양배추와 무의 동형 원형질체 융합을 이용한 식물체의 재분화

인동수,송민정,장인창,민병환,남석현,신종섭,이시우,한지학 한국식물생명공학회 2008 식물생명공학회지 Vol.35 No.2

Protoplasts from cabbage and radish were isolated and fused symmetrically by PEG treatment. The PEG treated mixture of high concentrated protoplasts produced lots of micro-calli after 2~3 weeks. The microcalli developed to normal calli and shoots were regenerated from the calli. A total of 218 shoots were regenerated, but none of them contained the NWB-CMS specific DNA marker, indicating that the transfer of the radish NWB-CMS character into cabbage did not occur. However, ISSR analysis revealed that the cell fusion between protoplasts from radish and cabbage was occurred (3 out of 208 plantlet). The fused regenerants possessed the characteristics of source plants used for protoplast fusion. After vernalization, three regenerants were flowered with white petal color as seen in radish. Only three seeds were able to obtain from one regenerant by backcrossing with the cabbage pollen.

Lycoris속 식물 4종의 핵형 분석

인동수,이민수,방재욱 충남대학교 기초과학연구소 1996 忠南科學硏究誌 Vol.23 No.2

Karyotypes of four Lycoris species, L. aurea, L. squamigera, L. koreana and L. radiata, was studied. The basic chromosome numbers of the diploid plants, L. aurea and L. koreana were x=8 and x=11 and those of triploids, L. squamigera and L. radiata were x=9 and x=11, respectively. The chromosome complement of L. aurea was composed of 3 pairs of large metacentrics, 1 pair of submetacentrics and 4 pairs of telocentrics. The karyotype of L. squamigera was 2 pairs of large metacentrics, 3 pairs of subtelocentrics and 4 pairs of telocentrics. The karyotype of L. koreana was 7 paris of subtelocentrics and 4 pairs of telocentrics and that of L. radiata was 3 pairs of subtelocentrics and 8 pairs of telocentrics. No large chromosome was found in L. aurea and L. koreana Supernumerary segment was found in chromosome 1 of L. koreana. Chromosome 4 of L. radiata carries satellite.

Cytochrome oxidase subunit I (COI) DNA sequence divergence between tow cryptic species of Oryzias in South Korea

인동수,장민호,최은숙,윤주덕,김정희,민준일,백승호 한국생태학회 2013 Journal of Ecology and Environment Vol.36 No.3

Oryzias latipes and Oryzias sinensis are indigenous species found in Japan, China, and other East Asian countries, including Korea. Based on morphological differences, the species have been classified distinctly. However, the range of morphological characters such as the number of gill rakers, vertebrae, and spots on the lateral body overlaps and is too vague for clear identification, so their classification based on their morphological characteristics remains uncertain. In this study, the mitochondrial cytochrome oxidase subunit I (COI) gene, which is used for DNA barcoding, was applied to clarify interspecific variation of O. latipes and O. sinensis. Intraspecific genetic diversity was calculated to identify correlations with geographic distributions. We studied two species collected from 55 locations in Korea. All individuals carried a 679-base pair gene without deletion or insertion. Between species, 525 base pairs of the gene were shared. The Kimura two parameter (K2P) distance of O. latipes and O. sinensis was 0.41% and 1.39%, respectively. Mean divergence within genera was 23.5%. Therefore, the species were clearly different. The distance between O. latipes and O. sinensis was 14.0%, which is the closest within genera. Interestingly O. latipes from the Japanese and Korean group represented 16.5% distant. These results were derived from geohistorical and anthropogenic environmental factors. The O. latipes haplotypes were joined in only one group, but O. sinensis was divided into two groups, one is found in the Han River and upper Geum River watershed; the other is found in the remaining South Korean watersheds. Further studies will address the causes for geographic speciation of O. sinensis haplotypes.

초저온 보존된 오이 배발생세포 현탁배양으로부터 식물체 재분화

김석원,인동수,정원중,우제욱,정민,유장렬,Kim, Seok-Won,In, Dong-Soo,Jung, Won-Joong,Woo, Je-Wook,Jung, Min,Yoo, Jang-Ryul 한국식물생명공학회 1998 식물생명공학회지 Vol.25 No.6

