• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.150-155
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• 1988

In order to produce L-lysine from cellulosic substrates by the intergeneric protoplast fusion between cellulolytic bacteria, Cellulomonas flavigena KFCC31221 and amino acid producing bacteria, Brevibacterium flavum ATCC14067, Corynebacteriurn glutamicum ATCC13032, conditions for protoplast formation and regeneration of these strains were investigated. After the strains were mutated with 500$\mu\textrm{g}$/$m\ell$ N-methyl-N'-nitro N-nitrosoguanidine for 30 min and the mutants were enriched by treating 300$\mu\textrm{g}$/$m\ell$ penicillin-G for 2 hrs, B. flavum Hse- Str$^{r}$ , C. glutamicum Met$^{-}$Thr$^{-}$ Rif$^{r}$ and Cellulomonas flavigena Thr$^{-}$Val$^{-}$Kan$^{r}$ were isolated. The rate of protoplast formation ranged from 95 to 98% when strains were treated at the concentration of 500$\mu\textrm{g}$/$m\ell$ of lysozyme, pH 6.5, 33$^{\circ}C$, for 6 hrs. in Tris- malate buffer supplemented with 0.4M sucrose as osmotic stabilizer. Approximately 30-33% protoplast was regenerated on the regeneration complete medium(RCM) containing 1.5% agar and 0.5M sodium succinate overlaid with the same medium except 0.7% agar.

• Journal of Life Science
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• v.15 no.4 s.71
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• pp.528-535
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• 2005

The non-enzymatic anti-oxidants and lunasin peptide from the extracts of the pepper were examined in order to utilize the discovery in natural products as cancer chemopreventive agents. The DPPH (1,1-diphenyl-2-picryl-hydrazyl) free radical scavenging activity on the fruit parts of the pepper was higher than that of the seed, but the difference was low. The Inhibition activity of xanthine/ xanthine oxidase in extracts of the seed was higher than that of the fruit and that of the seed on 20 days after flowering was the highest at the growing period. These were identified as fatty acids and phenolic compounds such as 1-eicosanol, palmitic acid, linoleic acid, linolenic acid and benzonitrile. The contents of fatty acids and phenolic compounds increased according to the time passing at the growing period. Peroxidase (POD) activity of the fruit at middle stage was high than that of other growing stages and that of the seed was the highest at later growing period. Though superoxide dismutase (SOD) activities in fruit were hish by passage of Slowing stage, the activity in seed was low. Lunasin was searched from seeds of the peppers by coomassie blue staining and western blot among them and we just found lunasin peptide from extracted protein of the pepper by western blot. In addition, we observed the contents of lunasin after flowering and confirmed to appear the lunasin at 35 days after flowering. We confirmed that lunasin is complex protein of maturing seeds. 100nM lunasin peptide in pepper showed inhibition effect on colony formation in $2\~12$ cells.

• Microbiology and Biotechnology Letters
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• v.16 no.4
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• pp.275-281
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• 1988

To develop cellulase and xylanase-producing strain by protoplast fusion, alkalophilic Bacillus sp. F204 and K17 were treated with NTG(N-methyl-N'-nitro-N-nitrosoguanidine) and isolated anti-biotics resistant strains of S20 (Km$^r$ , Cm$^r$) and G70 (Str$^r$). The frequency of protoplast formation was about 95% when cells of mid-log phase were treated with 200$\mu\textrm{g}$/ml Iysozyme at 37$^{\circ}C$ for 30-45 minutes. Under addition of 0.4-0.5M sodium succinate, 0.5% casamino acid, 1.5% polyvinylpyrrolidone, 25mM MgC1$_2$ and 50mM CaC1$_2$ to the regeneration medium, the regeneration frequency of Bacillus sp. F204 and K17 was 24.9% and 26.2%, respectively. The fusion frequency was 6.6$\times$10$^{-6}$ in the presence of 30% polyethylene glycol 6000 containing 50mM $Ca^{++}$ at 45$^{\circ}C$ for 5 minutes. Cellulase complex and xylanase activities of fusant were compared with parental strains.

• Journal of Life Science
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• v.7 no.4
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• pp.308-308
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• 1997

Role of plant hormones on the leaf senescence of Phaseolus vulgaris were investigated by measuring the disassembly of chlorophyll-protein complexes in detached leaves treated with NAA, GA$_{3}$ , or BA. The loss of chlorophyll that was characteristic of leaf senescence induced disassembly of chlorophyll-protein complexed. During dark-induced senescence, PSI complex was rapidly degraded after the early stage, whereas RC-Core3 was slightly increased until the middle stage and slowly decreased thereafter. And gradual degradation of trimeric LCHII progressed after the late stage of senescence. Exogenous application of NAA and GA$_{3}$ had little or no effect in protecting disassembly of chlorophyll-protein complexes during leaf senescence compared to control. However exogenous BA application strongly leaves. In the simultaneous treatment of plant hormones and light, BA application under illumination of light was most effective in the stability of chlorophyll-protein complexes, particularly PSI, LHCII, RC-Core2, RC-Core3 and SC-1. these results suggest, therefore, that simultaneous application of BA and light induced synergistic effect on the stability off chlorophyll-protein complexes during leaf senescence.

