摘要
分析了不同质量浓度的2,4-二硝基苯酚(简为2,4-D)对早熟禾愈伤组织诱导和愈伤组织分化成苗的影响,3 m g/L和0.1 m g/L 2,4-D分别是本试验所用早熟禾基因型(Poa pratensisL cv. nuble)诱导形成愈伤组织和愈伤组织分化成苗的最适质量浓度.利用基因枪将含GUS基因的pGA470和pAct1-D的质粒导入愈伤组织,通过组织染色法检测到了Gus基因的瞬时表达.含NPTⅡ基因的pGA470质粒和含bar 基因的pDM302质粒转化的愈伤组织,分别在含100 m g/L卡那霉素和含3 m g/Lphosphinothricin(PPT)的选择性培养基上培养,获得了抗卡那霉素和抗PPT的愈伤组织;
Factors affecting callus induction from mature seeds and plantlet regeneration in Kentucky Bluegrass ( Poa pratensis L) were optimised using a modified medium containing MS salts and B5 organic elements and the MS medium respectively. A 2,4 D concentration of 3 mg·L -1 was optimal for callus induction, and 0.1 mg·L -1 for plantlet regeneration. Transient expression of the GUS gene was observed on calli transformed with pGA470 which contains GUS gene driven by the 35S promoter, and pAct1 D which contains GUS gene driven by the rice actin 1 promoter. Putative stable transformants were selected from calli transformed with pGA470, containing the NPT Ⅱ gene driven by the 35S promoter, and pDM302, containing the bar gene driven by the rice actin 1 promoter, and cultured on media containing 100 mg·L -1 kanamycin and 3 mg·L -1 phosphinothicin respectively. A stable expression of NPT Ⅱ gene was detected in the 12th sub cultural callus.
出处
《复旦学报(自然科学版)》
CAS
CSCD
北大核心
1999年第5期540-544,共5页
Journal of Fudan University:Natural Science
基金
国家自然科学基金
上海市教育委员会资助
关键词
早熟禾
遗传转化体系
组织培养
基因枪
草坪草
Poa pratensis L
genetic transformation
tissue culture
micromprojectile bombardment