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

• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.431-437
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• 2017

Mulberry (Morus sp.) of the family Moraceae is very economically important in Asian countries including Korea, because its leaf and fruit have been commercially used in sericulture and horticultural industries. Therefore it is necessary to develop the optimal production system for rapid and cost-effective propagation of mulberry. Our studies focused on establishing an acclimatization method for the successful plantlet production of new cultivar 'Cheongsu' which was transferred ex vitro after in vitro culture. In particular, effect of abscisic acid (ABA) addition into the last subculture medium on plantlet response to subsequent ex vitro transfer and its growth was investigated. During acclimatization, stomatal conductance and transpiration rate of ABA-pretreated plantlets were significantly lower than those of non-treated plantlets. Net photosynthetic rate of ABA-pretreated plantlets decreased after ex vitro transfer but increased after 14 days, and it was mostly higher than that of non-treated plantlets. Moreover, relative water content as well as chlorophyll contents and its ratio were also higher in ABA-pretreated plantlets. On the other hand, proline was considerably higher than in control plantlets. After 1 month of ex vitro transfer, survival rate of ABA-pretreated plantlets was 85.6%, which increased by 29.1% in comparison with control (56.5%). More vigorous growth was also observed in ABA-pretreated plantlets. From these results, it was found that application of ABA to the last subculture medium could improve acclimatization and promote survival of mulberry plantlets after ex vitro transfer, inducing water stress tolerance and alleviating abiotic stresses.

• Korean Journal of Plant Resources
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• v.27 no.3
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• pp.256-262
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• 2014

Viscum album var. coloratum (mistletoe) is considered as one of the endangered plant species in Korea. Our objective is to restore its population and multiplication of plant by ex situ method. In this research we explored the maximum germination (in vitro) of freshly harvested and stored seeds of mistletoe collected in different time intervals. Ex vitro germination after artificial inoculation on the branches of Prunus mume in different physiological conditions was also monitored. The research revealed that the lately harvested seeds (Feb. and March 2014) were superior over early harvested seeds (Nov. 2013 and Jan. 2014) of mistletoe due to the higher percentage of germination (above 93%). According to the data, it is also revealed that the survival and germination rate of mistletoe seeds decreased with the increase in storage duration. In ex vitro germination, the fluctuated temperature of a glass house in natural condition enhanced (four fold) the rate of germination on the branches of Prunus mume than the constant temperature condition in the glass house.

• Korean Journal of Animal Reproduction
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• v.14 no.1
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• pp.17-22
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• 1990

The present study was carried to investigate the development potential of bovine follicular oocytes matured and fertilized in vitro. Bovine oocytes matured in vitro were fertilized, cultured, and transfered to the rabbit oviduct for in vitro culture to evaluate the development potential. The rates of in vitro maturation, fertilization and polyspermy were 96%(355/369), 90%(320/355) and 15%(17/320), respectively. The percentage of oocytes cleaved after culture for 48 hours, to 2 cell, 3~4 cell, 6~8 cell stage were 17%(60/356), 21%(75/356) and 19%(67/356), respectively and overall cleavage rate was 57%(202/356). The rate of recovered embryos after 5 days in rabbit oviduct was 56%(80/142), and 21%(17/80) of recovered embryos developed over morula stage.

• Journal of Embryo Transfer
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• v.8 no.2
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• pp.143-149
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• 1993

The objective of this study was to produce calves derived from in vitro fertilization of in vitro matured follicular oocytes. Oocytes aspirated from small antral folicles of ovaries obtained at a local slaughter house were matured and fertilized in vitro. At l8hrs after insemination with Korean native cattle semen, oocytes were co-cultured for 6~7 days by utilizing co-culture system with bovine oviduct epithelial cell. After co-culture, good or excellent quality late morulae or early blastocysts were selected by morphological criteria under stereo microscope. Selected embryos were transferred to recipients on day 6 or 7 (estrus = day 0). Recipients were monitored by observation for estrus and rectal palpation after 60 days from embryo transfer. One of them went to term with the birth of a calf. This case is the first production of calf derived from in vitro fertilization in Korea.

• Journal of Plant Biology
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• v.12 no.2
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• pp.23-27
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• 1969

The promotive effect of Ca in semi-vitro culture of Oenothera biennis pollen was by and large identical to that of in vitro systems. This promotive effect became additive when boron was supplemented to the media and even more so if K was further supplemented. No such Ca action was observed then performed under the conditions of relatively low temperature. An uncoupling agent of ATP in respiration, DNP, inhibited the promotive action of B, but not that of Ca. The inhibitory effect of DNP was greater at low temperatures. IAA was rather inhibitory on pollen growth in semi-vitro culture, even much greater than it if DNP was supplemented to the IAA-containing media.

