抗旱调控基因DREB2A转化辽荞5号的研究

Research on the Drought-Resistant Regulatory Genes DREB2A Transforming the ’Liao 5 Buckwheat’

通过农杆菌介导,将来源于干旱诱导下拟南芥的干旱调控基因DREB2A导入辽宁甜荞品种辽荞5号中,以提高其耐旱性。通过正交试验分析影响农杆菌转化的相关因素,建立并优化了转化体系。结果表明:(1)愈伤组织诱导过程中,选择子叶作为愈伤组织诱导受体,诱导愈伤培养基激素的配比为:2,4-D浓度为2 mg/L;6-BA浓度为1 mg/L;(2)农杆菌转化过程中,最适农杆菌OD600为0.5、侵染时间3 min、共培养3 d,乙酰丁香酮浓度100 mg/L、筛选培养羧苄霉素浓度50mg/L,草丁膦浓度100 mg/L;(3)通过对转化植株的PCR检测,印证了DREB2A基因已转化到辽荞5号中;(4)通过对转化植株的生理检测初步表明转基因荞麦比非转基因荞麦具有更高的抗旱性。

A drought regulatory gene from arabidopsis thaliana,DREB2 A,was transformed into the‘Liao 5 buckwheat’using agrobacterium-mediated method to improve its drought tolerance.The factors influencing agrobacterium transformation were analyzed by the orthogonal test.The transformation system was established and optimized.The results showed that:1.The cotyledons were selected as callus induction receptors during callus induction.The ratio of the hormones in the induced callus medium was that the concentration of 2,4-D was 2 mg/L,and the concentration of 6-BA was 1 mg/L.2.During the transformation of agrobacterium,the optimum agrobacterium OD600 was0.5,the infection time was 3 min,the co-culture time was 3 days,the concentration of acetosyringone was 100 mg/L,the concentration of carbenicillin was 50 mg/L,the concentration of phosphinothricin was 100 mg/L.3.The PCR results showed that the gene DREB2 A had been transformed into the‘Liao 5.buckwheat’.4.The physiological test of the transformed plants showed that the modified buckwheat had higher drought resistance than the non-transgenic.