摘要
为验证从狗蔷薇类原球茎克隆的RaWUS基因的功能,构建了组成型表达载体,并用根瘤农杆菌介导的叶盘转化法对烟草进行了遗传转化。通过除草剂草丁膦筛选,获得了转基因植株并对其进行了表型观察和RT-PCR分析。结果显示转基因烟草叶片形态和叶脉表现出形态变化,包括叶片浅裂、叶片边缘波浪状、叶脉紊乱,甚至叶片的主脉上有不定芽的产生等。RT-PCR分析表明,RaWUS基因仅仅在经过草丁膦筛选的抗性转基因烟草中强烈表达,在野生型烟草中没有表达。表明RaWUS基因已经被成功转入烟草中,并可能通过改变激素的水平和调控维管束的发育来影响叶片形状。
To analyze the function of RaWUS gene isolated from protocorm-like body of Rosa canina, constitutive expression vector was constructed and transformed into tobacco（Nicotiana tabacum） plants by using the leaf disc method mediated through Agrobacterium-tumefaciens. Screening with herbicide glufosinate resistance,transgenic tobacco plants were obtained. Their phenotypes were observed and RT-PCR was conducted. Transgenic tobacco plants exhibited changes in the leaf shape and veins, including lobed leaves,leaves with wary margin, disorder veins even ectopic bud formation on the leaves, and so on. The RT-PCR analysis indicated the RaWUS gene was expressed strongly in the regenerated plants which showed glyphosate resistance,but not in the wild-type tobacco. The results showed that RaWUS gene had been introduced into tobacco. Regulation of vascular tissue through changes in hormone levels was possible to alter leaf morphology. The effect of the gene on development of protocorm-like body and application in plant biotechnology were also discussed in this study.
出处
《核农学报》
CAS
CSCD
北大核心
2012年第2期262-269,F0003,共9页
Journal of Nuclear Agricultural Sciences
基金
国家自然科学基金(31171993)
农业部行业科研专项(200903020)
关键词
RaWUS基因
狗蔷薇的类原球茎
组成型表达
转基因烟草
叶片形态
维管束发育
RaWUS gene
protocorm-like body of Rosa canina
constitutive expression
transgenic tobacco
leafmorphology
vascular tissue development
