【作者】 安伟；

【导师】 李亚东；

【作者基本信息】 吉林农业大学 ， 果树学， 2002， 硕士

【摘要】 本实验以树莓6个栽培品种为试材，用叶片作为外植体进行离体培养，建立了树莓无性系B的高频、稳定、快速的离体再生体系。 通过实验发现基因型是影响树莓叶片离体再生的主要限制因子。在供试的6个品种中，无性系B的再生能力最强。在合适的培养基中其再生频率可达90％以上，平均每叶片的再生芽数可达12.0个。外源激素的种类、浓度及配比是影响树莓叶片离体再生的重要因素。在实验中激素配比BA2.0mg／L+IBA 0.1mg／L可获得最高的再生频率和最高的叶片再生芽数。实验中还发现21-30天苗龄的叶片的再生能力最强。MS培养基在供试培养基中是最适合树莓叶片离体再生的基本培养基。持续黑暗和较强的光照(3000Lux)均不利于树莓叶片的离体再生。 本研究采用叶盘法对树莓品种B、烟草品种A进行了植酸酶基因的遗传转化，并在筛选培养基中获得再生芽，对其中部分烟草的转化芽进行了PCR检测为阳性，说明植酸酶基因已整合到烟草植物基因组中。但对树莓的再生芽进行PCR检测却为阴性，本文分析了未成功进行转化的几点原因，为今后树莓转基因研究奠定了基础。更多还原

【Abstract】 Taking six raspberry cultivars as tested materials, this paper studied the factors affecting the adventitious shoot regeneration from leaf pieces in vitro, resulting in the establishment of a stable and efficient adventitious shoot regeneration system of raspberry clone B.It was discovered through experiment that genotype was the major restrictive factor affecting shoot regeneration from leaf disk in vitro . Among the 6 genotypes tested, clone B had the strongest ability in shoot regeneration with shoot induction frequency of 90% and mean number 12.0 shoots per leaf. Phytohormone type, concentration and their combinations in medium were the most important factors influencing shoot regeneration from leaf disk of raspberry . The phytohormone combination BA 2.0 mg/L and IBA 0.1 mg/L had the best effect on shoot induction frequency , mean number of shoots per leaf and shoot quality in our experiment. The 21-30 days old leaf had the highest shoot induction frequency and the highest mean number of shoots per leaf. MS medium was the most suitable basal medium for shoot induction. Dark and 3000 Lux light intensity were harmful to regenerate shoots .The paper also studied the factors affecting the phytase genetic transformation -mediated by Agrobacterium tumefaciens , we obtained the antibiotic plant of raspberry clone B and tobacco clone A . Results of PCR of the tobacco clone A showed that phytase gene had integrated into the genome of the tobacco clone A ,and was expressed properly. But raspberry clone B was failed . We gave several reasons about the failure of phytase genetic transformation of raspberry clone B.更多还原