亳菊EDT1抗旱基因遗传转化体系建立与耐旱性初步评价

发布时间：2018-03-26 03:15

  本文选题：亳菊　切入点：发根农杆菌　出处：《中药材》2017年01期

【摘要】：目的:建立亳菊EDT1抗旱基因遗传转化体系,并初步评价其耐旱性。方法:以亳菊茎段外植体为受体材料,抗草甘膦除草剂为筛选基因,通过发根农杆菌介导法转入EDT1抗旱基因。通过农杆菌介导,对影响亳菊遗传转化效率的四个因素进行讨论:侵染时间、农杆菌菌液浓度、乙酰丁香酮浓度和共培养时间,以及草甘膦最佳筛选浓度的研究。结果:建立了农杆菌介导的亳菊高效遗传转化体系,菌液浓度(A600)为0.6~0.8,乙酰丁香酮浓度150μmol/L,侵染时间25 min,共培养3 d,除菌培养后,可得到15 d最大转化效率95.6%。草甘膦对亳菊组培苗致死率的研究表明其最佳筛选浓度为100 mg/L。通过聚合酶链式反应(PCR)检测,初步得到5株转基因植株。结论:在干旱胁迫下,叶片中抗氧化酶活性和MDA含量测定表明,转基因植株具有较高的耐旱性。
[Abstract]:Objective: to establish a genetic transformation system for drought resistant genes of Boju EDT1 and to evaluate its drought resistance. Methods: the stem explants of Boju were used as acceptor materials and glyphosate resistant herbicides were used as screening genes. Through Agrobacterium tumefaciens mediated transformation of EDT1 drought-resistant gene, four factors affecting the efficiency of genetic transformation of Boju were discussed: infection time, concentration of Agrobacterium tumefaciens, concentration of acetyl eugenone and co-culture time. Results: Agrobacterium tumefaciens mediated high efficiency genetic transformation system of Boju was established. The concentration of A600) was 0.6 渭 mol / L, the concentration of acetyl eugenone was 150 渭 mol / L, the infection time was 25 min, and the culture time was 3 days. The maximum transformation efficiency of 15 days was 95.6%. The study of glyphosate on the mortality of the plantlets in tissue culture showed that the optimum concentration of glyphosate was 100 mg / L. five transgenic plants were preliminarily obtained by polymerase chain reaction (PCR). Conclusion: under drought stress, 5 transgenic plants were obtained. The determination of antioxidant enzyme activity and MDA content in leaves showed that transgenic plants had high drought tolerance.
【作者单位】： 安徽大学资源与环境工程学院;安徽省中草药产业化研发中心;
【基金】：国家自然科学基金(31471554) 安徽大学研究生创新强化项目(yqh100297)
【分类号】：S567.239
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本文编号：1666092