摘要
为了定向育种获得蓝色百合,该研究以百合Robina为蓝色基因最佳受体,以其花丝诱导产生的胚性愈伤组织和再生小植株小鳞片作为转化材料,利用农杆菌介导法,将蝴蝶兰F3′5′H基因导入百合Robina中。结果表明:以小鳞片为转化材料,预培养3d,OD_(600)为0.8,侵染10min,共培养3d,加入100μmol/L AS稳定转化率最高为12.78%;而以胚性愈伤为转化材料,预培养2d,OD_(600)为0.8,侵染10min,共培养3d,加入100μmol/L AS稳定转化率最高为12.22%。2种转化材料的最适潮霉素筛选浓度均为20mg/L。对抗性植株分别进行PCR和反转录PCR检测,获得9个阳性株系,Southern印记分析进一步确定了6株转基因百合中携带蓝色基因F3′5′H,为后续进一步获得蓝色百合奠定了基础。
To obtain bluish Liliumspp.in direct breeding,we screened the suitable variety-OT lily Robina for genetic transformation.Here,both embryonic callus induced from filament and regenerated bulb scales of OT lily Robina were used as the transformation material.Agrobacterium-mediated transformation of Phalaenopsis F3′5′H was studied.The results showed：with the pre-culture 3d,OD_（600）=0.8,infection time 10 min,co-cultured for 3d,100μmol/L acetosyringone conditions,the stable transformation rate of regenerated bulb scales could reach to the highest 12.78%;however,embryogenic callus pre-culture 2d,OD_（600）=0.8,infection time 10 min,co-cultured 3d,with 100μmol/L acetosyringone conditions,which the stable transformation rate was the highest 12.22%.In all the conditions,the best hygromycin-resistant screening concentrations always was 20mg/L.PCR and reverse transcription PCR assay showed 9putative transgenic plants were obtained.Southern hybridization analysisfurther proved 6plants that the transgenic lilium flowers carry blue gene F3′5′H.The results provided technical support and material basis for the continuing development of novel bluish Liliumflowers.
出处
《西北植物学报》
CAS
CSCD
北大核心
2016年第5期874-880,共7页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家自然科学基金(31471905)
