• Journal of Plant Biology
• /
• v.28 no.2
• /
• pp.141-148
• /
• 1985

The effects of sizes and densities of cells cultured, conditioned medium, and media pH on the somatic embryogenesis of carrot (Daucus carota L.) were examined. A large number of globular embryoids was formed after 4 days in cell culture, and later globular embryoids developed into heart and torpedo shape. High cell density resulted in higher number and better growth of embryos, especially on conditioned medium than Murashige-Skoog medium. The fresh weight and number of embryoids formed increased with the decrease in cell size. The significant reduction in fresh weight and number of embryoids was obtained when culturing cells with diameter of over 90 ${\mu}{\textrm}{m}$. Dry weight and number of embryoids were markedly reduced with medium pH of 4 or 7, but promoted with pH 6.0.

• Proceedings of the Korea Water Resources Association Conference
• /
• 2021.06a
• /
• pp.229-229
• /
• 2021

최근 이상기상으로 국지성 집중호우가 빈번하게 발생함에 따라 토사유출에 의해 피해가 증가하고 있다. 급경사 사면을 갖는 교란산지와 개발지의 경사면은 토양침식 위험에 노출되어 있다. 특히 지표유출수의 집중흐름에 의한 세류발달은 세류간침식에 비해 많은 토양침식과 토사유출을 발생시킨다. 이러한 급경사 사면에서의 세류발달에 대한 수리학적 메터니즘과 한계조건을 파악하고자 강우와 유입수 모의를 통한 토양침식 실험을 수행하였다. 강우와 유입수의 상호작용에 의한 세류발달의 특성과 토사유출량 증가를 파악하고자, 강우량의 증가와 유입수 유무에 따른 실험이 이루어졌다. 강우강도의 범위는 100~140mm/hr이고, 유입수는 170ml/s 이였다. 유입수량은 합리식으로 계산하였으며, 토양상자 상단에서 균등분사식으로 유입되었다. 세류발달의 수리학적 특성치를 취득하기 파악하기 위해, 염료주입 지표수 거동에 대한 동영상을 판독하였다. 세류발달은 유입수가 없는 경우 3분 이내 그리고 유입수가 있는 경우 30초 이내에 발생하였다. 토양상자내의 평균 유속범위는 7.83~29.27 cm/s 이였다. 실험에서 발달된 세류는 15분경과 후부터 평형상태를 유지하였다. 경사실험조건별로 세류 차수분석을 위해 각 세류의 최대길이, 폭, 넓이를 측정한 결과 세류의 차수는 강우만 있는 경우 1~2차로 발달하였으며, 유입수가 있을 때는 2~3차로 발달하였다. 지표유출이 증가하면서 세류발달의 개수는 줄었지만 규모는 더 크게 나타났다. 세류가 발달하는 시점의 한계유속은 10~20cm/s 범위인 것으로 파악되었다. 지표유출이 증가하면서 세류의 발달도 더 심화되지만, 동일한 강우에 일정시간동안 노출되면서 세류는 평형상태가 되어 더 이상 침식이 활발하게 진행되지는 않았다. 유입수가 있는 조건에서 수량은 약 2배가 증가하였지만, 토사유출량은 세류확장에 의해 약 4배가 증가하는 것으로 나타났다.

• Clinical and Experimental Reproductive Medicine
• /
• v.24 no.2
• /
• pp.241-251
• /
• 1997

