• Title/Summary/Keyword: dicots

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Technical Development for Large DNA Fragment Transformation in Plants

  • Park, Su-Ryun;Seo, Mi-Suk;Lee, Sang-Kug;Park, Jee-Young;Kim, Hye-Ran;Lee, Hyo-Yeon;Bang, Jae-Wook;Lim, Yong-Pyo
    • Journal of Plant Biotechnology
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    • v.2 no.2
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    • pp.89-96
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    • 2000
  • For large DNA fragment transformation in dicots and monocots, BIBAC2 vector system was applied to Arabidopsis thaliana and Oryza sativa L. cv. Jinmi as a model plant, respectively. For Arabidopsis, the Th1 gene in T23L3 BAC clone whose size is about 90 kb was used as the target gene source for transformation. Because T23L3 BAC clone was originally constructed in pBelloBAC11, the target gene was reconstructed into BIBAC2. As the results of reconstruction, 476 colonies were survived in selection medium containing 40 mg/L kanamycin. In colony hybridization analysis, 24 out of 476 colonies exhibited positive signals. In the pulsed-field gel electrophoresis analysis, 11 out of 24 positive clones exhibited the band at the location of 90 kb. In Southern hybridization, positive signal band at the location of 90 kb was observed in all 11 transformants. Using these verified clones, Agrobacterium-mediated transformation was applied to Arabidopsis thaliana th1-201 mutant for genetic complementation test. Twelve thousands T$_1$ seeds were harvested, and antibiotic selection test is being analyzed to verify whether these seeds were transformed. for rice, COR356 that contains 150 kb human genomic DNA in a BIBAC2 vector was used as the target gene. As the results of transformation, 151 out of 210 co-cultivated calli were survived in selection medium containing 5 mg/L hygromycin, and 45 out of 151 survived calli were regenerated into plants. Transformation efficiency was 21.6%. Progeny test using 71 seeds is being analyzed now. These results provide the potential that large DNA fragments can be transferred into both dicots and monocot by Agrobacterium-mediate d transformation system.

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Cloning and Characterization of the psbEF Gene Encoding Cytochrome b-559 of the Panax ginseng Photosystem II Reaction Center

  • Lee, Won-Kyu;Park, Dae-Sung;Tae, Gun-Sik
    • BMB Reports
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    • v.32 no.2
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    • pp.189-195
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    • 1999
  • From the Panax ginseng chloroplast, the psbE and psbF genes, encoding the $\alpha$- and $\beta$-subunits of cytochrome b-559 of the photosystem II reaction center, respectively, were cloned and characterized. The psbE and psbF genes were composed of 252 and 117 nucleotides, respectively. The deduced amino acid sequence of the $\alpha$-subunits showed 95%, 93%, and 91% homology to monocots, dicots, and liverwort, respectively, whereas the $\beta$-subunits showed approximately 98% to 95% homology to the same species. Southern blot analysis revealed that a single copy of the psbEF gene exists in the chloroplast plastid. Northern blot analysis indicated that the psbE and psbF genes are cotranscribed as a polycistron.

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Cloning and characterization of the psbA Gene from Panax ginseng(Characterization of the psbA Gene from P. ginseng)

  • Lee, Won-Kyu;Tae, Gun-Sik
    • Journal of Photoscience
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    • v.10 no.3
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    • pp.245-249
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    • 2003
  • The psbA gene of photo system II was cloned and characterized from the P. ginseng chloroplast. The psbA gene is composed of 1,062 nucleotides. The overall amino acid sequence shows 99% and 98% identities to dicots and monocots of higher plants, respectively. Southern blot analysis revealed that a single copy of the psbA gene existed in the chloroplast genome. Northern blot analysis of the in vivo accumulation of the psbA transcript, after being grown under the different intensities (5%, 10%, 20%, and 100%) of daylight, indicated that the steady-state level of the psbA transcript was not significantly affected by light intensity.

