• Title/Summary/Keyword: meristem

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Inhibition of Starch Biosynthesis by Antisense Expression of cDNAs Encoding ADP-Glucose Pyrophosphorylase Small Subunit in Sweetpotato (고구마에서 ADP-Glucose Pyrophosphorylase Small Subunit cDNA의 Antisense 발현에 의한 전분생합성 저해)

  • Min, Sung-Ran;Bae, Jung-Myung;Harn, Chee-Hark;Jeong, Won-Joong;Lee, Young-Bok;Liu, Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.277-283
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    • 2007
  • Embryogenic calluses derived from shoot apical meristem explants of sweetpotato were subjected to particle bombardment to generate transgenic plants for antisense expression of cDNAs encoding two different AGPase small subunit (ibAGP1 and ibAGP2). Plants were generated via somatic embryogenesis. PCR and Southern analysis demonstrated that the incorporation of ibAGP1 and ibAGP2 into the genome in an antisense orientation. Immunoblot analysis confirmed reduced levels of AGPase small subunit in transgenic plant leaves. Plants with both ibAGP1 and ibAGP2 produced a lower level of the protein than plants with ibAGP1 alone. iodine test demonstrated that transgenic plant leaves and storage root accumulated reduced amounts of starch. Iodine staining of leaf tissues indicated that transgenic plants accumulated less amount of starch than control. In accordance with western blot analysis, plants with both ibAGP1 and ibAGP2 accumulated a lower amount of starch than plants with ibAGP1 alone. Both transgenic plants exhibited a severely retarded growth, resulting in bare survival. It is suggested that disrupted expression of the gene encoding AGPase small subunit is lethal to the growth of sweetpotato contrast to other species including potato.

STUDIES ON THE TISSUE CULTURE OF PANAX GINSENG

  • Harn C
    • Proceedings of the Ginseng society Conference
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    • 1974.09a
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    • pp.9-22
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    • 1974
  • Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.

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Development of Leaf Mutant Cultivars of Cymbidium goeringii by Ethyl-methane-sulfonate (EMS) Treatment. (Ethyl-methane-sulfonate(EMS) 처리에 의한 춘란 잎 돌연변이 품종의 개발)

  • Shin, Yun-Ho;Song, In-Ja;Kang, Eun-Jung;Bae, Tae-Woong;Sun, Hyeon-Jin;Kang, Si-Young;Lim, Pyung-Ok;Lee, Hyo-Yeon
    • Korean Journal of Plant Resources
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    • v.24 no.1
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    • pp.17-22
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    • 2011
  • This study was for developing leaf chlorophyll mutant cultivars of Cymbidium goeringii by ethyl-methanesulfonate(EMS) treatment. Chlorophyll mutant rhizomes were easily produced by 0.2% EMS treatment in this genus. Among the mutants, they became dark brown about 50% of the rhizomes. When the dark-brown rhizomes were cultured in a solidified MS medium, new rhizomes were formed from part of the old ones. Chlorophyll mutant rhizomes were obtained from subcultured meristem tissues of newly-formed rhizomes. The rhizomes were cut and subcultured for a year and then became mutant plants. As the results, they produced 4 kinds of leaf mutant cultivars; zigzag-striped, comb-striped, net-striped, and dwarf types, indicating that the EMS treatment in the rhizome could produce versatile leaf chlorophyll regulating mutants. These results suggest that our method is useful for developing leaf mutant cultivars of this planta which they are estimated as higher commercial values.

