• Applied Microscopy
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• 제1권1호
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• pp.11-17
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• 1969

Conidio spores of Aspergillus niger (strain No. NRRL 330) cultured on potato dextrose agar media were studied by electron microscopy, using the thin sectioning techniques. Conidio spores to be sectioned were fixed by triple methods with $K_2Cr_2O_7$, Glutaraldehyde and $OsO_4$. After dehydrated with alcohol, the specimens were embedded in metacrylate and epon resin media, and thinly sectioned by Porter-Blum MT-2. After sectioned these specimens were negative-stained with uranyl acetate and observed. by Hitachi HS-6 electron microscope. The results of this experiment were summarized as follows. 1. The structures of spore ,wall system seem to be formed 4 layers; exosporium, basal layer, spore coat and unit cell membrane. The protuberance of spore surface that was looked like hair appears to be protrusived from the basal layer. 2. The 3 layers of unit cell membrane was constituted outer layer membrane, inner layer membrane and inter-mediate light layer. 3. The structures of intra cytoplasmic membrane appear as spiral form which was consisted of 3 layers membrane system; outer membrane, inner membrane, and intermediate layer, which has pits. 4. The cement substance of spore coat and cortex may be changed quantitatively by physiological state in cell. 5. In some cases, we observed that the ribosome was transformed into poly ribosome group, and the storage materials and the protein crystals were changed variously. It. has been suggested that the morphological change of some cytoplasmic materials may be caused by some specialized function of the physiological stage.

• Fisheries and Aquatic Sciences
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• 제19권2호
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• pp.9.1-9.7
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• 2016

This study was conducted to identify the general conditions for the isolation and in vitro culture of ovary-derived cells in rockfish (Sebastes schlegeli). The effects of three different enzymes on cell retrieval from ovarian tissues were evaluated first, and then the ovary-dissociated cells were cultured under various culture conditions, with varying basal media and culture temperatures, addition of growth factors, and/or culture types. We found that collagenase type I treatment was effective for cell isolation from ovarian tissues. From a total of 42 trials to evaluate the effects of basal media and culture temperatures on cell culture of ovary-dissociated cells, we observed that Leibovitz's L15 medium was more supportive than Dulbecco's modified Eagle's medium for culture, and the cells could grow at all three temperatures tested, 15, 20, and $25^{\circ}C$, at least up to passage 2. However, growth factor addition did not improve cell growth. Introduction of suspension culture after monolayer culture expanded the culture period significantly more than did monolayer culture alone. Our results may provide a basis for developing an in vitro system for S. schlegeli germline cell culture, which will ultimately lead to improvement of the species.

• Journal of Microbiology and Biotechnology
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• 제12권1호
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• pp.137-141
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• 2002

Anabaena variabilis ATCC 29413 grew in chromium (Cr) containing Chu-10 (basal) and nitrate-supplemented media, and the growth of the organism in $100{\mu}M$ chromium was found to be 50% of that in control medium. The growth in nitrate $({NO_3}^-)$ supplemented cultures was better as compared to cultures grown in basal medium. Free cells from basal and nitrate-supplemented media removed 5.2 and 7.4 nmol of chromium $mg^{-1}$protein in 8 h, respectively, from the medium containing $30{\mu}M$ chromium. The efficiency of chromium removal increased 7-fold in imidazole buffer (0.2 M, pH 7.0). A cell density equivalent to $100{\mu}g$ protein $ml^{-1}$ was found to be optimum for maximum Cr removal. Entrapment of cells in calcium-alginate beads did not affect the rate of Cr uptake by the cells. The efficiency of the laboratory-scale continuous flow bioreactor $(12.5{\times}2cm)$ loaded with alginate-immobilized cells (10 mg protein) and fed with $30{\mu}M$ chromium solution was compared at different flow rates. The efficiency of the bioreactor varied with flow rates. In terms of percent removal of Cr from influent, a flow rate of 0.1 ml $min^{-1}$ was found to be optimum for 6 h (54% Cr removal efficiency). Maximum amount of Cr (883 nmol) was removed by the cells in 3 h at a flow rate of 0.5 ml $min^{-1}$. The potential use of A. variabilis in removing Cr from industrial effluents is discussed.

