用农杆菌Ri诱导金银花发根培养的研究

A Study of Hairy Roots Culture of Flos lonicerae Induced by Agrobacterium rhizogenes Ri

通过预培养时间、菌液稀释倍数、浸染时间、共培养时间、抗生素质量浓度5因素4水平的正交实验,得出优化组合,认为组培苗叶片外植体预培养5 d,菌液稀释45倍,浸染4 min,共培养3 d,然后转入含500 mg/L羧苄青霉素的1/2M S培养基上培养,金银花的发根诱导率最高,达54.3%.随机抽取离体培养的发根经PCR检测,证实为转化发状根.

Orthogonal tests were conducted, on four levels,： over five factors, namely, pre-culture time, tested strain concentration, infection time, co-culture time and antibiotic concentration to obtain an optimum formulation for inducing Flos. Lonicera hairy roots. Results show that the induction rate of hairy roots is up to 54.3% when the OD60o of R1000 is about 0. 6-0. 8, the axenic plantlet leaves are pre-cultured for 5 days, and the Agrobacterium solution, immersed 4 rains, is diluted to 45 times, co-cultured for 3 days and transferred to culture in 1/2MS medium with 500 mg/L Carb. Hairy roots taken at random from the in vitro culture were tested by PCR to be transgenic.