혼반용 유색대두를 육성하고, 국내 대두품종을 이용한 재분화 체계를 확립하고자 하였으며, 대두의 자엽 절편에서 부정근형성시 품종반응과 절편내 전분분포 및 대량배양과 이들 기관형성체계를 이용하여 Agrobacterium에 의한 형질전환을 시도하였다. 고품질 유색대두를 육종하기 위하여 1994년 우수계통을 선발한 후 강원 2호로 계통명을 부여하였다. 1995년부터 1999년까지 계통의 생산력을 검정하였고, 1997년부터 1999년까지 수원 등 5개 지역에서 지역적응성을 검정하였다. 1999년 ‘흑청’이라는 품종명으로 육성되었다. ‘흑청콩’은 국내 최초로 육성된 흑색 종피의 녹색 자엽을 갖는 품종으로 괴저형 콩 모자이크 바이러스에 저항성이 높으며, 유한 신육형으로 취반 특성이 우수하였다. 신육형은 유한형이고 꽃은 자색, ...
혼반용 유색대두를 육성하고, 국내 대두품종을 이용한 재분화 체계를 확립하고자 하였으며, 대두의 자엽 절편에서 부정근형성시 품종반응과 절편내 전분분포 및 대량배양과 이들 기관형성체계를 이용하여 Agrobacterium에 의한 형질전환을 시도하였다. 고품질 유색대두를 육종하기 위하여 1994년 우수계통을 선발한 후 강원 2호로 계통명을 부여하였다. 1995년부터 1999년까지 계통의 생산력을 검정하였고, 1997년부터 1999년까지 수원 등 5개 지역에서 지역적응성을 검정하였다. 1999년 ‘흑청’이라는 품종명으로 육성되었다. ‘흑청콩’은 국내 최초로 육성된 흑색 종피의 녹색 자엽을 갖는 품종으로 괴저형 콩 모자이크 바이러스에 저항성이 높으며, 유한 신육형으로 취반 특성이 우수하였다. 신육형은 유한형이고 꽃은 자색, 모용은 황갈색, 꼬투리는 암갈색, 잎은 난형이다. 또한 불포화지방산의 종실내 함유율이 86.2%이고, lecithin 함유율이 48 mg/100 g으로 품종육성 표준품종인 ‘검정콩 1호’에 비해 높았으며, methionine과 cysteine 함유단백질인 11S globulin 함유율이 62.8%로서 11S globulin/7S globulin 비율이 1.74로 영양적으로 가치가 높은 품종으로 확인되었다. 국내 대두품종들을 이용하여 품종별 shoot 형성율, 적정배지 및 적정조직을 알아보았다. 유묘의 조직부위별 비교에서는 1개의 자엽을 포함한 절에서 3-5개의 shoot가 형성(multiple shoot)되는 비율이 높았고, 기본배지에서는 1/2 B5 배지가 효율적이었다. Cytokinin류에서는 benzyladenine(BA) 처리시에는 1-2개의 shoot가 형성(pair shoot)되는 비율이 높았고, thidiazuron(TDZ) 처리시에는 multiple shoot 형성율이 높았으며, 처리농도는 2 mg/L가 효율적이었다. 품종간 비교에서는 품종에 따라 shoot 형성양상이 구분되어 ‘흑청콩’ 등 11품종에서는 multiple shoot 형성율이 높았고, ‘흑청콩’ 등 16 품종에서는 pair shoot 형성율이 높았다. 대두의 자엽절편을 이용한 부정근 형성 양상과 품종반응 및 대량배양을 시도하였다. 품종별 부정근 형성 양상은 직접적인 부정근 형성 품종과 callus 형성이 동시에 이루어지는 품종들로 구분되었으며, callus 형성이 많은 품종에서는 부정근 형성수가 적은 결과를 보였다. 자엽 절편에서의 부정근 형성은 절편의 향축면 치상시에만 부정근이 형성되었고, 배축면과 향축면이 동시에 치상되어도 향축면에서만 부정근이 형성되며, 절편의 배축이 존재하였던 방향에서 부정근이 형성되는 것으로 보아 일정한 방향성이 존재함을 알 수 있었다. 부정근 형성시 자엽 절편내 전분은 형성초기보다 형성이 가장 많은 유기 4일에 가장 많이 관찰되었으며, 형성된 부정근의 생장이 이루어지는 7일 이후 관찰되지 않았다. 부정근의 대량배양은 기포발생 간이 생물반응기에서 배양 4주 정도에 대량증식 가능성이 확인되었으며, 1/2 B5 기본배지에 0.05 mg/L kinetin과 1.0 mg/L indolbutyric acid(IBA) 첨가시 생장량이 가장 높았다. 대두 자엽절에서의 shoot 형성시 Agrobacterium에 의한 형질전환후 shoot 형성율은 선발배지인 kanamycin 포함 배지에서 16%이었으며, 형성기간은 배양 6주로 나타났다. 대두 자엽절편에서 부정근시 Agrobacterium에 의한 형질전환 후 kanamycin 포함 배지에서의 절편당 부정근 형성수는 17개로 나타났으며, 형성기간은 배양 7일이었다. 부정근 형성 체계를 이용한 형질전환 효율은 자엽절편에 macerozyme 처리로 무처리시 절편당 7개의 부정근 형성에 비해 17개로 효율을 높일 수 있었다.
