摘要
选用对蚜虫具有明显抗性的基因———雪花莲外源凝集素酶基因 ,用宁杞 1号的幼叶、幼茎、幼根与携带质粒的农杆菌共培养后 ,将其转移到含有 6-BA0 .5mg/L +NAA 1mg/L +60mg/L卡那霉素的MS培养基中诱导愈伤组织 ,10～ 15d后将诱导产生的愈伤组织转移到含有 6-BA0 .5mg/L十NAA0 .0 1mg/L +60mg/L卡那霉素的MS分化培养基中 ,培养 2 0～ 30d ,在侵染外植体的愈伤组织上出现小芽 ,当芽长到 1.5～ 2cm时将其切下转入 6-BA0 .0 1mg/L +NAA0 .2mg/L +30mg/L卡那霉素的MS生根培养基中进一步筛选并使转化体生根 ,获得完整的抗蚜虫转基因枸杞。将获得的完整植株进行PCR分析检测 ,发现雪花莲外源凝集素酶基因已经整合到枸杞植株染色体上 ,片段大约在 50 0Pb ,其转化率为 65.1%。
Snowdrop foreign lectinase gene, which has obvious resistance to aphid, was selected for the test. The young leaf, young stem and young root of Chinese wolfberry Ningqi No.1 were cultrued together with Agrobacteriumtumefaciens carrying the plasmid. Then they were transferred onto MS medium containing 6-BA0.5mg/L+NAA 1mg/L+60mg/L Kanamycin for induction of wound callus. 10 to 15 days later the induced wound callus were transferred onto MS differentiation medium containing 6-BA0.5mg/L+60mg/L+NAA0.01mg/L+60mg/L Kanamycin. After 20 to 30 days of culture young buds emerged from the wound callus of infected external implanted tissue. When the buds grew to as long as 1.5～2cm they were cut off and transferred onto MS root-promoting medium containing 6-BA0.01mg/L+NAA0.2mg/L+30mg/L Kanamycin for further selecting and making the tissue grow root. By these transgenic Chinese wolfberry with resistance to aphid was obtained. The PCR analysis and detection with the obtained whole plant show that snowdrop foreign lectinase gene has been integrated with Chromosome of plant of Chinese wolfberry. The segment was about 500 pb and the transformation rate was 65.1% .
出处
《宁夏农林科技》
2001年第1期1-3,共3页
Journal of Ningxia Agriculture and Forestry Science and Technology