체외성숙과 수정된 돼지 난자의 체외발달능이 체외배발생 배양액인 NCSU 배양액에 0.4% BSA, 10% 혈청 혹은 아미노산 (2% BME 아미노산 용액과 1% MEM 아미노산 용액)을 첨가함으로서 조사되었다. 본 실험에 공시된 난자는 체외수정 추 30시간 (2-세포기)혹은 48 시간 ($2{\sim}4$-세포기)에 회수하였다. 실험I에서 0.4% BSA가 첨가된 NCSU 배양액에서 2-세포기 난자들의 배양경과시간에 따른 발달능을 조사한 결과, 배양 후 72 시간 (체외수정 후 102 시간)에 상실배기와 배반포기 배가 나타났으며, 배양 후 120 시간째 (체외수정 후 150 시간)에도 팽창된 배반포기 배까지만 발달하였다. 실험II는 체외수정 후 48 시간의 분할된 ($2{\sim}8$-세포기) 난자들의 핵과 외관적 분할구와의 수적 차이를 조사한 결과, $2{\sim}4$-세포기보다는 5-세포기 이상에서 핵과 분할구의 조화에 차이가 많았다. 실험III에서는 $2{\sim}4$-세포기 난자들을 배양후 5일째의 배반포들의 투명대의 두께, 난자 크기 그리고 inner cell mass (ICM)과 trophectoderm (TE)의 세포 배열을 조사한 결과, 난자의 크기가 커짐에 따라서 투명대가 얇아지고 전체 세포수가 증가하였지만, ICM의 비율은 차이가 없었다. 실험IV에서는 BSA, 혈청 혹은 아미노산이 첨가 혹은 무첨가된 배양액내에서 $2{\sim}4$-세포기 난자들의 배반포 후 부화능력을 조사한 결과, 모든 군에 있는 난자들은 팽창된 배반포기 배까지 발달할 수 있었던 반면, 난자의 부화는 아미노산 혹은 혈청이 포함된 배양액에서만 일어났다. 더우기 상실배기와 배반포기 시기에 혈청의 첨가는 부화 배반포기 배의 발달을 현저히 증가시켰다. 또한 아미노산과 혈청의 영향을 받은 팽창 배반포기 배는 얇은 투명대, 팽창된 난자의 크기 그리고 ICM과 전체 세포수의 증가를 보였다. 이상의 결과로 미루어 볼때, 배양액내에 대한 아미노산과 혈청의 첨가는 돼지 배반포기 배의 부화를 유도할 수 있다고 보며, 더우기 이들 요소들은 투명대의 두께, 난자의 크기 그리고 ICM과 전체 세포수에 영향을 미친다.

• Journal of Embryo Transfer
• /
• v.21 no.1
• /
• pp.35-43
• /
• 2006

Ions play important roles in various cellular processes including fertilization and differentiation. However, it is little known whether how ions are regulated during early embryonic development in mammalian animals. In this study, we examined changes in $Ca^{2+}\;and\;K^+$ concentrations in embryos and oviduct during mouse early embryonic development using patch clamp technique and confocal laser scanning microscopy. The intracellular calcium concentration in each stage embryos did not markedly change. At 56h afier hCG injection when 8-cell embryos could be Isolated from oviduct, $K^+$ concentration in oviduct increased by 26% compared with that at 14h after injection of hCG During early embryonic development, membrane potential was depolarized (from -38 mV to -16 mV), and $Ca^{2+}$ currents decreased, indicating that some $K^+$ channel might control membrane potential in oocytes. To record the changes in membrane potential induced by influx of $Ca^{2+}$ in mouse oocytes, we applied 5 mM $Ca^{2+}$ to the bath solution. The membrane potential transiently hyperpolarized and then recovered. In order to classify $K^+$ channels that cause hyperpolarization, we first applied TEA and apamin, general $K^+$ channel blockers, to the bath solution. Interestingly, the hyperpolarization of membrane potential still appeared in oocytes pretreated with TEA and apamin. This result suggest that the $K^+$ channel that induces hyperpolarization could belong to another $K^+$ channel such as two-pore domain $K^+(K_{2P})$channel that a.e insensitive to TEA and apamin. From these results, we suggest that the changes in $Ca^{2+}\;and\;K^+$ concentrations play a critical role in cell proliferation, differentiation and reproduction as well as early embryonic development, and $K_{2P}$ channels could be involved in regulation of membrane potential in ovulated oocytes.

• Korean journal of applied entomology
• /
• v.33 no.4
• /
• pp.242-249
• /
• 1994

