西兰花毛状根悬浮培养体系建立

为建立西兰花毛状根高增殖能力悬浮培养体系,实验对通过发根农杆菌ATCC15834浸染西兰花叶片获得的毛状根进行PCR检测,高增殖液体悬浮培养体系条件的优化,毛状根生长动态与萝卜硫素合成动力学研究。实验结果表明,诱导出的毛状根整合了发根农杆菌ATCC15834的rol B基因。毛状根液体悬浮培养的最佳条件为:接种量为1mg/mL、转速为110r/min、培养基体积100mL、培养温度为25℃,增殖倍数达26.47倍。毛状根悬浮培养生长分为迟滞期、分化期、增殖期、静止期、衰亡期五个时期,萝卜硫素的合成于培养的静止期,并向培养基中少量释放,且最佳收获时间为接种后的第21天。这一研究结果可为次生代谢产物萝卜硫素的生产提供一定的理论基础。

Establishment of Suspension Culture System for Broccoli Hairy Roots

To establish the suspension culture system with high proliferative capacity of broccoli hairy roots, the hairy roots, obtained by infecting broccoli leaves with Agrobacterium rhizogenes ATCC15834, were tested by PCR and optimized to the conditions of high-proliferation liquid suspension culture system, meanwhile the growth dynamics of hairy roots and the synthesis kinetics of sulforaphane were studied. The results showed that the induced hairy roots integrated the rol B gene of Agrobacterium rhizogenes ATCC15834. The optimal conditions for hairy roots were that inoculum size was 1 mg/mL, rotation speed was 110 r/min, medium volume was 100 mL, the culture temperature was 25℃. Under the optimized suspension culture system, the proliferation rate reached 26.47 times. The growth of hairy root suspension culture was divided into five periods: lag phase, differentiation phase,proliferative phase, stationary phase and decay phase. The synthesis of sulforaphane was in the stationary phase,meanwhile, there were a little sulforaphane been released into the medium at this stage. In addition, the optimal harvest time was 21 days after inoculation. This study could provide a theoretical basis for the production of secondary metabolite sulforaphane.