Growth Acceleration and Acclimatization of In Vitro Plantlets derived from Apical Meristem of Sweet Potato (고구마의 경정조직 유래 기내 소식물체의 생장촉진과 순화)

  • ;;Shiro Higashi
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.2
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    • pp.115-119
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    • 1999
  • The single node cuttings of sweet potato (cv. Mokpo #29) plantlets maintained in vitro were cultured with (MF+) or without membrane filter (MF-) under photomixotrophic (PM), hetrotrophic (HT) and autotrophic (AT) conditions. Shoot length was the greatest (11.9cm) in 3$0^{\circ}C$ (HT) treatment and it was the shortest (3.4 cm) in $25^{\circ}C$ (PM) treatment. Nodal explants cultured in 3$0^{\circ}C$ treatment looked more vigorous than those of $25^{\circ}C$ in appearance, and node number was the greatest (10.5 per plantlet) among the treatments. But plantlets grew in 3$0^{\circ}C$ (HT) treatment were observed all overgrown. The size in leaf area was about 2 times greater and shoot length was about 2 times shorter in PM than in HT condition. Percent dry matter of shoots was 5.9% (HT) and 7.4% (PM) in $25^{\circ}C$ treatment and 6.1% (HT) and 7.4% (PM) in 3$0^{\circ}C$ treatment. Plantlets cultured in the MF+ treatments were less succulent than those cultured in the MF- treatment. Vitrified plantlets were examinated 14.8% (both $25^{\circ}C$ and 3$0^{\circ}C$) in PM condition and 22.2% ($25^{\circ}C$) and 31.5% (3$0^{\circ}C$) in HT condition. Sucrose was necessary for the survival of in vitro plantlets. In the sucrose-free medium, explants cultured in the MF- had turned yellow and were dead after 30 days of culture. But explants cultured in the MF+ were alive and produced plantlets with shoot and root (AT). On the other hand, the survival of explants on the MS basal medium (sucrose-free and hormone-free) depended entirely upon the MF attachment.

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Effective Micropropagation of Pulsatilla cernua var. koreana through Apical Meristem Culture (할미꽃 정단 분열조직 배양을 통한 효율적 미세번식)

  • Ko, Jeong-Ae;Kim, Hyun-Soon
    • Korean Journal of Plant Resources
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    • v.21 no.5
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    • pp.362-367
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    • 2008
  • In order to investigate the effect of plant growth regulators on effective in vitro micropropagation, apical meristems of Pulsatilla cernua var. koreana were cultured on Murashige and Skoog's (MS) medium with 2,4-D, NAA, TDZ and BA. Media containing 2,4-D and kinetin, 2,4-D and TDZ, NAA and TDZ were not effective on callus induction. However, embryogenic or organogenic callus was obtained on media containing NAA and BA. Especially, on MS medium with 0.5mg/L NAA and 1.0mg/L BA was optimal for a high frequency (62%) of shoot or shoot bud obtained from callus. Callus proliferation, shoot multiplication and elongation were significantly increased by adding 10% coconut water on MS media with 0.5mg/L NAA and 1.0mg/L BA. Repeated subculturing of in vitro grown shoots resulted in propagation rate of 12.9 shoots per explant every 30 days. Root formation from the adventitious shoots was not easily achieved. However, roots were only produced through callus on MS medium with 2.0mg/L NAA alone or 0.5mg/L NAA and 1.0mg/L BA. These roots were used materials for callus and shoot production repetitively.

A Study of Mode of Action of Alachlor III. Effect of Alachlor on Cell Division, Cell Kinetics, Cell Elongation, and Cell Differentiation in Oat (Avena sativa L.) (Alachlor의 제초기구(除草機構)에 관(關)한 연구(硏究) III. Alachlor가 귀리의 세포분열(細胞分裂), Cell kinetics, 세포신장(細胞伸長) 및 분화(分化)에 미치는 영향(影響))

