基于甘薯毛状根的外源多基因表达系统的建立

Establishment of the Exogenous Gene Expression System Based on the Hairy Roots of Ipomoea batatas(L.) Lam.

用携带植物高效表达载体pCAMBIA1304+的"解除武装"的重组C58C1工程菌转化甘薯新品种"渝苏303"无菌苗的真叶,50个外植体中的36个长出了毛状根,诱导率达72%.PCR检测证实发根型质粒pRiA4和表达型质粒pCAMBIA1304+均能整合到甘薯毛状根基因组内,共转化率达36%.建立了基于发根农杆菌介导的甘薯毛状根的高效外源基因表达系统,为利用甘薯毛状根快速验证基因功能和将多个外源基因导入甘薯,实现其代谢工程及开展分子育种奠定了基础.

The disarmed Agrobacterium tumefaciens strain C58C1 harboring the plasmid pRiA4 was engineered by introducing the plant expressing vector pCAMBIA1304＋. The bacteria-free well-growing true leaves of ＇Yusu 303＇, a new cultivar of Ipomoea batatas, were infected with the engineered reeombined C58C1. Hairy roots were induced from the wounded sites of the leaves at a frequency of 72%. Then, genomic PCR was performed to detect rolC and antihygromycin gene in different monoclonal hairy root lines. Both genes were simultaneously detected in 36 % hairy root clones. These results demonstrated that exogenous genes could be expressed in hairy roots of L batatas at a relatively high frequency by the method in the present studies. It may be a fundamental work for rapid gene functional identification and gene cotransformation into I. batatas and will improve metabolic engineering and molecular breeding of this plant.