• Title/Summary/Keyword: in vitro cultivation

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Effects of Acclimatization to Different Light Colors on the Growth of Petunia (Petunia hybrida) in a Greenhouse (조직배양 페튜니아의 순화과정에서 광질에 따른 생장반응 특성)

  • Young-Sun Kim;Geung-Joo Lee
    • Korean Journal of Environmental Agriculture
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    • v.42 no.1
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    • pp.14-20
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    • 2023
  • Light is an important factor that influences the growth and development of flowering plants. The present study investigated the effects of in vitro acclimatization to different light colors (white light (WL; control), blue light (BL; 447 nm), green light (GL; 519 nm), and red light (RL; 667 nm)) on the growth of petunia (Petunia hybrida) and of hardening cultivation of plant transferred form in vitro to a greenhouse under sunlight. Compared to the control, the shoot length and leaf width of Petunia increased by 42% and 11.7%, respectively, after acclimatization to BL and the shoot growth increased by 29.3% after acclimatization to RL. The chlorophyll and carotenoid contents after acclimatization to BL and GL were 16.7% and 11.3% higher, respectively, and 14.4% and 11.9% higher, respectively, than those in the control. During greenhouse cultivation, the shoot length increased by 16.7% and 11.3%, respectively, after acclimatization to BL and RL, respectively, and the leaf length and leaf width increased by 14.4% and 11.9%, respectively, after acclimatization to GL. While dry weight of root of GL and BL was not significant difference in vitro, increased by 59.0% and 22.9% ex vitro than that of WL. Thus, acclimatization to BL increased the shoot growth and leaf chlorophyll contents, and acclimatization to GL and RL enhanced shoot and root growth, in petunia.

The screening test on the efficacy of anthelmintics by using third-stage larvae and adult of cultivation in vitro (시험관내에서 인공배양한 제 3기 자충 및 성충을 이용한 구충효능 선발시험)

  • Jee, Cha-ho;Park, Seung-jun
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.589-594
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    • 1998
  • The in vitro screening tests against the in vitro cultivated $L_3$ of Ascaris suum (in vitro $L_3$), which were cultivated from the embryonated egg to third-stage larva on 7 days in culture(DIC) and the in vivo rat's lung-derived $L_3$ of Ascaris suum (in vivo $L_3$), which were recovered from the lungs of rat on 7 days after infection, carried out in order to compare the anthelmintic efficacy of in vitro $L_3$ and that of in vivo $L_3$ in RPMI medium 1640 with 5% bovine calf serum. And also a screening test of efficacy against adult worms of Trichuris suis performed. The efficacies of screening tests were as follows : 1. The screening efficacies of abamectin and ivermectin against the in vitro $L_3$ were all 100% at the 10ppm concentration in RPMI medium 1640 on 5 DIC. 2. The screening efficacies of abamectin and ivermectin against the in vivo $L_3$ were all 100% at the 20ppm on 5 DIC or at 40ppm on 3 DIC. 3. The screening efficacies of abamectin and ivermectin against the adult worms of Trichuris suis were all 100% at 20ppm on 4 DIC. And therefore, the in vitro cultivated $L_3$ of Ascaris suum were used in the screening test as well as the in vivo rat's lung-derived $L_3$ of Ascaris suum. And also the adult worms such as Trichuris suis and filaroids which is small size and difficult to cultivate to vitro, were used in the screening test in vitro.

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Optimization of in vitro seed germination of Taraxacum platycarpum

  • Lee, Jung-Hwan;Kim, Young-Kwan;Oh, Eun-Yi;Jung, Kuk-Young;Ko, Ki-Sung
    • Korean Journal of Environmental Agriculture
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    • v.28 no.4
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    • pp.403-408
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    • 2009
  • Dandelion (Taraxacum platycarpum) has been widely utilized for medicinal purposes. However, the dandelion seeds are relatively difficult to germinate under cultivation conditions, which hampers seedling propagation of dandelion plants and reduces the opportunity of usage of such a useful medicinal plant. Thus, in this study, in vitro conditions for the dandelion seed germination were optimized to enhance the germination rate. In seed washing steps, the sequential treatments with 20% of ethanol, 20% of NaOCl, and distilled water avoided microbial contamination with the highest in vitro germination rate (67.5%) from seeds sown in germination media. The media supplemented with 1.4 g/L of MS salts and 1% of sucrose significantly enhanced the germination rate compared to the media with 4.4 g/L of MS and 3% of sucrose. Sowing the seeds vertically in the optimized media supplement conditions, 1.4 g/L of MS salts and 1% of sucrose, gave the maximum in vitro germination rate (61%), which was almost three times higher than sowing seeds on a soil pot (23%). Our results indicate that the seed washing and sowing methods including germination medium supplements can be optimized to enhance in vitro seed germination of dandelion.