• Journal of Life Science
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• v.29 no.4
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• pp.421-427
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• 2019

Mobilization of storage lipids is critical for the germination of oil seeds, as they supply carbon and energy until photosynthesis commences in cotyledons. In this study, we determined the levels of plant carnitine and associated changes in these levels from seed germination to cotyledon senescence. We also examined changes in the content of unsaturated fatty acids throughout seedling development. Carnitine levels peaked on day 3 at 14.5 nM in cotyledons and decreased sharply to 7.2 nM on day 4. On development day 3 carnitine levels were maintained at around 3 nM until day 7. The unsaturated fatty acid content dropped by half at the same time as carnitine peaked (day-3), and storage lipids were almost depleted by day 5. Thereafter, carnitine was hardly detected until the second stage of cotyledon senescence, at which stage the carnitine content was 6.8 nM, similar to that on day 4 at the time of fatty acid depletion in the cotyledons. Unsaturated fatty acids levels remained constant in green cotyledons but slightly increased in the senescing cotyledons. The latter can be explained by intracellular breakdown of membrane lipids. This is the first such discovery in developing cotyledons and may offer clues regarding other roles of the acetyl unit transport system in plants. The expression of BOU was closely associated with carnitine metabolism during seed germination and cotyledon development. The results provide support for the possibility of carbon re-routing during the glyoxylate cycle in the supply of energy for early germination and development.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.12 no.3
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• pp.225-233
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• 2007

To verify which loop regions of 18S rRNA gene are suitable as species-specific genetic markers in ciliates, we analyzed the genetic variation of 18S rRNA gene among 9 Euplotes species (Hypotrichia : Ciliophora). In our result, no inter-specific variation was detected from V1, V3 and V5 regions, and the length of V7 and V8 are 44 bp and 79 bp, respectively, which are too short to make genetic marker. In contrast, V2 and V4 may be good candidate segments of species-specific diagnostic molecular markers because these two regions are most variable ($1.75{\sim}20.61%$) and showed good inter-specific phylogeny. Furthermore, the sequences of V2 and V4 are 123 bp and 306 bp, respectively in length which are enough to make species-specific marker.

• Korean Journal of Ichthyology
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• v.36 no.1
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• pp.94-100
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• 2024

During ichthyoplankton survey in the southwestern sea of Korea, we collected six individuals of noodlefish larvae and juveniles between April and May 2023. They were identified as Neosalanx anderssoni by mitochondrial DNA cytochrome c oxidase subunit I or 16S ribosomal RNA sequences, and their external morphological traits were described for the first time. All six individuals have a slender and elongated body. When preflexion and flexion larval stages (10.24 mm notochord length, NL and 15.47 mm total length, TL, respectively), oval-shaped black melanophores were distributed in a row along the ventral side of the gut. However, when postflexion larval and juvenile stages (23.58~25.90 mm TL, and 29.20~31.26 mm TL, respectively), melanophores on the ventral side of the gut were disappeared, and dark spot-shaped melanophores appeared along the dorsal side of the gut in a single row. Also, from the postflexion larval stage (23.58 mm TL), two large black spots began to appear symmetrically on the caudal fin. Our results suggest that N. anderssoni may use coastal area as spawning and/or nursery ground unlike previous study (Kim and Park, 2002).

• Journal of Plant Biotechnology
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• v.39 no.1
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• pp.69-74
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• 2012

