中药材2017，Vol.40Issue(1)：1-6,6.DOI:13863/j.issn1001-4454.2017.01.001
亳菊EDT1抗旱基因遗传转化体系建立与耐旱性初步评价
Establishment Transformation System of Drought-resistant Transgenic Chrysanthemum morifolium with EDT1 Gene and Preliminary Evaluation of Drought Tolerance
摘要
Abstract
Objective:To establish transformation system of drought-resistant transgenic Chrysanthemum morifolium with EDT1 gene,and to evaluate the drought tolerance.Methods:Chrysanthemum morifolium with stem explant was used as receptor material,and resistanted to glyphosate herbicide as the filter gene and transfered the drought-resisitantgene(EDT1)into Chrysanthemum morifolium by using Agrobacterium rhizogenes.Four factors influence efficiency of gene transformation for Chrysanthemum morifolium,which included the time of infection,the concentration of Agrobacterium rhizogenes bacterial and acetosyringone,the time of co-cultured were studied,and screened the best concentration of glyphosate.Results:Transformation system of drought-resistant transgenic Chrysanthemum morifolium with EDT1 gene was established.The concentration of Agrobacterium rhizogenes bacterial (A60o) was 0.6 ～0.8,the concentration of acetosyringone was 150 μmol/L,the infection time was 25 min,and co-cultured time was 3 d.After 15 days's sterilization culture,the maximum conversion efficiency was 95.6％,the best selected concentration of glyphosateis was 100 mg/L.The regenerated plants by PCR testing,five transgenic plants were obtained.Conclusion:Antioxidant enzyme activity and MDA content measurement under drought stress indicate that the transgenic plants have higher drought tolerance.关键词
亳菊/发根农杆菌/抗旱基因/再生植株/抗氧化酶Key words
Chrysanthemum morifolium Ramat./Agrobacterium rhizogenes/Drought-resistant gene/Regenerated plants/Antioxidant enzyme分类
医药卫生引用本文复制引用
胡同华,陈达伟,王钰,刘芳源..亳菊EDT1抗旱基因遗传转化体系建立与耐旱性初步评价[J].中药材,2017,40(1):1-6,6.基金项目
国家自然科学基金(31471554) （31471554）
安徽大学研究生创新强化项目(wqh100297) （wqh100297）
