Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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In Vitro Culture and Transformation by Agroinfiltration of Lisianthus (Eustoma russellianus) Pollen

Lisianthus 화분의 기내배양 및 Agroinfiltration에 의한 형질전환

  • Park Hee Sung (Department of Plant Biotechnology, Catholic University of Daegu)
  • 박희성 (대구가톨릭대학교 식물유전공학과)
  • Published : 2004.12.01
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Abstract

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.

Agrobacterium을 이용한 lisianthus화분의 형질전환을 위하여 적정조건을 수립하였다. 화분의 발아 및 화분관의 발달은 화분발아배지(pollen germination medium; PCM)에 sucrose를 $7-15\%$ 첨가 시키고 pH조건을 5.5-7.0으로 조절하여 $20^{\circ}C-27^{\circ}C$에서 배양할 경우 성공적으로 이루어졌다. 형질전환을 위하여 Agrobacterium 현탁액을 lisianthus화분배양액에 첨가하여 진공침윤을 20 min실시하였으며 형질전환화분은 조직화학적 분석 그리고 발현되는 GUS mRNA를 이용한 RT-PCR 및 Southern hybridization에 의한 DNA산물 분석 등에 의하여 GUS발현을 확인하였다. 이러한 결과를 통하여 화분을 이용한 일시발현기술을 제시하게 되었다.

Keywords

References

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