In vitro Biological Control Against Trichoderma harzianum Using Antifungal Bacteria

  • Lee, Ho-Yong;Hyun, Soung-Hee
    • Korean Journal of Environmental Biology
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    • v.18 no.4
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    • pp.441-446
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    • 2000
  • Trichoderma harzianum is an aggressive causal agent of green mold disease on mushroom cultivation. Some bacterial strains isolated, from oyster mushroom compost in Wonju, were found to have in vitro antifungal activity against Trichoderma harzianum ATCC 6385, 6504, and our isolates Trichoderma spp. Y and G. Further in vitro antifungal studies on several strains of phytopathogenic fungi showed that all of 12 phytopathogenic fungal strains were significantly inhibited by the isolated antifungal bacteria in Petri dishes. Of these, KATB 99121 showed the broadest inhibiting effect and displayed as negative coagulase, negative sulfide production and rod shape. KATB 99121 was resistant to ampicillin, chlorampenicol, and kanamycin. Identification of isolates was determined by Biolog GN system, and KATB 99121 was identified as Photobacterium logei because of 96 probability, 0.65 similarity, and 4.97 disturbance. With electron microscopy, thin section of KATB 99121 strain revealed typical rod-like shaped cell (0.6-0.8${\mu}{\textrm}{m}$$\times$1.5-2.0${\mu}{\textrm}{m}$) with prokaryotic structure and organization.

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In Vitro Proliferation Model of Helicobacter pylori Required for Large-Scale Cultivation

  • Oh, Heung-Il;Lee, Heung-Shick;Kim, Kyung-Hyun;Paek, Se-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.367-374
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    • 2000
  • The composition of dissolved gases and nutrients in a liquid medium were determined for establishment of the optimum conditions for in vitro culture of Helicobacter pylori. A microaerobic condition facored by the organism was prepared by adjusting the partial pressure of the gas, agitation speed, and viscosity of the medium. The gaseous concentrations were controlled by utilizing CampyPak Plus that reduced oxygen while augmenting carbon dioxide. Agitation of the broth facilitated the oxygen transfer to the cells, yet inhibited the growth at high rates. An increase of viscosity in the medium repressed the culture although this variable was relatively insignificant. The chemical constituents of the liquid broth were examined to establish an economic model for H. pylori cultivation. The microbe required a neutral pH for optimum growth, and yet was also able to proliferate in an acidic condition, presumably by releasing the acidity-modulating enzyme, urease. Cyclodextrin and casamino acid were investigated as growth enhancers in place of serum, while yeast extract unexpectedly inhibited the cells. A low concentration of glucose, the unique carbon source for the organism, increased the cell density, yet high concentrations resulted in an adverse effect. Under optimally dissolved gas conditions, the cell concentration in brucella broth supplemented with serum substitutes and glucose reached $1.6{\times}10^8$ viable cells/ml which was approximately 50% higher than that obtained in the liquid medium added with only cyclodextrin or serum.

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The Effects of Aqueous Fibrolytic Enzyme Products Supplementation on In vitro Fermentation and Growth Performance of Hanwoo Steers (섬유소분해효소 발효액의 첨가가 In vitro 발효 및 거세한우의 성장에 미치는 영향)

  • Kook, Kil;Chung, Wan-Tae;Park, Hong-Seok;Jang, Ki-Yeoung;Lee, Ji-Wung;Moon, Seung-Ju;Kim, Kwang-Hyun
    • Journal of Animal Science and Technology
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    • v.51 no.1
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    • pp.39-44
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    • 2009
  • The objective of this study was to investigate the effects of an enriched diet supplemented with aqueous fibrolytic enzyme products (FEP) on In vitro fermentation and growth performance of Hanwoo Steers. Total mixed ration combined with fibrolytic enzyme products showed a slight increase in the pH level on In vitro fermentation. Acetate ratio in volatile fatty acids, and the concentration of total volatile fatty acids were significantly greater at 12hr and 24hr after cultivation (p<0.05). Dry matter digestibility was significantly increased in groups supplied with aqueous fibrolytic enzyme products at 24hr after cultivation (p<0.05). In a rearing trial, the average daily gain and feed efficiency improved in the treatment groups supplied with aqueous fibrolytic enzyme products. Nutrient digestibility such as DM, crude fiber, NDF, and ADF digestibility were also significantly increased with the addition of aqueous fibrolytic enzyme products (p<0.05). Therefore, the aqueous fibrolytic enzyme products supplementation on In vitro fermentation is effective in improving the condition of rumainal fermantation. Also two kinds of aqueous fibrolytic enzyme products were found to be effective in improving the average daily gain and feed efficiency in Hanwoo Steers.

