甘菊下胚轴遗传转化体系的建立

Establishment of Genetic Trasformation System for Chrysanthemum lavandulifolium

甘菊(Chrysanthemum lavandulifolium(Fisch.ex Trautv.)Makino)可作为菊花遗传育种研究模式植物,但目前尚未建立甘菊的遗传转化体系。本研究在甘菊下胚轴再生体系的基础上,以甘菊下胚轴为外植体,对影响转化体系的5个因素进行筛选,尝试利用农杆菌介导的方法获得最佳转化体系。研究结果表明:甘菊下胚轴在不经过预培养,农杆菌浓度OD600=0.4时浸染10 min,共培养2 d,延迟培养1 d时抗性芽分化率和转化率均最高。通过卡那霉素抗性筛选及PCR检测,获得45株PCR阳性植株,阳性转基因植株频率为5%。

Genetic transformation system of Chrysanthemum lavandulifolium which is the model plant of chrys- anthemum genetic engineering has not been established. Using hypocotyl of Chrysanthemum lavandulifolium as explants, an efficient and stable genetic transformation system mediated by Agrobacterium was established through investigating some factors influencing the transformation efficiency. The highest transformation frequency was obtained, with no pre-cultivate, the suitable infection duration 10 rain with bacterial concentration OD600=0.4, appropriate co-culture for 2 days. Several putative transformants were obtained after two round of selection proc- edure. PCR analysis indicated that target gene had been integrated into the genome of Chrysanthemum lavan- dulifolium and its transgenic frequency was 5%.