오이의 국내 F1 품종인 조생낙합의 하배축 유래 배발생 현탁배양 세포의 초저온 보존 시스템을 개발하였다. 액체질소 저장 후 캘러스 재생률은 2M DMNSO와 0.4 M sucrose를 혼용 처리하였을 때 캘러스 재생률이 85%로 가장 높았다. 그러나 glycerol 처리구에서는 처리농도에 상관없이 모든 처리구에서 캘러스 재생이 이루어지지 않았다. 또한 고농도의 삼투용액에서 배양세포의 전처리 과정은 필요하지 않았다. 재생된 캘러스를 1 mg/L 2,4-D가 첨가된 MS 배지로 이식하여 배양하였을 때 다수의 체세포배가 발달하였으며, 체세포배를 MS 기본배지로 옮겨 명배양한 결과 다수의 소식물체가 발달하였다. Conditions for high frequency plant regeneration from cryopreserved embryogenic cell suspension cultures derived from hypocotyl explants of cucumber (Cucumis sativus L.) are described. Cells cryoprotected with a mixture of 2 M DMSO and 0.4 M sucrose exhibited a regeneration frequency of 85%. However, cells cryoprotected with different concentrations of glycerol showed no regeneration after cryopreservation. Pretreatment of cells in a high osmotic medium was not necessary to the process. Upon transfer to MS medium supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid, regenerated calli gave rise to numerous somatic embryos, then underwent development into plantlets.

Agrobacterium 공동배양을 이용한 고추 소포자 유래 초기 배의GFP 발현

정민,인동수,김봉규,장인창,박은준,김문자,한지학 한국식물생명공학회 2008 식물생명공학회지 Vol.35 No.2

The aim of this research is to establish the conditions for Agrobacterium-mediated genetic transformation using microspore. The embryo induction from the microspore was examined under several Kanamycin concentration in media, and the induction rate decreased about 4, 8, 10 times when the Kanamycin concentration increased 10, 50, 100 mg/L, respectively. This indicates that the transformation rate would be much lower if the Kanamycin was used for selection marker. In order to apply the GFP gene as a reporter gene for Agrobacterium-mediated genetic transformation, GFP expression from the microspore- mediated embryos was observed using GFP filter under microscope. The GFP expression occurred when the microspore cultured toward the embryo development for 12, 24 and 48 days. The microspore formed a cluster by microspore division from 12 days culture and continuously became a bigger mass. We obtained a total of 8 GFP-expressing embryos suggesting that the transformation of microspore occurred. However, those young embryos were not fully developed. Further study pertinent to culture conditions is required to fulfill the Agrobacterium-mediated genetic transformation using microspore

Development of the plastic culture container to improve light transmission and stacking possibility

한성렬,인동수,조진래 대한기계학회 2017 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.31 No.9

This paper is concerned with the development of plastic culture container in order to resolve the problems of conventional culture bottle, such as heavy weight, reduction of transmitted light amount after sterilization operation, and impossibility of stacking of bottles. The plastic culture container was developed by seeking the optimum molding conditions for the gate diameter, the melting temperature and the coolant temperature. The optimum conditions were sought by making use of Taguchi design experiments and injection molding analyses. The performance of developed plastic culture container was evaluated in terms of the dimension accuracy and the transmitted light amount after sterilization operations, and the weight of container. It has been verified that all the performances are remarkably improved, when compared with conventional culture bottle.

금강수계에서 하상재료에 따른 어류의 종다양성 및 서식지 평가

허준욱,인동수,장민호,강형식,강경호 한국환경영향평가학회 2011 환경영향평가 Vol.20 No.6

In order to establish fundamental data for stream restoration and environmental flow, we investigated inhabitation and species diversity of fish to substrate size in the typical streams of Geum River Basin. Field monitoring including fish sampling was conducted from October 2007 to October 2009. Substrate size was determined according to six different : silt (Si), sand (Sa), fine gravel (Fg), coarse gravel (Cg), cobbles (Co) and boulders (Bo). A total number of fish caught in the 18 sites was 7,649 representing 10 families 50 species, and Si, Sa, Fg, Cg, Co and Bo stations occupied 30, 29, 38, 30, 27 and 17 species, respectively. The most frequently found species in number was pale chum (Zacco platypus, 29.7%, n=2,275) followed by Z. koreanus (22.5%, n=1,720) in total stations. Biological diversity with increase of substrate size from the dominance of part species showed higher values as dominance index, lower and diversity, richness and evenness index. Index of biological integrity (IBI) and qualitative habitat evaluation index (QHEI) scores decreased with increase of substrate size. Therefore, it is necessary to make an effort on stream rehabilitation with evaluation of physical habitat condition by indicator species in order to maintain biodiversity and perform ecological restoration.

기능획득 돌연변이 인삼 모상근의 대량생산

고석민,인동수,정화지,최동욱,유장렬,Ko, Suk-Min,In, Dong-Soo,Chung, Hwa-Jee,Choi, Dong-Woog,Liu, Jang-Ryol 한국식물생명공학회 2007 식물생명공학회지 Vol.34 No.4

