• Title/Summary/Keyword: K-L transformation

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Use of FT-IR to Identify Enhanced Biomass Production and Biochemical Pool Shifts in the Marine Microalgae, Chlorella ovalis, Cultured in Media Composed of Different Ratios of Deep Seawater and Fermented Animal Wastewater

  • Kim, Mi-Kyung;Jeune, Kyung-Hee
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1206-1212
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    • 2009
  • Growth rates, photosystem II photosynthesis, and the levels of chlorophyll $\alpha$ and secondary metabolites of Chlorella ovalis were estimated to determine if they were enhanced by the addition of swine urine (BM) or cow compost water (EP) that had been fermented by soil bacteria to deep seawater (DSW) in an attempt to develop media that enabled batch mass culture at lower costs. Growth of C. ovalis in f/2, f/2-EDTA+BM60%, DSW+BM30%, and DSW+EP60% was enhanced and maintained in the log phase of growth for 16 days. The cell densities of C. ovalis in DSW+EP60% ($4.1{\times}10^6$ Cells/ml) were higher than those of f/2 ($2.9{\times}10^6$ Cells/ml), f/2-E+BM60% ($3.7{\times}10^6$ Cells/ml), and DSW+BM30% ($2.7{\times}10^6$ Cells/ml). The growth rate was also more favorable for C. ovalis cultured in DSW+EP60% ($0.15\;day^{-1}$) than that of C. ovalis cultured in the control medium (f/2) ($0.12\;day^{-1}$). Furthermore, the chlorophyll a concentration of C. ovalis cultured in DSW+EP60% (4.56 mg/l) was more than 2-fold greater than that of C. ovalis cultured in f/2 (2.35 mg/l). Moreover, the maximal quantum yields of photo system II at 470 nm (Fv/Fm) were significantly higher in organisms cultured at f/2-E+BM60% (0.53) and DSW+EP60% (0.52) than in the other treatment groups. Finally, Fourier transformation infrared (FT-IR) spectroscopy revealed that C. ovalis grown in DSW+EP60% had more typical peaks and various biochemical pool shifts than those grown in other types of media. Taken together, the results of this study indicate that the use of DSW+EP60% to culture C. ovalis can reduce maintenance expenses and promote higher yields.

Efficient Kinect Sensor-Based Reactive Path Planning Method for Autonomous Mobile Robots in Dynamic Environments (키넥트 센서를 이용한 동적 환경에서의 효율적인 이동로봇 반응경로계획 기법)

  • Tuvshinjargal, Doopalam;Lee, Deok Jin
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.39 no.6
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    • pp.549-559
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    • 2015
  • In this paper, an efficient dynamic reactive motion planning method for an autonomous vehicle in a dynamic environment is proposed. The purpose of the proposed method is to improve the robustness of autonomous robot motion planning capabilities within dynamic, uncertain environments by integrating a virtual plane-based reactive motion planning technique with a sensor fusion-based obstacle detection approach. The dynamic reactive motion planning method assumes a local observer in the virtual plane, which allows the effective transformation of complex dynamic planning problems into simple stationary ones proving the speed and orientation information between the robot and obstacles. In addition, the sensor fusion-based obstacle detection technique allows the pose estimation of moving obstacles using a Kinect sensor and sonar sensors, thus improving the accuracy and robustness of the reactive motion planning approach. The performance of the proposed method was demonstrated through not only simulation studies but also field experiments using multiple moving obstacles in hostile dynamic environments.

