• Applied Biological Chemistry
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• 제47권4호
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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.

• 생명과학회지
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• 제14권6호
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• pp.1018-1022
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• 2004

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.

• Journal of Plant Biotechnology
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• 제7권4호
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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.