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甜菜质体基因组的遗传转化

Genetic transformation of the sugar beet plastome.

作者信息

De Marchis Francesca, Wang Yongxin, Stevanato Piergiorgio, Arcioni Sergio, Bellucci Michele

机构信息

Italian National Research Council, via della Madonna Alta, 130, 06128, Perugia, Italy.

出版信息

Transgenic Res. 2009 Feb;18(1):17-30. doi: 10.1007/s11248-008-9193-4. Epub 2008 Jun 13.

Abstract

It is very important for the application of chloroplast engineering to extend the range of species in which this technology can be achieved. Here, we describe the development of a chloroplast transformation system for the sugar beet (Beta vulgaris L. ssp. vulgaris, Sugar Beet Group) by biolistic bombardment of leaf petioles. Homoplasmic plastid-transformed plants of breeding line Z025 were obtained. Transformation was achieved using a vector that targets genes to the rrn16/rps12 intergenic region of the sugar beet plastome, employing the aadA gene as a selectable marker against spectinomycin and the gfp gene for visual screening of plastid transformants. gfp gene transcription and protein expression were shown in transplastomic plants. Detection of GFP in Comassie blue-stained gels suggested high GFP levels. Microscopy revealed GFP fluorescence within the chloroplasts. Our results demonstrate the feasibility of engineering the sugar beet chloroplast genome; this technology provides new opportunities for the genetic improvement of this crop and for social acceptance of genetically modified sugar beet plants.

摘要

扩展能够实现叶绿体工程技术的物种范围对于该技术的应用非常重要。在此,我们描述了通过对叶柄进行生物弹轰击来开发甜菜(Beta vulgaris L. ssp. vulgaris,糖用甜菜组)叶绿体转化系统的过程。获得了育种系Z025的同质性质体转化植株。使用一个将基因靶向甜菜质体基因组rrn16/rps12基因间隔区的载体实现了转化,采用aadA基因作为针对壮观霉素的选择标记,并使用gfp基因对质体转化体进行视觉筛选。在转质体植物中显示了gfp基因的转录和蛋白质表达。考马斯亮蓝染色凝胶中GFP的检测表明GFP水平很高。显微镜检查揭示了叶绿体内的GFP荧光。我们的结果证明了对甜菜叶绿体基因组进行工程改造的可行性;该技术为这种作物的遗传改良以及转基因甜菜植物的社会接受度提供了新的机会。

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