• Proceedings of the Plant Resources Society of Korea Conference
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• 2003.10a
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• pp.50-58
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• 2003

Isolation and culture of leaf protoplasts from two chilli cultivars (Capsicum annuum var. accumnatum and Bird chilli) were developed to enhance selection process in the somatic hybridization programmes. In order to isolate the protoplasts from leaves of these two chilli cultivars different incubation periods (3, 5 and 10 hours) were tested with combinations of enzyme mixtures containing cellulase and macerozyme. Leaves were incubated on three enzyme mixtures (2% cellulase + 0.4% macerozyme, 1% cellulase + 0.2% macerozyme and 0.5% cellulase + 0.1 % macerozyme in 13% mannitol) at 251oC in the dark. Three hours of incubation using 2% cellulase and 0.4% macerozyme was the best for the protoplast isolation of both chilli cultivars tested. The yield was 5 ${\times}$ 108protoplasts/ml/ g leaf tissue in both chilli varieties. It was found that in the mixed nurse method using Nagata and Takebe (NT) medium supplemented with 1.0mg/12,4-D, NAA and BAP with 0.5M mannitol and 1.2% Sea Plaque agarose is the best medium for protoplast culture. Protoplasts of Capsicum annum var. accumnatum were alive for 14 days forming cell walls and initiating cell division.

• Journal of Ginseng Research
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• v.25 no.3
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• pp.136-140
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• 2001

Effects of incubation temperature and pH on chlamydospore germination of Cylindrocarpon destrcutans (isolate CY-9802) causing root rot of Panax ginseng were studied. Germination rate of the chlamydospores on Czapek solution agar(CSA) was higher than on potato dextrose agar(PDA) at the incubation temperatures tested. The chlamydospores were able to be germinated at range of 5$\^{C}$ to 30$\^{C}$ after 48 hours incubation on CSA. Germination rate was 53.2∼6.27% at range of 15$\^{C}$ to 25$\^{C}$, and the optimum temperature was 20$\^{C}$, whereas they were very low at 30$\^{C}$ on PDA. Germination rate was 43.6% to 47.9% at range of 10$\^{C}$ to 20$\^{C}$, and the optimum temperature was 20$\^{C}$ as well. They were able to be germinated at pH of 5.2 to 8.1 on CSA and 5.2 to 7.2 on PDA. Optimum pHs for the germination on CSA and PDA were from 6.4 to 8.2 and from 5.2 to 6.0, respectively. Mycelial color of the fungus on CSA was pale brown at pH from 5.2 to 6.0 and white from pH 6.4 to 8.1, while it was typical dark brown ar range of pH 5.2 to 7.1 and brown at pH 7.2 on PDA after 21 days incubation.

• KSBB Journal
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• v.24 no.6
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• pp.556-562
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• 2009

The biological treatment of TNT-containing wastewater, known commonly as pink water, was investigated using a stirred tank reactor with Stenotrophomonas maltophilia OK-5 bacterial culture. S. maltophilia OK-5 exhibited effective degradation of TNT contained in pink water, completely degrading TNT (100 mg/L) within 6 days of incubation. The dark-red brown color derived from Hydride-Meisenheimer complex became more pronounced during the incubation period, which was determined quantitatively. High-pressure liquid chromatography was used to measure residual TNT, which also resolved the metabolic intermediates (i.e., 2,4-dinitrotoluene, 2,6-dinitrotoluene and 2,4-dinitro-6-hydroxytoluene). Gas chromatography-mass spectrometry was used to verify these intermediates. Quantification of the nitroreductase (pnrB) gene isolated from S. maltophilia OK-5 growing in pink water was performed with real-time PCR. The amount of pnrB gene copies increased to $10^3$-fold after 5 days of incubation time.

