农杆菌介导的雪花莲凝集素基因转入玉米骨干自交系

Introduction of snowdrop lectin gene into maize elite inbred lines via Agrobacterium tumefaciens

以农杆菌AGL0介导,将雪花莲凝集素基因转入玉米骨干自交系齐319和掖515胚性愈伤组织细胞,从筛选后的抗性愈伤组织获得再生植株.农杆菌浓度和共培养时间均能显著影响侵染后玉米愈伤组织的抗性频率.在农杆菌浓度OD6000.2～0.3,共培养时间3d时,侵染后玉米愈伤组织的抗性频率最高,平均约4%.对再生植株及其子代基因组DNA的PCR及Southern杂交分析表明雪花莲凝集素基因已经整合到玉米基因组中,并遗传给后代.在蚜虫人工接种试验中,转基因植株上蚜虫的繁殖力为非转基因对照植株上的50%,这表明转基因植株抗蚜性显著增强.

Snowdrop lectin (Galanthus nivalis agglutinin, GNA) gene was introduced into embryogenic calli of elite maize inbred lines Qi319 and Ye515 via Agrobacterium tumefaciens AGL_0, and transgenic plantlets were obtained from resistant maize calli. The resistant percentage of infected maize calli was significantly affected by the concentration of Agrobacterium and the duration of co-cultivation. The resistant percentage of infected maize calli was highest with an average of 4% while the concentration of Agrobacterium was OD_(600) 0.2~0.3 and the duration of co-cultivation was 3 days. PCR and Southern analysis of the genomic DNA from regenerated plants and filial generation showed that GNA gene was integrated into maize genome and inherited to filial generation. In aphid artificial feeding trial, the nymph production on transgenic plants was 50% of that on non-transgenic plants indicating that aphid-resistance was significantly improved in transgenic plants.