High-frequency plant regeneration from leaf-disc cultures of Jatropha curcas L.: an important biodiesel plant

  • Deore, Ajay C.;Johnson, T. Sudhakar
    • Plant Biotechnology Reports
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    • v.2 no.1
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    • pp.7-11
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    • 2008
  • A simple, high-frequency and reproducible protocol for induction of adventitious shoot buds and plant regeneration from leaf-disc cultures of Jatropha curcas L. has been developed. Adventitious shoot buds were induced from very young leaf explants of in vitro germinated seedlings as well as mature field-grown plants cultured on Murashige and Skoog's (MS) medium supplemented with thidiazuron (TDZ) ($2.27{\mu}M$), 6-benzylaminopurine (BA) ($2.22{\mu}M$) and indole-3-butyric acid (IBA) ($0.49{\mu}M$). The presence of TDZ in the induction medium has greater influence on the induction of adventitious shoot buds, whereas BA in the absence of TDZ promoted callus induction rather than shoot buds. Induced shoot buds were multiplied and elongated into shoots following transfer to the MS medium supplemented with BA ($4.44{\mu}M$), kinetin (Kn) ($2.33{\mu}M$), indole-3-acetic acid (IAA) ($1.43{\mu}M$), and gibberellic acid ($GA_3$) ($0.72{\mu}M$). Well-developed shoots were rooted on MS medium supplemented with IBA ($0.5{\mu}M$) after 30 days. Regenerated plants after 2 months of acclimatization were successfully transferred to the field without visible morphological variation. This protocol might find use in mass production of true-to-type plants and in production of transgenic plants through Agrobacterium/biolistic-mediated transformation.

Synthesis and Characterization of The Electrolessly Deposited Co(Re,P) Film for Cu Capping Layer (무전해 도금법으로 제조된 Co(Re,P) capping layer제조 및 특성 평가)

  • Han, Won-Kyu;Kim, So-Jin;Ju, Jeong-Woon;Cho, Jin-Ki;Kim, Jae-Hong;Yeom, Seung-Jin;Kwak, Noh-Jung;Kim, Jin-Woong;Kang, Sung-Goon
    • Korean Journal of Materials Research
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    • v.19 no.2
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    • pp.61-67
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    • 2009
  • Electrolessly deposited Co (Re,P) was investigated as a possible capping layer for Cu wires. 50 nm Co (Re,P) films were deposited on Cu/Ti-coated silicon wafers which acted as a catalytic seed and an adhesion layer, respectively. To obtain the optimized bath composition, electroless deposition was studied through an electrochemical approach via a linear sweep voltammetry analysis. The results of using this method showed that the best deposition conditions were a $CoSO_4$ concentration of 0.082 mol/l, a solution pH of 9, a $KReO_4$ concentration of 0.0003 mol/l and sodium hypophosphite concentration of 0.1 mol/L at $80^{\circ}C$. The thermal stability of the Co (Re,P) layer as a barrier preventing Cu was evaluated using Auger electron spectroscopy and a Scanning calorimeter. The measurement results showed that Re impurities stabilized the h.c.p. phase up to $550^{\circ}C$ and that the Co (Re,P) film efficiently blocked Cu diffusion under an annealing temperature of $400^{\circ}C$ for 1hr. The good barrier properties that were observed can be explained by the nano-sized grains along with the blocking effect of the impurities at the fast diffusion path of the grain boundaries. The transformation temperature from the amorphous to crystal structure is increased by doping the Re.

Improvement of Forage Crop Yield and Retardation of Leaf Senescence by Introduction of Gene for Cytokinin Synthetase into Plants (Cytokinin 합성효소의 도입에 따른 형질전환 식물체의 노화 지연 및 수량의 증가)

  • Lee, B.H.;Won, S.H.;Lee, H.S.;Kim, K.Y.;Kim, M.H.;Eun, S.J.;Jo, J.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.19 no.3
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    • pp.281-290
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    • 1999
  • The bacterial isopentenyl transferase (ipt) gene involved in cytokinin biosynthesis was fused with 35S promoter of cauliflower mosaic virus (CaMV) and introduced into tobacco plants (Nicotiana tabacum L. cv. Samsun) via Agrobacterium-mediated transformation. As expected, ipt gene was constitutively expressed in all tissues of transgenic plants. Several primary transgenic plants were obtained that expressed different level of transcripts for ipt gene. Three of transgenic plants with different expression level of ipt gene were selected and selfed to obtain homozygous line for further analysis. A number of interesting phenotypic changes such as viviparous leaves, delayed senescence, larger axillary shoots, an abundance of tiny shoots at the apex and a release of lateral buds, were observed in transgenic plants. Chlorophyll content was 1.5- t.o 4-fold higher in transgenic plants as compared with non-transformed plants. These results indicate that the cytokinin synthesized in transgenic plants could improve forage crop yield by delay of leaf senescence and increase of leaf number.

