포장 조건에서 CMVP0-CP 형질전환 고추 도입유전자의 지속성 조사
Assessment of the Persistence of DNA in Decomposing Leaves of CMVP0-CP Transgenic Chili Pepper in the Field Conditions
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초록
오이모자이크바이러스(cucumber mosaic virus, CMV)에 저항성을 가지도록 개발된 형질전환 CMVP0-CP 고추를 이용하여 포장 조건에서 CMVP0-CP 고추 도입유전자의 지속성을 조사하였다. CMVP0-CP 고추의 잎을 토양 표면에 올려놓거나 토양 내에 묻은 뒤 일정 시간 간격으로 PCR 및 실시간Table 2. Relative amount of persistent actin, 35S and nos genes in one to four months of incubation. Data are means±standard deviations (n=3).LinesGeneSampleconditions% of controls (0 month)1 month2 months3 months4 monthsControlactinBelow soil39.7±3.72.6±0.4ndandAbove soil42.3±4.43.3±0.6ndndCMVP0-CPactinBelow soil38.5±6.11.6±0.3ndndAbove soil36.1±3.22.9±0.4ndnd35SBelow soil30.1±4.51.3±0.1ndndAbove soil37.7±3.92.1±0.30.3±0.1ndnosBelow soil28.3±5.60.9±0.2ndndAbove soil31.8±7.42.0±0.2ndndand = not detected.PCR법을 이용하여 도입유전자를 정성, 정량 분석하였다. CMVP0-CP 고추에 도입된 유전자의 양은 10 cm 깊이의 토양 속 및 토양 표면에서 1개월 후 28.3-42.7%, 2개월 후 0.9-3.3%로 감소하였으며, 3-4개월 후에는 검출되지 않았다. 또한 이러한 유전자는 지중보다 지표에서 약 8% 더 오래 지속되었다. CMVP0-CP 고추의 잎이 토양 내에서 분해되는 과정에서 유출된 DNA가 주변 토양으로 전이되었는지를 조사하기 위해 낙엽주머니에 부착된 토양으로부터 DNA를 추출하여 PCR분석을 수행하였다. PCR 분석 결과 CMVP0-CP 고추에 도입된 유전자가 검출되지 않았다.
Abstract
This study was conducted to evaluate the persistence of DNA in the transgenic chili pepper resistant to cucumber mosaic virus (CMV) in the field condition. We analyzed the persistence of genes in the leaf samples obtained from two field conditions, below and above soil. Transgenic and non-transgenic leaf tissues were buried in the soil at a depth of 10 cm or placed on the soil surface. Qualitative and quantitative PCR analysis showed that the amount of transferred genes from the transgenic peppers below and above soil was dropped to 28.3-42.7% one month after buried and it was rapidly reduced to 0.9-3.3% after two months. The transgenes were not detected three to four month after buried. In addition, DNA of the leaves placed below soil decomposed about 8% more than those on soil surface. The gene transfer from decomposing leaves of the transgenic pepper to soil was investigated by PCR analysis with the soil attached to the samples. The PCR result indicated that the gene transfer from the transgenic pepper to soil was not occurred.
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