Using calli of $Muscari$ $armeniacum$ cv. 'Early Giant' that is monocotyledonous ornamental bulb crop with increasing demand in Korea, we carried out current studies to establish an in vitro multiple propagation protocol via somatic embryogenesis. We found that soft pale yellow green calli were induced from leaf explants cultured on all media containing 0.1~3.0 $mg{\cdot}L^{-1}$ auxins such as 1-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D). However, induced calli showed vigorous growth only when they further transferred on same media containing 2,4-D, 4-amino-3,5,6-tri-chloropicolinic acid (picloram), or 3,6-dichloro-o-anisic acid (dicamba). Although frequency of somatic embryo induction depended on callus source and PGR composition in somatic embryo induction media, somatic embryogenesis was initiated on surface of proliferated calli after transferring on media with no PGR or 0.01 $mg{\cdot}L^{-1}$ NAA co-supplemented with various cytokinins such as $N^6$-benzylaminopurine (BAP). Highest number of embryo at 9.3 per callus clump was obtained when calli which were grown under 0.1 $mg{\cdot}L^{-1}$ picloram supplementation were sub-cultured on medium with 0.01 $mg{\cdot}L^{-1}$ NAA and 0.5 $mg{\cdot}L^{-1}$ BAP. In addition, morphological characteristics of somatic embryo were categorized into following nine phases: globular, biased heart, biased torpedo, early cotyledonary, middle cotyledonary, late cotyledonary, early sprouting, middle sprouting, and late sprouting embryos.

• Applied Biological Chemistry
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• v.42 no.1
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• pp.20-28
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• 1999

To improve Bacillus strains producing biopolymer, conditions for protoplast formation and regeneration were investigated in biopolymer producing Bacillus subtilis K-1 and lactose utilizing Bacillus coagulans. Bacillus subtilis K-1 mutant (SM-2) and Bacillus coagulans mutants (CM-12) were marked auxotrophic and antibiotics-resistant (SM-2) and an antibiotics-resistant mutants, respectively. To formate protoplasts derived from the mutants, conditions were established as follows. For B. subtilis mutant SM-2, its culture in mid-logarithmic phase was added with penicillin G (1.0 unit/ml) and further reacted for 1.5 hr. Cells were collected and then treated in lysis fluid (pH 7.0) containing 0.4 M sucrose and lysozyme $25\;{\mu}g/ml$ for 40 min at $37^{\circ}$. Protoplast formation was very successful (99.6%) and the ratio of cell wall regeneration was 2.4%. For Bacillus coagulans mutant CM-12, its mid-logarithmic phase culture was treated with penicillin G (0.3 unit/ml) and glycine (0.5%) for 1hr. Cells were collected and then resuspended in lysis buffer (pH 7.0) containing 0.6 M lactose and lysozyme $(300\;{\mu}g/ml)$ for 30 min at $37^{\circ}$. Protoplast formation was also successful (90.8%) and cell wall regeneration ratio was similar to SM-2 (2.2%). To improve regeneration frequency, regeneration medium was obtained as followed condition,. Cell wall regeneration was improved 2-4 folds with 5.1% for B. subtilis SM-2 and 10.3% for B. coagulans CM-12 when protoplasts mixed with soft top agar(0.4%) was overlaid onto trypticase soy broth medium containing 0.4 M sucrose, 0.7% casamino acid, 1% PVP, 25 mM $MgCl_2,\;25\;mM\;CaCl_₂$ and 1.5% agar.

• Korean Journal of Ichthyology
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• v.35 no.2
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• pp.91-100
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• 2023

The early life history of Silurus microdorsalis living in Jahocheon Stream was studied by observing egg and morphological development. Live fish were captured in June 2018, then reared in a circulating filtration system under a 14L : 10D photoperiod with a water temperature of 18℃. To artificially induce spawning, females were injected with 0.5 mL of Ovaprim (Syndel, Nanaimo, BC, Canada) per kg of body weight, and males were injected with 10,000 IU/kg body weight of human chorionic gonadotropin. Approximately 15 h later, eggs were artificially inseminated by the dry method. Mature eggs were light pale yellow, which separated them from immature eggs. Fertilized eggs were 2.16±0.06 mm (n=8) in diameter and fully hatched at 181 h after fertilization. The fertilization rate was 63.1±2.2%, and 10.0±3.7% of the embryos were malformed at 18℃. The rates of development were 181 h at 18℃, 109 h at 21℃, and 76 h at 24℃. The larval size immediately after hatching was 4.64±0.22 mm (n=8), and the larvae displayed negative phototaxis at 1 day after hatching. The total larval length on 7 days after hatching was 12.47±0.53 mm, with 25~30 basal anal fin rays and 14~16 basal caudal fin rays observed. The total larval length was 14.13±0.51 mm on 9 days after hatching, and approximately 90% of the black endoplasmic reticulum was deposited on the head and body. The dorsal fin had formed, and a single basal body was observed. On 15 days after hatching, the total larval length was 16.69±0.31 mm; the number of basal caudal fin rays (18 poles) was an integer because 2 dorsal fin basal rays and 60~63 anal fin basal rays were observed. The total larval length was 28.96±1.10 mm on 50 days after hatching; the numbers of caudal fins (n=18), dorsal fins (n=3), pectoral fins (n=11), and anal fin basal rays (n=67~73) were integers.