• Title/Summary/Keyword: GUS histochemistry

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Utilization of Pollen Grain from Liluim cv. Casablanca as a Transient Gene Expression Host (유전자의 일시발현 분석용 숙주개발을 위한 카사블랑카백합(Lilium cv. Casablanca) 화분립의 이용)

  • Park, Hee-Sung
    • Applied Biological Chemistry
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    • v.47 no.4
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    • pp.430-433
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    • 2004
  • Lilium cv. Casablanca pollen grains stored at $-70^{\circ}C$ were grown in pollen germination medium with Agrobacterium tumefaciens LBA4404 cells harboring pBI121 for 18 hr at $27^{\circ}C$. Following this, cefotaxime (250 mg/L) was treated for 6 hr to eradicate the bacterial cells. Histochemical GUS analysis revealed that the transgenic pollen displayed deep blue color mostly from 12 hr after the co-cultivation. Presence of $200\;{\mu}M$ acetosyringone was determined not to be more effective for GUS transformation than its absence. GUS DNA integration in the transgenic pollen genomic DNA was clearly demonstrated by Southern blot analysis.

In Vitro Culture and Transformation by Agroinfiltration of Lisianthus (Eustoma russellianus) Pollen (Lisianthus 화분의 기내배양 및 Agroinfiltration에 의한 형질전환)

  • Park Hee Sung
    • Journal of Life Science
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    • v.14 no.6 s.67
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    • pp.1018-1022
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    • 2004
  • Optimized conditions for Agrobacterium-mediated lisianthus pollen transformation were adjusted using various factors such as temperature, pH and sucrose concentration. Pollen tube growth was successfully achieved in a medium (pollen germination medium; PGM) containing $7-15\%$ sucrose with pH in the range of 5.5-7.0 at temperature of $20-27^{\circ}C$. Lisianthus pollen was vacuum-infiltrated with Agrobacterium cell suspension for 20 min, and transformed pollen was confirmed by GUS histochemistry and Southern hybridization following RT-PCR. Transgenic pollen system may be utilized for establishing an area of plant transient expression systems based on the convenient pollen transformation procedure presented in here.

Effect of Sodium Hydrosulfite Solution on Agrobacterium-Mediated Chinese Cabbage Transformation and Transient Expression

  • Park Hee-Sung;Shin Dong-Il
    • Journal of Plant Biotechnology
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    • v.7 no.4
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    • pp.219-223
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    • 2005
  • We investigated chemical-wounding effect on Agrobacterium-mediated Chinese cabbage transformation via vacuum infiltration. Pre-germinated or germinating Chinese cabbage seeds were infiltrated with Agrobacterium tumefaciens LBA4404 cells carrying either GUS gene (pBI121) or hepatitis B virus surface antigen DNA (pBIHBsAg). Prior to agroinfiltration process, the seeds were soaked in sodium hydrosulfite (SHS) solution or just in sterile water as a control. Comparative transformation efficiency was determined by both of histochemistry and ELISA. We could demonstrate that SHS solution treatment especially to 1-day or 2-days old germinating seeds efficiently improved transformation process, and therefore, transient expression level. This strongly indicated that Agrobacterium infection could be facilitated indeed by SHS-causing wounds on Chinese cabbage seeds.