These studles were conducted to investigate the development of chem~cal resistance and cross.resista. nce m dlarnondback moth (Piuteiia xylostella L). The resistance development of diamondback moth greatly vaned under single selection of five insectlades. The tnflumuron and lambda cyhalothnn strains at 8th selected generation showed 374- and 29.l.fold resistant levels. respectively, as compared with the susceptible strain. However, the Baciilus thuringiensis-seleded at 8th selected generatLon exhibited 240-fold resistant level. and the prothiophos-selected at 8th generation revealed 14.3-fold resistant !eve1 while the cartap hydrochloride-selected at 8th generailon showed 9.1-fold resistant level Prothiophos- selected strain showed low cross-resistance level to cartap hydrochloride, while this strain exhibited no cross-resistance of 1.3 to 2.8-fold to other msectlcides. Cartap hydrochlonde- seleded strain showed 19.9-fold. a high cross-resistance to lambda cyhalothrin, but this strain showed 2.2-34 fold, no cross resistance to other insecticide. Lambda cyhalothnn-selected strain exhibited cross-resistance to cartap hydrochlolide and prothiophas. Triflumuron-seleded strain showed 1.3-4.9 fold. no cross-resistance to other ~nsectic~dTe he B. ihuring~ensis-selededs train showed no cross-resistance to other insecticides.

• Korean Journal of Plant Tissue Culture
• /
• v.24 no.2
• /
• pp.107-112
• /
• 1997

This study was carried out to establish a mass production of normal somatic embryos of Oenanthe iavanica (BL.) DC. including examination of nitrogen and sucrose sources, and ABA concentration. Embryogenic cell clumps and embryos were formed on the MS medium devoid of growth regulators. Proliferation of embryogenic cells and clumps was enhanced by 2, 4-D. Meanwhile embryo growth and development occurred on the media containing NAA and IBA. Growth of embryos was generally good in the media containing both 20 mM $KNO_3$ and 20 mM $NH_4NO_3$. The rate of shoot forming embryos was higher on the media containing on1y 20mM $NH_4NO_3$ than on the former. Addition of sucrose at 3-6% enhanced the embryo development, and normal embryos with short hypocotyl was observed on the medium containing $10\mu\textrm{M}$ ABA. Embryogenic cell clumps or globular embryos, when transferred to MS solid media devoid of growth regulators, developed into mature embryos and then into plantlets which had entire primary leaves like zygotic seedlings.

• Journal of Embryo Transfer
• /
• v.22 no.1
• /
• pp.75-80
• /
• 2007

The objective of this study was to examine the optimal concentration and the exposure time of ethanol, Ca-ionophore, and strontium to achieve massive recipient oocytes in porcine. The cleavage (51.4% vs. $21.3{\sim}44.3%$) and embryo development rates (45% vs. $13.3{\sim}29.9%$) were significantly higher (p.0.05) in oocytes treated with 10% ethanol for 10 min than other treatments. The oocytes treated with 25mM Ca-ionophore for a minimum of 2min and 20mM strontium for a minimum of 6h showed significantly higher cleavage and embryo development rates than those of other treatments (P<0.05). Cleavage rate with duplicated ethanol treatment was significantly lower than those with ethanol alone (P<0.05). The cleavage rate and embryo development rates were significantly lower in duplicated strontium treatment than those in both alone and combination (P<0.05). But the cleavage and embryo development rates in treatment with Ca-ionophore were significantly higher in combined treatment (Ca-ionophore and cycloheximide) than those in single or duplicated treatment (P<0.05). These results might induce establishment of the optimal concentration and the exposure time on activation media to build up activation condition of porcine oocytes.

• Reproductive and Developmental Biology
• /
• v.28 no.1
• /
• pp.29-36
• /
• 2004

The objective of this study was to investigate the effects of amino acid supplementation of oocyte maturation medium on 1st polar body(PB) extrusion, embryo development and blastocsyt cell number. In experiment 1, Cumulus oocyte complexes(COCs) were matured in in vitro maturation(IVM) medium supplemented with 1, 2, or 4-fold of 10 $\mu$l/ml MEM non-essential amino acid(NEAA) and 20 Park, $\mu$ l/ml BME essential amino acid(EAA). The PB extrusion rate of oocytes matured in 1-fold amino acid group was significantly higher than that matured in medium without amino acid (p＜0.05), but it was decreased by the increase of the dosage of amino acid. There were no difference in the percentage of embryos reaching 2-cell, 8-cell and blastocyst in all treatments. The number of trophectoderm(TE) cells and total cell number of blastocysts were highest in 2-fold amino acid group, and the number of inner cell mass(ICM) cells was increased by the increase of the dosage of amino acid. In experiment 2, COCs were matured in IVM medium with 1, 5, or 10 mg/ml lactalbumin hydrolysate(LAH). The PB extrusion rate of oocytes matured in medium with 5 mg LAH was significantly higher than that matured in medium with 1 mg LAH (p＜0.05). The development rate to the blastocyst stage was significantly higher in non-supplement and 1 mg LAH group than in 5 mg and 10 mg LAH group (p＜0.05). The number of TE cells and total cell number did not differ among treatment groups, but the number of ICM cells was increased by the increase of LAH supplement. These results suggested that the supplement of certain group of amino acid in IVM medium effective on the quality of blastocyst, and further studies will be accompany with the search of new sources of amino acid used for the use of in vitro embryo production.

