• Title/Summary/Keyword: genetic system

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Introduction of Bean Chitinase Gene into Korean Ginseng by Agrobaterium tumefaciens (Agrobacterium tumefaciens에 의한 강낭콩 키틴가수분해효소 유전자의 고려인삼으로의 도입)

  • 이행순;권석윤;백경희;김석원;이광웅;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.2
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    • pp.95-99
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    • 1995
  • We have previously established a system for plant regeneration through somatic embryogenesis and Agrobacterium-mediated transformation of Korean ginseng. In this study to produce a fungus-resistant plant, we introduced a bean chitinase gene into ginseng using the transformation system. A binary vector pChi/748 was constructed by introducing the bean basic chitinase gene into EcoRI site of pGA748 which carries the CaMV 35S promoter governing the introduced gene and neomycin phosphotransferase II(NPT-II)gene as a positive selection marker. Cotyledonary explants were cocultured with A. tumefaciens strain LBA4404 harboring the binary vertor pChi/748 for 48 h, and transferred to MS medium supplemented with l mg/L2,4-D,0.1mg/L kinetin, 100 mg/L kanamycin, and 500mg/L carbenicillin. Kanamycin-resistant calli were formed on the cut surface of cotyledonary explants after one month of culture, and subsequently they gave rise to somatic embryos. Upon transfer onto medium containing 1 mg/L each of BA and GA$_3$, most of them converted to plantlets after 5 weeks of culture. The genomic DNA of eight kanamycin-resistant regenerants was subjected to polymerase chain reaction (PCR) using two specific 21-mer oligonucleotides derived from the chitinase gene. PCR-Southern blot analysis confirmed that the chitinase gene was incorporated into six out of the eight regenerants..

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Plant Regeneration from Hypocotyl Explants of Several Species of Lycopersicon (토마토속 식물의 배축절편 배양에 의한 식물체 재분화)

  • 임학태;이건섭;용영록;송융남;김종화
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.137-143
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    • 1994
  • In an attempt to optimize the in vitro-regeneration conditions necessary for the genetic manipulation of tomato species, we examined several hybrid lines and wild species (peruvianum, pimpinellifolium, glandulosum) of Lycopersicon for. their, different regeneration ability. The basal medium used for callus growth and organogenesis was MSB (MS + B5) supplemented with three combinations of TDZ (Thidiazuron) 0.5mg/L+NAA 0.5mg/L, BA 2.0mg/L+NAA0.05 mg/L, and zeatin 3.0 mg/L + IAA 0.02 mg/L. In the genotype of Lycopenicon grandulosum, combination of TDZ and NAA was more effective in inducing shoot and root differentiation than those of BA and NAA or zeatin and IAA. When all genotypes tested were considered, however combination of zeatin and IAA was shown to be the best in shoot regeneration. Result indicate that callus and organogenesis of Lycopenicon species are dependent upon the hormone types and plant genotypes, but MSB medium with zeatin 3.0 mg/L + IAA 0.02 mg/L maybe appropriate for genotype-independent plant regeneration system of Lycopercicon species. We also tried TDZ as a cytokinin source in tomato tissue culture and found it highly significant in tomato regeneration system.

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Transformation of Populus Species by an Agrobacterium Binary Vector System (Agrobacterium Binary Vector에 의한 포플러 형질전환(形質轉換)을 위한 기초연구(基礎研究))

