• Title/Summary/Keyword: lily pollen

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Lily Pollen Growth in vitro and Agrobacterium-mediated GUS Gene Transformation via Vacuum-Infiltration

  • Park, In-Hae;Park, Hee-Sung
    • Journal of Plant Biotechnology
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    • v.4 no.4
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    • pp.151-154
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    • 2002
  • Conditions for lily pollen growth in vitro and transformation were optimized. Active pollen tube development was achieved effectively in a medium containing 7% sucrose with pH adjusted to 5.7 at the temperature of 27$^{\circ}C$ for about 16-24 hours. Pollen growth was little impaired by the presence of kanamycin at concentration up to 100 mg/L. Pollen rains near the beginning of germination stage were more reliable for Agrobacterium-mediated GUS DNA transformation via vacuum infiltration lasted for 20-40 minutes. GUS DNA integration and its expression in fully developed pollen tubes could be confirmed by Southern blot hybridization, RT-PCR and histochemical staining.

Isolation and Characterization of a Pollen-specific cDNA Clone from Easter Lily

  • Kim, Seong-Ryong;An, Gyu-Heung
    • Journal of Plant Biology
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    • v.39 no.3
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    • pp.197-202
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    • 1996
  • A pollen-specific cDNA clone, LMP50, was isolated from the mature pollen cDNA library of the Easter lily. The LMP50 transcript was highly abundant in mture pollen grains but not detectable in other organs. The LMP50 cDNA clone contains 1383 nucleotides and two open reading frames. The first codes for a peptide of 15 amino acid residues. The role of this peptide is nuclear. The second encodes a protein containing 329 amino acid residues. This protein exhibited a significant homology to human tartrate-resistant acid phosphatase and porcine uteroferrin. Both of these enzymes have been suggested to play a role in iron transport. Therefore, LMP50 may act as an iron carrier protein in mature pollen grains.

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Antioxidative Effect of Pine, Oak, and Lily Pollen Extracts (송화분, 참나무 및 백합화분 추출물의 항산화 효능)

  • Kim, Seok-Joong;Youn, Kwang-Sup;Park, Hee-Sung
    • Korean Journal of Food Science and Technology
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    • v.37 no.5
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    • pp.833-837
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    • 2005
  • Antioxidative activities of pine, oak, and lily pollen extracts were evaluated based on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging ability and inhibition of lipid peroxidation in animal tissues. Each pollen was extracted with 50% ethanol, 100% ethanol or water. DPPH radical-scavenging capacity of 50% ethanol extract ($EC_{50}$ 40.0 mg/mL) of pine pollen was higher than those of water (46.8 mg/mL) and 100% ethanol (131.2 mg/mL) extracts of pollen. Fifty percent ethanol (3,2 mg/mL) was also better than 100% ethanol (4.5 mg/mL) and water (8.3 mg/mL) for extraction of oak pollen. For preparation of lily pollen extracts, 100% ethanol was most effective (14.0 mg/mL), followed by water (18.8 mg/mL) and 50% ethanol (24.0 mg/mL). Oak pollen showed higher DPPH radical-scavenging activity than others. Lipid peroxidation in rat brain homogenate induced by ascorbate-Fe3+-EDTA and rat kidney homogenate were inhibited by water extracts of all pollens in dose-dependent manner. Extracts of oak and lily pollen showed higher lipid peroxidation inhibition than pine pollen extract. Polyphenol content was highest in oak pollen extract $(32.5{\pm}0.7\;{\mu}g/mg\;pollen)$, followed by lily extract $(25.9{\pm}1.4\;{\mu}g/mg\;pollen)$ and pine extract $(9.3{\pm}0.7\;{\mu}g/mg\;pollen)$.