본 연구는 아그로박테리움 공동배양법을 이용한 기능획득 인삼 모상근의 대량생산을 위한 조건 확립에 대한 것이다. 일반적으로, 인삼과 같이 형질전환을 통한종자의 확보가 어려운 식물에서는 loss-of-function을 이용한 기능유전체 연구에 한계가 있다. 한편, 유전자의 기능을 활성화시키는 방법 (gain-of-function)인 activation tagging 기술은 이러한 문제점을 극복할 수 있는 대안이 될 수 있으며, Agrobacterium rhizogenes를 이용한 모상근 생산 시스템은 대량의 돌연변이체를 안정적으로 용이하게 얻을 수 있다는 점에서 최적의 시스템이라고 할 수 있다. 본 연구에서는 activation-tagging된 효율적인 형질전환 모상근 생산에 있어서의 최적의 아그로박테리움 균주 및 인삼조직, 배지조성 등에 대한 조건을 확립하였으며, 다양한 배지에서의 형질전환 모상근의 생장률 및 분지율, 표현형 등을 조사하였다. 엽병 절편을 activation-tagging vector pKH01을 가지고 있는 A. rhizogenes R1000와 공동배양하였을 때 배양 4주후 85.9%의 빈도로 모상근이 생산되었다. 모상근의 최대 생장과 분지도를 나타내는 배양조건을 조사한 바 엽병절편을 1/2 SH 배지에서 4주 배양하였을 때 왕성하게 생장하였으며 2.6의 분지도를 보여주었다. 최종적으로 1,989개체의 독립적인 형질전환 모상근 line을 생산하였으며, 이들 모상근 line은 인삼 진세노사이드 생합성 관련 유전자의 발굴 및 기능해석에 유용하게 쓰일 것이다. This study describes conditions for the mass production of activation-tagged mutant hairy root lines of ginseng by cocultivation with Agrobacterium rhizogenes. Because it is not currently possible to produce progeny from transgenic ginseng, a loss-of-function approach for functional genomics cannot be appliable to this species. A gain-of-function approach is alternatively the choice and hairy root production by cocultivation of A. rhizogenes would be most practical to obtain a large number of mutants. Various sources of explants were subjected to genetic transformation with various strains of A. rhizogenes harboring the activation-tagging vector pKH01 to determine optimum conditions for the highest frequency of hairy root formation on explants. Petiole explants cocultivated with A. rhizogenes R1000 produced hairy roots at a frequency of 85.9% after 4 weeks of culture. Conditions for maximum growth or branching rate of hairy roots were also investigated by using various culture media. Petiole explants cultured on half strength Schenk and Hildebrandt medium produced vigorously growing branched roots at a rate of 2.6 after 4 weeks of culture. A total of 1,989 lines of hairy root mutants were established in this study. These hairy root lines will be useful to determine functions of genes for biosynthesis of ginsenosides.

기능획득 돌연변이 인삼 모상근의 대량생산

유장렬,고석민,인동수,정화지,최동욱 한국식물생명공학회 2007 JOURNAL OF PLANT BIOTECHNOLOGY Vol.34 No.4

This study describes conditions for the mass production of activation-tagged mutant hairy root lines of ginseng by cocultivation with Agrobacterium rhizogenes. Because it is not currently possible to produce progeny from transgenic ginseng, a loss-of-function approach for functional genomics cannot be appliable to this species. A gain-of-function approach is alternatively the choice and hairy root production by cocultivation of A. rhizogenes would be most practical to obtain a large number of mutants. Various sources of explants were subjected to genetic transformation with various strains of A. rhizogenes harboring the activation-tagging vector pKH01 to determine optimum conditions for the highest frequency of hairy root formation on explants. Petiole explants cocultivated with A. rhizogenes R1000 produced hairy roots at a frequency of 85.9% after 4 weeks of culture. Conditions for maximum growth or branching rate of hairy roots were also investigated by using various culture media. Petiole explants cultured on half strength Schenk and Hildebrandt medium produced vigorously growing branched roots at a rate of 2.6 after 4 weeks of culture. A total of 1,989 lines of hairy root mutants were established in this study. These hairy root lines will be useful to determine functions of genes for biosynthesis of ginsenosides.

Micropropagation of Hypericum erectum Thunbergby using Thidiazuron

김옥태,방경환,인동수,김태수,성낙술,차선우,안준철,황백 한국약용작물학회 2006 韓國藥用作物學會誌 Vol.14 No.5

The effect of plant growth regulators was investigated on in vitro shoot proliferation from axillary bud explantsof Hypericum erectum. To determine the optimal cytokinin for proliferation of axillay buds, we carried out screening four cytokinins(BA, kinetin, 2iP, TDZ). When nodal segments were cultured on MS medium supplemented with 4.5 μM TDZ (thidiazuron),a number of shoots were induced. Our results indicated that the addition of TDZ to culture medium resulted in theinduction of significantly more axillary buds than in the addition of other cytokinins. The optimal concentration of TDZ forproliferation of axillary buds was 10 μM. 92% of shoots spontaneously rooted without any plant growth regulator (PGR) andformed whole plantlets within one month. More than 95% of these regenerants survived and they did not show any detectablevariation in morphology or growth characteristics compared to their donor plants.