胡萝卜毛状根的诱导及条件优化

Induction of Hairy Roots from Carrot and Optimization of the Inducement Condition

以发根农杆菌A4、LBA9402、R1000为实验菌株,从胡萝卜中成功诱导出可在无激素条件下快速生长的毛状根,通过PCR方法对其进行了初步鉴定。该毛状根在MSR培养基上生长迅速,1个月左右即可长满整个平板,并经多次继代后仍保持其特性。研究了菌株类型、培养基、菌液量、菌液浓度和共培养天数对胡萝卜毛状根转化频率的影响,筛选出的较优胡萝卜发根条件为:使用A4发根农杆菌菌株,共培养培养基为添加50 mg/LVitamin C(Vc)的水琼脂,加菌量20μL,侵染液OD_(600)值0.6,共培养天数2 d。胡萝卜毛状根诱导体系的建立和条件优化为其他植物毛状根的诱导提供了技术参考,同时为进一步研究胡萝卜的次生代谢及建立菌根共生体系等奠定了基础。

This test successfully induced hairy roots from carrot which can grow rapidly on hormone-flee medium using the three kinds ofAgrobacterium tumsfeciens A4, R1600 and LBA9402 as test bacterium. The hairy roots were preliminarily identified by PCR technique. This kind of roots can cover the entire plate after one-month cuhrue and maintain the identities after subculture. Impacts of the strain type, medium type, the amount of bacterium, concentration of bacterium and time of co-culture on hairy roots transformation frequencies were studied and the optimal conditions for carrot hairy roots induction were screened ： use Agrobacterium tumsfeciens A4 for infection, water agar containing 50 mg/L Vitamin C （Vc） , 20μL for the amount of bacterium, 0.6 for OD600 of bacterium and 2 days for co-culture time. Establishment of induction system for carrot hairy roots and optimization of the inducement condition provided a technical reference for the induction of hairy roots of other plants, meanwhile, Laid the foundation for studying secondary metabolism of carrot and establishing mycorrhiza symbiotic system.