  • Kwon, S.W.;Kim, J.C.
    • Korean Journal of Weed Science
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    • v.12 no.2
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    • pp.102-109
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    • 1992
  • There was significant reduction in the mitotic indices of oat roots treated with alachlor. Uniform decrease in prophase, metaphase, anaphase, and telophase as treatment time increasing was observed. Alachlor did not disrupt mitosis, but rather inhibited the onset of mitosis. Labeled dividing cells were significantly inhibited, but the number of labeled interphase cells of all treatment were increased, as compared with control in 8 hr and 12hr period. Labeled dividing cells which entered mitosis thru $G_2$ were inhibited approximately 68% at 8hr after treatment with $1{\times}10^{-5}$ M of alachlor. Alachlor apparently inhibited from the $G_2$stage into mitosis of dividing cells. After 24 hr treatment, 12.1% abd 46.6% inhibition of coleoptile growth occurred at $1{\times}10^{-5}$ M and $1{\times}10^{-4}$ M, respectively. Cell elongation was inhibited by alachlor but was less sensitive than cell division. The longitudinal section cells of oat roots treated with $1{\times}10^{-4}$ M alachlor for 12 hr were observed to be enlarged central cylinder and also showed degradation of apical meristem zone, as compared with the untreated roots.

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Glyphosate Toxicity: II. EPSP-synthase Activity in Cell Suspension Culture of Corydalis Sempervirens and Lycopersicon Esculentum (Glyphosate 독성(毒性): II. corydalis Sempervirens와 토마토의 세포배양체(細胞培養體)에서 EPSP-synthase의 활성(活性)에 미치는 영향(影響))

  • Kim, Tae-Wan;Amrhein, Nikolaus
    • Korean Journal of Weed Science
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    • v.15 no.2
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    • pp.148-153
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    • 1995
  • Glyphosate (N-[phosphonomethyl]glycine) applied to the assimilate-exporting leaves or sprayed to the whole plants of tomato(Lycopersicon esculentum Mil var. Moneymaker) induced the rapid inhibition of 5-enolpyruvyl skimic acid 3-phosphate synthase(EPSP-synthase). It shows that EPSP-synthase activity precedes chlorophyll loss. There is no difference in EPSP-synthase activity between in vivo tomato meristem and cell suspension culture if glyphosate is not applied. The EPSP-synthase activity is in a range of 4 to 6 nkat per mg protein. The inhibition of EPSP-synthase action is induced within 36 h after glyphosate application while the Chl contents were reduced 48 h after the application. In cell suspension culture of tomato and Corydalis (Corydalis sempervirens), a sublethal concentration of glyphosate retards the fresh weight increase and prolonged lag phase. The fresh weight is reached maximal about 14 days after the subculture in the presence of glyphosate. The inhibitory effect of glyphosate on EPSP-synthase is remarkably induced in lag phase.

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Plant Regeneration of Major Cultivars of Sweetpotato (Ipomoea batatas) in Korea via Somatic Embryogenesis (체세포배발생을 통한 국내 주요 고구마 품종의 식물체 재분화)

  • Kwon, Eun-Jeong;Kwon, Suk-Yoon;Kim, Moon-Za;Lee, Joon-Seol;Ahn, Young-Sup;Jeong, Byeong-Choon;Kwak, Sang-Soo;Lee, Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.189-192
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    • 2002
  • An efficient plant regeneration system of major cultivars of sweetpotato (Ipomoea batatas (L.) Lam.) in Korea via somatic embryogenesis was established. Embryogenic calli were formed from shoot apical meristems of sweetpotato cultivars when cultured on LS medium supplemented with 1 mg/L auxin (2,4-D, picloram, dicamba). Among three kinds of auxin, 1 mg/L 2,4-D showed the highest embryogenic calli induction rate. After 4 weeks of cultures on LS medium supplemented with 1 mg/L 2,4-D, embryogenic calli induction rates of Sinhwangmi, Zami, Yulmi, and White Star were 86%, 78%, 76%, and 80%, respectively. Upon transfer onto LS basal medium, most of somatic embryos developed into plantlets. Regenerated plantlets were transplanted to potting soil and grown to mature plants in a greenhouse.