  • Chun, Young Woo;Klopfenstein, Ned B.;McNabb, Harold S. Jr.;Hall, Richard B.
    • Journal of Korean Society of Forest Science
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    • v.77 no.2
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    • pp.199-207
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    • 1988
  • Three clones of Populus alba ${\times}$ P. grandidentata have been tested for susceptibility to Agrobacterium tumefaciens strains A281 and A348. We determined the optimum concentration of kanamycin sulfate for effective selection of leaf disc-derived, transgenic tissues transformed using Agrobacterium binary vector pGA472 containing a neomycin phosphotransferase gene (NPT-II) which confers kanamycin resistance. Of the wild type Ti plasmids contained by the two Agrobacterium strains, pTiBo542 of strain A281 appears to be best suited to serve as a helper plasmid for binary vector systems. A relatively low concentration (10mg/l) of kanamycin sulfate inhibited adventitious shoot initiation from leaf discs on regeneration medium. Transformed kanamycin-resistant calli were obtained by culturing Agrobacterium inoculated leaf discs on selective regeneration medium. The transformed kanamycin-resistant calli continued to grow on regeneration media supplemented with kanamycin sulfate to levels of 50 and 200mg/l. The growth of non-co-cultivated control calli was severely inhibited on regeneration medium containing 50mg/l kanamycin sulfate.

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효율적인 생식선 카이메라 생산을 위한 최적 조건 확립에 관한 비교 연구

  • 김진남;박태섭;송권화;이영목;권혁모;한재용
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2001.11a
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    • pp.71-73
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    • 2001
  • In this study, we could improve transmission efficiency of germline chimeras by transfer of gonadal PGCs (gPGCs) cultured in vitro. Of hatched recipient chicks, 301 chickens (141 males and 160 females) were brought up to sexual maturity and these WLs (KOC) were mated with KOCs for testcross, resulting in 27 germline chimeras (15 males and 12 females) identified by black feather color of their progenies. The production efficiency of germline chimera production of experimental groups was observed (P=0.6831). The average transmission efficiency of proven germline chimeras was 0.6 ∼56.5% (15.0% on average). The transmission efficiency of experimental group which were transferred 10-days cultured gPGCs without Ficoll treatment was highest (49.7%) and that of experimental stock which transferred non-cultured gPGCs with Ficoll treatment was lowest (0.6%). The duration of in vitro culture before transferring was significantly important for the high efficiency of germline transmission. Transferring 10-days cultured gPGCs made the transmission efficiency higher rather than transferring non-cultured and 5-days cultured gPGCs, 50 times and 10 times, respectively (p<0.0001). However, Ficoll treatment for increasing the population ratio of gPGCs negatively affected the transmission efficiency and the effects of sexuality and the reciprocal interaction between treatments showed no significant differences. These findings demonstrated that the crucial factors for improving the germline transmission were the duration of in vitro culture prior to transfer. Thus, we developed the complete system for production of germline chimera using cultured gPGCs with highly improved efficiency and this system would be useful for genetic manipulation and obtaining the transgenic aves.

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Viroid-the Smallest Plant Pathogen (바이로이드-가장 작은 식물병원체)

  • Lee Jai Youl
    • Korean Journal Plant Pathology
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    • v.1 no.3
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    • pp.199-206
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    • 1985
  • Viroids are the smallest. well-characterized infectious agents presently known. and so far viroids have been found only in higher plants. The structures of viroid-molecules are single-stranded, covalently closed circular RNA molecules with a range of 240 to 380 nucleotides according to the various viroids. Viroids are remarkable not only as a new category of pathogen, which cause economically important diseases, but also as an excellent model system for biochemical and biophysical investigations because of their small size, relative stability and their self-replication. Four different patato spindle tuber viroid isolates, which express the different symptoms on the same host plant exchange only 2 to 6 nucleotides in the total number of 359 nucleotides, but now the mechanism of viroid pathogenicity is not explained fully. Viroid-melecules are replicated without any special viroid-associated proteins, and during the process of viroid replication oligomeric viroid-associated RNAs are detected at nuclei of viroid infected leaf tissue. The mechanism of viroid replication can now be illustrated according to a possible explanation of rolling-circle system. Although the rapid progress have been made in elucidation of the biochemical and biophysical properties of PSTV and other viroids, the mechanism of viroid replication and pathogenicity is less known and is still a matter of speculation. When these problems can be sufficiently explained, the viroid molecule could play an important role as an available vector in plant genetic engineering.

