• Korean Journal of Plant Resources
• /
• v.32 no.3
• /
• pp.237-243
• /
• 2019

Soybean is the one of recalcitrant legume species for shoot induction. Shoot regeneration via direct organogenesis was investigated in five soybean cultivars, 'Dawon', 'Pungsan', 'Daewon', 'Taekwang' and 'Chongdoo 1' by using cotyledonary node explants. Out of 5 soybean cultivars, an efficient shoot regeneration condition was developed in the two soybean cultivars, 'Dawon' and 'Pungsan'. When various kinds of plant growth regulators with different concentration were estimated, the optimum medium condition for shoot induction in both soybean cultivars was MS + B5 vitamin supplemented with BA at concentration 2 mg/L. In addition, shoot formation efficiency was increased with 97.09% and 93.88% by the pretreatment of BA onto the explants before in vitro culture in both cultivars. Shoot induction in 'Dawon' cultivar was originated from epidermal tissue and sub-epidermal layers when histological changes were investigated under shoot regeneration after culturing cotyledonary node segments on shoot induction medium for 0 to 21 days. Especially, cell dedifferentiation was observed from parenchyma cells to meristematic cell in 3-day cultured segments.

• Journal of Plant Biotechnology
• /
• v.49 no.3
• /
• pp.230-239
• /
• 2022

Holostemma annulare (Family Asclepiadaceae) is an invaluable vulnerable medicinal plant; the root tubers are used in Ayurveda medicine and by folk healers to treat various ailments. In this study, Schenk and Hildebrandt medium fortified with the cytokinins 6-benzyl adenine, kinetin, and auxins, including indole 3-butyric acid, indole 3-acetic acid, α-naphthaleneacetic acid, and 2,4-dichlorophenoxyacetic acid, were checked for their efficiency on root tuber induction from different explants. Adventitious root tubers were more successfully induced from in vitro leaf segments and shoots when cultured in Schenk and Hildebrandt medium supplemented with 0.5 mg/l of α-naphthaleneacetic acid. In addition, preliminary phytochemical analysis of in vitro root tubers and identification of different secondary metabolites were conducted. Thin layer chromatography and high performance thin layer chromatography analysis of the crude methanolic extracts of the in vitro root tuber identified the presence of lupeol, a bioactive triterpene. Adventitious root tuber induction offers a novel method for the in vitro production of bioactive metabolites that can be scaled up by bioreactors, thus ensuring the conservation and sustainable utilization of H. annulare. The study warrants further scale-up production and pharmacological investigation that can be extended for pharmaceutical needs.

• Plant Resources
• /
• v.2 no.1
• /
• pp.10-15
• /
• 1999

In vitro shoot regeneration from cotyledon and hypocotyl explants derived from germinating mature Impatiens seeds. The induction of organogenetic tissue was also influenced by the cotyledon and hypocotyl. Multiple shoot induction was higher in hypocotyl than in the cotyledon explant with Thidiazuron and a NAA medium.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2019.10a
• /
• pp.30-30
• /
• 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.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2022.09a
• /
• pp.63-63
• /
• 2022

Pear (Pyrus spp.) is a typical fruit and grown in the temperate climate regions throughout the world. Development of appropriate methods for in vitro propagation and root induction are important to increase the production rate and plant quality rapidly. This study was conducted to find the most appropriate media conditions for in vitro propagation and rooting of three pear cultivars, 'Barttlett', 'BaeYun No.3' and 'Oharabeni'. In vitro propagation was induced on Murashige and Skoog medium (MS) with 2.0 mg/L N6-benzyladenine (BA) and 0.2 mg/L indole-3-butyric acid (IBA) medium. For root induction of these cultivars, the shoot explants of the propagated plants were cultured on two different media containing 1/2 MS medium containing 0.2 mg/L IBA with 15 g/L Sucrose (Rooting Medium 1, RM1) and 1/4 Linsmaier and Skoog medium (LS) medium containing 1 mg/L IBA and 1 mg/L NAA hormone with 7.5 g/L glucose (Rooting Medium, RM2) and after 2 weeks, the plants on the RM2 medium are transferred on RM1 medium (RM2 condition). After nearly seven weeks, percentage of rooting formation were 22.2% in RM1 and 30% in RM2 conditions for Barttlett and 70% in RM1 and 60% in RM2 conditions for Oharabeni cultivars. No differences in these cultivars were observed between RM1 and RM2 conditions. However, BaeYun No.3 cultivar was observed 0% in RM1 and 72.7% in RM2 conditions. This study will help to propagation and root induction of in vitro plants for various pear cultivars.

• Journal of Plant Biotechnology
• /
• v.47 no.1
• /
• pp.26-39
• /
• 2020

In maize, immature embryos (IEs) are highly regenerative explants most suitable for producing high frequencies of plantlet regeneration in vitro. Apart from media, explants, and hormones, genotypic variation also influences in vitro characters to a great extent. In the present study, IEs were used to study the distinctive effect of variation of size/stage and hormones in different genotypes on five in vitro characters viz., frequency of callus induction, growth rate of total callus, frequency of E. callus induction, and volume and number of regenerated plantlets. LS medium with different concentrations of 2,4-D (0.5, 1.5, 2.5, 4.0 and 5.0 mg/L) were used to study the former four in vitro characters, and medium with 6-benzylaminopurine and kinetin (0.5 mg/L, each) was used for plantlet regeneration. IEs of 1.0, 1.5, 2.0, 2.5 and 3.0 mm in size were isolated from four inbred lines viz., NM74C, NM81A, NM5883 and NM5884. Two-way ANOVA revealed that explant size and genotypes, as well as hormonal concentrations showed significant effects on in vitro characters. Two millimeter IEs were found to be suitable for in vitro cultures. LS medium with 1.5 mg/L 2,4-D and LS with BAP and Kn (0.5 mg/L, each) were found to be the best hormonal concentrations for callus induction, maintenance, and regeneration, respectively. Among the four genotypes, NM81A and NM5883 yielded more non-embryogenic and Type I E. calli. In contrast, NM74C and NM5884 yielded more highly regenerative Type II calli. Inbred line NM5884 was found to be the best among these four genotypes.

