Identification of mRNAs that Move Over Long Distances Using an RNA-Seq Analysis of Arabidopsis/Nicotiana benthamiana Heterografts
Identification of mRNAs that Move Over Long Distances Using an RNA-Seq Analysis of Arabidopsis/Nicotiana benthamiana Heterografts
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DOI：
10.1093/pcp/pcu210
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发表时间：
2015-02-01
影响因子：
4.9
通讯作者：
Suzuki, Takamasa
中科院分区：
文献类型：
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作者：
Notaguchi, Michitaka;Higashiyama, Tetsuya;Suzuki, Takamasa
韧皮部是一种传导性组织，它将成熟的源叶中的营养物质分配到诸如年轻发育的组织中。韧皮部也传递蛋白质和RNA物种，如小RNA和mrna。对植物系统信号的深入研究揭示了蛋白质和RNA物种的重要作用。然而，它们的许多功能在很大程度上仍然是未知的，尤其是对转运mrna的作用知之甚少。一个主要的困难是缺乏准确和全面的移动抄本清单。在这项研究中，我们使用了一个以本烟为受体接穗，拟南芥为供体砧木的杂种嫁接系统，来鉴定在嫁接结合中长距离移动的转录本。我们鉴定出138个拟南芥转录本为可移动mRNA，我们将其统称为mRNA移动组。逆转录PCR、实时荧光定量PCR和微滴数字PCR分析证实了其迁移性。转录本包括潜在的信号因子，出乎意料的是，更一般的因素。在我们的研究中，我们没有发现偏好的转录物长度，在启动子或转录物序列中没有先前已知的序列基序，并且转录物水平与源叶片中的转录物水平没有相似性。对已鉴定的转录本ERECTA的功能进行嫁接实验，结果表明转录本的功能未被调动。据我们所知，这是第一次发现转录本在不同植物类群之间通过异种嫁接系统进行长距离迁移。
Phloem is a conductive tissue that allocates nutrients from mature source leaves to sinks such as young developing tissues. Phloem also delivers proteins and RNA species, such as small RNAs and mRNAs. Intensive studies on plant systemic signaling revealed the essential roles of proteins and RNA species. However, many of their functions are still largely unknown, with the roles of transported mRNAs being particularly poorly understood. A major difficulty is the absence of an accurate and comprehensive list of mobile transcripts. In this study, we used a hetero-graft system with Nicotiana benthamiana as the recipient scion and Arabidopsis as the donor stock, to identify transcripts that moved long distances across the graft union. We identified 138 Arabidopsis transcripts as mobile mRNAs, which we collectively termed the mRNA mobilome. Reverse transcription-PCR, quantitative real-time PCR and droplet digital PCR analyses confirmed the mobility. The transcripts included potential signaling factors and, unexpectedly, more general factors. In our investigations, we found no preferred transcript length, no previously known sequence motifs in promoter or transcript sequences and no similarities between the level of the transcripts and that in the source leaves. Grafting experiments regarding the function of ERECTA, an identified transcript, showed that no function of the transcript mobilized. To our knowledge, this is the first report identifying transcripts that move over long distances using a hetero-graft system between different plant taxa.