혼반용 유색대두를 육성하고, 국내 대두품종을 이용한 재분화 체계를 확립하고자 하였으며, 대두의 자엽 절편에서 부정근형성시 품종반응과 절편내 전분분포 및 대량배양과 이들 기관형성체계를 이용하여 Agrobacterium에 의한 형질전환을 시도하였다. 고품질 유색대두를 육종하기 위하여 1994년 우수계통을 선발한 후 강원 2호로 계통명을 부여하였다. 1995년부터 1999년까지 계통의 생산력을 검정하였고, 1997년부터 1999년까지 수원 등 5개 지역에서 지역적응성을 검정하였다. 1999년 ‘흑청’이라는 품종명으로 육성되었다. ‘흑청콩’은 국내 최초로 육성된 흑색 종피의 녹색 자엽을 갖는 품종으로 괴저형 콩 모자이크 바이러스에 저항성이 높으며, 유한 신육형으로 취반 특성이 우수하였다. 신육형은 유한형이고 꽃은 자색, 모용은 황갈색, 꼬투리는 암갈색, 잎은 난형이다. 또한 불포화지방산의 종실내 함유율이 86.2%이고, lecithin 함유율이 48 mg/100 g으로 품종육성 표준품종인 ‘검정콩 1호’에 비해 높았으며, methionine과 cysteine 함유단백질인 11S globulin 함유율이 62.8%로서 11S globulin/7S globulin 비율이 1.74로 영양적으로 가치가 높은 품종으로 확인되었다. 국내 대두품종들을 이용하여 품종별 shoot 형성율, 적정배지 및 적정조직을 알아보았다. 유묘의 조직부위별 비교에서는 1개의 자엽을 포함한 절에서 3-5개의 shoot가 형성(multiple shoot)되는 비율이 높았고, 기본배지에서는 1/2 B5 배지가 효율적이었다. Cytokinin류에서는 benzyladenine(BA) 처리시에는 1-2개의 shoot가 형성(pair shoot)되는 비율이 높았고, thidiazuron(TDZ) 처리시에는 multiple shoot 형성율이 높았으며, 처리농도는 2 mg/L가 효율적이었다. 품종간 비교에서는 품종에 따라 shoot 형성양상이 구분되어 ‘흑청콩’ 등 11품종에서는 multiple shoot 형성율이 높았고, ‘흑청콩’ 등 16 품종에서는 pair shoot 형성율이 높았다. 대두의 자엽절편을 이용한 부정근 형성 양상과 품종반응 및 대량배양을 시도하였다. 품종별 부정근 형성 양상은 직접적인 부정근 형성 품종과 callus 형성이 동시에 이루어지는 품종들로 구분되었으며, callus 형성이 많은 품종에서는 부정근 형성수가 적은 결과를 보였다. 자엽 절편에서의 부정근 형성은 절편의 향축면 치상시에만 부정근이 형성되었고, 배축면과 향축면이 동시에 치상되어도 향축면에서만 부정근이 형성되며, 절편의 배축이 존재하였던 방향에서 부정근이 형성되는 것으로 보아 일정한 방향성이 존재함을 알 수 있었다. 부정근 형성시 자엽 절편내 전분은 형성초기보다 형성이 가장 많은 유기 4일에 가장 많이 관찰되었으며, 형성된 부정근의 생장이 이루어지는 7일 이후 관찰되지 않았다. 부정근의 대량배양은 기포발생 간이 생물반응기에서 배양 4주 정도에 대량증식 가능성이 확인되었으며, 1/2 B5 기본배지에 0.05 mg/L kinetin과 1.0 mg/L indolbutyric acid(IBA) 첨가시 생장량이 가장 높았다. 대두 자엽절에서의 shoot 형성시 Agrobacterium에 의한 형질전환후 shoot 형성율은 선발배지인 kanamycin 포함 배지에서 16%이었으며, 형성기간은 배양 6주로 나타났다. 대두 자엽절편에서 부정근시 Agrobacterium에 의한 형질전환 후 kanamycin 포함 배지에서의 절편당 부정근 형성수는 17개로 나타났으며, 형성기간은 배양 7일이었다. 부정근 형성 체계를 이용한 형질전환 효율은 자엽절편에 macerozyme 처리로 무처리시 절편당 7개의 부정근 형성에 비해 17개로 효율을 높일 수 있었다.
