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油菜에서 細胞質雄性不稔(CMS)係 cytoplast와 耐寒性 雄性可稔系 原形質體間 融合親和性의 genotype間 差異
Yu Sun Nam(柳順南) 한국육종학회 1996 한국육종학회지 Vol.28 No.1
Subprotoplasts containing enuclic(cytoplasts) and nucleic(miniprotoplasts) subprotoplasts were isolated from freshly isolated protoplasts of Brassica napus ‘CMS Jet Neuf’ cotyledons. Protoplasts were separated by centrifuging at 30, 000g for 60min at 12℃ through specific iso-osmotic density gradient containing colloidial silica gel(Percell), CaCl₂ and mannitol. Using this method highly purified cytoplast fractions with less than 20% contamination with nucleased protoplasts were obtained. Cytoplasts prepared by this method were suitable for the use in fusion experiments aimed at transferring enucleated CMS-Donor-Cytoplast and nucleated Cold-Tolerant-CMS-Receptor-Protoplast seperately. Protoplastfusion culture between enucleated CMS-Donor-Cytoplast of ‘CMS Jet Neuf’ and nucleated Cold-Tolerant CMS-Receptor-Protoplast with PEG 6,000 showed the first cell division 9 days after the start of culture and survived for 26 days there after.
유순남(Sun Nam Yu),이명화(Myeung Hwa Lee),정지혜(Ji Hue Jung),장무웅(Moo Ung Chang) 한국원예학회 2006 원예과학기술지 Vol.24 No.4
2004년부터 2006년까지 한국에서 재배되고 있는 카네이션의 바이러스의 발생상황을 조사하였다. 카네이션 재배지역에서 채집된 67품종 575식물체로부터 바이러스를 검출하여 바이러스의 입자의 형태, 기주범위, 감염세포 내의 바이러스 입자 및 봉입체의 존재 형태, 혈청반응, coat protein(CP)유전자 등을 근거로 하여 바이러스를 동정한 결과, carnation mottle virus(CarMV), carnation latent virus(CLV), carnation necrotic fleck virus(CNFV) 등 3종의 바이러스가 확인되었다. 3종의 바이러스 감염률은 CarMV가 66.44%, CLV가 6.6%, CNFV가 6.78%로 각각 확인되었고, CLV와 CNFV는 전부 CarMV와 복합감염되어 있었다. 지역과 농가에 따라 바이러스 감염률에 차이가 있었다. 이들 바이러스에 감염되지 않은 카네이션, 즉 바이러스 무병독률(viruses- free plants)은 평균 33.56%로 확인되었다. 한국, 일본, 중국에서 채집한 카네이션에서 각각 분리된 CarMV-K, CarMV-J, CarMV-C의 CP 유전자를 특이적인 올리고뉴클레오티드 프라이머를 이용하여 역전사-중합호소반응(RT-PCR)으로 증폭시킨 후에 염기서열을 분석하였다. 일본과 중국의 카네이션에서 분리한 CarMV-K의 CP 유전자의 염기서열을 CarMV-J와 CarMV-C과 비교한 결과 99.6%의 상동성을 나타냈다. In the survey of virus diseases of carnations in Korea from 2004 to 2006, 3 viruses of carnation mottle virus (CarMV), carnation latent virus (CLV), carnation necrotic fleck virus (CNFV) were identified from the 575 carnation plants in 67 cultivars, based on shape and size of virus particles, host range, distribution pattern of virus particles and inclusion bodies in the infected cells, serological reaction, and coat protein gene. Viral infection rates were CarMV 66.44%, CLV 6.6%, CNFV 6.78% of the 575 samples surveyed. All CLV and CNFV were doubly infected with CarMV, respectively. The average of carnation plants free from these viruses was 33.56%. Carnations from different areas and propagators varied widely in the incidences of the tested viruses. Coat protein (CP) genes of CarMV-K, CarMV-J, CarMV-C isolated from carnations collected in Korea, Japan and China were amplified using specific oligonucleotide primer by RT-PCR. Sequence analysis of amplified CP gene of CarMV-K showed 99.6% nucleotide similarity to CarMV-J and CarMV-C.