• Journal of Embryo Transfer
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• v.7 no.2
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• pp.89-96
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• 1992

This research was conducted to investigate the interrelationship among methods of injection of PMSG-hCG to the number of ovulated eggs, percentage of matured oocytes and in vitro fertilization using out-bred ICR mice. The results obtained are as follows, 1) The optimurn dose was 5 IU for both PMSG and hCG, while the number of ovulated eggs was 42$\pm$8, percentage of M II was 73% and in vitro fertilization rate was 81 %. 2) The optimum injection interval of PMSG-hCG was 48 hours, while the number of ovulated eggs was 48 $\pm$ 8, percentage of M II was 80% and in vitro fertilization rate was 81%. 3) The optimum time for collecting eggs was between 16 and 18 hours after hCG injection, while the numbers of ovulated eggs were 44$\pm$8, 42$\pm$7 and 43$\pm$7 in 14,16 and 18 hours after hCG injection respectively, and percentages of M II were 79 and 81 %, and in vitro fertilization rates were 81 and 80% in 16 and 18 hours after hCG injection, respectively. 4) The repeat of superovulation decreased with the number of ovulated eggs, percentage of M II and in vitro fertilization rate, than in control. But it was recovered by increasing the repeat interval.

• Journal of Embryo Transfer
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• v.23 no.2
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• pp.77-80
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• 2008

In the present study, effects of concentration of cryoprotectant solutions on the nuclear maturation of vitrified-thawed porcine oocytes were examined. Also, the developmental capacity of vitrified-thawed immature porcine oocytes following ICSI was investigated. Oocytes were cultured in NCSU-23 medium supplemented with 5% FBS at $38^{\circ}C$ in 5% $CO_2$ and air. The in vitro maturation rate of vitrified-thawed oocytes ($24.1{\pm}2.5%$) was lower than that of the control ($46.0{\pm}3.2%$, p<0.05). The in vitro maturation rate of vitrified-thawed oocytes treated with $1.0{\sim}5.0\;ug$ CB + NCSU- 23 medium were $22.2{\pm}3.0%$, $30.7{\pm}3.2$, $46.3{\pm}3.1%$, $38.5{\pm}3.2%$, respectively. The in vitro maturation rate ($46.3{\pm}3.4%$) of the vitrified-thawed oocytes treated with $3.0\;{\mu}g$ CB for 30 min was the highest of all vitrification groups. When the in vitro developmental rates of the vitrified-thawed (with EDS and EDT) oocytes following ICSI were $18.5{\pm}2.5%$, $16.4{\pm}2.1%$, respectively. This results were lower than the control group ($24.0{\pm}2.5%$).

• Journal of Embryo Transfer
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• v.27 no.2
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• pp.71-80
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• 2012

Biotechnologies for cloning animals and in vitro embryo production have the potential to produce biomedical models for various researches. Especially, pigs are a suitable model for xenotransplantation, transgenic production and various areas of reproductive research due to its physiological similarities to human. However, utilization of in vitro-produced embryos for transfer remains limited. Despite improvement over past few decades, obstacles associated with the production of good quality embryos in vitro still exist which limit the efficiency of cloning. One of major problems includes improper in vitro maturation (IVM) and culture (IVC). Oxidative stress caused from in vitro culture conditions contributes to inadequate IVM and IVC which leads to poor developmental competence of oocytes, failure of fertilization and embryo development. To reduce the oxidative stress, various antioxidants have been used to IVM and IVC system. However, limited information is available on the effects of resveratrol on livestock reproductions. Resveratrol is a polyphenolic natural product and well known as an antioxidant in foods and beverages (e.g. in grapes and red wine). Resveratrol is known to be cardioprotective, anticarcinogenic, anti-inflammatory, antioxidant and antiapoptotic. This paper will review the effects of resveratrol on in vitro maturation of oocytes and embryo development.

• Asian-Australasian Journal of Animal Sciences
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• v.9 no.4
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• pp.375-379
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• 1996

The experiment investigated the possibility of using effluent from RUSITEC (rumen simulation technique) inoculated with rumen liquor or cow faeces as sources of micro-organisms for in vitro digestion of forages. Nine forages ${\times}3$ sources of inoculum were used in a factorial arrangement of treatments. Rumen liquor was collected from fistulated sheep and faeces was collected from cows. The RUSITEC apparatus consisted of 4 vessels, 2 vessels were charged with faecal liquor and 2 with rumen liquor. On the 8th day of the experiment RUSITEC effluent were collected to use in in vitro studies. In vitro OMD (g/kg) values using three sources of inoculum (fresh rumen liquor, RUSITEC effluent from rumen liquor or cow faeces) were statistically significant (p < 0.001). The regression relationships between OMD using fresh rumen liquor and RUSITEC effluent were highly significant ($R^2>0.90$). The results suggest that RUSITEC effluent either from rumen liquor or cow faeces can be used as a source of micro-organisms for in vitro digestion of forages.