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Rapid Agrobacterium-mediated genetic rice transformation method using liquid media (액체배양을 이용한 단기 벼 형질전환 방법)

  • Yang, Dae-Hwa;Chang, Ahn-Cheol;Ahn, Il-Pyung;Kim, Hae-Jung;Kim, Dong-Hern;Lee, Hyo-Yeon;Suh, Seok Cheol
    • Journal of Plant Biotechnology
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    • v.40 no.1
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    • pp.37-42
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    • 2013
  • Rice is one of the most important cereal crops as a model plant for functional genomics of monocotyledons and usually transformed using Agrobacterium tumefaciens. However, the transformation's process using previous method is still time consuming and uneconomical, low efficiency. In this study, we established a new method by modifying the general Agrobacterium protocol especially in the infection and co-cultivation, Agrobacterium elimination, infected calli's selection steps using liquid media. We directly inoculated Agrobacterium containing a ZjLsL gene under the control of constitutive promoter into the 1- to 3-week-old rice calli derived from mature seeds. After 3 days of co-cultivation, the infected calli were transferred onto liquid media of Agrobacterium elimination and calli's selection for 3 days. The calli were transferred to calli's growth solid media for 14 days and then the calli transferred to shoot induction and root induction media. Putative transformants were initially selected on the medium containing phosphinothricin, and the PAT protein verified by PAT strip test. This method in this study would lead to reduction of substantial labor and time to generate transgenic plants.

Characterization of Transgenic Lettuce (Lactuca sativa L.) Using a BL1 Gene Encoding Bromelain Isolated from Pneapple (제주산 파인애플 유래 Bromelain관련 유전자 (BL1)를 이용반 형질전환 상추의 특성)

  • Jung, Yu-Jin;Kim, Gi-Hyun;Choi, Jang-Sun;Lee, Soon-Youl;Nou, Il-Sup;Park, Jin-Heui;Kang, Kwon-Kyoo
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.27-32
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    • 2006
  • To clarify the roles of bromelain in plants, we isolated BL1 gene encoding bromelain from pineapple stem tissues and sequenced. The full length cDNA is 933 bp and encodes a polypeptide of 311 amino acid residues. The cDNA is most similar 94% at the amino acid level to bromelain previously isolated from pineapple (BAA21929). Explants of Lactuca sativa were co-cultivated with Agrobacterium tume-faciences LBA 4404 strains containing nptII and BL1 gene for transformation. Through initial selection of regenerated explants by culturing on a kanamycin and carbenicillin containing MS medium, multiple shoots were obtained after 2 months of culture. For a complementary step of selection, putative transgenic shoots were transferred to 1/2 Ms basal medium supplemented with 100 mg/L kanamycin and 500 mg/L carbenicillin. The selected shoots were obtained T1 generation seeds with emasculation, and tested with PCR analysis using 35S promoter and BL1 specific primers whether BL1 gene was introduced to genome of the plants. These results confirmed that produced the specific PCR bands in the putative transgenic lines. Additionally the Northern blot and endo protease activity showed that transcripts of BL1 gene were detected in transgenic lines. Theses results suggest that BL1 gene be successfully integrated and transcripted in the transgenic lettuce plants.