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Genetic Relationship between Seed size and Leaf Size in 66 $F_2$ Populations Derived from Mating of 12 Soybean Strains (대두 12 모본의 half diallel cross로부터 생성된 66 $F_2$ 분리집단에서의 종자크기와 잎 크기에 대한 관계)

  • 정종일
    • Journal of Life Science
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    • v.8 no.4
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    • pp.437-442
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    • 1998
  • Seed and leaf size is the important morphological traits considered by plant breeder and is the important yield components in soybean. The objective of this research was to know the relationship between seed size and leaf size in 66 $F_2$ populations derived from half diallel mating system with 12 soybean strains, representing distinct seed and leaf size. The range of seed size for 12 parents used was 6.7 to 43.8 g/100 seed. Leaf width leaf length ranged 5.7 to 8.6 cm and 9.4 to 12.9 cm, respectively. Leaf width was highly correlated with leaf length with an R square of 0.754 in the $F_2$ generation. The $F_2$ regression` coefficient indicated that leaves were, on average 1.4 times greater length than in width . Leaf size (width) was highly correlated (r.0.91) with seed size (g/100 seed) in the $F_2$ generation with an R square of 0.833. Our results indicate postive correlation within seed and leaf size is common in $F_2$ segregating populations derived from crossing with soybean. The strong liner relationship we observed between leaf size and seed size in $F_2$ segragating population is useful in that in that indirect selection for a secondary character may be superior to direct selection for the primary character.

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A Strain based Load Identification for the Safety Monitoring of the Steel Structure (철골 구조물의 안전성 모니터링을 위한 변형률 기반 하중 식별)

  • Oh, Byung-Kwan;Lee, Ji-Hoon;Choi, Se-Woon;Kim, You-Sok;Park, Hyo-Seon
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.18 no.2
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    • pp.64-73
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    • 2014
  • This study proposes a load identification for the safety monitoring of the steel structure based on measured strain data. Instead of parameterizing the stiffness of structure in the existing system identification researches, the loads on a structure and a matrix (the unit strain matrix) defined by the relationship between strain and load on structure are parameterized in this study. The error function is defined by the difference between measured strain and strain estimated by parameters. In order to minimize this error function, the genetic algorithm which is one of the optimization algorithm is applied and the parameters are found. The loads on the structure can be identified through the founded parameters and measured strain data. When the loads are changed, the unmeasured strains are estimated based on founded parameters and measured strains on changed state of structure. To verify the load identification algorithm in this paper, the static experimental test for 3 dimensional steel frame structure was implemented and the loads were exactly identified through the measured strain data. In case of loading changes, the unmeasured strains which are monitoring targets on the structure were estimated in acceptable error range (0.17~3.13%). It is expected that the identification method in this study is applied to the safety monitoring of steel structures more practically.

Genetic Transformation of Biocontrol Agent Bacillus sp, YBL-7 by Plasmid pE194 (생물방제균 Bacillus sp. YBL-7의 형질전환조건)

  • 한길환;정병곤;김상달
    • Microbiology and Biotechnology Letters
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    • v.20 no.4
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    • pp.384-389
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    • 1992
  • Bacillus sp. YBL-7 which had been isolated from ginseng root-rot suppressive soil was able to antagonize Fusarium solani causing ginseng root-rot by their antibiotic substance. In order to develop multifunctional antagonist on Bacillus sp. YBL-7 as a biocontrol agent against Fusarium salam', optimal conditions for protoplast transformation system of Bacillus sp.YBL-7 by the vector plasmid pE194 were investigated. The protoplasts of Bacillus sp. YBL-7 were obtained at best efficiency by treatment with 200${\mu}g$/ml of lysozyme in the pH 7.0 of SMM buffer for 90 minutes at $40^{\circ}C$. The cell wall of the protoplast was regenerated on the agar plate containing 1.2% agar and 0.7 M mannitol. Under the best condition for protoplast formation and regeneration, the optimal transformation was achieved with 40% polyethylene glycol (M.W. 4000) treatment for 10minutes. The vector plasmid pE194 showed the best transformation frequency at 5$\mu$g/ml of final concentration. The pE194 was very stable over 80% in the transformants.

