Adaptation of a seedling micro-grafting technique to the study of long-distance signaling in flowering of Arabidopsis thaliana
Adaptation of a seedling micro-grafting technique to the study of long-distance signaling in flowering of Arabidopsis thaliana
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DOI：
10.1007/s10265-008-0209-1
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发表时间：
2009-03-01
影响因子：
2.8
通讯作者：
Araki, Takashi
中科院分区：
文献类型：
--
作者：
Notaguchi, Michitaka;Daimon, Yasufumi;Araki, Takashi
通过韧皮部组织的长距离信号传递是器官间通讯的重要机制。这种信息传递使植物能够将环境信息整合到生理和发育反应中。嫁接提供了有说服力的证据，远距离信号参与各种过程，包括开花，块茎形成，开花，枝分枝，转录后基因沉默，抗病性。微嫁接技术，以产生两个芽嫁接可用于拟南芥幼苗，并适用于开花的研究。使用在韧皮部组织中表达β-葡萄糖醛酸酶（GUS）的转基因植物的组织学分析表明，在嫁接后7至10天之间建立了砧木和接穗之间的韧皮部连续性。利用示踪染料和增强型绿色荧光蛋白（EGFP）的实验表明，韧皮部连接是功能性的，能够影响大分子的传输。成功的移植物可以以高频率（10-30%）获得，并在术后生长2-3周后选择。该方法已成功地应用于植物开花的研究，这是长距离信号调控的重要事件之一。这种嫁接技术将有助于研究参与生长和发育各个方面以及信号分子运输的基因的远距离作用。
Long-distance signaling via phloem tissues is an important mechanism for inter-organ communication. Such communication allows plants to integrate environmental information into physiological and developmental responses. Grafting has provided persuasive evidence of long-distance signaling involved in various processes, including flowering, tuberization, nodulation, shoot branching, post-transcriptional gene silencing, and disease resistance. A micro-grafting technique to generate two-shoot grafts is available for young seedlings of Arabidopsis thaliana and was adapted for use in the study of flowering. Histological analysis using transgenic plants expressing beta-glucuronidase (GUS) in phloem tissues showed that phloem continuity between a stock and a scion was established between 7 and 10 days after grafting. Experiments using tracer dyes and enhanced green fluorescent protein (EGFP) showed that the phloem connection was functional and capable of effecting macromolecular transmission. Successful grafts can be obtained at high frequency (10-30%) and selected after 2-3 weeks of post-surgery growth. This method was applied successfully to the study of flowering, one of the important events regulated by long-distance signaling. This grafting technique will facilitate the study of the long-distance action of genes involved in various aspects of growth and development, and in transport of signal molecules.