【作者】 蔡文燕；

【导师】 肖华山； ?赜?；

【作者基本信息】 福建师范大学 ， 植物学， 2005， 硕士

【摘要】 中国水仙(Narcissus tazetta var.chinensis)是我国传统名花,具有重要的观赏价值和经济价值。但因其栽培历史悠久,病毒病危害有同趋严重之势,影响了中国水仙的商品化生产。中国水仙为三倍体植物,难以通过有性杂交方式选育新品种。基因工程技术的发展和应用为花卉新品种培育提供了_套全新思路。本文研究了中国水仙高效再生体系和遗传转化体系的建立,依照该体系,通过根癌农杆菌将具有抗细菌和病毒作用的人乳铁蛋白基因导入中国水仙,期望能获得转基因植株,为提高中国水仙的品质提供一条有效的途径。 以福建漳州水仙鳞茎盘组织切块为外植体,研究了低温预处理、不同植物生长调节剂对水仙小鳞茎诱导及再生、继代培养、生根及移栽的影响,建立了一套高频的中国水仙再生体系。在此基础上,以GUS基因为报告基因,通过根癌农杆菌介导,对农杆菌转化中国水仙的各项影响因素进行了详细研究,进行了抗生素基础抗性测定和抑菌剂的抑菌效果实验;比较了农杆菌菌株及共培养天数、农杆菌菌液浓度、预培养天数、AS添加浓度对GUS基因瞬时表达率的影响,建立和优化了遗传转化各环节的技术参数,为进行中国水仙的遗传操作奠定了基础。依照该体系,采用农杆菌介导法将人乳铁蛋白基因导入中国水仙中,经卡那霉素筛选和PCR检测,初步确定获得2株转基因植株。更多还原

【Abstract】 Narcissus tazetta var. chinensis is one of the Chinese traditional famous flowers with great economic and ornamental value. But its commercial production has been greatly affected by serious virus diseases. And it is hard to breed new cultivars of Narcissus by sexual hybridization since it is triploid plant. With the development of biotechnology, genetic engineering technology has become the focus of the ornamental plant breeding. In this study, we aimed at establishing a high-frequency regeneration system and a genetic transformation system of Narcissus tazetta, based on which hLFc with antimicrobial and antiviral function could be transformed into Narcissus tazetta mediated by Agrobacterium tumefaciens.A high-frequency regeneration system for Narcissus tazetta var. chinensis was established by studying the effects of low tempreture pretreatment, different types and concentration of plant growth regulator on induction and regeneration of narcissus bulbs. Factors affecting genetic transformation of Narcissus mediated by Agrobacterium tumefaciens were investigated in detail by using gusA as a reporter. The results showed that preculture days, bacterial strains and co-culture time, different bacterium concentration, infecting time, AS concentration all have certain relation with efficiency of transient expression of gusA. Basic resistance of bulbs to Kan and effect of antibiotics concentration on bacteria were also detected. The establishment of genetic transformation of Narcissus tazetta var. chinensis will provide a scientific basis for its further transformation. On the basis of above mentioned genetic transformation system, hLFc was transformed into narcissus bulb tissues mediated by Agrobacterium. Six transformed plants were obtained after resistance screening by Kan and PCR analysis showed that 2 of them were positive.更多还原