• Korean Journal of Plant Tissue Culture
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• v.25 no.5
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• pp.347-351
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• 1998

Plant regeneration via organogenesis from leaf and stipule explants of micropropagated shoots of strawberry (Fragaria $\times$ ananassa cv. Suhong) was achieved. Leaf and stipule explants were detached from shoot-tip cultured shoots and cultured on MS medium with various combinations of BA and NAA under light or dark condition. Shoot regeneration from leaf explant was observed after 3 weeks in culture and was good at the high ratio of BA and NAA among various combination treatments. The highest shoot regeneration frequency from leaf explants was obtained with 1.0 mg/L BA and 0.1 mg/L NAA, in which 31.1% shoot regeneration frequency(1.7 shoots per leaf explant) was yielded. In case of stipule explants, shoot regeneration was largely affected by plant growth regulators during incubation under dark condition for initial 4 weeks but not under continuous light condition. The combination treatment with 2.0 mg/L BA and 0.1 mg/L NAA showed the most excellent shoot regeneration from stipule explants, where 44.4% regeneration frequency(4.0 shoots per explants) obtained. Regenerated shoots were rooted on MS medium with 0.1 mg/L NAA after shoot elongation, and the plantlets regenerated were transferred to soil mixtures with vermiculite and perlite for acclimation.

• Journal of Crop Science and Biotechnology
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• v.10 no.2
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• pp.112-116
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• 2007

Kinetin(KN) is an inducer of rice(Oryza sativa L.) defense/stress responses, as evidenced by the induction of inducible secondary metabolite and defense/stress protein markers in leaf. We show a novel light-dependent effect of KN-triggered defense stress responses in rice leaf. Leaf segments treated with KN(100 ${\mu}M$) show hypersensitive-like necrotic lesion formation only under continuous light illumination. Potent accumulation of two phytoalexins, sakuranetin and momilactone A(MoA) by KN that peaks at 48 h after treatment under continuous light is completely suppressed by incubation under continuous dark. Using two-dimensional gel electrophoresis we identified KN-induced changes in ribulose-1, 5-bisphosphate carboxylase/oxygenase, energy- and pathogenesis-related proteins(OsPR class 5 and 10 members) by N-terminal amino acid sequencing and mass spectrometry. These changes were light-inducible and could not be observed in the dark(and control). Present results provide a new dimension(light modulation/regulation) to our finding that KN has a potential role in the rice plant self-defense mechanism.

• Journal of Photoscience
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• v.9 no.2
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• pp.427-429
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• 2002

We studied iron release from ferritin by irradiating the visible light, and then followed ferritin-mediated lipid peroxidation in the rod outer segment (ROS) fraction of the porcine retina. In the presence of several phosphorus compounds such as ADP and ATP, iron release from ferritin at pH 7.0 could be induced by irradiation of the visible light to the reaction mixtures. Furthermore, iron release from ferritin in the presence of ADP depended on the incubation time and the visible light irradiation. Moreover, we investigated lipid peroxidation level in the ROS fraction by two independent assay systems including the thiobarbituric acid (TBA) and ferrous oxidation/xylenol orange (FOX) methods. The visible light induced ferritin-mediated lipid peroxidation in the ROS fraction in time- and irradiance-dependent manners. In the dark condition, iron release and lipid peroxidation were not observed. Iron release from ferritin by irradiating the visible light may play an important role in the etiology of phototoxic injuries in vivo.

• The Plant Pathology Journal
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• v.17 no.4
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• pp.210-215
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• 2001

The phenotype of Rhizoctonia solani KR-13 was randomly segregated to both melanin-producing (M+) and non-producing (M-) types through successive cultures on PDA. M+type with dark melanin showed strong pathogenicity to rice and self-anastomosis. Meanwhile, M- type with white or less-melanized mycelia showed very weak pathogenicity and non-self-anastomosis. Melanin production of R. solani was affected by incubation temperature in both M+ and M- types, but not by light treatment. The application of tricyclazole, an inhibitor of fungal melanin biosynthesis, showed no controlling effect on R. solani causing rice sheath blight. Results of this study showed that melanization of mycelia of R. solani is an important pathogenicity factor in rice.