This research was carried out to breed a variety of colored soybean used for cooking with rice, and establish the efficient regeneration system using domestic soybeans. The following things were compared to each one such as the cultivar response, the starch distribution within the cotyledonary expla...
This research was carried out to breed a variety of colored soybean used for cooking with rice, and establish the efficient regeneration system using domestic soybeans. The following things were compared to each one such as the cultivar response, the starch distribution within the cotyledonary explants when adventitious roots were formed in the cotyledonary explants of soybean and the possibility of mass culture of roots segments. A transformation was also attempted with using an Agrobacterium. Breeding high-quality soybeans, a superior line was selected in 1994, named 'Kangwon 2'. The preliminary, advanced and regional yield trials for this cultivar were carried out from 1995 to 1997. It was named 'Heugcheong' which is the first cultivar that has the green cotyledon with black coat in Korea. It showed the high resistance to necrosis type of soybean mosaic virus. It has good adaptability for cooking with rice. It has determinate growth habit, purple flower, tawny pubescence and dark brown pod, and oval leaf shape. It contained the unsaturated fatty acid within the seed as high as 86.2%, with 48-mg/100 g of lecithin. There were 62.8% of 11S globulin that is the protein with methionine and cysteine. Furthermore, the nutritional value of the cultivar was greatly high, that is, the 11S globulin to 7S globulin ratio was 1.7. The shoot formation rate, optimal medium and optimal tissue were examined using domestic soybeans. Being compared with the parts of tissues in the seedling, a node that includes one cotyledon was higher to the 3 to 5 multiple shoot formation rate than other tissues. 1/2 B5 medium was efficient with the basal medium. The formation rates of Pair shoots, that is, 1 to 2 in shooting, were higher in the kind of cytokinin when benzyl adenine treatment was supplemented. The formation rates of Multiple shoots, i.e. 4 to 5 in shooting, were higher when thidiazuron treatment was done. The concentration of thidiazuron treatment was efficient at the 2 mg/L. Being compared with for the cultivars, multiple shoot formation rates were higher in the 11 cultivars including 'Heugcheongkong', and Pair shoot formation rates were higher in the 16 cultivars including 'Geomjeongolkong'. The patterns of adventitious root formation for each cultivar were compared, and mass culture was attempted with using the formed roots at cotyledonary explants of soybean. The results of adventitious root formation by cultivars are classified as two groups; the first group showed the direct adventitious root formation, and the second group resulted in the callus and adventitious root formation. The cultivars that have much callus formation had less the adventitious root formation. The adventitious root formation in the cotyledonary explants was occured only at the inoculation of adaxial side. When adaxial and abaxial side was inoculated simultaneously, the adventitious roots were formed at the adaxial side. Thus, it suggests that there must be direction to some extent. Starch in the cotyledonary explants were more abundant at the 4 days after induction than at the early stage of the adventitious root formation, but the starch was not observed after 7 days, that the growth stage of adventitious roots. The possibility of mass culture was certified in culture period of 4 weeks, in the aerated simple bioreactor. The growth rates of roots segments were the highest when 0.05 mg/L kinetin and 1.0 mg/L indolbutyric acid was supplemented at the 1/2 B5 basal medium. When the adventitious shoots was formed at the cotyledonary nodes, the adventitious shoot formation rate was 16% at the medium contained with kanamycin, which is the selection medium, after transformation by the Agrobacterium. The period of adventitious shoot regeneration was 6 weeks after culture. The adventitious root formation rates were 17% at the medium contained kanamycin after the transformation by the Agrobacterium in cotyledonary explants. The period of the adventitious root formation was 7 days after culture. Macerozyme treatment improved the efficiency of the transformation by Agrobacterium using the system of adventitious root formation. These results showed that breeding the colored new cultivars could be well adaptable for cooking with rice, and multiple shoots were induced from cotyledonary node cultured on 1/2 B5 medium supplemented with 2 mg/L thidiazuron in a few cultivars. Also, adventitious roots were formed at adaxial side of cotyledonary explants, and mass cultured on 1/2 B5 basal medium supplemented with 0.05 mg/L kinetin and 1.0 mg/L indolbutyric acid using the aerated simple bioreactor. These organogenesis systems can be advantageously utilized in the production of transgenic plants and roots by Agrobacterium.
