5MT concentration. A feedback insensitive form of anthranilate synthesis, the pathway specific control enzyme for tryptophan synthesis, was detected from the 5MT resistant lines. Contents of the free amino acids in five resistant lines (MR12-1 to MR12-5) showed a 7.4 to 46.6 times greater level than that in the control culture. Tryptophan, phenylalanine, and tyrosine levels in the shikimate pathway were 28.1 and 22.5 times higher in MR12-3 and MR12 4, respectively, than that measured in the control cells. Four AEC resistant cell lines were isolated from cultures grown on medium containing 1 mM AEC, They were able to grow stably with 2 mM AEC, while sensitive calli were inhibited by 0.5 mM AEC. Aspartate kinase activities of the resistant lines were insensitive to the natural inhibitor, Iysine, and accumulated 2.2 to 12.9-fold higher levels of free Iysine than that of the control cells. Especially, the levels of aspartate, asparagine, and methionine in the aspartate pathway showed higher accumulation in the AEC resistant lines than that in the control cells.

• Korean Journal Plant Pathology
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• v.14 no.5
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• pp.445-451
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• 1998

A severe Phytophthora rot of strawberry caused by a species of Phytophthora has been widely occurred at major cultivation areas of Kimhae on August in 1997. Incidence of the disease was obtained in the range of 69.2~83.6% in surveyed 4 fields and showed an average of 75.2%. A species of Phytophthora was mostly isolated from the crown of infected strawberry plants and all the isolates were identified as P. nicotianae var. nicotianae (=P. parasitica). The fungus showed strong pathogenicity on strawberry by inoculation test. As a result of the leaf inoculation using mycelial disks of the fungus, both leaves and petioles were darkly browned, and were finally blighted. As a result of the root inoculation of zoospore suspension, both roots and crowns were rotten with dark brown. Although the fungus produced sporangia either on V-8 juice agar medium or liquid medium, the sporangia observed on the liquid medium appeared to be broadly turbinate and noncaducous. Moreover the fungus cultured on the liquid medium often produced sporangia having two papilla. The number of zoospores in sporangia was found to be ranged from 3 or 4 to as many as 20 or 25. In addition, the released zoospore from the sporangium became the cystospore during the prolonged culture of the fungus. The sporangia were measured as av. 49$\times$35 ${\mu}{\textrm}{m}$ with l/b ratio of 1.43. All isolates from crowns were heterothallic and A1 mating type since oospores were abundantly formed on clarified V-8 juice agar by dual culture with P. capsici A2 mating type. Aplerotic oospores were sized 24-26 ${\mu}{\textrm}{m}$. Antheridia were always amphigynous and recoreded an average of 12$\times$10 ${\mu}{\textrm}{m}$. Hyphal swlling were easily observed, and terminal or intercalary chlamydospores were abundantly formed on V-8 juice agar as well as in C/Z solution and sized av. 28.2 ${\mu}{\textrm}{m}$. This is the first report of Phytophthora rot of strawberry in Korea.

• Korean Journal of Plant Resources
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• v.19 no.6
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• pp.706-715
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• 2006

Nitrate reductase deficient (NR) mutant lines were selected indirectly by their resistance to 100mM chlorate in cell cultures of P. parviflora. A total of 585 chlorate resistant lines were confirmed by a second passage on a high concentration of chlorate. Frequency of spontaneous mutation was $9.7{\times}10^{-7}$ in 3 month old suspension-cultured cells, and in non-selective media containing amino acids as sole nitrogen source. The frequency of mutation could be increased up to 11-fold by culture for 12 months. Out of 40 randomly selected calli, 22 were fully deficient in NR. The rest of the clones contained a decreased level of NR activity. Further characterization was carried out in 13 mutant lines which were fully deficient in NR and in 5 mutant lines containing residual (0-7.0%) NR activity, as compared to wild-type cells cultured on the same medium. The $NR^-$ mutants were tentatively classified as defective in the NR apoenzyme (nia-type; 11 mutant lines including the 5 with residual NR activity) or in the molybdenum cofactor (cnx-type; 7 mutant lines) by the XDH activity. The cnx-type could be further classified into two groups. In one group (5 mutant lines) of these, the NR activity could be partially restored by nonphysiologically high (1.0mM) molybdate in the culture medium. Both types of $NR^-$ mutants were unable to grow on minimal medium containing nitrate as sole nitrogen source, but grew well on amino acids. They also proved to be extremely sensitive to the standard medium ($MSP_1$) containing nitrate and ammonium. Shoot regeneration was obtained only in the $NR^-$ mutants, which contained residual NR activity, but they so far have failed to grow into plants.

