茶条槭愈伤组织的再生体系建立及其没食子酸含量的测定

Establishment of Callus Regeneration System for Acer ginnala Maxim and Determination of Gallic Acid in Callus

通过愈伤组织诱导途径,建立了快速高效的茶条槭再生体系。成熟种子在MS+1.0mg·L-16-BA的培养基中萌发,以茎段作为外植体,在WPM+0.002-0.01mg·L-1TDZ+0.1mg·L-16-BA中培养3周诱导形成愈伤组织,诱导频率平均为98.0%。愈伤组织转入WPM+0.01mg·L-1TDZ+0.1mg·L-16-BA培养基中得到再生芽,分化频率为42.0%,平均每块愈伤产生再生芽10个左右。转到WPM+0.3mg·L-1IBA的培养基上的再生芽均可生根并长成完整植株,小苗移栽成活率达到89.0%。实验还建立了愈伤组织中没食子酸的提取和HPLC检测方法。对深绿色愈伤组织连续培养2个继代后,没食子酸含量达到2.8%。

A reproducible system for callus induction of Acer ginna/a Maxim is described. The system involves mature seeds sprouted on MS medium supplemented with 0.1 mg·L^-1 6-BA, the stems of young seedling as explants, and calli easily induced on woodyplant medium （WPM） supplemented with 0.002-0.01 mg·L^-1 TDZ and 0.1 mg·L^-1 6-BA 3 weeks following induction. The frequency of callus induction can be as high as 98.0%. On WPM containing 0.01 mg·L^-1 TDZ and 0.1 mg·L^-1 6-BA, 42% of calli differentiated, and the mean number of buds per piece of callus was about 10. The buds developed roots on WPM medium with 0.3 mg·L^-1 IBA and formed plantlets, 89% of which survived on transplantation to the greenhouse. Methods to extract and determine gallic acid from callus were established and showed 2.8% content of gallic acid in bottles of green callus subcultured twice.