摘要
利用来自小麦花药、幼胚愈伤组织和带有GUS基因的Ti质粒,对农杆菌转化小麦的条件进行了初步研究,证实适当浓度的乙酰丁香酮是诱发T-DNA转移的必需条件,其最低有效浓度为50μmol/L.在20~35℃,pH5.2~6.8条件下,均可有效诱导T-DNA的转移.农杆菌菌液密度以108ml-1细胞为宜,添加适当的表面活性剂有促进T-DNA转化的作用.对4种小麦材料的研究证明,T-DNA的转移效率常因受体材料基因型的不同而有很大差异;愈伤组织的活力与T-DNA转移的效率有密切关系.显微观察发现,胚状体细胞比非胚状体细胞群更容易发生转化.
The conditions for wheat genetic transformation mediated by Agrobacterium tumefaciens were optimized using calli derived from anthers and immature embryos and a Ti plasmid containing the GUS gene. The presence of acetosyringon in co-culture medium was essential for the efficient transfer of T-DNA into callus cells,with the lowest effective concentration of 50 μmol/L. A high incidence of T-DNA transfer was closely associated to the physiological state of callus. The coculture temperature from 20℃ to 35℃ and the medium pH from 5. 2 to 6. 8 have the same T-DNA transfer efficiency. The inoculated bacterium cell intensity was optimized as 108 cells/ml. In addition, it was found that surfactant could enhance T-DNA transfer.
出处
《复旦学报(自然科学版)》
CAS
CSCD
北大核心
1998年第4期569-572,共4页
Journal of Fudan University:Natural Science