农杆菌介导的花生遗传转化体系的优化  

OPTIMIZATION OF Agrobacterium tumefaciens-MEDIATED TRANSFORMATION SYSTEM OF PEANUT

作　　者：乔利仙[1] 于新玲 隋炯明[1] 范乾程[1] 孙世孟[1] 王晶珊[1] 

机构地区：[1]青岛农业大学生命科学学院/山东省高校植物生物技术重点实验室,山东青岛266109 [2]烟台工贸技师学院,山东烟台264003

出　　处：《核农学报》2012年第9期1244-1248,F0003,共6页Journal of Nuclear Agricultural Sciences

基　　金：国家自然基金青年基金(31101178);山东省自然基金(ZR2012CM014);山东省自然基金青年基金(ZR2011CQ026);山东省中青年科学家奖励基金(BS2009NY028)

摘　　要：以花生胚小叶及其诱导形成的愈伤组织为转化受体,采用农杆菌介导法,通过评估外植体GUS基因瞬时表达率,优化花生遗传转化条件。结果表明,侵染液中添加表面活性剂2-氮吗啉乙烷磺酸(MES)150mg/L或烟草提取物0.5ml/30ml有利于提高胚小叶外植体GUS瞬时表达率;采用针刺法辅助农杆菌介导转化,其胚小叶外植体多点GUS瞬时表达率(29.1%)明显高于未针刺的对照(12.1%);利用愈伤组织作为转化受体,其多点GUS瞬时表达率高达70.1%。利用此优化体系进行花生遗传转化,获得的抗性苗2011年经嫁接、移栽于试验田,PCR检测获得了T0代转基因植株,2012年部分T1代植株也检测出目的条带。By evaluating GUS transient expression frequency using embryonic leaflet and callus as explants, several factors which may affect genetic transformation mediated by Agrobaeterium tumefaciens were studied. The results showed that Agrobacterium suspension supplemented with 150mg/L MES or 0.50ml/30ml tobacco extract could significantly improve the transformation efficiency. Acupuncture treatment significantly increased the GUS transient expression frequency by 29.1% with multi-dot staining compared with CK（ 12. 1% ）. High-frequency GUS transient expression of muhi-dot staining （70.1%） was obtained using callus as the transformation receptor. Resistant plantlets obtained using this transformation system -were grafted and transplanted to the field, then the TO positive plants were identified by PCR analysis and the target gene was inherited steadily in to some of T1 plants.

关 键 词：花生(Arachis HYPOGAEA L ) 遗传转化体系 针刺法 胚小叶 愈伤组织 GUS瞬时表达 

分 类 号：S565.2[农业科学—作物学]