Analysis of UreB Protein Synthesis from Transgenic Lily Pollen (형질전환 백합화분을 이용한 UreB단백질의 발현분석)

  • 박희성;박인혜
    • KSBB Journal
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    • v.17 no.6
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    • pp.577-581
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    • 2002
  • In an attempt to produce recombinant proteins using the pollen enriched in some plant species, a 1.7 kb DNA encoding urease subunit B (UreB) amplified by PCR from Helicobacter pylori urease gene cluster in pH808 plasmid was cloned to be expressed under CaMV35S promoter in lily (Lilium longiflorum) pollen tubes elongated in vitro. Lily pollen at early germinating stage was transformed with the ureB DNA using Agrobacterium via vacuum infiltration and, incubated for a full pollen tube growth 16 - 24 h in the dark in the presence of kanamycin. DNA integration and expression in the transgenic pollen were analyzed by the standard molecular techniques and the results suggest that the pollen in vitro may be employed as a protein factory in a disposable fashion.

Impact of Physical, Chemical and Biological Factors on Lily (Lilium longiflorum cv. Georgia) Pollen Growth and GUS Expression Via Agro-infiltration (물리적, 화학적, 생물적 요인에 의한 백합 (Lilium longiflorum cv. Georgia) 화분의 생장 및 Agro-Infiltration을 이용한 GUS 발현)

  • Park, Hee-Sung
    • Journal of Plant Biotechnology
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    • v.31 no.4
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    • pp.279-283
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    • 2004
  • To lily (Lilium longflorum cv. Georgia) pollen, impacts by some physical, chemical and biological factors were examined in respects of its growth and transient gene expression via agro-infiltration. Rolling movement in liquid medium or vacuum pressure during Agro-infiltration was regarded as a impact that should be minimized for normal pollen growth. Pollen growth was maintained well in relatively broad range of temperature (19 to 27$^{\circ}C$) or pH (5.0 to 8.0). Chemical factors such as cefotaxime (up to 300mg/L), acetosyringone (up to 800 $\mu$M) and syringealdehyde (up to 800 $\mu$M) did not show any harmful effects but kanamycin severely did even at concentration as low as 25mg/L in some cases. For GUS gene expression, acetosyringone at 200 to 400 $\mu$M slightly improved the efficiency while syringealdehyde did not. Brief agro-infiltration followed by 18 hr of co-incubation of pollen along with Agrobacterium was suggested as a condition basically required for the transient expression system using lily pollen regardless of the presence of acetosyringone.

Analysis of Tissue Plasminogen Activator Expression using Pollen Culture in vitro (기내 화분배양을 이용한 Tissue Plasminogen Activator 발현분석)

  • 박인혜;박희성
    • KSBB Journal
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    • v.17 no.6
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    • pp.582-585
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    • 2002
  • In an effort to use plant biotechnology for the production of biopharmaceuticals, pollens collected from lily (Lilium longiflorum) were grown in vitro and transformed with a PCR-amplified 1.7 kb cDNA encoding human tissue plasminogen activator (tPA) using Agrobacterium via a vacuum infiltration process. Western blotting showed that transgenic lily pollen tubes selected on kanamycin for 16 hrs expressed a tPA protein with a size similar to the human standard, suggesting their possible use as a disposable host for rapid foreign protein production.

Immunotoxicological Evaluation of Pollen Intake Using Mice Model (실험동물을 이용한 화분섭취의 면역안전성 평가)

  • Park Hee Sung;Heo Young Jeu;Byun Jung-A;Heo Yong
    • Journal of Environmental Health Sciences
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    • v.31 no.4 s.85
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    • pp.287-293
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    • 2005
  • Pollen has been used for Prevention or treatment of certain diseases such as diabetes, arthritis, or cancer in traditional medicine. In addition, pollen is under investigation as a host cell for a gene expression. This study was undertaken to evaluate the immunologic safety of pollen intake. BALB/c mice were administered with 500, 50,5, or 0.5 mg/kg bw of lily pollen for five times a week for four weeks through gastric intubation. Comparing the control mice administered with distilled water, no significant changes were observed in body weight gain, weight of liver, spleen, lung, and his-topathological findings of liver and kidney of the mice groups administered with the pollen. Plasma level of IgG1, IgG2a, and IgE was not different among the groups. When splenic B lymphocytes were stimulated in vitro with lipopolysaccharides for 7 days, level of IgGl and IgGwa produced in the culture supernatants was not significantly different among the groups. Furthermore, no significant alteration was observed in IL-4 and $IFN{\gamma}$ producing ability with splenic T lymphocytes stimulated in vitro with phytohemagglutinins for 48 hours between the pollen-administered and the control mice. Overall, this study suggests that the lily pollen intake is Inducing no significant modulation of humoral and cell-mediated immunity in mice.