• Korean Journal of Plant Resources
• /
• v.35 no.6
• /
• pp.770-777
• /
• 2022

The main objective of this study was to identify the most appropriate condition for root formation of in vitro micropropagated pear (Pyrus spp.) plants. In vitro propagation was induced on Murashige and Skoog (MS) medium with 2.0 mg/L of N6-benzyladenine (BA) and 0.2 mg/L of Indole-3-butyric acid (IBA) medium. The short pre-treatment of explants with a high concentration (1 mg/L) of NAA and IBA (R0 medium) in dark for three days, followed by transfer to five different media (R1 to R5) resulted in good rooting responses in the pear 'Oharabani (P. pyrifolia × P. communis)' genotype. For the rooting experiments, the highest rooting percentage (83.3 ± 8.3%), average root length (3.6 ± 1.9 mm), total root number (31 ± 4.0), and average root number per plant (2.6 ± 2.1) were obtained on half strength (1/2) of MS medium supplemented with 30 g/L sucrose without hormones and activated charcoal (AC) (R1 medium). The highest rooting percentage was obtained at 83.3% from explants on R1 and R3 media. The rooting procedure described in this study resulted in good root formation and significantly shorting the root induction time to within 14 days of culture. Further studies are underway to test the suitability of the protocol developed in this study for other pear genotypes.

• Korean Journal of Medicinal Crop Science
• /
• v.21 no.2
• /
• pp.142-147
• /
• 2013

Gastrodia elata has been cultivated as an important medicinal resources to treat various human diseases. One of the major problems associated with its field production is the degeneration of seed tubers, which is mainly caused by soil-borne pathogens. This study was conducted to produce disease-free seed tubers by the development of in vitro micropropagation method. First, tubers of G. elata were treated with $HgCl_2$ prior to culturing in vitro. Among various culture medium tested, water agar (WA) and WPM medium were the most effective on the induction and growth of vegetative stems. NAA ($0.1mg/{\ell}$) or TDZ ($1.0mg/{\ell}$) in WA medium showed better growth of vegetative stems compared to other plant hormones. Finally the induction and growth of vegetative stems were better in the dark compared to the light condition. In this study, we established an in vitro micropropagation system of G. elata, which might be an efficient way to increase the yield and quality of G. elata tubers in the field production.

• Journal of Pharmaceutical Investigation
• /
• v.38 no.3
• /
• pp.183-189
• /
• 2008

The present study compared the feasibility of Caco-2 and MDCK cells as an efficient in-vitro model for the drug classification based on Biopharmaceutics Classification System (BCS) as well as an in-vitro model for drug interactions mediated by P-gp inhibition or P-gp induction. Thirteen model drugs were selected to cover BCS Class I{\sim}IV$ and their membrane permeability values were evaluated in both Caco-2 and MDCK cells. P-gp inhibition studies were conducted by using vinblastine and verapamil in MDCK cells. P-gp induction studies were also performed in MDCK cells using rifampin and the P-gp expression level was determined by western blot analysis. Compared to Caco-2 cells, MDCK cells required shorter period of time to culture cells before running the transport study. Both Caco-2 and MDCK cells exhibited the same rank order relationship between in-vitro permeability values and human permeability values of all tested model compounds, implying that those in-vitro models may be useful in the prediction of human permeability (rank order) of new chemical entities at the early drug discovery stage. However, in the case of BCS drug classification, Caco-2 cells appeared to be more suitable than MDCK cells. P-gp induction by rifampin was negligible in MDCK-cells while MDCK cells appeared to be feasible for P-gp inhibition studies. Taken all together, the present study suggests that Caco-2 cells might be more applicable to the BCS drug classification than MDCK-cells, although MDCK cells may provide some advantage in terms of capacity and speed in early ADME screening process.

• Toxicological Research
• /
• v.3 no.1
• /
• pp.1-8
• /
• 1987

In vitro induction of cytochrome 450 associated monooxygenase activities by phenobarbital (PB) and 3-methylcholanthrene (MC) was investigated in primary cultures of adult rat hepatocytes. PB and MC were added to the culture 24 hr after the initial plating of hepatocytes. A signiftcant increase of the activities of 7-ethoxycoumarin 0-deethylase and aryl hydrocarbon hydroxylase were observed in MC and PB treated culture. MC caused about 500% induction of the initial oxidation rates of both enzymes in 48 hr. However the PB maintained both enzyme activities close to the level of freshly isolated hepatocytes. Biphenyl 4-hydroxylase and aminopyrine N-demethylase activities were also induced by MC and PB. But the level of induction was less than that occuring with 7-ethoxycoumarin 0-deethylase and aryl hydrocarbon hydroxylase. When aflatoxin $B_1$ was added to the hepatocyte cultures which have been treated with MC or PB, it caused a significant increase of the unscheduled DNA synthesis at higher dose of aflatoxin $B_1$ as compared to those of untreated control hepatocyte cultures. The results suggest that microsomal enzyme activities can be selectively controlled preferably in hepatocyte cultures by the in vitro induction method. This principle may be useful for studying the metabolism and other toxicological studies.