Genetic Transformation of Chrysanthemum with Cold Regulated Gene (BN115) (저온저항성 유전자를 이용한 국화 형질전환)

  • Han, Soo-Gon;Choi, In-Young;Kang, Chan-Ho;Ko, Bok-Rai;Choi, Joung-Sik;Lee, Wang-Hyu
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.19-25
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    • 2006
  • With the use of Agrobacterium and gene-gun, cold regulated gene (BN115) has been injected in Chrysanthemum leaf disc and transgenic plants have been produced successfully on the selection media containing phytohormone. To determine the presence of the transferred cold regulated gene (BN115) in the transgenic Chrysanthemum, PCR-amplification indicated the presence of that gene. Real-Time PCR for confirmation of the putative transgenic plants was established. The copy number of cold regulated gene (BN115) is extrapolated on the basis of a standard curve. Serial dilutions of known number of gene copies were in triplicates. In this diagram, PCR cycles are plotted against the fluorescence intensity. The cycle at which the fluorescence reaches a threshold cycle is inversely proportional to the starting amount of target DNA.

Establishment of discrimination system using multivariate analysis of FT-IR spectroscopy data from different species of artichoke (Cynara cardunculus var. scolymus L.) (FT-IR 스펙트럼 데이터 기반 다변량통계분석기법을 이용한 아티초크의 대사체 수준 품종 분류)

  • Kim, Chun Hwan;Seong, Ki-Cheol;Jung, Young Bin;Lim, Chan Kyu;Moon, Doo Gyung;Song, Seung Yeob
    • Horticultural Science & Technology
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    • v.34 no.2
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    • pp.324-330
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    • 2016
  • To determine whether FT-IR spectral analysis based on multivariate analysis for whole cell extracts can be used to discriminate between artichoke (Cynara cardunculus var. scolymus L.) plants at the metabolic level, leaves of ten artichoke plants were subjected to Fourier transform infrared(FT-IR) spectroscopy. FT-IR spectral data from leaves were analyzed by principal component analysis (PCA), partial least square discriminant analysis (PLS-DA) and hierarchical clustering analysis (HCA). FT-IR spectra confirmed typical spectral differences between the frequency regions of 1,700-1,500, 1,500-1,300 and $1,100-950cm^{-1}$, respectively. These spectral regions reflect the quantitative and qualitative variations of amide I, II from amino acids and proteins ($1,700-1,500cm^{-1}$), phosphodiester groups from nucleic acid and phospholipid ($1,500-1,300cm^{-1}$) and carbohydrate compounds ($1,100-950cm^{-1}$). PCA revealed separate clusters that corresponded to their species relationship. Thus, PCA could be used to distinguish between artichoke species with different metabolite contents. PLS-DA showed similar species classification of artichoke. Furthermore these metabolic discrimination systems could be used for the rapid selection and classification of useful artichoke cultivars.

Procambium differentiation and shoot apical meristem development in somatic embryos of soybean (Glycine max L.) (대두 체세포배에서 전형성층 분화와 경단분열조직의 발달)

  • Choi, Pil Son;Kwon, Suk Yoon
    • Journal of Plant Biotechnology
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    • v.40 no.1
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    • pp.55-58
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    • 2013
  • Immature embryos of Glycine max L. was cultured on Murashige and Skoog's (MS) medium supplemented with 1 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D). After 6 to 8 weeks of culture, immature embryos produced somatic embryos. Of somatic embryos, two cotyledonary embryo (14%), one cotyledonary embryo (37%), fused cotyledonary embryo (43%), and stunted globular embryos (6%) were observed. The procambial strand of cotyledons originated from circular procambial tissues of lower hypocotyl. The circular procambial tissues were independently divided into one or two procambial strand at the edge of cotyledonary-node, and then connected to each cotyledon to form somatic embryos with one or two cotyledons. When cotyledon was a fused type, the circular procambial strand in lower hypocotyl was continuously connected to the cotyledon. Also, somatic embryos with two cotyledons developed a functional shoot apex with the tunica-corpus structure. In contrast, somatic embryos with one or fused cotyledon formed an abnormal shoot apex without the tunica-corpus structure or with non-dome shape in the inter-cotyledonary area. These results indicated that the variation of cotyledon in somatic embryos is closely related to procambial differentiation and shoot apical meristem development.