胡萝卜下胚轴遗传转化体系的优化

Optimization of Genetic Transformation System of Carrot Hypocotyl

【研究目的】优化胡萝卜下胚轴转化体系,为胡萝卜基因工程研究提供一个简便而快捷的方法;【方法】以胡萝卜下胚轴为外植体,对农杆菌介导的的遗传转化体系进行优化;【结果】实验结果表明:1)将MS培养基的有机成分进行调整配制成基本培养基MST,有利于愈伤组织的诱导,但不利于苗的分化与再生培养;2)农杆菌侵染外植体的时间为15 min,共培养时间为50 h,获得抗性愈伤组织诱导率达78%;3)羧苄青霉素的浓度从愈伤组织诱导阶段的500 mg/L下降到苗再生阶段的100 mg/L时,提高抗性苗再生率到88%;【结论】通过体系的优化,极大的提高了胡萝卜下胚轴的转化效率,为胡萝卜基因工程的研究提供有效的技术平台。

【OBJECTIVE】The objective was to optimize a Agrobacterium tumefaciens-mediated carrot hypocotyl transformation system,to offer an easy and rapid protocol for carrot genetic engineering research;【METHOD】The system of carrot hypocotyl transformation by Agrobacterium tumefaciens was optimized.【RESULTS】The results showed that： 1） the basic medium MST from the MS medium adjusted by us in the organic components,is conducive to callus induction,but not conducive to callus differentiation and shoot regeneration;2） when carrot hypocotyls was infected for 15 minutes and was co-cultivated for 50 hours with Agrobacterium tumefaciens,the callus induction rate of resistance could be raised up to 78%;3） If the carbenicillin concentration of 500 mg/L was lowered to 100 mg/L in the the callus induction phase,the rate of resistant shoot regeneration could be raised up to 88%.【CONCLUSION】Through the optimization of Agrobacterium tumefaciens-mediated carrot hypocotyl transformation system,transformation efficiency was greatly improved,and then an effective technology platform have been provided for carrot genetic engineering research.