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通过根癌农杆菌介导的共转化,用 gusA 报告基因在黄麻(Corchorus capsularis L.)中开发转基因毛状根系统。

Development of a transgenic hairy root system in jute (Corchorus capsularis L.) with gusA reporter gene through Agrobacterium rhizogenes mediated co-transformation.

机构信息

Advanced Laboratory for Plant Genetic Engineering, Advanced Technology Development Center, Indian Institute of Technology Kharagpur, Kharagpur 721302, India.

出版信息

Plant Cell Rep. 2011 Apr;30(4):485-93. doi: 10.1007/s00299-010-0957-y. Epub 2010 Dec 9.

Abstract

Transgenic hairy root system is important in several recalcitrant plants, where Agrobacterium tumefaciens-mediated plant transformation and generation of transgenic plants are problematic. Jute (Corchorus spp.), the major fibre crop in Indian subcontinent, is one of those recalcitrant plants where in vitro tissue culture has provided a little success, and hence, Agrobacterium-mediated genetic transformation remains to be a challenging proposition in this crop. In the present work, a system of transgenic hairy roots in Corchorus capsularis L. has been developed through genetic transformation by Agrobacterium rhizogenes harbouring two plasmids, i.e. the natural Ri plasmid and a recombinant binary vector derived from the disarmed Ti plasmid of A. tumefaciens. Our findings indicate that the system is relatively easy to establish and reproducible. Molecular analysis of the independent lines of transgenic hairy roots revealed the transfer of relevant transgenes from both the T-DNA parts into the plant genome, indicating the co-transformation nature of the event. High level expression and activity of the gusA reporter gene advocate that the transgenic hairy root system, thus developed, could be applicable as gene expression system in general and for root functional genomics in particular. Furthermore, these transgenic hairy roots can be used in future as explants for plantlet regeneration to obtain stable transgenic jute plants.

摘要

转基因毛状根系统在一些抗性植物中很重要,在这些植物中,根癌农杆菌介导的植物转化和转基因植物的产生是有问题的。黄麻(Corchorus spp.)是印度次大陆的主要纤维作物之一,是其中一种抗性植物,体外组织培养取得了一些成功,因此,根癌农杆菌介导的遗传转化在这种作物中仍然是一个具有挑战性的命题。在本工作中,通过携带两个质粒的发根农杆菌(Agrobacterium rhizogenes)的遗传转化,在 Corchorus capsularis L. 中建立了转基因毛状根系统,即天然 Ri 质粒和源自根癌农杆菌去武装 Ti 质粒的重组二元载体。我们的研究结果表明,该系统相对容易建立且具有可重复性。对独立的转基因毛状根系的分子分析表明,相关的转基因从 T-DNA 部分转移到植物基因组中,表明该事件具有共转化的性质。gusA 报告基因的高水平表达和活性表明,所开发的转基因毛状根系统可普遍适用于基因表达系统,特别是适用于根部功能基因组学。此外,这些转基因毛状根可以在未来用作获得稳定的转基因黄麻植物的植物再生的外植体。

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