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Stress as a Trigger of Pollen Embryogenesis

  • Zarsky, Viktor;Soukupova, Hana
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.5
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    • pp.411-413
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    • 2000
  • The ability of microspores or young pollen grains (male gametophytes) to undergo developmetal switch to embryogenic (sporophytic) pathway exemplifies the concept of totipotency as applied to haploid posmeiotic cells. As a first step pollen is devoid of positional information provided in situ by the intact anther - by isolation and cultivation in vitro in artificial media. This is inevitably accompanied by some degree of stress response in microspore/pollen. It has been shown in both monocots and dicots that intentional stress treatment (mostly starvation or heat shock) greatly stimulates embryo induction rate. Using transgenic sHSP antisense Nicotiana tabacum we show that expression of small heat shock proteins is an integral part of successful embryo and later haploid plant production from pollen grains. Our recently published data show that sHSP chaperone function is optimal in the absence of ATP.

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A Simple and Reliable Method for Preparation of Cross-Contamination-Free Plant Genomic DNA for PCR-Based Detection of Transgenes

  • Hwang, Seon-Kap;Kim, Young-Mi
    • BMB Reports
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    • v.33 no.6
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    • pp.537-540
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    • 2000
  • A simplified but reliable method was developed for the polymerase chain reaction (PCR)-based detection of genetically modified (GM) plants. The modified CTAB (mCTAB) method enabled us to prepare a high quality of genomic DNA from several hundred plant leaf samples in one day. Using DNA samples prepared from seven dicots and two monocots, approximately 1.75-kb regions spanning 17 S to 25 S ribosomal RNA genes were successfully amplified in a 2X PCR pre-mix containing BLOTTO. Further fidelity assessment of the mCTAB method by PCR analysis with Roundup Ready soybean (RRS) and non-RRS plants showed that the DNA samples prepared alternately from each of two lines were evidently free of cross-contamination. These results demonstrate that the mCTAB method is highly recommended for the rapid detection of transgenes in large numbers of leaf samples from diverse transgenic plants.

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Misconceptions and Truths of Morphological Characteristics in Plant Stomata (식물에서 기공 형태에 대한 오해와 진실)

  • Kim, Dae Jae;Lee, Joon Sang
    • Journal of Life Science
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    • v.27 no.2
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    • pp.241-246
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    • 2017
  • The walls of guard cells have many different specialized features. Guard cells are present in leaves of bryophytes, ferns and almost all of the vascular plants. Guard cells show considerable morphological diversities. It is understood that the stomata show two types in terms of morphological characterizations of guard cells. The first type is only found in a few monocots including Poaceae and Cyperaceae. In rice and corn, guard cells have the morphological characteristics of dumbbell shape. The morphological characteristics of dumbbell shape always have subsidiary cells. The other type is found in every dicots and many monocots and they are kidney-shaped guard cells. The plants of kidney-shaped guard cells rarely have subsidiary cells except Commelina communis L. Therefore, it could be concluded that two types of the morphological characteristics of guard cells cannot divide according to monocots or dicots. Every plants in which stomatal characteristic features were all different, most of them belong to kidney-shaped guard cells. However in case of Sedum sarmentosum, guard cells were shown to be long and narrow lips type. In Tradescantia virginiana, the shape of guard cells could be called perfectly to half-moon type. Therefore, it could be concluded that kidney-shaped types are all different in some way, but dumbbell-shaped types are almost constant.

Family of floral homeotic genes (MADS-box genes) expressed in early flower Panax genseng

  • Yoon, Sunha;Yoon, Euisoo
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2002.11b
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    • pp.15-15
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    • 2002
  • In higher dicotyledonous plants, the floral organs are arranged in four different whorls, containing sepals, petals, stamens and carpels. petals, stamens and carpels. The specification of floral organ identity is explained by the ABC model (Weigel and Meyerowitz 1994). Expression of an A-function gene specifies sepal formation in whorl 1. the combination of A-and B-function genes specifies the formation of petals in whorl 2, B-and C-function genes spesify stamen formation in whorl 3, and expression of the C-function alone determines the formation of carpels in whorl 4. A-. B-, C-function genes have been isolated from many plant species and most of them belong to the family of MADS-box genes encoding transcription factor. In contrast to the flower of higher dicots, the perianths of genseng plants have three whorls of almost identical petaloid organs. van Tunen et al. (1993) proposed a modified ABC model, exemplified with tulip. In this model, B-function genes are expressed in whorl 1 as well as whorl 2 and 3, theefore the organs of whorl 1 and whorl 2 have the same petaloid structure. They proposed this model with the molphological data of wild type and mutant flowers of tulip, however, there are no molecular data.(중략)

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