Effects of lactic acid bacteria and molasses on fermentation dynamics, structural and nonstructural carbohydrate composition and in vitro ruminal fermentation of rice straw silage

  • Zhao, Jie;Dong, Zhihao;Li, Junfeng;Chen, Lei;Bai, Yunfeng;Jia, Yushan;Shao, Tao
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.6
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    • pp.783-791
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    • 2019
  • Objective: This study was to evaluate the fermentation dynamics, structural and nonstructural carbohydrate composition and in vitro gas production of rice straw ensiled with lactic acid bacteria and molasses. Methods: Fresh rice straw was ensiled in 1-L laboratory silos with no additive control (C), Lactobacillus plantarum (L), molasses (M) and molasses+Lactobacillus plantarum (ML) for 6, 15, 30, and 60 days. After storage, the silages were subjected to microbial and chemical analyses as well as the further in vitro fermentation trial. Results: All additives increased lactic acid concentration, and reduced pH, dry matter (DM) loss and structural carbohydrate content relative to the control (p<0.05). The highest organic acid and residual sugar contents and lignocellulose reduction were observed in ML silage. L silage had the highest V-score with 88.10 followed by ML silage. L and ML silage improved in vitro DM digestibility as compared with other treatments, while in vitro neutral detergent fibre degradability (IVNDFD) was increased in M and ML silage (p<0.05). M silage significantly (p<0.05) increased propionic acid (PA) content and decreased butyric acid content and acetic acid/PA as well as 72-h cumulative gas production. Conclusion: The application of ML was effective for improving both the fermentation quality and in vitro digestibility of rice straw silage. Inclusion with molasses to rice straw could reduce in vitro ruminal gas production.

Shoot Induction and Genetic Stability of in vitro Cultured Pea

  • Kantayos, Vipada;Bae, Chang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.30-30
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    • 2019
  • Pea (Pisum sativum) is one of important legume crops in the world. It is commonly used as a protein source for animal and human diet, and also used as a natural nitrogen source which is produced by a symbiotic bacterium in their root nodule and helpful for terrestrial ecosystem. The successful in vitro manipulation is depended on three main factors including physiology of plant donor, in vitro manipulation approach, and stress physiology during plant cultivation. Moreover, genotype is an important for plant manipulation; different genotype gives the different response to regeneration efficiency. An efficient condition of shoot induction for pea (Pisum sativum cv. 'Sparkle') was developed by using optimum explant, plant growth regulator concentrations, and pretreatment of BA onto explant. The average shoot number per explant showed the highest on two kinds of shoot induction media (MSB5 media containing 2 mg/L BA and a combination of 2 mg/L BA and 1 mg/L TDZ) with cotyledonary node explants culture. Moreover, the pretreatment of explant in 200 mg/L BA solution was found to be more effective in shoot induction than that of non-pretreatment. The analysis of genetic stability of regenerants by using 13 ISSR markers presented that in vitro regenerated plants showed polymorphism with 8.3% compared with their mother plants.

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Bipolar Heterothallism, a Principal Mating System of Cordyceps militaris In Vitro

  • Shrestha Bhushan;Kim Ho Kyung;Sung Gi Ho;Spatafora Joseph W.;Sung Jae Mo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.6
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    • pp.440-446
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    • 2004
  • Interest in in vitro study of entomopathogenic fungi, including Cordyceps species, has been increasing due to their valuable bioactive compounds and biocontrol effects. Among Cordyceps species, in vitro stromata of C militaris has been successfully produced and cultivated for industrial purposes. However, genetic study on in vitro stromata formation of C militaris has not been carried out yet. Here, relationship between mating system and perithecial stromata formation of C militaris is reported. Mating system was determined by observing perithecial stromata formation from mono-ascospore cultures and their pair-wise combinations. Certain combinations of mono-ascospore strains produced perithecial club-shaped stromata, whereas other combinations produced either no stromata or only abnormal non-perithecial stromata. Similarly, mono­ascospore cultures without combination produced either no stromata or only abnormal non­perithecial stromata. Despite obvious heterothallism, self-fertility was occasionally observed in few strains of C militaris. These observations indicated that C militaris behaves as a bipolar het­erothallic fungus and requires two mating compatible strains in order to produce regular club­shaped perithecial stromata, a fundamental requirement for its industrial cultivation.

Effect of Rabbit Peritoneal Fluid(PF) on in vitro Maturation of Porcine Follicular Oocytes (가토복수가돼지 난포란의 체외성숙에 미치는 효과)

  • 정형민;박세필;오종훈;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.15 no.3
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    • pp.221-224
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    • 1991
  • This study was undertaken to evaluate the effect of rabbit peritioneal fluid(rPF) on in vitro maturtion of porcine follicular oocytes. From does 20h after hCG injection, rPF was aspirated aseptically at laparatomy, and then centrifuged, filtrated, and preincubated immediately for 12h. Porcine follicular oocytes isolated from ovaries of slaughtered animals were incubated in TCM-HEPES+10% FCS, TCM-HEPES+rPF(v/v, 50/50), or rPE only and examined the nuclear maturation after aceto-orcein or hochest staining. After identifying the optimal incubation time, this experiment was repeated for 5 times. Under the TCM-HEPES containing hormones and serum codition, the time range of porcine follicular oocyte maturation was 38 to 44 hours and the optimal time of maturation of follicular oocyte in vitro was 42 hour cultivation, respectively. The maturatin rates(89.4% and 92.7%) of porcine follicular oocytes cultured in the media with 50% rPF or only rPF were signifciantly higher thanthat (84.6%) of oocytes cultured with TCM-HEPES, respectively. These results suggest that the unknown components(s) of rPF promoted in vitro maturation of porcine follicular oocytes.

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