Transformation and regeneration of the holoparasitic plant Phelipanche aegyptiaca.
Transformation and regeneration of the holoparasitic plant Phelipanche aegyptiaca.
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DOI：
10.1186/1746-4811-7-36
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发表时间：
2011-11-08
期刊：
影响因子：
5.1
通讯作者：
Westwood JH
中科院分区：
文献类型：
--
作者：
Fernández-Aparicio M;Rubiales D;Bandaranayake PC;Yoder JI;Westwood JH
全寄生植物的转化和随后的再生从未被报道过，部分原因是在开发转化方案方面的挑战，但也因为专性寄生物的再生是困难的，因为它们的存活完全取决于宿主的吸器成功穿透和维管连接的形成。最近完成的大规模转录组测序项目（寄生植物基因组计划）将推动基因组工具在寄生植物研究中的应用。需要一个可靠的系统，全寄生虫转化，实现反向遗传学和功能基因组学研究的资源的全部价值。在这里，我们证明了埃及菲利班车的转化是通过用含有黄色荧光蛋白（YFP）的发根农杆菌感染3个月大的体外生长的埃及菲利班车愈伤组织来实现的。四个月后，将YFP阳性再生愈伤组织接种到在微流控系统中生长的番茄植株上。接种后8天，转基因寄生虫组织形成侧向吸器，其穿透宿主，并且可以在UV照射下通过完整的宿主根组织可视化。接种后一个月观察到YFP阳性的芽。这项工作构成了全寄生植物研究方法的突破。这里描述的方法是一个强大的系统的转化和再生的全寄生植物，并将促进独特的寄生植物的能力，如寄主植物识别，吸器的形成，渗透和维管连接的研究。
Transformation and subsequent regeneration of holoparasitic plants has never been reported, in part due to challenges in developing transformation protocols, but also because regeneration of obligate parasites is difficult since their survival depends completely on successful haustorium penetration of a host and the formation of vascular connections. The recent completion of a massive transcriptome sequencing project (the Parasitic Plant Genome Project) will fuel the use of genomic tools for studies on parasitic plants. A reliable system for holoparasite transformation is needed to realize the full value of this resource for reverse genetics and functional genomics studies. Here we demonstrate that transformation of Phelipanche aegyptiaca is achieved by infection of 3 month-old in vitro grown P. aegyptiaca calli with Agrobacterium rhizogenes harboring the yellow fluorescent protein (YFP). Four months later, YFP-positive regenerated calli were inoculated onto tomato plants growing in a minirhizotron system. Eight days after inoculation, transgenic parasite tissue formed lateral haustoria that penetrated the host and could be visualized under UV illumination through intact host root tissue. YFP-positive shoot buds were observed one month after inoculation. This work constitutes a breakthrough in holoparasitic plant research methods. The method described here is a robust system for transformation and regeneration of a holoparasitic plant and will facilitate research on unique parasitic plant capabilities such as host plant recognition, haustorial formation, penetration and vascular connection.
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影响因子：
3.7
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