• Korean Journal of Plant Tissue Culture
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• v.24 no.5
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• pp.279-284
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• 1997

The extracellular sesquiterpenoids were accumulated in cell and hairy root cultures of Hyoscyamus muticus by elicitation using extracts of Rhizoctonia solani. The vetispiradiene synthase (VS) which is the first committed step in biosynthetic pathway leading to formation of solavetivone, lubimin, and rishitin from isoprenoid intermediate farnesyl pyrophosphate was induced upon elicitation, whereas no sesquiterpenoids and VS activity were detected in both control cell and hairy root cultures. VS activity increased rapidly and reached its maximum 12 h in both cell and hairy root cultures upon elicitor treatment. VS activities were paralleled with the absolute levels of VS polypeptide(s). Interestingly, the profiles of sesquiterpenoid accumulation in hairy root cultures were different from those in cell cultures. The hairy root culture seemed to fail to metabolize solavetivone further to lubimin.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.2
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• pp.155-161
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• 2005

The inherent instability of metabolite production in plant cell culture-based bioprocessing is a major problem hindering its commercialization. To understand the extent and causes of this instability, this study was aimed at understanding the variability of anthocyanin accumulation during long-term subcultures, as well as within subculture batches, in Vitis vinifera cell cultures. Therefore, four cell line suspensions of Vitis vinifera L. var. Gamay Freaux, A, B, C and D, originated from the same callus by cell-aggregate cloning, were established with starting anthocyanin contents of $2.73\;\pm\;0.15,\;1.45\;\pm\;0.04,\;0.7\;\pm\;0.024\;and\;0.27\;\pm\;0.04$CV (Color Value)/g-FCW (fresh cell weight), respectively. During weekly subculturing of 33 batches over 8 months, the anthocyanin biosynthetic capacity was gradually lost at various rates, for all four cell lines, regardless of the significant difference in the starting anthocyanin content. Contrary to this general trend, a significant fluctuation in the anthocyanin content was observed, but with an irregular cyclic pattern. The variabilities in the anthocyanin content between the subcultures for the 33 batches, as represented by the variation coefficient (VC), were 58, 57, 54, and $84\%$ for V. vinifera cell lines A, B, C and D, respectively. Within one subculture, the VCs from 12 replicate flasks for each of 12 independent subcultures were averaged, and found to be $9.7\%$, ranging from 4 to $17\%$. High- and low-producing cell lines, VV05 and VV06, with 1.8-fold differences in their basal anthocyanin contents, exhibited different inducibilities to L-phenylalanine feeding, methyl jasmonate and light irradiation. The low-producing cell line showed greater potential in enhanced the anthocyanin production.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.spc1
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• pp.237-241
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• 2006

This experiment was carried out comparison with haploid plants productivity by microspore culture among domestic cultivars of Brassica napus L. Isolated microspore from flower buds were cultured on NLN medium supplemented with 13% sucrose, $0.05mg/{\ell}$ BA and $0.5mg/{\ell}$ NAA. Genotype was important factor in haploid embryo productivity 'Tamlayuchae' showed the highest haploid embryo production frequency (176 embryos formed from 1 flower bud). But, 'Hallayuchae' and 'Youngsanyuchae' were not generated embryo even cell division. When suspension culture on NLN liquid medium at 100 rpm, embryos were developed multilobe abnormal embryo cluster. Multilobe abnormal embryos on MS medium basal solid medium were regenerated multiple shoots. Regenerated haploid plant with well developed shoots and roots on MS basal medium were successfully transferred to pots.