Storage of Pollen Biofactory for in vitro Growth and rProtein Synthesis (기내배양과 재조합 단백질 생산을 위한 화분 Biofactory의 저장기술의 개발)

  • 박희성;고재철
    • KSBB Journal
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    • v.19 no.3
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    • pp.174-177
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    • 2004
  • A method of collection and long-term storage of viable lily (Lilium longiflorum) pollen grains were developed for their in vitro growth and transformation in consistency. Petroleum ether, n-heptane, cyclohexane and benzene, as pollen collection medium, were determined less toxic to pollen growth in vitro than others tested. Pollen grains, however, lost their growth activity if stored in these solvents more than a week, So, a serial performance, that is, pollen grain collection in these solvents, air-drying and immediate transfer to low temperature condition was determined desirable for keeping the viability much longer. Pollen grains from this storage showed a successful transformation in vitro with a cDNA encoding tissue plasminogen activator (TPA) protein using Agrobacterium via vacuum infiltration according to western blotting analysis.

Characteristics of Self-compatible Variety from Native Lilium tigrinum Thunberg (우리나라 자생 참나리에서 선발된 자가결실성 2배체 품종의 특성(特性))

  • Ha, Yoo-Mi;Kim, Dong Yeob;Han, In Song
    • FLOWER RESEARCH JOURNAL
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    • v.18 no.4
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    • pp.284-290
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    • 2010
  • This study was carried out to breed a self-compatible variety of Tiger Lily from the seedlings originated from Chinju city, Gyeongsangnam-do. The morphological characteristics, ploidy, and the resistance of seedlings to virus infection were investigated. A progeny test was also conducted to examine whether the propagated progenies had the same characteristics as the mother plant. The self-compatible diploid lily variety developed in this study showed a tall type like native triploid lily, Lilium tigrinum, and bulbils were formed on both lily varieties. The morphological characteristics of the flowers and leaves were not much different between the varieties, while the size of the bulbils and bulbs showed significant differences. The percentage of seed generation by self-pollination was 72.6% for the self-compatible variety, while there was no seed generated for native triploid lily. The number of chromosome was 2n = 26 (x = 13, diploid) for the self-compatible variety, while 2n = 39 (x = 13, triploid) in native lily variety. The progenies of the self-compatible diploid lily variety showed the same characteristics as those of its mother plant in morphology, seed germination, and polyploidy. The mother plant of the self-compatible diploid lily variety showed 58% pollen germination and the 2-year-old and 3-year-old progenies showed similar germination percentages. The pollen grains of Korean native triploid lily, however, never germinated.

Utilization of pollen grains for the expression of epidermal growth factor (Epidermal growth factor 발현을 위한 화분립의 이용)

  • Choi, Byung-Jin;Park, Hee-Sung
    • KSBB Journal
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    • v.23 no.5
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    • pp.460-462
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    • 2008
  • Pollens grains collected from fully dehisced lily (Lilium longiflorum) anthers were given wounds by means of shaking in the presence of aluminum oxide particles. They were transformed by infiltration with Agrobacterium cells harboring a synthetic DNA encoding signal peptide-fused epidermal growth factor (EGF). After incubation for 24 hr in vitro, the pollen culture showed that EGF mRNAs and proteins were successfully expressed in the analysis of cDNA blot hybridization and immuno-blotting.