CAJ | 학술논문

以双亚7号亚麻为试验材料,通过农杆菌介导法,将抗性基因AtNDPK2导入亚麻,并对影响遗传转化的主要因素进行了优化研究,结果表明：双亚7号培养5d苗龄的亚麻,下胚轴切成0.5 cm左右,预培养2 d后,取出放于OD 0.6-0.8的农杆菌菌液中摇动10-15 min。然后用灭过菌的滤纸吸干下胚轴表面的农杆菌,置于不含抗生素的共培养基上共培养3 d,共培养结束后将下胚轴转入含50 mg·L-1 G418和300 mg·L-1 Cef 的下胚轴分化增殖的筛选培养基中进行分化增殖。获得抗性植株6株,PCR检测表明有4株阳性植株,转化率为66.67%,初步表明AtNDPK2基因已导入亚麻中。
이쌍아7호아마위시험재료,통과농간균개도법,장항성기인AtNDPK2도입아마,병대영향유전전화적주요인소진행료우화연구,결과표명：쌍아7호배양5d묘령적아마,하배축절성0.5 cm좌우,예배양2 d후,취출방우OD 0.6-0.8적농간균균액중요동10-15 min。연후용멸과균적려지흡간하배축표면적농간균,치우불함항생소적공배양기상공배양3 d,공배양결속후장하배축전입함50 mg·L-1 G418화300 mg·L-1 Cef 적하배축분화증식적사선배양기중진행분화증식。획득항성식주6주,PCR검측표명유4주양성식주,전화솔위66.67%,초보표명AtNDPK2기인이도입아마중。
The AtNDPK2 gene was transferred into flax cultivar“ShuangYa-7” by Agrobacterium mediated transforma-tion and the principal factors affecting transformation system was optimized. The results showed that the hypocotyledonary axis of cultivar“ShuangYa-7” was cut into about 0. 5 centimeter after being cultured for 5 days, then were preincubated for 2 days, and were immersed into the Agrobacterium Tumefaciens liquid which OD value is 0. 6-0. 8 and were shaken for 10-15 min. the Agrobacterium Tumefaciens on hypocotyledonary axis was sipped up by sterilized filter paper, the hy-pocotyledonary axis were cocultured in the medium without antibiotics for 3 days and then were transferred into the selec-tive medium with 50 mg·L-1 G418 and 300 mg·L-1 Cef. 6 putative positive plants were obtained. Detection of PCR showed that 4 plants were positive, and transformation efficiency was 66. 7%, the success of AtNDPK2 gene being trans-ferred into flax was preliminarily proved.