【作者】 罗莉；

【导师】 韩烈保；

【作者基本信息】 北京林业大学 ， 森林培育， 2005， 硕士

【摘要】 本研究对草地早熟禾的6个栽培品系DP-37-61,DP192-10,82-37,DP-LF-300,2W42-117,PRO-4287进行了组培筛选,以茎尖分生组织和成熟种子作为外植体诱导愈伤组织,建立了草地早熟禾组培再生体系。挑选胚性愈伤率和再生率较高的品系DP-37-61和PRO-4287诱导胚性愈伤组织,利用基因枪介导法成功地将含有抗潮霉素基因(hpt)的质粒pAct1IHPT-4和DREB转录因子编码的质粒M14转入草地早熟禾胚性愈伤组织,轰击后的愈伤组织在添加100mg/L潮霉素的选择性培养基上进行筛选。品系DP-37-61经过转化共筛选出84个抗性愈伤,其中59个成功再生出抗性植株,PCR检测证明获得的59棵抗性植株全部为转基因植株,转化效率为13%;品系PRO-4287共筛选出8个抗性愈伤,其中6个抗性愈伤成功再生,PCR检测证明获得的6棵抗性植株全部为转基因植株,转化效率为3%。本项研究建立了完善成熟的草地早熟禾组培再生体系和高效的基因转化体系。更多还原

【Abstract】 Six breeding lines of DP-37-61, DP192-10, 82-37, DP-LF-300, 2W42-117, PRO-4287 were cultured using meristem and mature seed as explants to establish the tissue culture system of Kentucky bluegrass (Poapratensis L.) . The embryogenic calli of breeding line DP-37-61 and PRO-4287 which had higher embryogenic-calli induction rate and regeneration rate were co-transformed with plasmid pAct1IHPT-4 containing hpt gene and plasmid M14 containing transcription factor DREB. Bombarded calli were selected on the selection medium supplemented with 100 mg/L hygromycin. Eighty-four resistant calli of breeding line DP-37-61 survived after 12-20 weeks selection on the selected medium, of which 59 resistant calli regenerated successfully with the very high transformation rate of 13%. Eight resistant calli of breeding line PRO-4287 survived after 10-16 weeks selection on the selected medium, of which 6 resistant calli regenerated successfully with the high transformation rate of 3%. The independent nature of transformed plants was verified by PCR analysis. An efficient and reproducible transformation system for Kentucky bluegrass (Poa pratensis L.) was set out with a microprojectile bombardment delivery system.更多还原