• Journal of Plant Biotechnology
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• 제35권3호
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• pp.203-208
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• 2008

Philodendron cannifolium을 기내배양하여 일시에 균일한 식물체를 대량생산하기 위하여 실험을 실시하였다. Philodendron의 경정에서 부정 다아체 형성은 BA $2.0{\sim}5.0$ mg/L 첨가배지와 TDZ $0.05{\sim}1.0$ mg/L 첨가배지에서 양호하였다. 그러나 TDZ이 첨가된 배지에서 생육한 식물체는 모두 엽록소가 결핍되었다. 신초경정에서 형성된 부정 다아체를 $5{\sim}7$ mm 정도로 종으로 절단하여 배양한 결과 BA $1.0{\sim}3.0$ mg/L가 첨가된 배지에서 신초수가 30개 이상으로 신초의 증식이 양호하였다. 그러나 BA가 고농도(10.0 mg/L)로 첨가된 배지 및 TDZ이 첨가된 배지는 기부의 callus가 이상 비대하였다. 증식된 신초의 색도 cream색으로 엽록소가 부족하였다. 증식된 신초의 생육 및 발근은 IBA $0.5{\sim}2.0$ mg/L가 첨가된 MS 배지에서 양호하였다. 발근된 신초의 순화는 vermiculite와 perlite 및 vermiculite의 1:1 혼합용토가 적합하였다. MS배지에 활성탄 10 g/L와 sucrose 30 g/L가 첨가된 액체배지 15 mL를 첨가하여 신초를 생육시킨 다음, IBA $500{\sim}2,000$ ppm이 첨가된 용액에서 10 초간 침지하여 perlite와 vermiculite가 1:1로 혼합된 용토에 재식하였을 때, 신초의 생육 및 발근이 양호하였다.

• Journal of Plant Biotechnology
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• 제31권2호
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• pp.115-119
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• 2004

본 실험은 Philodendron wend-imbe를 기내배양하여 일시에 균일한 식물체를 대량생산하기 위하여 실시하였다. Philodendron의 경정에서 다아체 형성은 BA 5.0-10.0mg/L 또는 TDZ 0.05-0.1 mg/L가 첨가된 MS 배지에서 양호하였다. 형성된 다아체 절편체 (5-7mm)를 BA 5.0 mg/L와 sucrose 20g/L가 첨가된 MS 배지에서 배양한 결과, 신초 및 다아체의 증식이 매우 양호하였으며, 신초기부에서 callus 발생이 억제되었다 다아체 절편체에서 신초의 발생 및 발근은 IBA 1.0-2.0mg/L 또는 NAA 0.1 mg/L가 첨가된 배지가 효과적이었으며, 발근된 신초의 순화는 perlite와 peat moss의 1:1 혼합용토 또는 peat moss가 적합하였다.

• Journal of Plant Biotechnology
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• 제48권2호
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• pp.86-92
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• 2021

The present study aimed to evaluate the initiation, proliferation potential, and mass clonal production ability of a micropropagation system for banana through tissue culture. A total of 60 explants were cultured on basal media supplemented with various concentrations of BAP and NAA. Banana plants regenerated on MS basal medium (control) without the addition of BAP + NAA showed a significantly (P < 0.05) lower survival rate with no signs of shoots up to the end of the experimental period. The results further revealed that the performance in MSS-XI medium was almost 89%, followed by MSS-IX and MSS-X media, both of which showed performance up to 88%. In contrast, the performance in the MSS-XVI medium was less than 60%, at the less duration of time and highly shoot induction detected at MSS-XIII medium. The maximum number of shoots (4.9) was observed in the medium supplemented with growth adjuster MSS-XI, followed by the MSS-XII medium (4.5). Surprisingly, the best performance was observed for the MSR-VII medium approximately 16 days after initiation, while the lowest performance was observed with MSR-XI (approximately 31 days). The maximum rooting percentage (98%) was observed in the MSR-V to MSR-VIII media (98%), while the minimum rooting percentage was observed in MSR-XI (approximately 45%).