• Microbiology and Biotechnology Letters
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• v.22 no.2
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• pp.115-119
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• 1994

White mutant of Ganoderma lucidum(G4142) induced the non-basediocarpous basidiospores(NBB) from the aerial mycelia on agar media by the light illumination. Light was found to be necessary for NBB formation, but it also inhibited the growth of mycelium. The best sporulation was obtained at the periodic exposure of 16 hour light and 8 hour dark. Blue and yellow light were the most effective on sporulation, however, near UV and red light did not induce any spores. Effective light intensity for NBB bearing was about 1,000 lux as white light. Even after 16 days of culture, this strain did not form the pinhead nor chlamydospore. Optimum temperature for the mycelial growth and NBB formation were 30$\circ $C. Ganoderma lucidum G4142 exhibited the formation of stroma after five days of incubation at 30$\circ $C.

• Journal of Korea Foresty Energy
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• v.23 no.2
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• pp.21-28
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• 2004

Three different strains of Lentinula edodes, Sanlim 5-Ho, Sanlim 6-Ho and Nongki 3-Ho, were cultured in the sawdust media of Mongolian oak(Quercu mongolica Fisch) for 90 days under dark and light conditions(each 30 days) and fruiting period(30 days). Weight loss of sawdust media was determined after fungal cultures and the contents of ergosterol in fungal mycelia were quantified by HPLC analysis followed by solvent extraction. Compared with the two other fungal strains$(8\%)$, weight loss of Sanlim 5-Ho was slightly lowered to $7\%$. The level of ergosterol content, a parameter for fungal growth, was continuously enhanced in Sanlim 5-Ho for dark and light incubation periods. However, Sanlim 6-Ho and Nongki 3-Ho recorded the maximized fungal growth under light condition. In fruiting periods the ergosterol contents were lowered in the three strains. Intra- and extracellular enzymes during cultural and fruiting periods were also characterized. The activity of Mn-peroxidase and laccase, which are characteristics enzymes for white rot fungi as lignin degrading enzymes, were determined as a high level overall the periods. As cellulose degrading indicators, the activity of CMCase, avicelase, xylanase and glucanase were detectable in initial incubation period.

• Korean Journal of Microbiology
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• v.31 no.3
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• pp.218-223
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• 1993

Conditions for the release and regeneration of protoplasts form Rhizopus oryzae and intergeneric protoplast fusion between Rhizopus oryzae and Aspergillus oryzae were studied. High yields of protoplast fusion between Rhizopus oryzae and Aspergillus oxyzae were studied. High yield of protoplasts from young germilings of R. oryzae were obtained by using lytic enzymes containing chitosanase (3 mg/ml), chitinase (3 mg/ml) and Novozym 234 (5 mg/ml). 0.5M glucose was used as the osmotic stabilizer and optimum pH of buffer was determined to be pH 7.5-8.0. Under these conditions, protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts regenerated on solid medium with a soft agar overlay. We have also carried out protoplasts fusion between R. oryzae and A. oryzae and have succeeded in obtaining three types of intergeneric fusants. In these experiments, 35% PEG-4000 and 10 mM CaCl$_{2}$ were used as fsogenic agents, and auxotrophic properties were used as a genetic marker to select fusants. Complementation frequency be protoplasts fusion of A. oxyzae and R. oryzae was 4.4% * 10$^{-5}$ . The fusant strains of the first type were prototrophs showing an Aspergillus type morphology with dark-yellow sporulation, those of the second type were also Apergillus type morphology but showed no sporulation. And the strains of the third type stopped growing when fusion products grown on regeneration minimal medium were transferred to fresh minimal medium. The formation of fusion products was observed by fluorescent vital stains for complementary labelling of protoplats from R. oryzae and A. oryzae. Rhodamine 6G and fluorescein diacetate wer useful complementary vital stains of Rhizopus and Aspergillus protoplasts for visualization of requency and type (dicell, multicell) of fusion.