发根农杆菌诱导花生发根的条件优化  

OPTIMIZATION OF THE HAIRY ROOT INDUCTION BY Agrobacterium rhizogene IN PEANUT(Arachis hypogaea L.)

作　　者：任艳[1,2] 李磊[3] 崔潇[3] 王辉[1] 石延茂[1] 李双铃[1] 刘杰[1] 袁美[1] 

机构地区：[1]山东省花生研究所/农业部花生生物学与遗传育种重点实验室,山东省花生重点实验室,山东青岛266100 [2]中国海洋大学,山东青岛266100 [3]青岛市农业科学院,山东青岛266100

出　　处：《核农学报》2012年第3期439-443,共5页Journal of Nuclear Agricultural Sciences

基　　金：山东省农科院博士基金(2006YBS021);青年基金(2005YQ010);科技部863项目(2006AA100106),科技部作物种业科技工程(2011BAD35B04)

摘　　要：以花生无菌苗的不同部位为外植体,用发根农杆菌(Agrobacterium rhizogene)GIM1.141进行侵染,诱导发根,以为花生的转基因研究和花生发根提取白藜芦醇奠定基础。通过对重悬菌液乙酰丁香酮的添加、不同外植体、菌液浸泡时间以及发根培养基4种因素的研究,优化发根农杆菌的转化体系。结果表明,花生的叶片、子叶、下胚轴均能被GIM1.141诱导出发根,相同诱导条件下,各外植体的发根诱导率顺序为叶片>子叶>下胚轴;适宜的菌液浸泡时间为10min;附加乙酰丁香酮(100μmol/L),发根诱导率明显提高,花生叶片、子叶、下胚轴的最高发根诱导率分别为87.29%、75.8%、23.33%,因此,花生发根诱导的合适条件为以叶片和子叶为外植体,在添加乙酰丁香酮(100μmol/L)的菌液中浸泡10min诱导出来的发根经过除菌,能在MSB5培养基上自主生长。In order to set up peanut genetic transformation system for the production of resveratrol from hairy roots of peanut,different parts of sterile seedling of peanut were infected with Agrobacterium rhizogene GIM1.141 to induce hairy roots.Four factors including addition of Acetosyringone（AS） in bacterium suspension,different explants,soaking period of bacterium suspension and hairy root-inducing medium were investigated.The hairy roots could be observed at each type of explants（leaflet,cotyledon and hypocotyl）.Leaflet explant produced the high proportion of hairy roots followed by cotyledon and hypocotyl.Suitable soaking period of bacterium suspension for 10min or adding 100μmol/L AS increased the rate of hair roots induction significantly.The highest rate of hairy roots induction from leaf,cotyledon and hypocotyls was 87.29%,75.8%,23.33%,respectively.So hairy roots of leaflet and cotyledon induced by Agrobacterium rhizogenes with 100μmol/L AS for 10min were the optimum methods.After eliminating bacteria with cefotaxime,hairy roots could grow independently on MSB5 medium without auxin.

关 键 词：发根农杆菌 花生 发根诱导 乙酰丁香酮 

分 类 号：S565.2[农业科学—作物学]