• Journal of Plant Biotechnology
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• 제49권2호
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• pp.124-130
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• 2022

The application of traditional taro propagation methods for large-scale cultivation would be insufficient to meet the high demand for quality planting materials. Therefore, this study aimed to develop an in vitro micro-propagation technique for two local taro cultivars (cv.), Wangi and Putih. Taro cormels were collected from the Malaysian Agricultural Research and Development Institute (MARDI) germplasm (Serdang, Malaysia). Explants were taken from the shoot tip of cormels and initially cultured on Murashige and Skoog (MS) basal media for four weeks. The explants were then transferred to different multiplication media supplemented with different types and concentrations of cytokinins such as 6-benzylaminopurine (BAP ) and Thidiazuron (TDZ). Shoot production was quantified after six weeks of culture. The highest mean number of new shoots was produced by the Wangi cultivar on MS medium supplemented with 2.0 mg/l BAP (2.10 shoots), MS medium supplemented with 0.5 mg/l TDZ (2.18 shoots), and Gamborg B5 medium supplemented with 6.0 mg/l BAP (2.43 shoots). The maximum average number of the Putih cultivar shoots was obtained on MS supplemented with 2.0 mg/l BAP (3.57 shoots). MS basal media was used for root initiation, as it produced an average of 25 roots with an 11-cm length. Various types of substrate mixtures were used during acclimatization. The best acclimatization substrate for the Wangi cultivar was 100% peat soil, whereas the Putih cultivar grew optimally in a combination of peat and perlites at a 1:1 ratio. Taro plantlets require approximately 4 to 6 weeks to acclimatize before they can be transferred to the field.

• Journal of Life Science
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• 제9권1호
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• pp.50-53
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• 1999

Effects of growth regulators and medium components were tested for shoot multiplication and callus growth from shoot explants of black locust. During shoot multiplication, callus growth at the cut end of shoot explants proceeded shoot development. The basal callus growth seemed to be a function of both mineral components and cytokinin supplemented in the medium. Maximum callus growth was induced by 0.5${\mu}$M BAP and the callus growth decreased as the level of BAP increased. Positive correlations were found between basal callus growth, and shoot multiplication and growth. Shoot multiplication was greatest on BSM medium (black locust shoot culture medium) supplemented with 1 $\mu$M BAP. With medium containing high nitrogen content, both shoot multiplication and growth were significantly enhanced. A new BRM medium was the most effective for rooting of black locust among three rooting media tested.

• 생약학회지
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• 제25권1호
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• pp.28-30
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• 1994

The rate of growth and production of bioactive substances from Rehmannia glutinosa Liboschitz (Scrophulariaceae) were studied with the variation on the constituents of the culture media. The best growth was observed from MS basal medium containing 3.0 ppm NAA and 2.0 ppm kinetin. Carbohydrates (fructose, glucose and sucrose), phytosterols(${\beta}-sitosterol$, campesterol and stigmasterol) and carotenoid like substances were identified by GC-MS and TLC from the callus mass. However, catalpol was not detected from both solid and cell suspension cultures containing geraniol. Callus cultured Rehmannia glutinosa in the MS basal medium containing 0.1 ppm NAA and 0.1 ppm kinetin become differentiated to root.

• Plant Biotechnology Reports
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• 제2권2호
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• pp.153-161
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• 2008

The factors influencing somatic embryo maturation, high frequency somatic embryo germination, and plantlet formation were studied in Terminalia chebula Retz. Maturation of somatic embryo were influenced by a number of factors such as in vitro culture passage, concentrations of sucrose, levels of abscisic acid (ABA), basal media and media additive combinations. Maximum frequency of somatic embryo maturation ($57.22{\pm}2.02$), was obtained on MS medium supplemented with 50 g/l sucrose. Different factors such as strengths of MS nutrients, plant growth regulators, media additives and their combinations controlling somatic embryo germination and plantlet formation were studied. High frequency of germination and plantlet formation ($58.80{\pm}1.47$) were achieved by subsequent subculture of mature somatic embryos on MS medium containing 30 g/l sucrose and 0.5 mg/l benzyl-adenine (BA). However, although duration of in vitro passage of the callus tissue was critical, contribution of the combinations of plant growth regulators and media additives showed nugatory effect on somatic embryo maturation and germination as evident from variable responses.