根癌农杆菌介导北海道黄杨遗传转化体系的建立

 在建立北海道黄杨(Euonymus japonicus 'Cu zhi')再生体系的基础上，建立其遗传转化体系，以获得转雪花莲外源凝集素(Galanthus nivalis agglutinin，GNA)基因植株。采用根癌农杆菌(Agrrobacterium tumefaciens)　( LBA4404菌株)介导法，研究影响北海道黄杨遗传转化的若干因素。结果表明，预培养与否、菌液浓度、侵染时间、共培养时间等对转化频率都有一定的影响。在遗传转化过程中，不经预培养，根癌农杆菌浓度OD₆₀₀ = 0.6，侵染下胚轴40 min，再共培养3 d，有利于获得最高遗传转化效率。抗性植株经PCR和DNA-Southern blot检测表明，目的基因已整合到北海道黄杨基因组中。

Establishment of Genetic Transformation System of Euonymus japonicus 'Cu Zhi' Mediated by Agrobacterium tumefaciens

In order to provide genetically improved new germplasm resources and achieve the GNA transgenic plants of Euonymus japonicus 'Cu zhi', genetic transformation system was established based on the system of regeneration. Several factors affected genetic transformation of Euonymus japonicus 'Cu zhi' mediated by Agrobacterium tumefaciens (LBA4404) were studied. The results showed that preculture or not, bacterial concentrateions, the time of infection and co-cultivation had some effect on transformation frequency. The highest transformation frequency was obtained through the following transformation procedure after preculture for 0 d, the explants were infected for 40 min with the concentration of Agrobacterium tumefaciens OD600=0.6 , and then co-cultivated for 3 d. The PCR and PCR-Southern blotting analysis showed the snowdrop lection gene gene was integrated into the genome of Euonymus japonicus 'Cu zhi'. The system of genetic transformation of Euonymus japonicus 'Cu zhi' could provide new germplasm resource used in genetic improvement of Euonymus japonicus 'Cu zhi' and the other plants of Euonymus.