Characterization of Oszinc626, knock-out in zinc finger RING-H2 protein gene, in Ac/Ds mutant lines of rice(Oryza sativar L.) (Zinc finger RING-H2 protein관련 Ac/Ds전이인자 삽입 변이체 Oszinc626 유전자의 특성 분석)

  • Park, Seul-Ah;Jung, Yu-Jin;Ahn, Byung-Ohg;Yun, Doh-Won;Ji, Hyeon-So;Park, Yong-Hwan;Eun, Moo-Young;Suh, Seok-Cheol;Lee, Soon-Youl;Lee, Myung-Chul
    • Journal of Plant Biotechnology
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    • v.35 no.3
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    • pp.177-183
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    • 2008
  • Ac/Ds mutant lines of this study were transgenic rice plants, each of which harbored the maize transposable element Ds together with a GUS coding sequence under the control of a promoterless(Ds-GUS). We selected the mutants that were GUS expressed lines, because the GUS positive lines will be useful for identifying gene function in rice. One of these mutants was identified knock-out at Oszinc626(NP_001049991) gene, encoding a RING-H2 zinc-finger protein, by Ds insertion. In this mutant, while primary root development is normal, secondary root development from lateral root was very poor and seed development was incomplete compare with normal plant. RING zinc-finger proteins play important roles in the regulation of development in a variety of organisms. In the plant kingdom, a few genes encoding RING zinc-finger proteins have been documented with visible effects on plant growth and development. The consensus of the RING-H2(C3-H2-C3 type) domain for this group of protein is $Cys-X_2-Cys-X_{28}-Cys-X-His-X_2-His-X_2-Cys-X_{14}-Cys-X_2-Cys$. Oszinc626 encodes a predicted protein product of 445 amino acids residues with a molecular mass of 49 kDa, with a RING-zinc-finger motif located at the extreme end of the C-terminus. RT-PCR analysis indicated that the expression of Oszinc626 gene was induced by IAA, cold, dehydration, high-salinity and abscisic acid, but not by 2,4-D, and the transcription of Oszinc626 gene accumulated primarily in rice immature seeds, root meristem and shoots. The gene accumulation patterns were corresponded with GUS expression.

Growth, Reproduction, Mortality, and Production of Laminaria japonica Areschoug on the Southeastern Coast of Korea (한국 동남해안에서 다시마(Laminaria japonica Areschoug)의 성장, 성숙, 사망률 및 생산)

  • Kang, Rae-Seon;Koh, Chul-Hwan
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.4 no.3
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    • pp.226-236
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    • 1999
  • Growth, reproduction, mortality, and production of Laminaria japonica were experimentally studied at a cultivation ground on the coast of llkwang, where the largest amount of cultivated Laminaria has been produced in Korea. For this experiment, young sporophytes (0.33 cm in mean length) grown in the laboratory were transplanted at the depth of 3 m and field surveys on them were conducted twice a month from December, 1995 to August, 1996. Plants exhibited an annual life span; they were completely dead by August. Frond width, thickness, and wet weight showed similar pattern of seasonal growth and reached their maxima in July, but frond length showed no more increment after May. Maximum mean frond length and weight were 199.8 cm and 333.0 g wet wt., respectively. Overall meristematic growth in length and weight were 384.0 cm and 393.6 g wet wt., respectively. Absolute growth rates (AGR) which were calculated from the length of tissue developed from meristem varied seasonally; AGR of length and weight reached maxima in March (3.6 $cm{\cdot}d^{-1}$) and May (3.8 g wet $wt{\cdot}d^{-1}$), respectively. Absolute attrition rates gradually increased from February to July. Seasonal differences in growth and attrition rates appeared to be related to seawater temperature and nitrogen concentration in seawater. Reproductive sporophytes bearing sprorangium sorus began to occur from April, and the ratio of sorus area to blade area reached its maximum in July (0.034). Survival rate was exponentially decreased; more than 90% of plants decayed within 56 days after outplanting. After February, mortality was size-specific; mortality of smaller plants less than 30 cm in length were relatively higher. Maximum biomass occured in July (285.6 kg wet $wt{\cdot}m^{-2}$) and annual production was 758.7 kg wet $wt{\cdot}m^{-2}$.

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