This research was carried out to breed a variety of colored soybean used for cooking with rice, and establish the efficient regeneration system using domestic soybeans. The following things were compared to each one such as the cultivar response, the starch distribution within the cotyledonary explants when adventitious roots were formed in the cotyledonary explants of soybean and the possibility of mass culture of roots segments. A transformation was also attempted with using an Agrobacterium. Breeding high-quality soybeans, a superior line was selected in 1994, named 'Kangwon 2'. The preliminary, advanced and regional yield trials for this cultivar were carried out from 1995 to 1997. It was named 'Heugcheong' which is the first cultivar that has the green cotyledon with black coat in Korea. It showed the high resistance to necrosis type of soybean mosaic virus. It has good adaptability for cooking with rice. It has determinate growth habit, purple flower, tawny pubescence and dark brown pod, and oval leaf shape. It contained the unsaturated fatty acid within the seed as high as 86.2%, with 48-mg/100 g of lecithin. There were 62.8% of 11S globulin that is the protein with methionine and cysteine. Furthermore, the nutritional value of the cultivar was greatly high, that is, the 11S globulin to 7S globulin ratio was 1.7. The shoot formation rate, optimal medium and optimal tissue were examined using domestic soybeans. Being compared with the parts of tissues in the seedling, a node that includes one cotyledon was higher to the 3 to 5 multiple shoot formation rate than other tissues. 1/2 B5 medium was efficient with the basal medium. The formation rates of Pair shoots, that is, 1 to 2 in shooting, were higher in the kind of cytokinin when benzyl adenine treatment was supplemented. The formation rates of Multiple shoots, i.e. 4 to 5 in shooting, were higher when thidiazuron treatment was done. The concentration of thidiazuron treatment was efficient at the 2 mg/L. Being compared with for the cultivars, multiple shoot formation rates were higher in the 11 cultivars including 'Heugcheongkong', and Pair shoot formation rates were higher in the 16 cultivars including 'Geomjeongolkong'. The patterns of adventitious root formation for each cultivar were compared, and mass culture was attempted with using the formed roots at cotyledonary explants of soybean. The results of adventitious root formation by cultivars are classified as two groups; the first group showed the direct adventitious root formation, and the second group resulted in the callus and adventitious root formation. The cultivars that have much callus formation had less the adventitious root formation. The adventitious root formation in the cotyledonary explants was occured only at the inoculation of adaxial side. When adaxial and abaxial side was inoculated simultaneously, the adventitious roots were formed at the adaxial side. Thus, it suggests that there must be direction to some extent. Starch in the cotyledonary explants were more abundant at the 4 days after induction than at the early stage of the adventitious root formation, but the starch was not observed after 7 days, that the growth stage of adventitious roots. The possibility of mass culture was certified in culture period of 4 weeks, in the aerated simple bioreactor. The growth rates of roots segments were the highest when 0.05 mg/L kinetin and 1.0 mg/L indolbutyric acid was supplemented at the 1/2 B5 basal medium. When the adventitious shoots was formed at the cotyledonary nodes, the adventitious shoot formation rate was 16% at the medium contained with kanamycin, which is the selection medium, after transformation by the Agrobacterium. The period of adventitious shoot regeneration was 6 weeks after culture. The adventitious root formation rates were 17% at the medium contained kanamycin after the transformation by the Agrobacterium in cotyledonary explants. The period of the adventitious root formation was 7 days after culture. Macerozyme treatment improved the efficiency of the transformation by Agrobacterium using the system of adventitious root formation. These results showed that breeding the colored new cultivars could be well adaptable for cooking with rice, and multiple shoots were induced from cotyledonary node cultured on 1/2 B5 medium supplemented with 2 mg/L thidiazuron in a few cultivars. Also, adventitious roots were formed at adaxial side of cotyledonary explants, and mass cultured on 1/2 B5 basal medium supplemented with 0.05 mg/L kinetin and 1.0 mg/L indolbutyric acid using the aerated simple bioreactor. These organogenesis systems can be advantageously utilized in the production of transgenic plants and roots by Agrobacterium.
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