• Journal of Plant Biotechnology
• /
• v.32 no.1
• /
• pp.37-44
• /
• 2005

Microspores were isolated from pepper (Capsicum annuum L.) anthers by using a micro-blender and cultured in modified NLN medium at $25^{\circ}C$. The influence of pretreatment period at $32^{\circ}C$, adding the 2-hydroxynicotinic acid to a pretreatment medium, and co-pretreatment anthers with microscopes on the induction of embryo were examined. Globular and torpedo embryos were observed from 3 weeks after culture. Embryo development was not synchronized within culture. After 4 weeks in culture, in addition to globular and torpedo embryos, cotyledonary embryos were observed. Normal cotylodonary embryos developed into plantlets when transferred to a solid hormone free B5 medium containing $2\%$ sucrose. Embryo yields were significantly higher after 1- and 2-day pretreatment at $32^{\circ}C$. However the development of embryo ceased at the globular or heart stage. In contrast, embryo yields were lower after 3- to 6-day pretreatment at $32^{\circ}C$ and embryo developed at the cotyledonary stage. After adding the 2-hydroxynicotinic acid to anther pretreatment solution, embryo yields were slightly increased. However most embryos occurred were at the globular or heart stage. Co-pretreatment of microspores with anthers was deleterious for embryo induction and development. AS far as we know, this is the first report of success in obtaining high frequency of embryogenesis and plantlets formation from isolated microspores of pepper. Although the culture conditions have to be optimized further, this promising microspore culture system can be used for genetic transformation, selection for dominant and recessive traits as well as for the production of homozygous doubled haploid plants.

• Proceedings of the Korea Water Resources Association Conference
• /
• 2023.05a
• /
• pp.194-194
• /
• 2023

세류침식은 급경사 나지사면에서 증가하는 지표흐름에 의해 빈번하게 발생하고, 과도한 토사유출로 인해 홍수 및 토사재해 위험 증가와 수질오염 등의 문제를 야기한다. 본 연구는 개발지역의 마사토를 활용하여 1.2 m × 5.5 m 규모의 3개 중규모 플롯에서 세류발달 특성, 유출 및 토사유출량을 파악하고자 강우와 유입수 모의실험을 수행하였다. 경사 조건 15°와 20°에서 유입수 유무에 따른 4회의 반복실험이 진행되었으며 마사토의 평균입경은 0.89 mm이다. 강우강도 범위는 90~140 mm/hr이며, 유입수 유량은 합리식으로 계산하였으며 100~130 ml/sec이다. 하천 차수분석방법인 Horton방법을 사용하여 세류별 차수를 나누었다. 세류절개는 유입수가 없는 경우 실험 시작 약 1분 후에 발생하였고, 최대 2차수까지 세류가 발달하였으며, 유입수가 있는 경우 약 30초 후 발생하였고, 최대 3차수까지 세류가 발달하였다. 세류발달에 대한 수리학적 특성을 파악하기 위하여 염료 추적방법에 의한 동영상 이미지 분석결과 유속은 0.06~0.43 m/s의 범위를 보였다. 유입수와 강우가 함께 공급되는 경우 강우모의 공급수량에 비해 1.32~1.69배 증가했고, 이에 따라 지표유출수는 1.13~3.93배로 증가폭의 범위가 넓었다. 세류발달에 의한 토사유출량은 유입수 유무에 따라 6.7~32.3배로 증가하였다. 결론적으로 강우와 유입수가 상호작용하는 경우 강우에 의한 박리현상보다 유입수에 의한 측벽붕괴 활동이 활발하게 진행되었고 이는 세류 발달 과정에서 지배적으로 이루어졌기 때문으로 판단된다.