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Spatial and temporal variation on fruit set in Epipactis thunbergii (Orchidaceae) from southern Korea (한국남부 자생 닭의난초 (난초과)의 시 공간에 따른 결실률 변이)

  • Chung, Mi Yoon;Chung, Myong Gi
    • Korean Journal of Plant Taxonomy
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    • v.45 no.4
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    • pp.353-361
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    • 2015
  • Spatio-temporal variation in fruit set in orchids would affect long-term population viability and will influence genetic diversity over many generations. The aim of this study was to examine the breeding system of the nectariferous terrestrial orchid Epipactis thunbergii, to specifically determine levels of fruit set in terms of time and space under natural conditions. We examined pollination under natural conditions and conducted hand pollination experiments during a 2-year survey in four populations located along 1.5 km of coastal line in Jinguiri (rual village) [Jeollanam-do (province), southern Korea]. We found that, over a 2-year period, levels of percentage of fruit set were similar within patches of the four populations. By contrast, we detected significant differences in the percentage of fruit set among patches. We also found that plants with larger inflorescence size produced significantly more fruits than plants with fewer flowers. Over a 2-year period, the percentage of fruit set for E. thunbergii was similar but low (14.1%) compared to that averaged for eighty-four rewarding species (37.1%). However, an increase in fruit set was achieved by hand-pollinations: artificial self-pollination (90.5-95.2%), artificial geitonogamy (94.7-95.0%), and cross-pollination (artificial xenogamy, 91.3-91.4%). No emasculated flowers produced fruits and no automatic pollination was found in E. thunbergii. Our findings suggest that E. thunbergii is a self-compatible terrestrial orchid that depends on pollinators (insects) to achieve fruit set in natural habitats, and that local environmental conditions were similar over a period of 2 years in the study area. Our results also highlight the cryptic variation of fruit production in time, but more pronounced variability in space.

Inhibitory Effects of Human Glutamate Dehydrogenase Isozymes by Antipsychotic Drugs for Schizophrenia (정신분열증 치료제에 의한 사람 글루탐산염 탈수소효소 동종효소의 억제효과)

  • Nam, A-Reum;Kim, In-Sik;Yang, Seung-Ju
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.17 no.1
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    • pp.152-158
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    • 2016
  • Glutamate is one of the major excitatory neurotransmitters in the central nervous system of vertebrates. Human GDH (hGDH) is the enzyme that regulates the glutamate metabolism and its expression is higher in the brains of schizophrenia patients than in normal subjects. This study examined the changes in the hGDH enzymatic activity caused by antipsychotic drugs (haloperidol, risperidone, (${\pm}$)-sulpride, chlopromazine hydrochloride, melperone, (${\pm}$)butaclamol, domperidone, clozapine) related to schizophrenia. First of all, hGDH isozymes (hGDH1, hGDH2) were synthesized by genetic recombination. As a result of the enzyme assay, haloperidol, (${\pm}$)-sulpride, melperone and clozapine had an inhibitory effect on the hGDH isozymes. In addition, haloperidol showed a non-competitive inhibition against the substrate, 2-oxoglutarate. In contrast, it showed an uncompetitive inhibition against another substrate, NADH. The inhibitory effect of haloperidol on hGDH2 was abolished by the presence of L-leucine, an allosteric effector of hGDH, but by not other antipsychotic drugs. These results revealed the inhibition of enzyme activity by psychotropic drugs in hGDH isoenzymes (hGDH1 and hGDH2) and the possibility that haloperidol may be used to regulate the GDH activity and glutamate concentration in the central nervous system.